記述言語：英語   掲載種別：研究論文（学術雑誌）  

BACKGROUND:
The common carp (Cyprinus carpio) is the oldest, most domesticated and one of the most cultured fish species for food consumption. Besides its economic importance, the common carp is also highly suitable for comparative physiological and disease studies in combination with the animal model zebrafish (Danio rerio). They are genetically closely related but offer complementary benefits for fundamental research, with the large body mass of common carp presenting possibilities for obtaining sufficient cell material for advanced transcriptome and proteome studies.
RESULTS:
Here we have used 19 different tissues from an F1 hybrid strain of the common carp to perform transcriptome analyses using RNA-Seq. For a subset of the tissues we also have performed deep proteomic studies. As a reference, we updated the European common carp genome assembly using low coverage Pacific Biosciences sequencing to permit high-quality gene annotation. These annotated gene lists were linked to zebrafish homologs, enabling direct comparisons with published datasets. Using clustering, we have identified sets of genes that are potential selective markers for various types of tissues. In addition, we provide a script for a schematic anatomical viewer for visualizing organ-specific expression data.
CONCLUSIONS:
The identified transcriptome and proteome data for carp tissues represent a useful resource for further translational studies of tissue-specific markers for this economically important fish species that can lead to new markers for organ development. The similarity to zebrafish expression patterns confirms the value of common carp as a resource for studying tissue-specific expression in cyprinid fish. The availability of the annotated gene set of common carp will enable further research with both applied and fundamental purposes.

DOI： 10.1186/s12864-016-3038-y

その他リンク： http://bmcgenomics.biomedcentral.com/articles/10.1186/s12864-016-3038-y

リポジトリ公開URL： http://hdl.handle.net/2324/4784016

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Complements have long been considered to be a pivotal component in innate immunity. Recent researches, however, highlight novel roles of complements in T-cell-mediated adaptive immunity. Membrane-bound complement regulatory protein CD46, a costimulatory protein for T cells, is a key molecule for T-cell immunomodulation. Teleost CD46-like molecule, termed Tecrem, has been newly identified in common carp and shown to function as a complement regulator. However, it remains unclear whether Tecrem is involved in T-cell immune response. We investigated Tecrem function related to T-cell responses in ginbuna crucian carp. Ginbuna Tecrem (gTecrem) proteins were detected by immunoprecipitation using anti-common carp Tecrem monoclonal antibody (mAb) and were ubiquitously expressed on blood cells including CD8α(+) and CD4(+) lymphocytes. gTecrem expression on leucocyte surface was enhanced after stimulation with the T-cell mitogen, phytohaemagglutinin (PHA). Coculture with the anti-Tecrem mAb significantly inhibited the proliferative activity of PHA-stimulated peripheral blood lymphocytes, suggesting that cross-linking of Tecrems on T-cells interferes with a signal transduction pathway for T-cell activation. These findings indicate that Tecrem may act as a T-cell moderator and imply that the complement system in teleost, as well as mammals, plays an important role for linking adaptive and innate immunity.

DOI： 10.1016/j.molimm.2015.11.010

その他リンク： http://www.sciencedirect.com/science/article/pii/S016158901530122X

リポジトリ公開URL： http://hdl.handle.net/2324/4776957

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Thrombocytes are nucleated blood cells in non-mammalian vertebrates, which were recently focused on not only as hemostatic cells but also as immune cells with potent phagocytic activities. We have analyzed the phagocytic activation mechanisms in common carp (Cyprinus carpio) thrombocytes. MACS-sorted mAb+ thrombocytes showed no phagocytic activity even in the presence of several stimulants. However, remixing these thrombocytes with other anti-thrombocyte mAb− leukocyte populations restored their phagocytic activities, indicating that carp thrombocyte phagocytosis requires an appropriate exogenous stimulation. Culture supernatant from anti-thrombocyte mAb− leukocytes harvested after PMA or LPS stimulation, but not culture supernatant from unstimulated leukocytes, could activate thrombocyte phagocytosis. This proposed mechanism of thrombocyte phagocytosis activation involving soluble factors produced by activated leukocytes suggests that thrombocyte activation is restricted to areas proximal to injured tissues, ensuring suppression of excessive thrombocyte activation and a balance between inflammation and tissue repair.

DOI： 10.1016/j.dci.2015.05.002

リポジトリ公開URL： http://hdl.handle.net/2324/4776937

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Recent studies on fish immunity highlighted the significance of gills as mucosal immune tissues. To understand potential of gills as vaccination sites for inducing adaptive systemic immunity, we investigated virus-specific cell-mediated and humoral immune responses following a “per-gill infection method”, which directly exposes virus only to gills. The viral load in crucian carp hematopoietic necrosis virus (CHNV)-infected gills decreased after peaking at a particular time point. Furthermore, the viral titers in the gills following the secondary infection were lower than that after the primary infection, indicating that local adaptive immunity helped the elimination of virus. Gene expression analysis demonstrated that IFN-γ in gills and perforin in kidney were increased after the gill infection. CD8+ cells in kidney leukocytes increased after the secondary infection, whereas IgM+ cells decreased. These results suggest that IFN-γ and CTL contribute in controlling CHNV-replication in gills and kidney. Gill infection could induce specific cell-mediated cytotoxicity of peripheral blood leukocytes (PBL) and secretion of CHNV-specific IgM in serum, indicating that local priming of the gill site can generate adaptive systemic immunity. Thus, the gills could be prospective antigen-sensitization sites for mucosal vaccination.

DOI： 10.1016/j.dci.2015.04.016

リポジトリ公開URL： http://hdl.handle.net/2324/4776936

記述言語：英語   掲載種別：研究論文（学術雑誌）  

In the complement system, the regulators of complement activation (RCA) play crucial roles in controlling excessive complement activation and in protecting host cell from misdirected attack of complement. Several members of RCA family have been cloned from cyclostome and bony fish species and classified into soluble and membrane-bound type as in mammalian RCA factors. Complement-regulatory functions have been described only for soluble RCA of lamprey and barred sand bass; however, little is known on the biological function of the membrane-bound RCA proteins in the lower vertebrates. In this study, a membrane-bound RCA protein, designated teleost complement-regulatory membrane protein (Tecrem), was cloned and characterized for its complement-regulatory roles. Carp Tecrem, an ortholog of a zebrafish type 2 RCA, ZCR1, consists of four short consensus repeat modules, a serine/threonine/proline-rich domain, a transmembrane region, and a cytoplasmic domain, from the N terminus, as does mammalian CD46. Tecrem showed a ubiquitous mRNA expression in carp tissues, agreeing well with the putative regulatory role in complement activation. A recombinant Chinese hamster ovary cell line bearing carp Tecrem showed a significantly higher tolerance against lytic activity of carp complement and less deposition of C3-S, the major C3 isotypes acting on the target cell, than control Chinese hamster ovary (mock transfectant). Anti-Tecrem mAb enhanced the depositions of carp C3 and two C4 isotypes on autologous erythrocytes. Thus, the present findings provide the evidence of complement regulation by a membrane-bound group 2 RCA in bony fish, implying the host-cell protection is an evolutionarily conserved mechanism in regulation of the complement system.

DOI： 10.4049/jimmunol.1303179

リポジトリ公開URL： http://hdl.handle.net/2324/4777997

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Thrombocytes, nucleated hemostatic blood cells of non-mammalian vertebrates, are regarded as the functional equivalent of anucleated mammalian platelets. Additional immune functions, including phagocytosis, have also been suggested for thrombocytes, but no conclusive molecular or cellular experimental evidence for their potential ingestion and clearance of infiltrating microbes has been provided till date. In the present study, we demonstrate the active phagocytic ability of thrombocytes in lower vertebrates using teleost fishes and amphibian models. Ex vivo, common carp thrombocytes were able to ingest live bacteria as well as latex beads (0.5-3 μm in diameter) and kill the bacteria. In vivo, we found that thrombocytes represented nearly half of the phagocyte population in the common carp total peripheral blood leukocyte pool. Phagocytosis efficiency was further enhanced by serum opsonization. Particle internalization led to phagolysosome fusion and killing of internalized bacteria, pointing to a robust ability for microbe elimination. We find that this potent phagocytic activity is shared across teleost (Paralichthys olivaceus) and amphibian (Xenopus laevis) models examined, implying its conservation throughout the lower vertebrate lineage. Our results provide novel insights into the dual nature of thrombocytes in the immune and homeostatic response and further provide a deeper understanding of the potential immune function of mammalian platelets based on the conserved and vestigial functions.

DOI： doi: 10.3389/fimmu.2014.00445

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Zymosan, a yeast cell wall preparation that binds activated forms of complement C3, is a useful model target to activate the complement system. In our trial to analyze C3 diversity in Nile tilapia at the protein level using zymosan, we found that a novel 240-kDa serum protein (ZBP-240) also bound to zymosan in addition to C3-derived fragments. In the present study, we aimed to characterize tilapia C3 and ZBP-240, focusing on their immune-related functions. Four distinct C3 isoforms were purified from tilapia serum and shown to possess an intrachain thioester bond. ZBP-240 was also isolated from tilapia serum and examined for its binding properties to various microbial targets. As a result, ZBP-240 showed a wide spectrum of binding to Gram-positive and Gram-negative bacteria and yeasts. Amino acid sequence analysis of CNBr fragments of ZBP-240 suggested that this is a novel protein with no homologous sequence in protein databases. It was also suggested that the binding of ZBP-240 to microbes largely depends on hydrophobic interactions in a divalent-cation-independent manner, and that there may be a divalent-cation-dependent factor that enhances the binding of ZBP-240 in tilapia serum. Interestingly, ZBP-240 showed opsonic activity for tilapia kidney phagocytes at a level comparable to that of C3, implying that ZBP-240 is a novel teleost opsonic serum protein.

DOI： 10.1007/s12562-014-0700-7

リポジトリ公開URL： http://hdl.handle.net/2324/4776908

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Regulators of complement activation (RCA) play a role in protecting cells from excessive complement activation in humans. cDNA corresponding to three isoforms of teleost membrane-bound RCA protein (gTecrem) have been identified in the ginbuna crucian carp. gTecrem-1 consists of seven short consensus repeats (SCRs), whereas gTecrem-2 and gTecrem-3 have four SCRs. While gTecrem-1 possesses a tyrosine phosphorylation site in its cytoplasmic region, gTecrem-2 and gTecrem-3 lack the site. Tissue distribution analysis showed that gTecrem-1 and gTecrem-2 mRNAs were expressed in almost all tissues examined, whereas gTecrem-2 expression was not significantly detected in gill, liver, or intestine. Furthermore, analysis showed that gTecrem-1 was expressed in both peripheral blood leukocytes (PBLs) and erythrocytes and was also expressed in T cell subsets such as CD4+, CD8+ T cells, and IgM+ B cells. gTecrem-2 expression was not detected in either PBLs or erythrocytes, whereas gTecrem-3 was expressed only in erythrocytes. These results suggested that gTecrem isoforms may serve different functional roles; gTecrem-1, which is expressed in T cells and possesses a tyrosine phosphorylation site, may act as a complement regulator and a cellular receptor in adaptive immunity.

DOI： 10.1016/j.fsi.2013.08.002

リポジトリ公開URL： http://hdl.handle.net/2324/4776942

記述言語：英語   掲載種別：研究論文（学術雑誌）  

The central component of complement, C3, plays a versatile role in innate immune defense of vertebrates and some invertebrates. A notable molecular characteristic of this component is an intra-chain thioester site that enables C3 to bind covalently to its target. It has been reported that the binding preference of the thioester to hydroxyl or amino groups is primarily defined by presence or absence of the catalytic histidine residue at position 1126 in human C3. In teleosts, a unique C3 (non-His type) has been found, in addition to the common His type C3. These distinct C3 isoforms may provide diversity in the target-binding attributable to the different binding specificities of their thioesters. In the present study, we examine the hypothesized correlation of the catalytic histidine with the binding spectra of two major C3 isotypes of carp towards various model and natural targets. The results reveal that non-His type C3, rather than His type C3, has a wider range of binding spectrum, despite the binding specificity of its thioester being limited to amino groups. It is therefore hypothesized that the binding spectra of C3 isotypes are not defined by the binding specificity of the thioester but is more affected by differences in microbe-associated molecular patterns that activate complement. Overall, the present data imply that non-His type C3 plays a significant role against bacterial infections in the fish defense system.

DOI： 10.1016/j.dci.2012.03.004.

リポジトリ公開URL： http://hdl.handle.net/2324/4776913

記述言語：英語   掲載種別：研究論文（大学，研究機関等紀要）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Trypanoplasma borreli is an extracellular blood parasite of carp belonging to the same Order (Kinetoplastida) as African trypanosomes. These mammalian parasites have developed different strategies to evade the host immune system including antigenic variation, immunosuppression and clearance of surface-bound antibodies. The latter mechanism allows trypanosomes to use their swimming movement to cause surface-bound antibodies to 'sail' and accumulate at the posterior end of the parasite, to be internalized via the flagellar pocket and be degraded. There is no evidence that T. borreli shows antigenic variation, but during the late phases of infection NO-mediated immunosuppression is observed. High levels of nitric oxide (NO) lead to extensive tissue nitration whereas the parasite itself is not affected. Therefore, the induction of NO has thus far been considered a parasite-driven response with immunosuppressive effects. In the present study, we show that the induction of NO, particularly during the early phase of T. borreli infections, should be re-considered an effective part of the host immune response. We show that T. borreli rapidly removes surface-bound IgM. In addition, moderate concentrations of NO, by hindering surface antibody clearance, maintain high the concentrations of membrane-bound IgM, thereby favoring antibody-dependent complement-mediated parasite lysis. We performed a comprehensive quantitative gene expression analysis of in total seven different complement factors involved in all three activation pathways, differentiating between 1 and 4 isoforms for each complement gene. Our gene expression analysis supports an important role for antibody-dependent complement-mediated lysis of T. borreli in vivo. To our knowledge, NO-dependent inhibition of antibody clearance from the surface of kinetoplastid parasites has not been investigated. Our data support a role for NO as an important player in host-parasite interactions, not only as immune suppressor (late response) but also as immune effector (early response) in infections with bloodstream parasites such as T. borreli.

DOI： 10.1016/j.molimm.2009.08.011

リポジトリ公開URL： http://hdl.handle.net/2324/4776956

記述言語：英語   掲載種別：研究論文（大学，研究機関等紀要）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Interleukin-8 (IL-8) is a CXC-type chemokine with a chemotactic activity mainly on neutrophils and plays a key role in promoting inflammation. In teleosts, several CXC-chemokines have been cloned and characterized as being IL-8-like. Phylogenetic data however indicate that the reported teleost IL-8-like chemokines are substantially remote from mammalian IL-8, forming a fish-specific clade of IL-8-like chemokines distinct from that of tetrapod IL-8. In the present study, a novel IL-8-like chemokine, designated CaIL-8, has been found in the expressed sequence tags of carp gills and identified as an orthologue of mammalian IL-8. The CaIL-8 transcript encodes 99 amino acids containing a typical CXC motif but lacks an ELR motif, as in most teleost IL-8-like chemokines. Phylogenetic tree constructed by the maximum likelihood method suggests a closer relationship of CaIL-8 with mammalian IL-8 than with other teleost CXC-chemokines reported to be IL-8-like. In a normal unstimulated carp, CaIL-8 mRNA was detected by RT-PCR only in gills, kidney, spleen, heart and peripheral blood leukocytes, in contrast to a previously reported carp IL-8-like chemokine CXCa, which shows ubiquitous basal expression. The results, taken together, are strongly indicative of the presence of two major IL-8-like lineages of CXC-chemokines in teleost.

