Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Miki Nakao Last modified date:2019.07.25

Professor / Division of Molecular Bioscience / Department of Bioscience and Biotechnology / Faculty of Agriculture


Papers
1. Seisuke Tajimi, Masakazu Kondo, Teruyuki Nakanishi, Takahiro Nagasawa, Miki Nakao, Tomonori Somamoto, Generation of virus-specific CD8
+
T cells by vaccination with inactivated virus in the intestine of ginbuna crucian carp, Developmental and Comparative Immunology, 10.1016/j.dci.2018.12.009, 93, 37-44, 2019.04, 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..
2. Somamoto, T., Maruyama, S., Nagasawa, T., Nakao, M., Sato, A., Hatta, H., Sato, M., Murakami-Yamaguchi, Y., Kizu-Mori, K., Hirakawa, Y., Narita, H. , Development of anti-atypical Aeromonas salmonicida monoclonal antibodies for diagnosis of “new ulcer disease" in koi carp, Fish Pathology, 10.3147/jsfp.53.36, 53, 1, 36-39, 2018.06.
3. Yumie Tokunaga, Masamichi Shirouzu, Ryota Sugahara, Yasutoshi Yoshiura, Ikunari Kiryu, Mitsuru Ototake, Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao, Comprehensive validation of T- and B-cell deficiency in rag1-null zebrafish
Implication for the robust innate defense mechanisms of teleosts, Scientific Reports, 10.1038/s41598-017-08000-2, 7, 1, 2017.12, rag1 -/- zebrafish have been employed in immunological research as a useful immunodeficient vertebrate model, but with only fragmentary evidence for the lack of functional adaptive immunity. rag1-null zebrafish exhibit differences from their human and murine counterparts in that they can be maintained without any specific pathogen-free conditions. To define the immunodeficient status of rag1 -/- zebrafish, we obtained further functional evidence on T- and B-cell deficiency in the fish at the protein, cellular, and organism levels. Our developed microscale assays provided evidence that rag1 -/- fish do not possess serum IgM protein, that they do not achieve specific protection even after vaccination, and that they cannot induce antigen-specific CTL activity. The mortality rate in non-vaccinated fish suggests that rag1 -/- fish possess innate protection equivalent to that of rag1 -/- fish. Furthermore, poly(I:C)-induced immune responses revealed that the organ that controls anti-viral immunity is shifted from the spleen to the hepatopancreas due to the absence of T- and B-cell function, implying that immune homeostasis may change to an underside mode in rag-null fish. These findings suggest that the teleost relies heavily on innate immunity. Thus, this model could better highlight innate immunity in animals that lack adaptive immunity than mouse models..
4. Ratchanu Meidong, Kulwadee Khotchanalekha, Sompong Doolgindachbaporn, Takahiro Nagasawa, Miki Nakao, Kenji Sakai, Saowanit Tongpim, Evaluation of probiotic Bacillus aerius B81e isolated from healthy hybrid catfish on growth, disease resistance and innate immunity of Pla-mong Pangasius bocourti, Fish and Shellfish Immunology, 10.1016/j.fsi.2017.11.032, 73, 1-10, 2018.02, 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..
5. Kolder IC, van der Plas-Duivesteijn SJ, Tan G, Wiegertjes GF, Forlenza M, Guler AT, Travin DY, Miki Nakao, Moritomo T, Irnazarow I, den Dunnen JT, Anvar SY, Jansen HJ, Dirks RP, Palmblad M, Lenhard B, Henkel CV, Spaink HP, A full-body transcriptome and proteome resource for the European common carp., BMC Genomics, 10.1186/s12864-016-3038-y, 17, 701-1-701-12, 2016.09, [URL], 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..
6. Nur I, Abdelkhalek NK, Motobe S, Nakamura R, Tsujikura M, Tomonori Somamoto, Miki Nakao, Functional analysis of membrane-bound complement regulatory protein on T-cell immune response in ginbuna crucian carp, Molecular Immunology, 10.1016/j.molimm.2015.11.010, 70, 1-7, 2016.02, [URL], 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..
7. Tomonori Somamoto, Yuhei Miura, Teruyuki Nakanishi, Miki Nakao, Local and systemic adaptive immune responses toward viral infection via gills in ginbuna crucian carp, Developmental and Comparative Immunology, 10.1016/j.dci.2015.04.016, 52, 1, 81-87, 2015.07, 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.