DOI： 10.1016/j.fsi.2009.08.004

リポジトリ公開URL： http://hdl.handle.net/2324/4776939

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Complement factor I is a crucial regulator of mammalian complement activity. Very little is known of complement regulators in non-mammalian species. We isolated and sequenced four highly similar complement factor I cDNAs from the liver of the nurse shark (Ginglymostoma cirratum), designated as GcIf-1, GcIf-2, GcIf-3 and GcIf-4 (previously referred to as nsFI-a, -b, -c and -d) which encode 689, 673, 673 and 657 amino acid residues, respectively. They share 95% (≤) amino acid identities with each other, 35.4-39.6% and 62.8-65.9% with factor I of mammals and banded houndshark (Triakis scyllium), respectively. The modular structure of the GcIf is similar to that of mammals with one notable exception, the presence of a novel shark-specific sequence between the leader peptide (LP) and the factor I membrane attack complex (FIMAC) domain. The cDNA sequences differ only in the size and composition of the shark-specific region (SSR). Sequence analysis of each SSR has identified within the region two novel short sequences (SS1 and SS2) and three repeat sequences (RS1-3). Genomic analysis has revealed the existence of three introns between the leader peptide and the FIMAC domain, tentatively designated intron 1, intron 2, and intron 3 which span 4067, 2293 and 2082 bp, respectively. Southern blot analysis suggests the presence of a single gene copy for each cDNA type. Phylogenetic analysis suggests that complement factor I of cartilaginous fish diverged prior to the emergence of mammals. All four GcIf cDNA species are expressed in four different tissues and the liver is the main tissue in which expression level of all four is high. This suggests that the expression of GcIf isotypes is tissue-dependent.

DOI： 10.1016/j.molimm.2009.04.002

リポジトリ公開URL： http://hdl.handle.net/2324/4776954

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Background: The chemokine family plays important roles in cell migration and activation. In humans, at least 44 members are known. Based on the arrangement of the four conserved cysteine residues, chemokines are now classified into four subfamilies, CXC, CC, XC and CX3C. Given that zebrafish is an important experimental model and teleost fishes constitute an evolutionarily diverse group that forms half the vertebrate species, it would be useful to compare the zebrafish chemokine system with those of mammals. Prior to this study, however, only incomplete lists of the zebrafish chemokine genes were reported. Results: We systematically searched chemokine genes in the zebrafish genome and EST databases, and identified more than 100 chemokine genes. These genes were CXC, CC and XC subfamily members, while no CX3C gene was identified. We also searched chemokine genes in pufferfish fugu and Tetraodon, and found only 18 chemokine genes in each species. The majority of the identified chemokine genes are unique to zebrafish or teleost fishes. However, several groups of chemokines are moderately similar to human chemokines, and some chemokines are orthologous to human homeostatic chemokines CXCL12 and CXCL14. Zebrafish also possesses a novel species-specific subfamily consisting of five members, which we term the CX subfamily. The CX chemokines lack one of the two N-terminus conserved cysteine residues but retain the third and the fourth ones. (Note that the XC subfamily only retains the second and fourth of the signature cysteines residues.) Phylogenetic analysis and genome organization of the chemokine genes showed that successive tandem duplication events generated the CX genes from the CC subfamily. Recombinant CXL-chr24a, one of the CX subfamily members on chromosome 24, showed marked chemotactic activity for carp leukocytes. The mRNA was expressed mainly during a certain period of the embryogenesis, suggesting its role in the zebrafish development. Conclusion: The phylogenic and genomic organization analyses suggest that a substantial number of chemokine genes in zebrafish were generated by zebrafish-specific tandem duplication events. During such duplications, a novel chemokine subfamily termed CX was generated in zebrafish. Only two human chemokines CXCL12 and CXCL14 have the orthologous chemokines in zebrafish. The diversification observed in the numbers and sequences of chemokines in the fish may reflect the adaptation of the individual species to their respective biological environment.

DOI： 10.1186/1471-2164-9-222

リポジトリ公開URL： http://hdl.handle.net/2324/4778000

記述言語：英語   掲載種別：研究論文（学術雑誌）  

The clonal triploid ginbuna crucian carp Carassius auratus langsdorfii, a naturally occurring gynogenetic fish, is a useful model for studying T-cell-mediated immunity. CD4, a T-cell receptor (TCR) coreceptor, is a membrane-bound glycoprotein found on helper T-cells, and assists in the binding of major histocompatibility complexes. In the present study, full-length cDNAs encoding the CD4 molecule from the S3n strain of ginbuna crucian carp were cloned and characterized. 5′-rapid amplification of cDNA ends (RACE) and 3′-RACE yielded two distinct cDNA clones of CD4 homolog from the ginbuna, and these sequences share 95% identity at the amino acid level. These ginbuna CD4 molecules consisted of a signal peptide, immunoglobulin superfamily (IgSf) like domains, a transmembrane domain, and a cytoplasmic domain similar to other known CD4. A tyrosine protein kinase p56lck binding motif is conserved in the cytoplasmic tail of ginbuna CD4. Phylogenetic tree analysis indicated that ginbuna CD4 sequences are closely related to CD4L-1 from other fish species. Expression of ginbuna CD4 mRNA was detected in the gill, thymus, head kidney, trunk kidney and peripheral blood leukocytes, indicating that its expression pattern is similar to that of ginbuna TCRβ mRNA. The results suggest that ginbuna CD4 sequences are useful as molecular probes for helper T-cells.

DOI： 10.1111/j.1444-2906.2008.01530.x

リポジトリ公開URL： http://hdl.handle.net/2324/4778001

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Factor B and C2 are serine proteases that provide the catalytic subunits of C3 and C5 convertases of the alternative (AP) and classical (CP) complement pathways. Two Bf/C2 cDNAs, GcBf/C2-1 and -2 (previously referred to as nsBf/C2-A and nsBf/C2-B), were isolated from the nurse shark, Ginglymostoma cirratum. GcBf/C2-1 and -2 are 3364 and 3082 bp in length and encode a leader peptide, three CCPs, one VWFA, the serine protease domain and have a putative factor D/C1s/MASP cleavage site. Southern blots show that there might be up to two Bf/C2-like genes for each of the two GcBf/C2 isoforms. GcBf/C2-1 and -2 are constitutively expressed, albeit at different levels, in all nine tissues examined. Expression in erythrocytes is a novel finding. Structural analysis has revealed that the localization of glycosylation sites in the SP domain of both putative proteins indicates that the molecular organization of the shark molecules is more like C2 than factor B. Phylogenetic analysis indicates that GcBf/C2-1 and -2 and TrscBf of Triakis scyllia (another shark species) originated from a common ancestor and share a remote ancestor with Bf and C2 of mammals and bony fish.

DOI： 10.1016/j.dci.2007.03.001

リポジトリ公開URL： http://hdl.handle.net/2324/4776912

記述言語：英語   掲載種別：研究論文（学術雑誌）  

The lectin pathway of complement is considered to be the most ancient complement pathway as inferred from identification of ancient homologs of mannose-binding lectin (MBL) and MBL-associated serine proteases (MASPs) in some invertebrates. MBL homologs with galactose selectivity and an MASP3-like sequence also occur in bony fish, linking the evolution of the lectin complement pathway from invertebrates to higher vertebrates. However, these cannot be considered authentic complement components until confirmatory functional evidence is obtained. Here, we report the isolation and characterization of two MBL homologs from a cyprinid teleost, the common carp, Cyprinus carpio. One, designated GalBL, corresponds to the MBL-like molecule with the galactose specificity. The other is an authentic MBL with mannose specificity. Both were found to associate with a serine protease that cleaves native human C4 into C4b but not C4i with a hydrolyzed thioester. Molecular cloning and phylogenetic analysis revealed this C4-activating protease to be carp MASP2, indicating that MASP2 arose before the emergence of bony fish. Database mining of MBL-like genes reveals that MBL and GalBL genes are arranged in tandem in the zebrafish genome and that both lectins are conserved in the distantly related puffer fish. These results imply that bony fish have developed a diverged set of MBL homologs that function in the lectin complement pathway.

DOI： 10.4049/jimmunol.177.8.5471

リポジトリ公開URL： http://hdl.handle.net/2324/4778002

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Our previous studies have demonstrated that virus-specific cell-mediated cytotoxicity of sensitized leukocytes can be induced using clonal ginbuna crucian carp and their syngeneic cell lines. In the present study, we attempt to determine if virus-specific cytotoxic cell populations of fish express CD8α and TCRβ genes. Leukocytes from ginbuna crucian carp were separated into four fractions by immunomagnetic separation and density gradient centrifugation: Fraction A, leukocytes with a density of 1.08 g/ml (primarily lymphocytes); Fraction B, sIg-negative leukocytes with density of 1.08 g/ml; Fraction C, sIg-positive cells (primarily B-lymphocytes); Fraction D, leukocytes with a density of 1.08-1.09 g/ml (primarily neutrophils). Leukocytes in all fractions from uninfected fish do not exhibit cytotoxic activity against virus-infected syngeneic cells and weakly express CD8α and TCRβ mRNAs. In contrast, leukocytes in fractions A and B from virus-infected fish exhibit a high level of cytotoxic activity and strongly express CD8α and TCRβ mRNAs. In addition, mRNA expressions of CD8α and TCRβ in effector cells are upregulated by cocultivation with virus-infected target cells but not uninfected ones. The present study suggests that fish possess virus-specific cytotoxic cells with phenotype and gene expression pattern similar to those of CTLs in mammals.

DOI： 10.1016/j.virol.2006.01.019

リポジトリ公開URL： http://hdl.handle.net/2324/4777909

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Duplication and diversification of several complement components is a striking feature of bony fish complement systems. It gives an interesting insight into an evolutionary strategy for the possible enhancement of the repertoire of innate immunity. The present study is aimed at examining diversity in bony fish C4, a member of the thioester-containing complement components. Two diverged cDNA sequences sharing only ∼32% identity at the amino acid level were isolated from the common carp and designated C4-1 and C4-2. C4-1 and C4-2 share a number of C4-like structural signatures, such as the thioester site and a disulfide-linked three-chain structure. Interestingly, they differ at the residue corresponding to the thioester-catalytic histidine, as seen in the human C4A and C4B isotypes, suggesting their distinct substrate specificities in the binding reaction of the thioester. Phylogenetic analysis indicates that the divergence of C4-1 and C4-2 predated the separation of the cartilaginous and bony fish lineages. Genomic Southern hybridization suggests the presence of single copy genes each encoding C4-1 and C4-2 in the carp genome. An activation fragment, C4a, was shown to be released from each isotype in carp serum activated via the classical aad/or lectin pathways. Synthetic peptides representing a putative C2 binding site on C4-1 and C4-2 inhibited the classical pathway-mediated hemolytic activity of carp seram in a dose-dependent manner. The results suggest that C4-1 and C4-2 represent two major lineages of C4 that are present in carp serum, have distinct binding specificities, and are functional in the classical/lectin pathways of complement activation.

リポジトリ公開URL： http://hdl.handle.net/2324/4778003

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Three isotypes of immunoglobulin (Ig) light (L) chain, designated L1A, L1B, and L3, have been characterised in the common carp (Cyprinus carpio L.) to date. In this paper the molecular cloning of a fourth IgL isotype in carp, designated L2, is described. A reverse transcription-polymerase chain reaction (RT-PCR) method including 5′- and 3′-RACE was used to isolate carp L2 cDNA clones. The VL sequences could be divided into two distinct VL families, designated VL2-1 and VL2-2, most similar to rainbow trout (68% similarity) and zebrafish (78%) VL2 amino acid sequences, respectively. The CL amino acid sequences showed the highest similarity to zebrafish L2 (80%), and contained the characteristic cysteines necessary for intradomain or interchain disulphide bridges as did the VL sequences. Neither the VL nor CL sequences demonstrated such a high similarity to the other carp IgL isotypes, L1A, L1B, and L3. For JL segments, sequence variations appeared to be confined to a few positions. In the course of 5′- and 3′-RACE, cDNA clones containing recombination signal sequence (RSS), representatives of IgL sterile transcripts, were obtained. Southern blot analyses suggested that the locus of carp L2 has a cluster-like organisation. Phylogenetic analyses showed that both carp VL2 and CL2 amino acid sequences highly clustered with other teleost L2 sequences.

DOI： 10.1016/j.fsi.2003.06.002

リポジトリ公開URL： http://hdl.handle.net/2324/4776938

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

The complement component C5 plays important roles in inflammatory responses and complement-mediated cytolysis. In bony fish, although C5 has been identified at the DNA or the protein level in trout, carp and gilthead seabream, only partial C5 sequences are available. The present study was designed to obtain the complete primary structure of C5 from the common carp (Cyprinus carpio) and to examine its possible structural diversity. Reverse-transcribed polymerase chain reaction amplification from carp hepatopancreatic RNA resulted in isolation of six distinct C5-like cDNA segments, which were grouped into two divergent types (type I and type II). Using two sequences representative of the two types as probes, two distinct full-length cDNA clones (C5-1 and C5-2) were isolated, in addition to a truncated isoform of C5-1 (C5-1'). The deduced amino acid sequences of C5-1 and C5-2 share 83% identity and predict a typical two-chain structure of the mature protein that lacks the thioester bond, as in C5 from other animals. Southern hybridization of genomic DNA suggested the presence of multiple genes encoding C5-type I and a single gene encoding C5-type II. Interestingly, carp C5-type I contains novel subtypes like C5-1 that have a histidine instead of the well-conserved arginine at the cleavage site for the C5 convertase, both in the complete and truncated forms. Northern blotting analysis suggested that C5-type I and C5-type II are mainly expressed in hepatopancreas, and the expression levels are significantly increased by stimulating carp with lipopolysaccharide or β-1,3-glucan. Possible functional divergence among the C5 isotypes in carp is discussed.

DOI： 10.1007/s00251-002-0528-7

リポジトリ公開URL： http://hdl.handle.net/2324/4776906

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Factor I is a novel serine protease that regulates complement activation. Here we report the complete primary structure of two isotypic factor Is isolated from the common carp (Cyprinus carpio), a pseudotetraploid teleost. A carp hepatopancreas cDNA library was screened using two RT-PCR-amplified cDNA fragments encoding part of the carp factor I-like serine protease domain. Two distinct cDNA clones, designated FI-A and FI-B, were isolated. Their deduced amino acid sequences share 75.2% identity with each other. FI-A has a typical factor I-like domain organization composed of two disulfide-linked polypeptides (H-chain and L-chain). On the other hand, FI-B contains a novel sequence of 115 amino acids inserted at the N-terminus of the H-chain. Genomic Southern hybridization suggests that FI-A and FI-B are encoded by distinct genes in the carp genome. Expression analysis by RT-PCR revealed that the major site of FI-A expression is the ovary, whereas FI-B expression is detected mainly in the hepatopancreas at a level higher than that of FI-A. The present data, taken together, suggest that carp have duplicated genes coding for factor I, and FI-B with the novel insertion plays a dominant role in the complement system. In addition, homology search of the fugu genome database using the carp FI-A and FI-B sequences identified a putative fugu factor I gene, which has an exon/intron organization different from that of the human orthologue.

DOI： 10.1007/s00251-002-0518-9

リポジトリ公開URL： http://hdl.handle.net/2324/4776904

記述言語：英語   掲載種別：研究論文（学術雑誌）  

In the course of suppression subtractive hybridisation between sodium alginate-induced peritoneal cells (SA-PC) and normal head kidney cDNAs in common carp (Cyprinus carpio), a cytokine-like cDNA clone was found. The clone, named M17, contains a 1600bp nucleotide sequence that encodes a 215 amino acid putative protein that would have a pI of 9.01 and would include a 33 amino acid signal peptide. The 3′ untranslated region has seven ATTTA mRNA destabilising motifs that are common in cytokines and oncogenes. In a BLASTP search, M17 was most similar to chicken ciliary neurotrophic factor (CNTF) with 25% amino acid identity, followed by mammalian CNTF, cardiotrophin-1 and leukemia inhibitory factor (LIF) all of which belong to the IL-6 subfamily. However, M17 has some differences with CNTF in that CNTF has no signal sequence, the gene organisation of M17 is three exons and two introns, whereas that of CNTF is two exons and one intron, M17 has seven cysteines while CNTF has one cysteine, and M17 mRNA is detected in peripheral blood leukocytes as well as brain, whereas CNTF is expressed only in the nervous system. Compared to other members in the IL-6 subfamily cytokines, M17's cysteine positions and gene organisation are similar to those of oncostatin M and LIF, although amino acid identities are only 15-17%. Southern hybridisation suggested that M17 is a single copy gene. SA-PC showed significantly higher M17 mRNA levels than normal head kidney cells, which are considered to be a source of the SA-PC, indicating that M17 is inducible by inflammatory stimulation.

DOI： 10.1016/S0145-305X(02)00074-5

リポジトリ公開URL： http://hdl.handle.net/2324/4778008

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Opsonization is a crucial function of the complement system in immune clearance. To identify receptors responsible for the complement-mediated opsonization in teleost, head kidney leukocytes from β-1,3-glucan-injected and non-injected common carp (Cyprinus carpio) were subjected to a rosette-forming assay with fixed sheep erythrocytes (Ef). Leukocyte fractions rich in granulocytes and macrophages from the glucan-injected carp had an ability to form the rosette with complement-coated Ef ut the corresponding fractions from non-injected fish did not, suggesting that β-1,3-glucan elicits expression of complement receptors on those cells. While sensitization of Ef with antibodies slightly elevated the rosette formation, complement-coated Ef showed a significant increase in rosette formation and this formation was inhibited with anti-carp C3. Rosette formation was also inhibited by EDTA but not by EGTA. These results suggest that complement-dependent opsonization in carp is mediated by CS-receptors analogous to mammalian complement receptor type 3 (CD11b/CD18 or integrin αM/β2), which recognizes the iC3b fragment in a divalent cation-dependent manner.

DOI： 10.1046/j.1444-2906.2003.00709.x

リポジトリ公開URL： http://hdl.handle.net/2324/4778007

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s00251-002-0447-7

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1089/10799900252952190

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1897/1551-5028(2002)021<1229:PAIOAT>2.0.CO;2

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/S0145-305X(01)00083-0

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/S0145-305X(00)00057-4

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1006/fsim.2000.0331

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s002510000216

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   出版者・発行元：Fish and Shellfish Immunology  

There are numerous fish diseases that affect the central nervous system. However, few studies have investigated the immune cells and immunological responses of fish brains. Meanwhile, microglial cells, as the brain's first line of defense, play a vital role in neuroimmunology. Furthermore, CD83 is a co-stimulatory protein that regulates immunological responses and the activation of dendritic cells and macrophages. Although CD83 expression has been linked to the initial activation of microglia in various disease scenarios in mammals, its role in teleost microglial biology remains unclear. In a recent investigation, we discovered that Ginbuna crucian carp (Gb) contains two CD83 homologs (GbCD83 and GbCD83-L). In this study, we used modified procedures of mouse-based macrophage culture from the brain and kidney to identify that GbCD83-L is highly expressed by the brain microglia-like cells and kidney-resident macrophages (KRMs) at both the protein and gene levels. Interestingly, GbCD83-L was considerably elevated in the microglia-like cells and KRMs after 24 h of lipopolysaccharide stimulation. These findings provide the first evidence of CD83 as a potential marker for active microglia and KRMs in teleosts, thus making it a crucial regulator in fish neuroimmunology and a candidate for future immunomodulatory applications in aquaculture.

DOI： 10.1016/j.fsi.2024.110038

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PubMed

記述言語：英語   出版者・発行元：Developmental and Comparative Immunology  

CD83 is a costimulatory molecule of antigen-presenting cells (APCs) that plays an important role in eliciting adaptive responses. It is also a well-known surface protein on mature dendritic cells (DCs). Furthermore, monocytes have been reported to differentiate into macrophages and monocyte-derived dendritic cells, which play an important role in innate immunity. CD83 expression affects the activation and maturation of DCs and stimulates cell-mediated immune responses. This study aims to reveal the CD83 expression during monocyte differentiation in teleosts, and the CD83 homologs evolutionary relationship. This study found two distinct CD83 homologs (GbCD83 and GbCD83-L) in ginbuna crucian carp (Gb) and investigated the evolutionary relationship among GbCD83 homologs and other vertebrates and the gene and protein expression levels of the homologs during 4 days of monocyte culture. The phylogenetic tree showed that the two GbCD83 homologs are classified into two distinct branches. Interestingly, only ostariophysians (Gb, common carp, rohu, fathead minnow and channel catfish), but not neoteleosts, mammals, and others, have two CD83 homologs. Morphological observation and colony-stimulating factor-1 receptor (CSF-1R), CD83, CD80/86, and CCR7 gene expressions illustrated that there is a differentiation of monocytes isolated from peripheral blood leukocytes after 4 days. Specifically, gene expression and immunocytochemistry revealed that GbCD83 is mainly expressed on monocytes at the early stage of cell culture, whereas GbCD83-L is expressed in the latter stage. These findings provided the first evidence of differential expression of CD83 homologs during monocytes differentiation in teleost.

DOI： 10.1016/j.dci.2024.105212

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PubMed

出版者・発行元：東京医学社  

DOI： 10.24479/kd.0000001385

CiNii Research

記述言語：英語   出版者・発行元：Zoological Science  

Sexual dimorphism allows species to meet their fitness optima based on the physiological availability of each sex. Although intralocus sexual conflict appears to be a genetic constraint for the evolution of sex-specific traits, sex-linked genes and the regulation of sex steroid hormones contribute to resolving this conflict by allowing sex-specific developments. Androgens and their receptor, androgen receptor (Ar), regulate male-biased phenotypes. In teleost fish, ar ohnologs have emerged as a result of teleost-specific whole genome duplication (TSGD). Recent studies have highlighted the evolutionary differentiation of ar ohnologs responsible for the development of sexual characteristics, which sheds light on the need for comparative studies on androgen regulation among different species. In this review, we discuss the importance of ar signaling as a regulator of male-specific traits in teleost species because teleost species are suitable experimental models for comparative studies owing to their great diversity in male-biased morphological and physiological traits. To date, both in vivo and in vitro studies on teleost ar ohnologs have shown a substantial influence of ars as a regulator of male-specific reproductive traits such as fin elongation, courtship behavior, and nuptial coloration. In addition to these sexual characteristics, ar substantially influences immunity, inducing a sex-biased immune response. This review aims to provide a comprehensive understanding of the current state of teleost ar studies and emphasizes the potential of teleost fishes, given their availability, to find molecular evidence about what gives rise to the spectacular diversity among fish species.

DOI： 10.2108/zs230098

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PubMed

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Web of Science

記述言語：英語   出版者・発行元：Elsevier  

CD8+ cytotoxic T cells (CTLs) are a main cellular component of adaptive immunity. Our previous research has shown that CD8+ cells demonstrate spontaneous cytotoxic activity against the parasite Ichthyophthirius multifiliis in ginbuna crucian carp, suggesting that CD8+ cells play an important role in innate immunity. Herein, we investigated the molecules and cellular signal pathways involved in the cytotoxic response of ginbuna crucian carp. We considered non-specific cytotoxic receptor protein-1 (NCCRP-1) as candidate molecule for parasite recognition. We detected NCCRP-1 protein in CD8+ cells and the thymus as well as in other cells and tissues. CD8+ cells expressed mRNA for NCCRP-1, Jak2, and T cell-related molecules. In addition, treatment with a peptide containing the presumed antigen recognition site of ginbuna NCCRP-1 significantly inhibited the cytotoxic activity of CD8+ cells against the parasites. The cytotoxic activity of CD8+ cells was significantly inhibited by treatment with the JAK1/2 inhibitor baricitinib. These results suggest that teleost CTLs recognize I. multifiliis through NCCRP-1 and are activated by JAK/STAT signaling.

DOI： 10.1016/j.fsi.2023.108904

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CiNii Research

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Scientific Reports  

Infectious disease is one of the most serious problems in the aquaculture industry for ornamental or
edible fsh. This study attempted to develop a new device for preventing an aquatic bacterial disease,
ulcer disease, caused by Aeromonas salmonicida (As), using “afnity silk”. Afnity silk is a silk proteincontaining fbroin L-chain (FibL) fused to the single-chain variable fragment (scFv). It can be easily
processed into diferent formats such as fbers, gels, sponges, or flms. A transgenic silkworm that
could express a cDNA construct containing FibL fused to an scFv derived from a monoclonal antibody
(MAb) against As was successfully generated. An enzyme-linked immunosorbent assay was used to
detect As by employing 96-well plates coated with scFv-conjugated afnity silk. As could be captured
efciently by glass wool coated with afnity silk in the column. Furthermore, the air-lift water flter
equipped with the afnity silk-coated wool could considerably reduce the concentration of As in water
and was estimated to have sufcient ability to trap a lethal dose of As. These fndings show that the
“afnity silk flter” is a potential device for the prophylaxis of aquatic animal diseases.

DOI： 10.1038/s41598-022-13408-6

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CiNii Research

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Ichthyophthirius multifiliis is a ciliated protozoan parasite and is known to infect many freshwater teleosts. Characterizing the immune system in epithelial tissues, where the parasites penetrate and settle, is key to understanding host–parasite interactions. This study examined local immune responses in vivo to the infective stage (theront and trophont) of the parasites using intra-fin administration, which has been developed to analyze in vivo immune responses using fish fin. CD8α+ and CD4+ T-cell compositions were increased significantly in the fin cavity injected with theront or trophont antigens. The expression of GATA-3 and T-bet mRNA, which regulate differentiation of helper T-cells, was upregulated significantly in leukocytes from the trophont antigen–injected site. In contrast, the percentages of macrophages and neutrophils, which are innate immunity components, were decreased significantly in the injection sites. These results suggest that I. multifiliis antigens inhibit the migration of macrophages and neutrophils, and T-cells are the first responders to I. multifiliis. Thus, to better understand the interaction of host immunity and I. multifiliis, further studies should focus on exploring the inhibitory factors from I. multifiliis or examining innate functions of teleost T-cells.

DOI： 10.1016/j.fsi.2021.08.013

記述言語：英語   掲載種別：研究論文（学術雑誌）  

A mixed culture of probiotics, one from the genus Bacillus and one lactic acid bacterium (LAB), was developed to be used as a feed additive for enhancing growth, innate immunity and disease resistance in Pangasius bocourti. From our earlier work, three probiotic Bacillus species, Bacillus siamensis B44v, Bacillus sp. B51f and Bacillus aerius B81e, and three probiotic LABs, Streptococcus lutetiensis L7c, Lactiplantibacillus paraplantarum (synonym. Lactobacillus paraplantarum) L34b-2 and Lactiplantibacillus plantarum (synonym. Lactobacillus plantarum) L42g, were selected for comparison. These bacteria, which express probiotic properties including bacteriocin-like activity against Aeromonas hydrophila, were subjected to in vivo screening in hybrid catfish (Clarias macrocephalus × Clarias gariepinus). A 30-day feed-trial followed by a challenge test in screening experiments resulted in the prominent B. aerius B81e and L. paraplantarum L34b-2 being selected. A mixture of these bacteria was added to a diet for P. bocourti. After 60-day feeding, the fish fed with mixed probiotics had weight gain, specific growth rate and feed conversion ratio improved significantly (p < 0.01) when compared to the control. Both humoral and cellular immunity were significantly higher in probiotic-fed fish. Following the 60-day feeding experiment, P. bocourti fed with the diet containing mixed probiotics had a higher survival rate than the control fish after injection with a virulent A. hydrophila. It can be concluded that a combination of B. aerius strain B81e and L. paraplantarum strain L34b-2 markedly improved growth performance, innate immunity and disease resistance of P. bocourti.

DOI： 10.1016/j.jbiosc.2021.06.006

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Cytotoxic T cells are known to have the ability to kill microbe-infected host cells, which makes them essential in the adaptive immunity processes of various vertebrates. In this study, we demonstrated innate cell-mediated cytotoxicity of CD8+ T cells against protozoan parasites found in the ginbuna crucian carp. When isolated effector cells such as CD8+, CD4+ (CD4-1+), or CD8− CD4− (double-negative, DN), from naïve ginbuna crucian carp were co-incubated with target parasites (Ichthyophthirius multifiliis), CD8+ cells from the kidney and gill showed the highest cytotoxic activity. On the other hand, DN cells, which include macrophages and CD4− CD8− lymphocytes, showed the lowest cytotoxic activity against I. multifiliis. Additionally, the cytotoxic activity of CD8+ cells was found to significantly decrease in the presence of a membrane separating the effector cells from I. multifiliis. Furthermore, the serine protease inhibitor 3,4-dichloroisocoumarin and perforin inhibitor concanamycin A significantly inhibited the cytotoxic activity of CD8+ cells. These results demonstrate that CD8+ T cells of ginbuna crucian carp can kill extracellular parasites in a contact-dependent manner via serine proteases and perforin. Therefore, we conclude that CD8+ T cells play an essential role in anti-parasite innate immunity of teleost fish.

DOI： doi.org/10.1016/j.dci.2020.103886

リポジトリ公開URL： http://hdl.handle.net/2324/4113184

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Probiotics have increasingly gained interest as alternatives to antibiotics in controlling infectious diseases in aquaculture. This research aimed to isolate and evaluate lactic acid bacteria (LAB) for potential use as probiotics for Pangasius bocourti. A total of 656 LAB isolates were obtained from fish gut and fermented beef samples, of which 367 isolates displayed antagonistic activity against Aeromonas hydrophila FW52 and/or Streptococcus agalactiae F3S which were employed as indicator fish pathogens. Among these antagonistic LAB, only 18 isolates produced bacteriocin-like activity. These were further evaluated in vitro for other probiotic properties. After in vitro screening, the isolate L34b-2 obtained from Thai indigenous fermented beef was chosen as a suitable probiotic LAB for in vivo studies based on its remarkable probiotic characteristics. These included broad-spectrum bacteriocin-like activity, tolerance to simulated gastrointestinal tract conditions, mucin adhesion ability, co-aggregation ability with fish pathogens tested, non-blood hemolysis, antibiotic susceptibility and protease enzyme production. Identification of the isolate L34b-2 using conventional methods together with 16S rDNA analysis revealed that it belonged to Lactobacillus paraplantarum. Then, the effects of 60-day-dietary administration of L. paraplantarum L34b-2 on fish growth, innate immune responses and disease resistance were investigated in Pangasius bocourti. In feed-trial studies, after feeding for both 30 and 60 days it was found that the Pangasius fish fed with basal diet containing 107 CFU g−1 L. paraplantarum L34b-2 had significantly (p < 0.05) higher weight gain and specific growth rate, and the lowest feed conversion ratio, compared to the control fish that received only basal diet. For immune response studies, it was found that the fish fed with the probiotic strain L34b-2, when compared to the control fish, had significantly higher lysozyme, alternative complement and bactericidal activities but no significant differences (p > 0.05) in phagocytic and respiratory burst activities. After 60-day feeding, fish were subjected to challenge tests by intraperitoneal injection with 106 CFU of a virulent Aeromonas hydrophila FW52 and rearing was continued for two more weeks. As expected, fish that received L34b-2 had a significantly higher (p < 0.05) post challenge survival rate than the control group. These results clearly show that L. paraplantarum L34b-2 displayed beneficial effects on P. bocourti as a probiotic by which it not only promoted growth but also enhanced innate immunity and disease resistance against A. hydrophila.