.
8. Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao, Carp thrombocyte phagocytosis requires activation factors secreted from other leukocytes, Developmental and Comparative Immunology, 10.1016/j.dci.2015.05.002, 52, 2, 107-111, 2015.07, 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..
9. Gan H, He H, Sato A, Hatta H, Nakao M, Somamoto T, Ulcer disease prophylaxis in koi carp by bath immersion with chicken egg yolk containing anti-Aeromonas salmonicida IgY, RESEARCH IN VETERINARY SCIENCE, 10.1016/j.rvsc.2015.01.016, 99, 82-86, 2015.04.
10. Masakazu Tsujikura, Takahiro Nagasawa, Satoko Ichiki, Ryota Nakamura, Tomonori Somamoto, Miki Nakao, A CD46-like Molecule Functional in Teleost Fish Represents an Ancestral Form of Membrane-Bound Regulators of Complement Activation, JOURNAL OF IMMUNOLOGY, 10.4049/jimmunol.1303179, 194, 1, 262-272, 2015.01, 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..
11. Takahiro Nagasawa, Chihaya Nakayasu, Aja M. Rieger, Daniel R. Barreda, Tomonori Somamoto, Miki Nakao, Phagocytosis by thrombocytes is a conserved innate immune mechanism in lower vertebrates. , Frontiers in Immunology, doi: 10.3389/fimmu.2014.00445, 5, 445, Article 445, 2014.09, 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..
12. Tomonori Somamoto, Masakazu Kondo, Teruyuki Nakanishi, Miki Nakao, Helper function of CD4⁺ lymphocytes in antiviral immunity in ginbuna crucian carp, Carassius auratus langsdorfii, Developmental and Comparative Immunology, 10.1016/j.dci.2013.12.008., 44, 1, 111-115, 2014.05.
13. Abdel-Salam, Soha G. R., Masakazu Kondo, Masakazu Tsujikura, Tomonori Somamoto, Miki Nakao, Purification and functional characterization of complement C3 and a novel zymosan-binding protein in tilapia serum, Fisheries Science, 10.1007/s12562-014-0700-7, 80, 2, 301-310, 2014.03, 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..
14. Tomokazu Yamaguchi, Kazufumi Takamune, Masakazu Kondo, Yukinori Takahashi, Yoko Kato-Unoki, Miki Nakao, Tamotsu Fujii, Hagfish C1q: Its Unique Binding Property, Developmental and Comparative Immunology, 39, in press, 2013.10.
15. Indriyani Nur, Hikari Harada, Masakazu Tsujikura, Tomonori Somamoto, Miki Nakao, Molecular characterization and expression analysis of three membrane-bound complement regulatory protein isoforms in the ginbuna crucian carp Carassius auratus langsdorfii., Fish Shellfish Immunol., 10.1016/j.fsi.2013.08.002, 35, 4, 1333-1337, 2013.10.
16. Tomonori Somamoto, Masakazu Kondo, Teruyuki Nakanishi, Miki Nakao, Identification of anti-viral cytotoxic effector cells in the ginbuna crucian carp, Carassius auratus langsdorfii, Developmental and Comparative Immunology, 10.1016/j.dci.2012.11.001, 39, 4, 111-115, 2013.04.
17. Yamauchi T, Takenaka K, Urata S, Shima T, Kikushige Y, Tokuyama T, Iwamoto C, Nishihara M, Iwasaki H, Miyamoto T, Honma N, Nakao M, Matozaki T, Akashi K., Polymorphic Sirpa is the genetic determinant for NOD-based mouse lines to achieve efficient human cell engraftment., Blood, 10.1182/blood-2012-06-440354, Epub ahead of print, 2013.01.
18. Ichiki S, Kato-Unoki Y, Somamoto T, Nakao M., The binding spectra of carp C3 isotypes against natural targets independent of the binding
specificity of their thioester., Developmental and Comparative Immunology, 10.1016/j.dci.2012.03.004., 38, 1, 10-16, 2012.08.
19. Sameshima S, Nakao M, Somamoto T., Diversity of CD2 subfamily receptors in cyprinid fishes., Result Immunol, 2, 25-34, 2012.02.