DOI： doi.org/10.1016/j.aquaculture.2020.735878

リポジトリ公開URL： http://hdl.handle.net/2324/4776909

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Although a previous study using ginbuna crucian carp suggested that cell-mediated immunity can be induced by the oral administration of inactivated viruses, which are exogenous antigens, there is no direct evidence that CD8
⁺
cytotoxic T cells (CTLs) in teleost fish are generated by vaccination with exogenous antigens. In the present study, we investigated whether antigen-specific CD8
⁺
CTLs in ginbuna crucian carp can be elicited by intestinal immunization with an exogenous antigen without any adjuvant. The IFNγ-1 and T-bet mRNA expressions were up-regulated in intestinal leukocytes following the administration of formalin-inactivated crucian hematopoietic necrosis virus (FI-CHNV), whereas the down-regulation of these genes was observed in kidney leukocytes. Furthermore, an increase in the percentage of proliferating CD8
⁺
cells was detected in the posterior portion of the hindgut, suggesting that the virus-specific CTLs are locally generated in this site. In addition, cell-mediated cytotoxicity against CHNV-infected syngeneic cells and the in vivo inhibition of viral replication were induced by immunization with FI-CHNV. Unexpectedly, intraperitoneal immunization with FI-CHNV induced a type I helper T cell (Th1)-response in the intestine, but not in the kidney; however, its effect was slightly lower than that reported after intestinal immunization. These findings suggest that the posterior portion of the intestine is an important site for generating virus-specific CTLs by vaccination with the inactivated vaccine.

DOI： 10.1016/j.dci.2018.12.009

リポジトリ公開URL： http://hdl.handle.net/2324/2552964

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3147/jsfp.53.36

リポジトリ公開URL： http://hdl.handle.net/2324/4784368

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Infectious diseases have been found to be a major cause of mortality in fish hatcheries. Probiotics have been introduced to replace antibiotics commonly used for treatment of bacterial infection in aquaculture. This study was conducted to isolate, screen, and evaluate the probiotic Bacillus spp. for potential use as a feed supplement to enhance fish growth, disease resistance and innate immunity of Pla-mong Pangasius bocourti. Bacillus aerius strain B81e was selectively isolated from the intestine of healthy catfish and chosen based on its probiotic properties both in vitro and in vivo. This bacterium produced a bacteriocin-like substance and exhibited a broad-spectrum antibacterial activity inhibiting both Gram-positive and Gram-negative bacteria especially the fish pathogens Aeromonas hydrophila and Streptococcus agalactiae. The susceptibility to all 8 antibiotics tested implies that it is unlikely to be an antibiotic-resistant bacterium. B. aerius strain B81e possessed interesting adhesion properties as shown by its high percentages of hydrophobicity, auto-aggregation, co-aggregation with fish pathogens A. hydrophila FW52 and S. agalactiae F3S and mucin binding. The strain B81e survived simulated gastrointestinal conditions, producing protease and lipase but not β-haemolysin. The study also evaluated the effects of dietary supplementation with strain B81e on growth performance, innate immunity, and the disease resistance of P. bocourti against A. hydrophila infection. Fish with a mean body weight of 69 g were fed strain B81e at 0 (control) and 10⁷ CFU g⁻¹ feed (test) for 60 days. Various growth and immune parameters were examined at 30 and 60 days post-feeding. Fish were challenged with A. hydrophila 60 days post-feeding and mortalities were recorded over 14 days post-infection. Results showed that the administration of strain B81e for 60 days had significant effects (p < 0.05) on weight gain, specific growth rate and feed utilization efficiency of P. bocourti. Dietary administration of strain B81e increased the serum lysozyme and bactericidal activities of P. bocourti significantly throughout the experimental period whereas the alternative complement, phagocytic and respiratory burst activities were significantly (p < 0.05) higher in the test fish compared to the control fish after 60 days of feeding. In addition, the fish fed a strain B81e supplemented diet had a significantly higher (p < 0.05) post-challenge survival rate than the control fish. The results in this study indicate that B. aerius B81e has beneficial effects on growth performance, innate immunity and disease resistance of P. bocourti. This is the first report on the probiotic roles of B. aerius in aquaculture.

DOI： 10.1016/j.fsi.2017.11.032

リポジトリ公開URL： http://hdl.handle.net/2324/4776943

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.rvsc.2015.01.016

リポジトリ公開URL： http://hdl.handle.net/2324/4777907

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.dci.2013.12.008.

リポジトリ公開URL： http://hdl.handle.net/2324/4776935

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Hagfish C1q (HaC1q) was identified and characterized as a pattern-recognition molecule (PRM) in the hagfish complement system. The serum from hagfish, Eptatretus burgeri, was applied to a GlcNAc-agarose column and eluted sequentially with GlcNAc and EDTA. Four (31, 27, 26, and 19 kDa) and one (26 kDa) proteins were detected as bound molecules in the GlcNAc- and the EDTA-eluates, respectively. Among these, the 26 kDa protein from the EDTA eluate was found to be a homologue of mammalian C1q through cDNA analysis. HaC1q had an ability to bind to various microbes in a Ca2+-dependent manner and its target ligands on the microbes were lipopolysaccharide, lipoteichoic acid, and peptidoglycan. The binding of HaC1q to GlcNAc-agarose was not inhibited by an excess amount of monosaccharide such as GlcNAc. While HaC1q bound to Sepharose 6B with a matrix of GlcNAc-agarose (polymer of agarobiose), it did not bind to Sepharose 4B that contained lower concentration of agarobiose than Sepharose 6B. Therefore, the target of HaC1q on GlcNAc-agarose was concluded to be agarobiose and high density of the target moiety seemed to be required for the stable binding. This finding was in accordance with the known behavior of other lectins involved in the complement system. We have concluded that HaC1q recognizes agarobiose-like structures present on the surface of microbes and acts as a pattern-recognition molecule in the process for elimination of invading microbes.

DOI： https://doi.org/10.1016/j.dci.2013.10.009

リポジトリ公開URL： http://hdl.handle.net/2324/4776934

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Previous studies have suggested that anti-viral cytotoxic effector cells induced by infection with a sublethal dose of crucian carp hematopoietic necrosis virus (CHNV) correspond to mammalian cytotoxic T-lymphocytes (CTLs), because the mRNA expression patterns of the effector cells are similar to those of mammalian CTLs. To further characterize the effector population in cell-mediated cytotoxic (CMC) activity, we isolated the effector cells using an anti-CD8α monoclonal antibody, a density gradient and plastic adherence. As expected, the purified CD8α-positive cells killed CHNV-infected cells, indicating that the fish CTLs are one of anti-viral effector cells similar those to in mammals. However, it appeared that cytotoxic cells other than CTLs were the dominant effectors, because CTL-depleted peripheral blood leukocytes (PBL) exhibited significant cytotoxic activity against CHNV-infected cells. In addition, the adoptive transfer of CTL-depleted PBL provided as efficient protection against CHNV-infection as the transfer of PBL containing CTLs. Further analyses showed that sIg/CD8α-negative cells and monocyte-enriched effectors possessed activities that were comparable to or were higher than that of CD8α-positive cells, suggesting that natural killer (NK)-like cells and monocytes are among the dominant effector cells. CMC inhibition assays with concanamycin A suggested that CTLs and CD8α-negative lymphocytes lysed virus-infected cells by a perforin-based cytotoxic pathway. These results indicate that CMC induced by viral-infection is executed by not only CTLs but monocytes and CD8/IgM-negative lymphocytes.

DOI： 10.1016/j.dci.2012.11.001

リポジトリ公開URL： http://hdl.handle.net/2324/4776915

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Current mouse lines efficient for human cell xenotransplantation are backcrossed into NOD mice to introduce its multiple immunodeficient phenotypes. Our positional genetic study has located the NOD-specific polymorphic Sirpa as a molecule responsible for its high xenograft efficiency: it recognizes human CD47 and the resultant signaling may cause NOD macrophages not to engulf human grafts. In the present study, we established C57BL/6.Rag2nullIl2rgnull mice harboring NOD-Sirpa (BRGS). BRGS mice engrafted human hematopoiesis with an efficiency that was equal to or even better than that of the NOD.Rag1nullIl2rgnull strain, one of the best xenograft models. Consequently, BRGS mice are free from other NOD-related abnormalities; for example, they have normalized C5 function that enables the evaluation of complement-dependent cytotoxicity of antibodies against human grafts in the humanized mouse model. Our data show that efficient human cell engraftment found in NOD-based models is mounted solely by their polymorphic Sirpa. The simplified BRGS line should be very useful in future studies of human stem cell biology.

DOI： 10.1182/blood-2012-06-440354

リポジトリ公開URL： http://hdl.handle.net/2324/4777998

記述言語：英語   掲載種別：研究論文（学術雑誌）  

CD2 family receptor (CD2f) is evolutionarily conserved and is widely expressed by various types of leukocytes. To elucidate the phylogenetic diversity of the CD2f, we characterized CD2f in teleosts using ginbuna crucian carp and zebrafish. The identified CD2f isoforms of the ginbuna carp (caauCD2f) exhibited high sequence similarity to the mammalian CD2 subsets CD48, CD244, and CD319, but it was difficult to classify them into their respective mammalian CD2f based on sequence similarity, the presence of an immunoreceptor tyrosine-based switch motif (ITSM), and phylogenetic tree analysis. Although the four caauCD2f isoforms share an extracellular domain with quite high identity (83-94% identity at the nucleic acid level), they differ in the number of ITSM motifs in their cytoplasmic tail. RT-PCR and in situ hybridization analyses showed that the caauCD2f isoforms are expressed by different cell populations, suggesting that they, like mammalian CD2f, have diverse roles. Interestingly, immunoglobulin (Ig) domain-like sequences with high identity to caauCD2fs are clustered close together within 0.6. Mbp on zebrafish chromosomes 1 and 2 (at least 8 and 35 sequences, respectively), and many pairs of the Ig domains share more than 90% identity at the amino acid level. Therefore, the teleost CD2fs with considerably high identity have been probably generated from a common ancestral Ig-domain gene by a very recent gene duplication event. These findings suggest that the identified CD2f acquired functional diversification through successive duplications together with the acquisition of ITSM.

DOI： 10.1016/j.rinim.2012.01.003

リポジトリ公開URL： http://hdl.handle.net/2324/4776958

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Clonal ginbuna crucian carp is, a naturally gynogenetic fish, and is a useful model animal for studying T-cell-mediated immunity. To gain molecular information on MHC class I molecules from this species, we have identified four types of MHC class I (caauUA-S3n, caauUF-S3n, caauZE-S3n, and caauZB-S3n) and five beta 2-microglobulin (β2m) (caauβ2m-1a, caauβ2m-1b, caauβ2m-2, caauβ2m-3a and caauβ2m-3b) by an expressed sequence tag (EST) analysis and using homology cloning with degenerated primers. Like UA class I genes in other cyprinid fish, the caauUA-S3n shows features of classical MHC class I, such as conservation of all key amino acids interacting with antigenic peptides, and ubiquitous tissue expression. A phylogenetic analysis shows that the β2m-1 and β2m-2 isoforms are clustered with those of other cyprinid fishes, while β2m-3 isoforms make a cluster that is separated from a common ancestor of salmonid and cyprinid fishes. This finding suggests that the β2m isoforms of ginbuna cruician carp comprise two lineages and may possess different functions. The MHC class I and β2m sequences from one clonal strain will facilitate our understanding of the interaction of MHC class I with β2m in teleosts.

DOI： 10.1016/j.fsi.2011.06.004

リポジトリ公開URL： http://hdl.handle.net/2324/4776940

記述言語：英語   掲載種別：研究論文（学術雑誌）  

We successfully purified Trub.TBT-bpα, a tributyltin (TBT) binding protein (bp) of the tiger puffer, Takifugu rubripes. Tiger puffer was injected intraperitoneally with TBT (1.0 mg/kg body weight) and Trub.TBT-bpα was purified from serum by ammonium sulfate fractionation, gel filtration chromatography and polyacrylamide gel electrophoresis. Gel electrophoresis revealed that the Trub.TBT-bpα has a molecular mass of approximately 48.5 kDa and contains at least 40% N-glycan. The deduced 212 amino acid sequence of the protein showed the highest identity (41%, 212 amino acid overlap and E-value: 9e−42) with TBT-binding protein type 1 (TBT-bp1) of Paralichthys olivaceus (Japanese flounder). Analysis of the gene structure of Trub.TBT-bpα suggests that this protein belongs to the lipocalin superfamily, which may be important in the accumulation and elimination of TBT. Phylogenetic analysis suggests that functionalization of TBT-bps has occurred during evolution, and that the functions of this group of proteins might be important for fish survival.

DOI： http://dx.doi.org/10.1016/j.cbpc.2010.07.009

リポジトリ公開URL： http://hdl.handle.net/2324/4776910

記述言語：英語   掲載種別：研究論文（学術雑誌）  

To characterise fish haematopoietic stem/progenitor cells, it is necessary to develop a culture system that supports proliferation and differentiation of these cells. In the present study, we established cell lines from various tissues of carp (Cyprinus carpio) and ginbuna (Carassius auratus langsdorfii). By using these cell lines, we developed a culture system in which carp haematopoietic cells proliferated and were successively passaged. Cell lines from carp thymus (KoT), carp fin (KoF1) and ginbuna thymus (GTS6 and GTS9) were newly established. In addition to these cell lines, ginbuna fin (CFS) cell lines were also used as feeder layers. Kidney haematopoietic cells co-cultured with these feeder layers proliferated rapidly and were passaged over 20 times for more than 60 days. To characterise the proliferating cells, expression of marker genes for blood cell development were analysed. In the primary culture, marker genes for myeloid/erythroid progenitors (gata1), haematopoietic stem cells (gata2), neutrophils (mpx/mpo), B-cells (IgH) and T-cells (lck, TCRβ and gata3) were detected by reverse transcriptase polymerase chain reaction (RT-PCR). Expression of most of the genes disappeared after the third passage, only T-cell marker genes were highly expressed after passages. These results indicate that multiple blood cells developed in the primary culture and then T-cell lineages dominantly proliferated after several passages.

DOI： 10.1016/j.vetimm.2009.03.007

リポジトリ公開URL： http://hdl.handle.net/2324/4777908

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Little is known about antigen-specific T-cell responses to viruses in teleosts due to a lack of a suitable experimental system using inbred or clonal animals. In the present study we have successfully induced an in vitro generation of virus-specific cytotoxic T-cells (CTLs) from isogeneic ginbuna crucian carp. Responder cells (primarily lymphocytes) from crucian carp haematopoietic necrosis virus (CHNV)-infected fish were capable of proliferating after stimulation in vitro with CHNV-infected syngeneic stimulator cells (primarily lymphocytes and macrophages). The effector cells collected 8 and 12 days after the in vitro stimulation efficiently lysed CHNV-infected syngeneic cells, but not CHNV-infected allogeneic cells or different virus (EVA)-infected syngeneic cells. Furthermore, in situ hybridization analysis showed that some effector cells binding to a CHNV-infected target were TCRβ or CD8α positive. These results provide evidence that the teleost effector cells generated in vitro correspond to virus-specific CTL and they recognize virus-infected target cells in a similar manner of mammalian counterparts.

DOI： 10.1016/j.virol.2009.04.008

リポジトリ公開URL： http://hdl.handle.net/2324/4777910

記述言語：英語   掲載種別：研究論文（学術雑誌）  

The goal of this study was to examine the significance of allelopathy by the raphidophyte Heterosigma akashiwo in a multispecies phytoplankton community in the field. Towards this aim, we sought allelochemicals of H. akashiwo, which had allelopathic effect both in laboratory experiments and in the field. As an initial step, we showed that the allelopathic effects of H. akashiwo filtrate were both species-specific and dependent upon the cell density of the target species. Secondly, we found for the first time that extracellular, high-molecular weight allelochemicals that is, polysaccharide-protein complexes (APPCs) were produced by a marine phytoplankton species, H. akashiwo. Thirdly, we indicated that the purified APPCs selectively inhibited the growth of the diatom Skeletonema costatum that is a major competitor of H. akashiwo, and thereby tended to promote the formation of monospecific H. akashiwo blooms. Furthermore, we demonstrated that the inhibitory effect of APPCs on the growth of the diatoms was determined by binding to the cell surface of the target species. Finally, we succeeded in the detection of APPCs in the field samples at concentrations exceeding their experimentally determined action threshold during the H. akashiwo bloom. Strategies for ecosystem control, including mitigation of harmful algal blooms (HABs), should take into account that red-tide organisms like H. akashiwo are already part of complex webs involving inter-specific allelopathic inhibition and ecosystem control during their dense blooms.

DOI： 10.1038/ismej.2009.24

リポジトリ公開URL： http://hdl.handle.net/2324/4777999

記述言語：英語   掲載種別：研究論文（学術雑誌）  

In fish, two types of immunoglobulin heavy chain (IGH) genes, namely, IgM and IgD, have been cloned and characterized. Recently, a new IGH isotype specific to teleosts had been identified from zebra fish, rainbow trout, and fugu. In zebra fish, the domains of this new gene are present upstream of the μ region along the IGH locus. During this study, a novel IGH chimera (IgM-IgZ) has been discovered from common carp. The cloned cDNA encodes a typical leader peptide, a variable region, two constant regions, and a secretory tail. The first constant region is made up of the CH1 domain of carp IgM, while the second constant region shares a high similarity to the C H4 domain of the IgZ from zebrafish. Southern hybridization studies of the μ and ζ domains, conducted separately, revealed the presence of at least three copies of the respective genes, and μ and ζ domains might be present on the same loci, although far apart. Expression studies of the IGH genes suggest that there is an increase in chimeric immunoglobulin gene transcription when stimulated with lipopolysaccharide.

DOI： 10.1007/s00251-005-0015-z

リポジトリ公開URL： http://hdl.handle.net/2324/4776907

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Molecular approaches were used to study thiolester-containing genes in the ascidian, Ciona intestinalis. RT-PCR, RACE and genome mining revealed that this animal expresses not only conventional alpha-2-macroglobulin (α2m) and two forms of C3 but also a gene encoding a glycosylphosphatidylinositol (GPI)-anchored α2m. Previously, GPI-anchored α2ms have been reported only for humans and mice. We propose that GPI-anchored α2ms constitute a third subgroup of the α2m superfamily and may represent an important evolutionary stage in the phylogeny of the thiolester containing proteins. Its occurrence in an ascidian shows its origin pre-dates the evolution of the vertebrates. In C. intestinalis this GPI-anchored α2m, designated Ciona α2m-GPI, is expressed in the hepatopancreas, circulating coelomic blood cells and the gut of adults. It is also expressed in 3-5 days old larvae. Its tissue distribution coupled with its sequence characteristics and unusual domain structure indicate that the encoded protein probably assists in host defence by entrapping and inhibiting proteases from micro-organisms.

DOI： 10.1016/j.molimm.2004.09.017

リポジトリ公開URL： http://hdl.handle.net/2324/4776949

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Rainbow trout (Onchorhynchus mikiss) possess two genes encoding putative leucine-rich repeat (LRR)-containing proteins similar to human TLR5. Molecular cloning of these two LRR proteins suggested the presence of a TLR5-like membrane form (rtTLR5M) and a soluble form (rtTLR5S). Here we elucidated the primary structures and the unique combinational functions of these fish versions of TLR5. The LRR regions of rtTLR5S and rtTLR5M exhibited 81% homology and relatively high (35.6 and 33.7%) homology to the extracellular domains of human TLR5 (huTLR5). Thus, two distinct genes encode the TLR5 orthologs in fish, one of which has a consensus intracellular domain (TIR). In order to test their functions, we constructed fusion proteins with the LRR region of rtTLR5S (S-chimera) or thai of rtTLR5M and the TIR of huTLR5 (M-chimera). The S- and M-chimeras expressed in HeLa or CHO cells signaled the presence of Vibrio anguillarum flagellin, resulting in NF-κB activation. rtTLR5M was ubiquitously expressed, whereas rtTLR5S was predominantly expressed in the liver. In the hepatoma cell lines of the rainbow trout RTH-149, stimulation of rtTLR5M with V. anguillarum or its flagellin allowed the up-regulation of rtTLR5S. Flagellin-mediated NF-κB activation was more significant in the presence of or simultaneous expression of rtTLR5S. Therefore, a two-step flagellin response occurred for host defense against bacterial infection in fish: (a) flagellin first induced basal activation of NF-κB via membrane TLR5, facilitating the production of soluble TLR5 and minimal acute phase proteins, and (b) the inducible soluble TLR5 amplifies membrane TLR5-mediated cellular responses in a positive feedback fashion.

DOI： 10.1074/jbc.M407634200

リポジトリ公開URL： http://hdl.handle.net/2324/4778004

記述言語：英語   掲載種別：研究論文（学術雑誌）  

A terminal degradation product (C3d) of mammalian complement component C3 plays an important role in modulation of the adaptive immune response through the interaction with complement receptor type 2 (CR2) on B cells. The present study is aimed at determining whether this is a functional bridge between the innate and adaptive immune systems in bony fish. The fragmentation of the complement component C3 in carp (Cyprinus carpio) serum, activated with zymosan, was analysed to ascertain if carp C3 also generates a mammalian C3d-like fragment under physiological conditions. A 35 kDa peptide reactive to the anti-carp C3 α-chain was detected on the zymosan particles and in the activated serum. Its N-terminal amino acid sequence identified it as carp C3d derived from the C3-H1 isoform. Another C3 isoform, C3-S, of carp was found to yield a C3d fragment at lower efficiency than C3-H1. Recombinant C3d fragments derived from C3-H1 and C3-S were produced in Escherichia coli as fusion proteins with glutathione-S-transferase (GST), and used for ligands to examine the presence of a possible CR2-like C3d receptor on carp lymphocytes. An enzyme-linked immunoadsorbent assay system, using the recombinant C3d proteins and anti-GST on a microplate to which was attached carp peripheral lymphocytes, detected a significant binding of carp C3d to the lymphocyte. The degree of binding of C3-H1-derived C3d was higher than that of C3-S-derived C3d. In addition, the binding of both ligands was inhibited by anti-C3 α-chain, but not by EDTA or EGTA, indicating that the putative C3d receptor does not require divalent cation. These properties agree well with those reported for mammalian CR2.

DOI： 10.1016/S1050-4648(03)00057-3

リポジトリ公開URL： http://hdl.handle.net/2324/4778006

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Promotion of inflammatory response is an important role of the complement system, but this kind of function is poorly documented for the lower vertebrates. Here we report chemotactic activity of purified anaphylactic fragments derived from the complement components C3, C4 and C5 of the common carp. The purified anaphylatoxins are two C5a-desArg peptides derived from the C5-I isotype, an intact form and a desArg form of C4a from C4-2 isotype, and an intact form and a desArg form of C3a from C3-H1 isoform. These were identified by N-terminal sequencing, mass spectrometry, and peptide mass fingerprinting. In the chemotaxis assay using carp kidney neutrophils, the two C5a-desArg fragments, which are probably allotypic variants, showed a potent chemotactic activity at 0.5-1nM, whereas C3a or C4a showed no significant activity. The results suggest that C3a, C4a and C5a of bony fish have functionally diverged to the state similar to their mammalian homologs.

DOI： 10.1016/j.dci.2004.01.006

リポジトリ公開URL： http://hdl.handle.net/2324/4776911

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/S0145-305X(01)00021-0

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s002510050031

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1006/fsim.2000.0294

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1002/1521-4141(200003)30:3<858::AID-IMMU858>3.3.CO;2-D

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1006/fsim.2000.0263

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1006/fsim.1999.0253

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1007/s002510050573

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/S0141-1136(97)00082-2

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1006/fsim.1998.0149

担当ページ：pp 51-61.   記述言語：日本語   著書種別：学術書

Properties of bony fish complement system.

担当ページ：Vol. 3, Part I" . pp 15-32,   記述言語：英語   著書種別：学術書

開催年月日： 2023年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：九州大学伊都キャンパス   国名：日本国  

開催年月日： 2023年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：九州大学伊都キャンパス   国名：日本国  

開催年月日： 2023年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：九州大学伊都キャンパス   国名：日本国  

開催年月日： 2023年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：別府国際コンベンションセンター   国名：日本国  

開催年月日： 2022年10月

記述言語：英語   会議種別：口頭発表（招待・特別）  

開催地：Dalian Ocean University   国名：中華人民共和国  

Mucosal immunity is regarded as a crucial first-line defense against microbial infection in teleost fish, as supported by identification of well-developed mucosa-associated lymphoid organs, secretary forms of IgM and IgT, mucus lectins and antimicrobial peptides. Innate immune effector mechanisms, however, remain to be elucidated by functional evidence. We have identified CD8+ T cells of ginbuna crucian carp as effector cells to kill extracellular parasites, Ichthyophthirius multifiliis, in a contact-dependent and MHC-independent manner via serine proteases and perforin, suggesting the ancestral innate function of teleost T cells. On the other hand, presence of components of the complement system, a major humoral innate immune factor, in teleost skin has been reported with less information on their local activation and effector functions. Recently we have detected activation of the classical complement pathway towards Aeromonas hydrophila and A. salmonicida in carp skin mucus. This activation proceeds at least until the reaction step of C3, suggesting that the mucus complement is functional as an opsonic system. Interestingly, epithelial cells express a CD46-like regulator of complement activation (Tecrem) and stimulation of Tecrem seems to enhance robustness of epithelial sheet by up-regulating tight junction proteins in carp and ginbuna crucian carp. Since C3b, an activated form of C3, is a major ligand of Tecrem, mucosal complement activation may also contribute to strengthen the epithelial barrier against microbial infection.

開催年月日： 2022年8月 - 2022年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：酪農学園大学   国名：日本国  

開催年月日： 2022年8月 - 2022年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：酪農学園大学   国名：日本国  

開催年月日： 2021年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北海道大学函館キャンパス   国名：日本国  

開催年月日： 2021年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北海道大学函館キャンパス   国名：日本国  

開催年月日： 2021年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北海道大学函館キャンパス   国名：日本国  

開催年月日： 2021年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：千里ライフサイエンスセンター（オンライン）   国名：日本国  

開催年月日： 2021年8月 - 2019年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：水産大学校（オンライン）   国名：日本国  

開催年月日： 2021年8月 - 2019年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：水産大学校（オンライン）   国名：日本国  

開催年月日： 2021年8月 - 2019年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北里大学海洋生命科学部（オンライン）   国名：日本国  

開催年月日： 2021年1月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：九州大学大学院農学研究院（オンライン）   国名：日本国  

開催年月日： 2020年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学海洋科学部   国名：日本国  

開催年月日： 2019年11月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Dalian Zhongxia Garden Hotel, Dalian   国名：中華人民共和国  

その他リンク： http://asdci.dlou.edu.cn/2019/1025/c6432a102342/page.htm

開催年月日： 2019年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：九州大学　稲盛財団記念館   国名：日本国  

開催年月日： 2019年8月

記述言語：日本語   会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：コングレスクエア日本橋   国名：日本国  

開催年月日： 2018年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Fukuoka Recent Hotel   国名：日本国  

開催年月日： 2018年8月 - 2018年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北九州国際会議場（北九州市）   国名：日本国  

開催年月日： 2018年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：日本大学生物資源科学部   国名：日本国  

開催年月日： 2018年6月 - 2018年9月

記述言語：英語  

開催地：La Fonda Hotel (Santa Fe, New Mexico)   国名：アメリカ合衆国  

Bony fish is one of the ancestral vertebrates that possess immunoglobulins, on which the classical pathway of complement system relies for target recognition. To access the role of the classical pathway in bony fish complement, we have analyzed the hemolytic reaction and C3b-deposition on the target surface of the common carp (Cyprinus carpio) complement, using carp serum immunochemically depleted of factor D (Df), a serine protease responsible for activation of the alternative pathway, an activation and amplification cascade that bypasses the classical pathway. To our surprise, the Df-depletion abolished hemolytic activities of the serum both through the classical and alternative pathways, when assayed using antibody-sensitized and non-sensitized sheep and rabbit erythrocytes. However, a low level of C3b-deposition, probably resulted from the classical pathway activation in the absence of Df was demonstrated on various target surfaces by flow cytometry and ELISA. In addition, Df-depletion inhibited C5-deposition essential for the cytotoxic complex formation on the target cells. The C3b-bound erythrocytes were hemolyzed by serum but easily lysed by Mg-EGTA-serum, probably triggering the alternative pathway amplification more efficiently than unbound erythrocytes. These results, taken together, suggest that an ancestral classical pathway alone lacks C5-activating ability but tags target cells with a small amount of C3b, which triggers enhanced C3-activation by the alternative pathway, leading to completion of the terminal cytolytic pathway.

開催年月日： 2018年6月 - 2018年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：La Fonda Hotel (Santa Fe, New Mexico)   国名：アメリカ合衆国  

開催年月日： 2018年5月 - 2017年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：コラッセ福島（福島市）   国名：日本国  

開催年月日： 2018年5月 - 2017年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：コラッセ福島（福島市）   国名：日本国  

開催年月日： 2018年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Cytotoxicity of T cells against a prasite Ichthyophthirius multiliis in ginbuna crucian carp

開催年月日： 2018年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Cytotoxicity of T cells against a prasite Ichthyophthirius multiliis in ginbuna crucian carp

開催年月日： 2018年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

【目的】魚類における補体の自然免疫機能を正確に理解するには、タンパク質レベルでの機能解析が必要である。しかし、補体成分の多くは、魚類では特に不安定で、活性を保持した補体成分の精製が困難である。本研究では、特異抗体のアフィニティーカラムを用い、活性を保持した補体成分C5を高純度に精製するための、迅速法の確立を目指した。 【方法】(1)抗コイC5の作成：コイC5-1アイソタイプのnetrinドメインを、N末端に6xHisタグを付加した組換タンパク質として大腸菌で発現させ、Ni-カラムで精製後、ウサギに免疫注射して抗血清を得た。 (2)アフィニティーカラムの調製： L-lysineおよび抗コイC5-1ウサギIgGをNHS-活性化HiTrapカラム（各1 ml）にそれぞれ固定化後、TBSで平衡化した。(3)溶血活性測定：抗体感作ヒツジ赤血球を標的としたCH50補体価測定法に準拠した。 【結果】コイ血清（10 ml）を、直列に接続したLysine-HiTrap→抗C5-HiTrapカラムにシリンジで負荷した。この直列カラムを20 mlのTBSで洗浄後、Lysine-HiTrapカラムを外し、抗C5-HiTrapカラム出口をHiTrap Desaltingカラム入口に接続した。5 mlの0.1 M glycine-HCl (pH 2.5)を送って抗C5-HiTrapカラムにトラップされたC5を脱離させると同時に、TBSへの緩衝液交換を済ませた。本法により1.5時間以内で精製されたC5は、SDS-PAGEでα鎖(110 kDa)とβ鎖(70 kDa)の典型的な二本鎖構造を示し、C5除去コイ血清と組み合わせると溶血活性を保持していることが確認された。

開催年月日： 2018年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

開催年月日： 2018年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Praparation of anti-zebrafish C5a to elucidate roles of complement C5a in inflammatory reaction of fish

開催年月日： 2017年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Tokyo University of Marine Science and Technology (Tokyo)   国名：日本国  

開催年月日： 2017年9月

記述言語：英語  

開催地：Hotel Scandic Copenhagen (Copenhagen)   国名：デンマーク王国  

その他リンク： http://emchd2017.dk/

開催年月日： 2017年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北海道大学   国名：日本国  

開催年月日： 2017年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北海道大学   国名：日本国  

開催年月日： 2017年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Development of KHV　disease in ginbuna crucian carp by suppression of cellular immunity.