20. Urabe S, Somamoto T, Sameshima S, Unoki-Kato Y, Nakanishi T, Nakao M., Molecular characterization of MHC class I and beta-2 microglobulin in a clonal strain of ginbuna crucian carp, Carassius auratus langsdorfii., Fish Shellfish Immunol., 31, 469-474, 2011.09.
21. Oba Y, Yamauchi A, Hashiguchi Y, Satone H, Miki S, Nassef M, Shimasaki Y, Kitano T, Nakao M, Kawabata S, Honjo T, Oshima Y, Purification and characterization of tributyltin-binding protein of tiger puffer, Takifugu rubripes, Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, http://dx.doi.org/10.1016/j.cbpc.2010.07.009, 153, 1, 17-23, 2011.01.
22. Forlenza M, Nakao M, Wibowo I, Joerink M, Arts JA, Savelkoul HF, Wiegertjes GF, Nitric oxide hinders antibody clearance from the surface of Trypanoplasma borreli and increases susceptibility to complement-mediated lysis., Molecular Immunology, 46, 16, 3188-3197, 2009.10.
23. VO Kha Tam, Masakazu TSUJIKURA, Tomonori SOMAMOTO, and Miki NAKAO, Expression responses of the complement components in zebrafish organs after stimulation with poly I:C, mimicry of viral infection, Journal of Faculty of Agriculture, Kyushu University, 54, 2, 389-395, 2009.10.
24. VO Kha Tam, Masakazu TSUJIKURA, Tomonori SOMAMOTO, and Miki NAKAO, Identification of cDNA sequences encoding the complement components of zebrafish (Danio rerio), Journal of Faculty of Agriculture, Kyushu University, 54, 2, 373-387, 2009.10.
25. Nevien K. Abdelkhalek, Asuka Komiya, Yoko Kato-Unoki, Tomonori Somamoto, Miki Nakao, Molecular evidence for the existence of two distinct IL-8 lineages of teleost CXC-chemokines, Fish & Shellfish Immunology, 27, 6, 763-767, 2009.09.
26. Katakura F, Takizawa F, Yoshida M, Yamaguchi T, Araki K, Tomana M, Nakao M, Moritomo T, Nakanishi T, Co-culture of carp (Cyprinus carpio) kidney haematopoietic cells with feeder cells resulting in long-term proliferation of T-cell lineages., Vet Immunol Immunopathol., 10.1016/j.vetimm.2009.03.007, 131, 1-2, 127-136, 2009.09.
27. Tomonori Somamoto, Nobuaki Okamoto, Teruyuki Nakanishi, Mitsuru Ototake, and Miki Nakao, In vitro generation of viral-antigen dependent cytotoxic T-cells from ginbuna crucian carp, Carassius auratus langsdorfii, Virology, 389, 1-2, 26-33, 2009.06.
28. Dong-Ho Shin, Barbara M. Webb, Miki Nakao, Sylvia L. Smith, Characterization of shark complement factor I gene(s): Genomic analysis of a novel shark-specific sequence, Molecular Immunology, doi:10.1016/j.molimm.2009.04.002, 2009.05.
29. Yasuhiro Yamasaki, Tomoyuki Shikata, Atsushi Nukata, Satoko Ichiki, Sou Nagasoe, Tadashi Matsubara, Yohei Shimasaki, Miki Nakao, Kenichi Yamaguchi, Yuji Oshima, Tatsuya Oda, Makoto Ito, Ian R Jenkinson, Makio Asakawa and Tsuneo Honjo, Extracellular polysaccharide-protein complexes of a harmful alga mediate the allelopathic control it exerts within the phytoplankton community, The ISME Journal, doi:10.1038/ismej.2009.24, 2009.03.
30. Nomiyama H, Hieshima K, Osada N, Kato-Unoki Y, Otsuka-Ono K, Takegawa S, Izawa T, Yoshizawa A, Kikuchi Y, Tanase S, Miura R, Kusada J, Nakao M, Yoshie O, Extensive Expansion and Diversification of the Chemokine Gene Family in Zebrafish: Identification of a Novel Chemokine Subfamily CX, BMC Genomics, 9: 222., 2008.05.