開催年月日： 2017年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Functional analysis of two C7 isotypes of carp complement system.

開催年月日： 2017年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Cytotoxicity of T cells against a prasite Ichthyophthirius multiliis in ginbuna crucian carp

開催年月日： 2016年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Kanazawa   国名：日本国  

Mammalian CD46 has been reported as a multitasking immune modulator, which regulates complement activation on host cells, T cell-mediated adaptive responses, and wound repair by involving epithelial cells. It is of particular interest to explore the evolutionary path of such versatile functions of CD46. Our group has identified a CD46-like molecule, termed teleost complement regulatory membrane protein or Tecrem, in a few cyprinid fish species and has shown its regulatory function on complement activation at the protein level. Furthermore, we have also found that Tecrem expressed on T cells modulates mitogen-induced T cell proliferation in ginbuna crucian carp, indicating that modulation of adaptive immunity is one of the evolutionarily conserved functions of the CD46-like complement regulator. In the present study, we have explored a homeostatic role of Tecrem in the maintenance of fish epithelium, by analyzing expression behavior of Tecrem on an epithelial cell line (KF-1) derived from carp fin. Flow cytometric analysis of Tecrem expression on KF-1 using anti-carp Tecrem monoclonal antibody (MAb) (1F12) suggested that Tecrem expression may be affected by cell aggregation and adhesion. Fluorescent microscopic observation and ELISA-based assay for Tecrem also indicated a role of Tecrem in the adhesion of KF-1 to the surface of culture media. Furthermore, 1F12 MAb deposited on the culture plate significantly enhanced an early stage of cell adhesion process of KF-1. Towards further functional analyses of carp Tecrem, we have prepared recombinant Tecrem proteins in the bacterial expression system. Among the four short consensus repeat (SCR) modules making up the extracellular domains of Tecrem, the N-terminal two SCRs (rSCR1-2) and the C-terminal two SCRs (rSCR3-4) were separately expressed as 6xHis-tagged soluble protein using pCold-I vector and Origami B strain. Interaction of the two recombinant domains with carp C3 isotypes and their inhibition of carp complement activation cascades are currently analyzed in parallel with raising polyclonal antibodies, for the clarification of functional modules and their mode of action.

開催年月日： 2016年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Kanazawa   国名：日本国  

The alternative pathway (ACP) of the complement system is an antibody-independent activation pathway, in which properdin (Pf) has been known as a positive regulator of the activation and a possible pattern-recognition molecule to trigger ACP activation. Teleost complement system has a striking feature that some of its components are diversified into multiple isoforms with different functions. However this diversity is less characterized for teleost Pf, especially at the protein level. The present study was aimed at elucidating isotypic diversity and functional differentiation of Pf in the common carp. Molecular cloning of carp Pf revealed two distinct full-length cDNA sequences, CaPf1 and CaPf2, that predicts mature proteins composed of seven thrombospondin type1 domains (TSP0-TSP6), sharing 77% amino acid sequence identity. Genomic Southern hybridization suggested that CaPf1 and CaPf2 are encoded by single each gene in carp genome. Real-time quantitative PCR indicated that expression level of CaPf1 is most abundant in the spleen, whereas CaPf2 was detected mainly in head kidney and renal kidney. Rabbit antibodies were raised against their recombinant proteins corresponding to TSP4-6 domains. Western blotting using anti-CaPf1 and anti-CaPf2 revealed that CaPf1 and CaPf2 are mainly present as a hexamer of polypeptide with molecular weights of 49,000 and 48,000, respectively, in carp serum. Interestingly, CaPf1 and CaPf2 showed different spectra of binding to various microbes, suggesting their functional diversity.

開催年月日： 2016年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京医科歯科大学   国名：日本国  

開催年月日： 2016年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Gene knockout of myeloperoxidase in zebrafish that lacks adaptive immunity

開催年月日： 2016年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Effects of dietary soybean meal protein on antibody response in fish

開催年月日： 2015年6月 - 2015年7月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Murcia   国名：スペイン  

The alternative pathway (ACP) of the complement system is an antibody-independent activation pathway, in which properdin (Pf) has been known as an essential positive regulator of the activation and possibly as a pattern-recognition molecule to trigger ACP activation. Teleost complement system has a striking feature that some of its components are diversified into multiple isoforms with different functions. However this diversity is less characterized for teleost Pf, especially at the protein level. The present study was aimed at elucidating isotypic diversity and functional differentiation of Pf in the common carp. Molecular cloning of carp Pf revealed two distinct full-length cDNA sequences, CaPf1 and CaPf2, that predicts mature proteins composed of seven thrombospondin type1 domains (TSP0-TSP6), sharing 77% amino acid sequence identity. Genomic Southern hybridization suggested that CaPf1 and CaPf2 are encoded by single each gene in carp genome. Real-time quantitative PCR indicated that expression level of CaPf1 is most abundant in the spleen, whereas CaPf2 was detected mainly in head kidney and renal kidney. Rabbit antibodies were raised against their recombinant proteins corresponding to TSP4-6 domains. Western blotting using anti-CaPf1 and anti-CaPf2 revealed that CaPf1 and CaPf2 are mainly present as a hexamer of polypeptide with molecular weights of 49,000 and 48,000, respectively, in carp serum. Interestingly, CaPf1 and CaPf2 showed different spectra of binding to various microbes, suggesting their functional diversity.

開催年月日： 2015年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

開催年月日： 2015年3月 - 2014年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

開催年月日： 2015年3月 - 2014年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

開催年月日： 2014年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：九州大学箱崎キャンパス   国名：日本国  

Immune-regulatory mechanism in rag1-deficient zebrafish

開催年月日： 2014年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：神戸常盤大学   国名：日本国  

開催年月日： 2014年7月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東北大学　片平さくらホール   国名：日本国  

開催年月日： 2014年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北海道大学水産学部   国名：日本国  

開催年月日： 2013年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：三重大学   国名：日本国  

Effect of soy bean meal feeding on the complement system of rainbow trout

開催年月日： 2013年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：三重大学   国名：日本国  

開催年月日： 2013年8月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：岡山理科大学   国名：日本国  

開催年月日： 2013年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Carp thrombocytes are activated by other leukocytes to show phagocytosis.

開催年月日： 2013年1月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：九州大学中央図書館視聴覚ホール   国名：日本国  

開催年月日： 2013年1月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：九州大学中央図書館視聴覚ホール   国名：日本国  

開催年月日： 2013年1月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：九州大学中央図書館視聴覚ホール   国名：日本国  

開催年月日： 2013年1月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：九州大学中央図書館視聴覚ホール   国名：日本国  

開催年月日： 2012年8月

会議種別：口頭発表（一般）  

開催地：大阪府立成人病センター   国名：日本国  

開催年月日： 2012年7月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Hilton Fukuoka Sea Hawk Hotel   国名：日本国  

Thrombocytes have been recognized as haemostatic cells in non-mammalian vertebrates. Unlike mammalian platelets, thrombocytes are nucleated cells with a lymphocyte-like morphological feature, and possible involvement of thrombocytes in innate immune function has been considered in addition to the haemostatic function. In the present study, we report phagocytic abilities of some teleost thrombocytes. Using a monoclonal antibody specific for thrombocytes of common carp (Cyprinus carpio), thrombocytes were isolated from peripheral blood and examined for expression of various immune-related genes, resulting in detection of significant level of lysozyme, iNOS and MHC class II using RT-PCR. Upon flow cytometry-based phagocytosis assay, a number of thrombocytes ingested fluorescent latex particles (0.5 μm, 1 μm, 2 μm and 3 μm), bacteria (Escherichia coli) and zymosan particles. Phagocytosis by the thrombocytes was also confirmed by fluorescent microscopy and transmission electron microscopy, which revealed internalization of these particles into thrombocytes. We also observed that thrombocytes of the olive flounder (Paralichthys olivaceus) had similar phagocytic behavior. These data indicate that thrombocytes of those species are potent phagocytes, suggesting that those phagocytic characteristics of thrombocytes are widely conserved in teleosts. We also assessed a phagolysosome formation ability of teleost thrombocytes against the ingested pathogens, for detecting intracellular bactericidal activities of those thrombocytes. By using an ImageStream multi-spectral flow cytometer, we assessed phagolysosome fusion of goldfish (Carassius auratus) thrombocytes. On this analysis we detected that lysosomes of these thrombocytes visualized with fluorescent dextran were co-localized with the ingested small beads and formed a ring around large beads like other typical phagocytes. Overall, the results indicate that teleost thrombocytes have dual functions as not only haemostatic cells but also as phagocytic immune cells against microbial infections.

開催年月日： 2012年7月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Hilton Fukuoka Sea Hawk Hotel   国名：日本国  

Isotypic diversity of complement components is a striking feature of the teleost complement system. As a first line of innate defence, the complement system has been considered as an important clue to humoral defence in early development however functional diversity of the isotypes during teleost ontogeny is poorly understood. The present study aimed at clarifying comprehensive picture of ontogenetic expression of the diversified complement component isotypes in carp. Real-time quantitative PCR detected embryonic expression of C1r/s, MASPs, factor B/C2, C3, C4, C5, C6, C7, C8, C9, and factor I. A remarkable difference in the expression time course was noted between the isotypes in C3, C4, and C5. Especially, teleost-specific isotypes of C3 and C4 (non-histidine-type) started around hatching, in contrast to evolutionarily common isotypes (histidine-type), which showed much earlier expression. Whole-mount in situ hybridization of carp embryos revealed some difference in embryonic expression sites of two major C3 isotypes (C3-H1 and C3-S) in addition to common expression sites such as the yolk syncytial layer. The temporal and spatial differences in expression among the isotypes suggest that the isotypes are functionally differentiated in teleost early development.

開催年月日： 2012年7月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Hilton Fukuoka Sea Hawk Hotel   国名：日本国  

All extant jawed vertebrates share a common adaptive immune system in which immunoglobulin domain-based molecules act as antigen receptors. On the other hand, jawless vertebrates, lamprey and hagfish, use variable lymphocyte receptors composed of leucine-rich repeat cassettes for antigen recognition. Since invertebrates and primitive chordates do not have an adaptive immune system, the immune system seems to change dramatically in the course of evolution from primitive chordates to gnathostomes. From a phylogenic perspective of defence mechanisms, previously we found complement C3 and mannose-binding lectin-associated serine protease 1 (MASP-1) in hagfish and suggested the involvement of lectin pathway in hagfish innate immune system. In this study, we focused on the pattern recognition molecules in the hagfish, Eptatretus burgeri, and tried to purify them from serum by affinity chromatography. When the serum was treated with GlcNAc-agarose, followed by successive elution of the binding molecules with GlcNAc and EDTA, mainly four proteins (31 kDa, 27 kDa, 26 kDa, and 19 kDa) and 26 kDa protein were detected in the GlcNAc-eluate and EDTA-eluate, respectively. Collagenase treatment showed the presence of collagen-like domain only in the 26 kDa protein in the EDTAeluate. Since common pattern recognition molecules such as mannose-binding lectins possess collagen-like domains, we examined the entity of the 26 kDa protein by sequencing its N-terminal amino acids and cDNA obtained by 3’ and 5’ RACE methods, and identified it as a member of C1q family. Herein, it will be referred to as hagfish C1q (hagC1q). Western blot analyses using anti-hagC1q, MASP-1, and complement C3 antibodies showed that hagC1q associated with MASP-1 and complement C3 in the serum and had binding ability to Escherichia coli as a divalent cation-dependent manner. These results suggest that hagC1q plays an important role in hagfish innate immune system.

開催年月日： 2012年7月

記述言語：英語  

開催地：Hilton Fukuoka Sea Hawk Hotel   国名：日本国  

Two serotypes of Streptococcus parauberis, pathogens of streptococcosis in Japanese flounder (Paralichthys olivaceus), have caused severe losses of the livestock, due to the lack of effective vaccine and its resistance to antibiotics approved for flounder culture. These strains show different virulence, but their mechanisms of their pathogenicity are totally unknown, and identification of virulence factors are needed for effective vaccine development. The present study, therefore, was aimed at clarifying effect of S. parauberis culture supernatant on flounder. Since the diseased flounder shows anaemia, we examined the effects of S. parauberis on flounder peripheral blood. As a result, S. parauberis -injected flounder showed decreased numbers of peripheral erythrocytes, suggesting that S. parauberis may produce a haemolytic factor. To characterize the haemolytic factor, a supernatant of S. parauberis culture (25°C for 48h in Todd Hewitt broth) was mixed with sheep erythrocytes, resulting in significant level of haemolysis. The haemolytic factor was stable on heating at 100°C for 10 min, and passed through an ultrafiltration membrane with a 5 kDa cut off limit (Amicon Ultra15), indicating that the factor is not proteinaceous haemolysin but a heat-resistant low molecular mass substance. We also investigated the effects of the culture supernatant on flounder leukocytes. Peripheral leukocytes were separated from flounder and carp using Percoll discontinuous density gradient centrifugation and cultured with the supernatant in 96-well plate. The stimulated flounder cells showed agglutination and significant proliferation as assayed by BrdU uptake, while carp cells did not. These results suggest that S. parauberis secrets a mitogen specific for flounder leucocytes. Characterization of the mitogen and resulting leucocyte response is in progress.

開催年月日： 2012年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

開催年月日： 2011年9月 - 2011年10月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：長崎大学水産学部   国名：日本国  

開催年月日： 2011年9月

記述言語：日本語   会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：旭川市大雪クリスタルホール   国名：日本国  

開催年月日： 2011年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：横浜市   国名：日本国  

開催年月日： 2011年8月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：横浜市   国名：日本国  

開催年月日： 2011年5月 - 2011年6月

記述言語：英語   会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：Nagoya Congress Center, Nagoya   国名：日本国  

開催年月日： 2011年5月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：University of Alberta, Edmonton   国名：カナダ  

開催年月日： 2011年5月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：University of Alberta, Edmonton   国名：カナダ  

開催年月日： 2011年3月

会議種別：口頭発表（一般）  

開催地：日本大学生物資源科学部   国名：日本国  

Phagocytic function of fish thrombocytes

開催年月日： 2010年9月

会議種別：口頭発表（一般）  

開催地：コラッセ福島（福島市）   国名：日本国  

Diversity in target-recognition of carp serum collectins related to the lectin pathway of the complement Akito Ichiki, Haruka Tsukamoto, Tomonori Somamoto, and Miki Nakao

開催年月日： 2010年7月

会議種別：口頭発表（一般）  

開催地：Universitat Autònoma de Barcelona   国名：スペイン  

Functional diversity of the complement component isotypes in bony fish innate immunity: a model study using the common carp. Nakao, M., Ichiki, S., Mutsuro, J., Tsujikura, M., and Somamoto, T.

開催年月日： 2010年3月

会議種別：口頭発表（一般）  

開催地：日本大学生物資源科学部   国名：日本国  

Functional differentiation of C7 isotypes in the common carp complement system

開催年月日： 2010年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：日本大学生物資源科学部   国名：日本国  

Recognition specificity of carp serum lectins associated with the complement system

開催年月日： 2009年8月

会議種別：口頭発表（一般）  

開催地：九州大学西新プラザ（福岡市）   国名：日本国  

Structural diversity and functional differentiation of the complement components in bony fish. Miki Nakao

開催年月日： 2009年8月

会議種別：口頭発表（一般）  

開催地：九州大学西新プラザ（福岡市）   国名：日本国  

Functional differentiation of complement C3 isotypes in the teleost immune defense. Satoko Ichiki, Yoko Kato-Unoki, Tomonori Somamoto, Miki Nakao

開催年月日： 2009年8月

会議種別：口頭発表（一般）  

開催地：九州大学西新プラザ（福岡市）   国名：日本国  

Expression pattern of complement component C3 isotypes during early development of the common carp. Vo Kha Tam, Chie Okura, Tomonori Somamoto, Miki Nakao

開催年月日： 2009年8月

会議種別：口頭発表（一般）  

開催地：九州大学西新プラザ（福岡市）   国名：日本国  

Subcomponent structure of the first complement component (C1) in the common carp. Takashi Ichii, Masakazu Tsujikura, Tomonori Somamoto, Yoko Kato-Unoki, Shin-ichi Kato, Michiyasu Yoshikuni, Miki Nakao

開催年月日： 2009年8月

会議種別：口頭発表（一般）  

開催地：日本大学生物資源科学部   国名：日本国  

cDNA cloning and expression analysis of Fas ligand in ginbuna crucian carp

開催年月日： 2009年8月

会議種別：口頭発表（一般）  

開催地：日本大学生物資源科学部   国名：日本国  

Subcomponent structure of the first complement component (C1) in the common carp

開催年月日： 2009年8月

会議種別：口頭発表（一般）  

開催地：日本大学生物資源科学部   国名：日本国  

Phagocytic ability of carpo thrombocytes

開催年月日： 2009年8月

会議種別：口頭発表（一般）  

開催地：九州大学西新プラザ（福岡市）   国名：日本国  

Cloning, purification and characterization of zymosan-binding proteins in Nile Tilapia (Oreochromis niloticus) Soha Gomaa, Tomonori Somamoto and Miki Nakao

開催年月日： 2009年7月

会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：Aquatic Biotechnology Center, Gyeongsang National University, Jinju   国名：大韓民国  

開催年月日： 2009年6月 - 2009年7月

開催地：Praque   国名：チェコ共和国  

開催年月日： 2009年6月 - 2009年7月

会議種別：口頭発表（一般）  

開催地：Praque   国名：チェコ共和国  

開催年月日： 2009年6月 - 2009年7月

会議種別：口頭発表（一般）  

開催地：Praque   国名：チェコ共和国  

開催年月日： 2009年5月

会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：早稲田大学理工学部   国名：日本国  

Complement components constituting the classical activation pathway, an antibody-dependent activation cascade of the complement system.

開催年月日： 2008年10月

会議種別：口頭発表（一般）  

開催地：Pacifico Yokohama   国名：日本国  

開催年月日： 2008年10月

会議種別：口頭発表（一般）  

開催地：Pacifico Yokohama   国名：日本国  

開催年月日： 2008年10月

会議種別：口頭発表（一般）  

開催地：Pacifico Yokohama   国名：日本国  

開催年月日： 2008年10月

会議種別：口頭発表（一般）  

開催地：Pacifico Yokohama   国名：日本国  

開催年月日： 2008年10月

会議種別：口頭発表（一般）  

開催地：Fukuoka Recent Hotel   国名：日本国  

開催年月日： 2008年8月

会議種別：口頭発表（一般）  

開催地：東京医科歯科大学   国名：日本国  

cDNA cloning of four lineages of MHC class I genes in a cell line from clonal ginbuna crucian carp, and their expression in the cell line infected with CHNV

開催年月日： 2008年8月

会議種別：口頭発表（一般）  

開催地：東京医科歯科大学   国名：日本国  

Diversity of carp serum lectins involved in the lectin pathway of the complement activation.

開催年月日： 2008年8月

会議種別：口頭発表（一般）  

開催地：東京医科歯科大学   国名：日本国  

Identification of a novel membrane-bound regulator of complement activation in carp and zebrafish

開催年月日： 2008年7月

会議種別：口頭発表（一般）  

開催地：St. Ann's College, University of Oxford, Oxford   国名：グレートブリテン・北アイルランド連合王国(英国)  

開催年月日： 2008年7月

会議種別：口頭発表（一般）  

開催地：北海道大学医学部   国名：日本国  

Identification and expression analysis of two isotypes of carp complement component C7

開催年月日： 2008年5月

会議種別：口頭発表（一般）  

開催地：National Taiwan Ocean University   国名：台湾  

会議種別：口頭発表（一般）  

開催地：National Taiwan Ocean University   国名：台湾  

Structural and functional diversity of the complement components in bony fish: implication for innate immune defense

会議種別：口頭発表（招待・特別）  

開催地：Charleston, SC   国名：アメリカ合衆国  

THE COMPLEMENT SYSTEM IN INVERTEBRATES AND LOWER VERTEBRATES

会議種別：口頭発表（一般）  

開催地：Charleston, SC   国名：アメリカ合衆国  

Molecular cloning and characterization of the complement C1q A, B, and C chains in common carp

会議種別：口頭発表（一般）  

開催地：Charleston, SC   国名：アメリカ合衆国  

MOLECULAR CLONING AND CHARACTERIZATION OF CD4 IN GINBUNA CRUCIAN CARP

会議種別：口頭発表（一般）  

開催地：Charleston, SC   国名：アメリカ合衆国  

DIVESIFICATION OF COMPLEMENT COMPONENT ISOTYPES IN THE COMMON CARP: EXPRESSION PATTERN AND SOME FUNCTIONAL ASPECTS

会議種別：口頭発表（一般）  

開催地：九州大学   国名：日本国  

Molecular cloning of C1q A, B and C chains from common carp

会議種別：口頭発表（一般）  

開催地：県立広島大学   国名：日本国  

Molecular cloning of a novel complement-regulatory factor from the common carp

会議種別：口頭発表（一般）  

開催地：長崎大学   国名：日本国  

Molecular cloning of a complement-regulatory factor from zebrafish

会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：Hawaii   国名：アメリカ合衆国  

Molecular cloning of C3d, a fragment from complement component C3 with possible adjuvant activity, from marine cultured fish

会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：Hawaii   国名：アメリカ合衆国  

Identification of the genes encoding complement components from the zebrafish genome database

会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：Hawaii   国名：アメリカ合衆国  

EST Analysis of mRNAs Expressed in Gill and Intestine of Carp (Cyprinus carpio. L)

会議種別：口頭発表（一般）  

開催地：高知グリーン会館   国名：日本国  

Inference of bio-defensive functions of the complement system based on its gene expression pattern.

会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Identification and expression analysis of zebrafish complement component genes

会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Recombinant carp complement component C4 and C5: Expression and functional analysis

会議種別：口頭発表（一般）  

開催地：東海大学工学部   国名：日本国  

Identification of novel regulators of complement activation in fish

会議種別：口頭発表（一般）  

開催地：東海大学工学部   国名：日本国  

Functional analysis of carp C3 isoforms using monoclonal antibodies

会議種別：口頭発表（一般）  

開催地：東海大学工学部   国名：日本国  

Identification of Carp Complement Component C1

会議種別：口頭発表（一般）  

開催地：浜松市舞阪文化センター   国名：日本国  

Expression analysis of complement component isotypes during early development of the common carp

会議種別：口頭発表（一般）  

開催地：浜松市舞阪文化センター   国名：日本国  

cDNA cloning of MHC class I and MHC-related genes in a cell line from clonal ginbuna crucian carp

会議種別：口頭発表（一般）  

開催地：浜松市舞阪文化センター   国名：日本国  

Expressed sequence tag analysis of kidney and spleen from ginbuna crucian carp, Carassius auratus langsdorfii

会議種別：口頭発表（一般）  

開催地：Kyushu University   国名：日本国  

Identification and expression analysis of a novel interleukine 8 (IL-8)-like CXC chemokine in carp (Cyprinus carpio)

開催地：Kyushu University   国名：日本国  

Functional analysis of carp C3 isotypes using monoclonal antibodies

会議種別：口頭発表（一般）  

開催地：Sampaguita Garden, New Washington, Aklan   国名：フィリピン共和国  

Antibacterial substances from bony fish: structure, function, and application

会議種別：口頭発表（一般）  

開催地：東海大学海洋学部   国名：日本国  

Structure and regulation of the isotypic genes of the complement factor B in the common carp

会議種別：口頭発表（一般）  

開催地：東海大学海洋学部   国名：日本国  

Identification of a novel complement regulatory factor in bony fish

会議種別：口頭発表（一般）  

開催地：東海大学海洋学部   国名：日本国  

Structure and regulation of the isotypic genes of the complement factor B in the common carp

会議種別：口頭発表（一般）  

開催地：東海大学海洋学部   国名：日本国  

開催年月日： 2024年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

開催年月日： 2024年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

開催年月日： 2024年3月

記述言語：英語  

開催地：東京海洋大学   国名：日本国  

開催年月日： 2023年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：日本大学生物資源科学部   国名：日本国  

開催年月日： 2022年3月

記述言語：日本語  

開催地：日本大学生物資源科学部（オンライン開催）   国名：日本国  

【背景・目的】 微生物による感染防御に重要な免疫系には、獲得免疫と自然免疫の2つの機構がある。脊椎動物における両免疫機構の進化を深く理解するためには、より原始的な動物の免疫系を調査することが重要である。本研究の目的は、原始的な脊椎動物モデルとして軟骨魚類、ドチザメ（Triakis scyllium）を用い、その血清中の微生物結合因子および抗菌因子の検出・同定を試みた。 【材料・方法】ドチザメ血清300 µLをSuperdex 200 increaseカラム（1 x 30 cm）を使用したゲル濾過で分画し、各フラクション（Fr.）の1) Micrococcus luteusに対する溶菌活性、2) M. luteus, Aeromonas hydrophilla, Staphylococcus aureus, Edwardsiella tarda, Streptococcus parauberis, およびFlavobacterium sp.に対する凝集活性を測定した。また、各フラクションに含まれるタンパク質をSDS-PAGEで分析した。 【結果・考察】 Superdex 200 increaseの限界排除分子量に相当するFr. 7〜8に、S. parauberis を除く全菌種に対する凝集活性が検出された。また、E. tardaおよびS. aureusに対しては、分子量数十万〜数万に相当する広い範囲のFr.が凝集活性を示した。一方、分子量約1〜２万に相当するFr. 24〜27に、M. luteusに対する溶菌活性が認められた。各フラクションのSDS-PAGEによって、Fr. 7〜8には、IgM抗体のH鎖と考えられる分子量約75,000のバンドが認められた。また、Fr. 24〜27は、分子量14,000の1本のバンドを示した。このポリペプチドのN末端アミノ酸配列は、YVYSKXELARTLRRNGLDGYと解読され、これはジンベイザメのLysozyme C-1-likeと有意な相同性（80%）を示した。したがって、分子量約14,000の溶菌因子は、ドチザメのリゾチームと同定された。

開催年月日： 2021年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北海道大学水産学部（オンライン）   国名：日本国  

開催年月日： 2021年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：北海道大学水産学部（オンライン）   国名：日本国  

開催年月日： 2019年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：フェニックスプラザ（福井県福井市）   国名：日本国  

開催年月日： 2019年9月 - 2019年10月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：福井県立大学永平寺キャンパス   国名：日本国  

開催年月日： 2019年6月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Grand Canaria   国名：スペイン  

A line of studies has shown that several humoral immune factors includingcomplement, lectins and antibodies are involved in protection fromparasite infections. However, cell-mediated immunity against parasiteshas poorly been understood in teleostfish. In the present study, directcytotoxic activity of leukocytes againstIchthyophthirius multifiliishas beendemonstrated in ginbuna crucian carp. Leukocytes labeled by eachmonoclonal antibody (2C3: anti-CD8, 6D1:anti-CD4, GB20: anti-macro-phages/neutrophils) were co-incubated withI. multifiliis. Thefluorescentmicroscopic observation showed that CD8+T cells from naïve ginbunacarp, but not other leucocytes, contactedI. multifilii. The cytotoxic activityof CD8+T cells was significantly higher than that of other leucocytes,indicating that CD8+T cells are dominant effector cells againstI. multifiliis.The cytotoxic assay using a trans-well insert suggested that CD8+T cellsrequire to contact the parasites for the direct killing. Furthermore, a serineprotease inhibitor 3, 4-dichloroisocoumarin (DCI) inhibited the cytotoxicactivity of CD8+Tcells, but a perforin inhibitor Concanamycin A (CMA) didnot. These results indicate that teleost CD8betaT cells have natural cell-mediated cytotoxicity against extracellular parasite by utilizing serineproteases, such as granzyme, suggesting that CD8+T cells play animportant role in innate immunity against extracellular protozoanparasites.

開催年月日： 2019年6月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：Palacio de Congresos de Canarias   国名：スペイン  

In early stage of viral infection, type-I interferons (IFNs) are produced by signaling from innate immune receptors such as Toll-like receptors (TLRs), which recognize virus-specific molecular patterns including nucleic acids. The type-I IFN transcriptions induced by TLRs are regulated via nuclear factor kB (NF-kB) complex and interferon regulatory factors (IRFs), but these details are still unclear in fish. In the present study, we show that thrombocytes in common carp (Cyprinus carpio) have a potent ability to produce large amount of IFNs in response to TLR signaling. Magnetic- sorted HB-8 mAbþ carp thrombocytes and negatively sorted other pe- ripheral blood leukocytes (PBLs) were incubated with resiquimod (also called R848, a potent agonist of TLR7/8), followed by qPCR analysis. The expression levels of the common carp type-I IFNs (ccIFN1 and ccIFN2) in thrombocytes were considerably higher compared with that of in other PBLs. Whereas the ccIFN1 expression was relatively lower than the ccIFN2, the R848 stimulant highly upregulated the ccIFN1 expression than ccIFN2. Although typical inflammatory cytokines including interleukin-6 were also upregulated in thrombocytes, the expression levels were still lower than those in other PBLs. These results indicate that activation of carp thrombocytes by R848 inclines immune system toward antiviral response, rather than inflammation. Expression levels of IRF3 and IRF7 were also upregulated by R848, implying that the IFN transcriptions were activated by these IRFs. The expression of the IFNs and inflammatory cytokines were decreased by several NF-kB signaling inhibitors such as BAY11-7082 or phenethyl caffeate, however, sensitivities to each inhibitor were different between the IFNs and other cytokines. In the presence of those inhibitors, the ccIFN2 expression was correlated with the level of IRF3. In contrast, ccIFN1 expressions seem to be linked to IRF7, suggesting that these two IRFs regulates different IFN genes separately. Our finding suggests that fish thrombocytes are important components for antiviral immunity and can be a new target for the strategy of disease control and vaccine development.

開催年月日： 2018年9月

記述言語：英語   会議種別：口頭発表（一般）  

開催地：広島大学   国名：日本国  

開催年月日： 2018年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：広島大学   国名：日本国  

開催年月日： 2018年9月

記述言語：英語   会議種別：口頭発表（招待・特別）  

開催地：Fukuoka Recent Hotel   国名：日本国  

開催年月日： 2017年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：ホテルメリージュ（宮崎市）   国名：日本国  

開催年月日： 2017年9月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：ホテルメリージュ（宮崎市）   国名：日本国  

開催年月日： 2015年1月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：宮崎大学   国名：日本国  

開催年月日： 2013年3月

記述言語：日本語   会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Monoclonal antibody preparation and expression analysis of ginbuna CD83, a dendritic cell marker.

開催年月日： 2012年1月

会議種別：口頭発表（一般）  

開催地：鹿児島大学   国名：日本国  

開催年月日： 2011年8月

会議種別：口頭発表（一般）  

開催地：横浜市   国名：日本国  

会議種別：口頭発表（招待・特別）  

開催地：Charleston, SC   国名：アメリカ合衆国  

JADCI: EVOLVING THROUGH ANNUAL MEETINGS

会議種別：口頭発表（一般）  

開催地：Helsinki   国名：ノルウェー王国  

会議種別：口頭発表（一般）  

開催地：大阪   国名：日本国  

Nakao M, Mutsuro J, Fujiki K, Yano T. Molecular cloning of multiple C3 cDNA from the common carp. 35th Complement Symposium, Osaka, July 1998.

会議種別：口頭発表（一般）  

開催地：栃木   国名：日本国  

Nakao M, Smith SL, Nakazawa M, Mutsuro J, Fujiki K, Yano T. Diversity of the complement factor B/C2 from carp and shark. Japanese Association for Developmental and Comparative Immunology, Tochigi, August 1998.

会議種別：口頭発表（一般）  

開催地：栃木   国名：日本国  

Shin DH, Fujiki K, Nakao M, Yano T. cDNA cloning of NKEF from the commmon carp. Japanese Association for Developmental and Comparative Immunology, Tochigi, August 1998.

会議種別：口頭発表（一般）  

開催地：栃木   国名：日本国  

Fujiki K, Shin DH, Nakao M, Yano T. cDNA cloning of cytokines from the common carp using the SSH technique. Japanese Association for Developmental and Comparative Immunology, Tochigi, August 1998.

開催地：Rhodos   国名：ギリシャ共和国  

開催地：Rhodos   国名：ギリシャ共和国  

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M, Mutsuro J, Totsuka S, Tanaka N, Kato Y, Yano T. Purification and cloning of the complement component C4 from the common carp. 38th Complement Symposium, Tokyo, June 1999.

会議種別：口頭発表（一般）  

開催地：St. Andrews   国名：グレートブリテン・北アイルランド連合王国(英国)  

会議種別：口頭発表（一般）  

開催地：福岡   国名：日本国  

Mutsuro J, Nakao M, Kato Y, Fujiki K, Yano T. Isolation and identification of carp complement C3 isoforms. Japanese Association for Developmental and Comparative Immunology, Fukuoka, Augusut 1999.

会議種別：口頭発表（一般）  

開催地：福岡   国名：日本国  

Nagai T, Nakao M, Kato Y, Mutsuro J, Kimura M, Fujiki K, Yano T. Molecular cloning and gene analysis of the MASP isoforms from the common carp. Japanese Association for Developmental and Comparative Immunology, Fukuoka, Augusut 1999.

会議種別：口頭発表（一般）  

開催地：福岡   国名：日本国  

Kimura M, Fujiki K, Nakao M, Shin DH, Yano T. cDNA cloning and gene analysis of the C3 receptor beta-subunit from the common carp. Japanese Association for Developmental and Comparative Immunology, Fukuoka, Augusut 1999.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Kato Y, Mutsuto J, Nakao M, Yano T. Monoclonal antibodies against carp complement component C5. Japanese Society for Fish Pathology, Tokyo, April 2000.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Tanaka N, Mutsuro J, Nakao M, Yano T. cDNA cloning of the complement C4 isotypes (C4A, C4B) from the common carp. Japanese Society for Fisheries Science, Tokyo, April 2000.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M, Osaka K, Mutsuro J, Kimura M, Yano T. cDNA cloning of the complement component C1r/C1s from the common carp. Japanese Society for Fisheries Science, Tokyo, April 2000.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M, Matsumoto M, Mutsuro J, Kimura M, Miura C, Fujiki K, Yano T. Cloning and expression analysis of a novel complement factor B/C2 isotype from the common carp. Japanese Society for Fish Pathology, Tokyo, April 2000.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M, Shimozono M, Kodama H, Moritomo T, Nakanishi T, Yano T. Purification and functional analysis of carp IgM isoforms. Japanese Society for Fisheries Science, Tokyo, April 2000.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M, Osaka K, Kato Y, Yano T. Molecular cloning of the complement component C1r/C1s from the common carp. Japanese Association for Develpmental and Complarative Immunology, Tokyo, July 2000.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Kimura M, Fujiki K, Nakao M, Shin DH, Yano T. cDNA cloning of carp leukocyte integrin molecules. Japanese Association for Developmental and Comparative Immunology, Tokyo, July 2000.

開催地：Snowbird   国名：アメリカ合衆国  

会議種別：口頭発表（一般）  

開催地：Cairns   国名：オーストラリア連邦  

会議種別：口頭発表（一般）  

開催地：Cairns   国名：オーストラリア連邦  

会議種別：口頭発表（一般）  

開催地：Cairns   国名：オーストラリア連邦  

開催地：Cairns   国名：オーストラリア連邦  

開催地：Snowbird   国名：アメリカ合衆国  

会議種別：口頭発表（一般）  

開催地：Cairns   国名：オーストラリア連邦  

会議種別：口頭発表（一般）  

開催地：大阪   国名：日本国  

Miura C, Nakao M, Kimura M, Yano T. Detection of carp CR3 beta-subunit using anti-peptide antibodies. 37th Complement Symposium, Osaka, August 2000.

会議種別：口頭発表（一般）  

開催地：大阪   国名：日本国  

Nakao M, Matsumoto M, Nakazawa M, Fujiki K, Yano T. Diversity of carp complement factor B/C2. 37th Complement Symposium, Osaka, August 2000.

会議種別：口頭発表（一般）  

開催地：Saga   国名：日本国  

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M, Nakahara M, Bayne CJ, Fujiki K, Yano T. cDNA cloning of the complement component C2 from bony fish. Japanese Society for Fisheries Science, Tokyo, April 2001.

会議種別：口頭発表（一般）  

開催地：札幌   国名：日本国  

Nakao M, Mutsuro J, Tanaka N, Kato Y, Yano T. Diversity and evolution of the thioester-containing proteins in the common carp. Japanese Association for Developmental and Comparative Immunology, Sapporo, July 2001.

会議種別：口頭発表（一般）  

開催地：京都   国名：日本国  

Shimozono M, Kodama H, Moritomo T, Nakao M, Yano T, Tomana M, Nakanishi T. Effect of IgM isoforms on phygocytic activity of carp neutrophils. Japanese Society for Bio-Defense, Kyoto, August 2001.

会議種別：口頭発表（一般）  

開催地：京都   国名：日本国  

Kato Y, Nakao M, Mutsuro J, Yano T. cDNA cloning of carp complement C5. 38th Complement Symposium, Kyoto, August 2001.

会議種別：口頭発表（一般）  

開催地：宮崎   国名：日本国  

Miura C, Nakao M, Mutsuro J, Yano T. Generation of a C3d fragment from carp C3. Japanese Society of Fish Pathology, Miyazaki, October 2001.

会議種別：口頭発表（一般）  

開催地：宮崎   国名：日本国  

Nakahara M, Nakao M, Yano T. Expression of recombinant protein of carp complement factor B/C2. Japanese Society of Fish Pathology, Miyazaki, October 2001.

会議種別：口頭発表（一般）  

開催地：Yokohama   国名：日本国  

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M. A strategy for host defense of bony fish: diversity of the complement system. A Joint Symposium of the Develpmental and Comparative Immunology, Comparative Endocrinology, and Comparative Biochemistry, Tokyo, December 2001.

会議種別：口頭発表（一般）  

開催地：奈良   国名：日本国  

Yano T, Nakao M. The complement system of bony fish. Symposium of Japanese Society for Fisheries Science, Nara, April 2002.

会議種別：口頭発表（一般）  

開催地：奈良   国名：日本国  

Nakao M, Yano T. Cytokine receptors and complement receptors in bony fish. Symposium of Japanese Society for Fisheries Science, Nara, April 2002.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M, Nakahara M, Harada H, Mutsuro J, Kato Y, Yano T. Linkage analysis and functional diversity of the complement factor B/C2 isotypes in the common carp. 39th Complement Symposium, Tokyo, July 2002.

会議種別：口頭発表（一般）  

開催地：名古屋   国名：日本国  

Nakao M, Hisamatu S, Nakahara M, Mutsuro J, Kato Y, Yano T. Molecular cloning of the complement regulatory factor I from the common carp. Japanese Association of Develpmental and Comparative Immunology, Nagoya, August 2002.

会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：長崎   国名：日本国  

Nakao M, Mutsuro J, Yano T. Diversity of the complement-encoding genes in bony fish and its biological implication. Symposium of Japanese Society for Aqua-Genome, Nagasaki, September 2002.

会議種別：口頭発表（一般）  

開催地：長崎   国名：日本国  

Nakahara M, Nakao M, Mutsuro J, Yano T. Functional analysis of carp complement factor B/C2 isotypes. Symposium of Japanese Society for Aqua-Genome, Nagasaki, September 2002.

会議種別：口頭発表（一般）  

開催地：Fukuoka   国名：日本国  

開催地：Palermo   国名：イタリア共和国  

開催地：Palermo   国名：イタリア共和国  

会議種別：口頭発表（一般）  

開催地：熊本   国名：日本国  

Nakao M, Miura C, Ito S, Nakahara M, Okumura K, Yano T Generation of a complement C3 fragment equivalent to mammalian C3d and detection of its receptor in the common carp (Cyprinus carpio). 35th Complement Symposium, Kumamoto, July 2003.

会議種別：口頭発表（一般）  

開催地：St. Andrews   国名：グレートブリテン・北アイルランド連合王国(英国)  

会議種別：口頭発表（一般）  

開催地：St. Andrews   国名：グレートブリテン・北アイルランド連合王国(英国)  

会議種別：口頭発表（一般）  

開催地：St. Andrews   国名：グレートブリテン・北アイルランド連合王国(英国)  

会議種別：口頭発表（一般）  

開催地：St. Andrews   国名：グレートブリテン・北アイルランド連合王国(英国)  

開催地：St. Andrews   国名：グレートブリテン・北アイルランド連合王国(英国)  

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Katayose Y, Nakao M, Matsushita M, Fujita T, Yano T. cDNA cloning of a C1q-like gene from the common carp. Japanese Association of Developmental and Comparative Immunology, Tokyo, August 2003.

会議種別：口頭発表（一般）  

開催地：東京   国名：日本国  

Nakao M, Maeda H, Hatanaka D, Matsushita M, Nakata M, Fujita T, Yano T. Purification and cloning of the mannose-binding lectin from the common carp. Japanese Association of Developmental and Comparative Immunology, Tokyo, August 2003.

会議種別：口頭発表（一般）  

開催地：鹿児島市  

Diversity of the mannose-binding lectin family in the lectin pathway of carp complement

会議種別：口頭発表（一般）  

開催地：Honolulu   国名：アメリカ合衆国  

An MBL-MASP2 complex that functions in the lectin pathway of a bony fish, the common carp (Cyprinus carpio)

会議種別：口頭発表（一般）  

開催地：東京大学  

Two MBL-like lectins functioning in the lectin pathway of carp complement

会議種別：口頭発表（一般）  

開催地：琉球大学  

cDNA cloning of grass carp complement components

会議種別：口頭発表（一般）  

開催地：函館  

Effect of the complement C3d fragment on the antibody response in the common carp

会議種別：口頭発表（一般）  

開催地：東京医科大学  

In silico cloning of zebrafish complement components

会議種別：口頭発表（一般）  

開催地：東京医科大学  

Diversity of alpha-2-macroglobulin in rainbowtrout

会議種別：シンポジウム・ワークショップ パネル（公募）  

開催地：福岡市  

Cyprinid fish as model animals for fish immunological research

会議種別：口頭発表（一般）  

開催地：高知大学   国名：日本国  

Functional analysis of factor I from carp complement

会議種別：口頭発表（一般）  

開催地：東京海洋大学   国名：日本国  

Complement component-encoding genes identified from the zebrafish genome database.

記述言語：日本語  

記述言語：日本語  

記述言語：日本語  

記述言語：日本語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

記述言語：日本語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

Phylogeny and evolution of activation pathways of the complement system

記述言語：英語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

記述言語：英語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

記述言語：日本語   出版者・発行元：(株)東京医学社  

＜文献概要＞はじめに　哺乳類の補体系は,抗体依存的・非依存的な3つの活性化経路と標的細胞を直接傷害する溶解経路,過剰な活性化あるいは自己細胞上での活性化を抑制する制御因子群,そして活性化成分を認識して免疫細胞のエフェクター機能を媒介する補体レセプター群から構成される。その機能は,自然免疫としての異物認識・排除のほかに,T・B細胞の活性化による獲得免疫応答の制御,さらに血液凝固系の活性化,創傷治癒,免疫複合体や死細胞の撤去処理などの恒常性維持をも担う。補体系の構成成分は,多様な蛋白質ファミリーに属し,各ファミリー内で遺伝子重複とドメインのシャッフリングによってさらに多様化し,現在われわれに備わっているような,高度に発達した補体系が完成したと考えられる。

記述言語：英語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

Sexual dimorphism allows species to meet their fitness optima based on the physiological availability of each sex. Although intralocus sexual conflict appears to be a genetic constraint for the evolution of sex-specific traits, sex-linked genes and the regulation of sex steroid hormones contribute to resolving this conflict by allowing sex-specific developments. Androgens and their receptor, androgen receptor (Ar), regulate male-biased phenotypes. In teleost fish, ar ohnologs have emerged as a result of teleost-specific whole genome duplication (TSGD). Recent studies have highlighted the evolutionary differentiation of ar ohnologs responsible for the development of sexual characteristics, which sheds light on the need for comparative studies on androgen regulation among different species. In this review, we discuss the importance of ar signaling as a regulator of male-specific traits in teleost species because teleost species are suitable experimental models for comparative studies owing to their great diversity in male-biased morphological and physiological traits. To date, both in vivo and in vitro studies on teleost ar ohnologs have shown a substantial influence of ars as a regulator of male-specific reproductive traits such as fin elongation, courtship behavior, and nuptial coloration. In addition to these sexual characteristics, ar substantially influences immunity, inducing a sex-biased immune response. This review aims to provide a comprehensive understanding of the current state of teleost ar studies and emphasizes the potential of teleost fishes, given their availability, to find molecular evidence about what gives rise to the spectacular diversity among fish species.

DOI： DOI: 10.2108/zs230098

記述言語：日本語   出版者・発行元：(一社)日本補体学会  

記述言語：日本語   出版者・発行元：(株)日本臨床社  

記述言語：英語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

This guideline was provided by the Japanese Association for Complement Research targeting clinicians for making an accurate diagnosis of hereditary angioedema (HAE), and for prompt treatment of the HAE patient in Japan. This is a 2010 year version and will be updated according to any pertinent medical advancements.

記述言語：英語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

記述言語：日本語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

Protective effects of polysaccharides for infectious fish diseases.

掲載種別：記事・総説・解説・論説等（学術雑誌）  

A unique strategy of the immune system of bony fish based on diversity of complement components.

記述言語：英語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

種別：査読等 

外国語雑誌　査読論文数：22

種別：大会・シンポジウム等 

参加者数：80

種別：大会・シンポジウム等 

参加者数：100

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参加者数：1,000

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外国語雑誌　査読論文数：20

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外国語雑誌　査読論文数：22

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参加者数：80

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参加者数：120

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外国語雑誌　査読論文数：24

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参加者数：150

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外国語雑誌　査読論文数：30

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参加者数：100

種別：大会・シンポジウム等 

参加者数：800

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

参加者数：300

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

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種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

参加者数：70

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

参加者数：70

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

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種別：大会・シンポジウム等 

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種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

種別：大会・シンポジウム等 

種別：大会・シンポジウム等