31. Nonaka S, Somamoto T, Kato-Unoki Y, Ototake M, Nakanishi T, Nakao M, Molecular cloning of CD4 from ginbuna crucian carp Carassius auratus langsdorfii , Fisheries Science, 74(2): 341-346, 2008.04.
32. Shin DH, Webb B, Nakao M, Smith SL, Molecular cloning, structural analysis and expression of complement component Bf/C2 genes in the nurse shark, Ginglymostoma cirratum
, Dev Comp Immunol, 31(11):1168-1182, 2007.04.
33. Nakao M, Kajiya T, Sato Y, Somamoto T, Kato-Unoki Y, Matsushita M, Nakata M, Fujita T, Yano T, Lectin pathway of bony fish complement: Identification of two homologues of the mannose-binding lectin associated with MASP2 in the common carp (Cyprinus carpio), Journal of Immunology, 177(8): 5471-5479, 2006.10.
34. Somamoto T, Yoshiura Y, Sato A, Nakao M, Nakanishi T, Okamoto N, Ototake M., Expression profiles of TCRbeta and CD8alpha mRNA correlate with virus-specific cell-mediated cytotoxic activity in ginbuna crucian carp., Virology, 348(2): 370-377, 2006.05.
35. Mutsuro J, Tanaka N, Kato Y, Dodds AW, Yano T, Nakao M, Two divergent isotypes of the fourth complement component from a bony fish, the common carp (Cyprinus carpio)., Journal of Immunology, 175, 7, 4508-4517, 175(7): 4508-4517, 2005.10.
36. Savan R, Aman A, Nakao M, Watanuki H, Sakai M, Discovery of a novel immunoglobulin heavy chain gene chimera from common carp (Cyprinus carpio L.), Immunogenetics, 10.1007/s00251-005-0015-z, 57, 6, 458-463, 57(6): 458-463, 2005.07.
37. Hammond JA, Nakao M, Smith VJ, Cloning of a glycosylphosphatidylinositol-anchored alpha-2-macroglobulin cDNA from the ascidian, Cionaintestinalis, and its possible role in immunity, Molecular Immunology, 10.1016/j.molimm.2004.09.017, 42, 6, 683-694, 42(6): 683-694, 2005.04.
38. Tsujita T, Tsukada H, Nakao M, Oshiumi H, Matsumoto M, Seya T, Sensing bacterial flagellin by membrane and soluble orthologs of Toll-like receptor 5 in rainbow trout (Onchorhynchus mikiss), Journal of Biological Chemistry, 10.1074/jbc.M407634200, 279, 47, 48588-48597, 279 (47) 48588-48597, 2004.11.
39. Ishikawa J, Imai E, Moritomo T, Nakao M, Yano T, Tomana M, Characterisation of a fourth immunoglobulin light chain isotype in the common carp., Fish & Shellfish Immunology, 10.1016/j.fsi.2003.06.002, 16, 3, 369-379, 16 (3), 369-379, 2004.03.
40. Nakao M, Miura C, Itoh S, Nakahara M, Okumura K, Mutsuro J, Yano T, A complement C3 fragment equivalent to mammalian C3d from the common carp (Cyprinus carpio): Generation in serum after activation of the alternative pathway and detection of its receptor on the lymphocyte surface., Fish & Shellfish Immunology, 10.1016/S1050-4648(03)00057-3, 16, 2, 139-149, 16 (2), 129-149, 2004.02.
41. Kato Y, Nakao M, Shimizu M, Wariishi H, Yano T, Purification and functional assessment of C3a, C4a and C5a of the common carp (Cyprinus carpio) complement, Developmental and Comparative Immunology, 10.1016/j.dci.2004.01.006, 28, 9, 901-910, 28, 901-910, 2004.01.
42. Nakao M, Nakahara M, Yano T, Isolation of carp complement factor B and formation of the C3-convertase, Journal of Faculty of Agriculture, Kyushu University, 49, 1, 101-110, 49 (1), 101-110, 2004.01.
43. Kato Y, Nakao M, Mutsuro J, Zarkadis IK, Yano T (2003), The complement component C5 of the common carp (Cyprinus carpio) : cDNA cloning of two distinct isotypes that differ in a functional site., Immunogenetics, 10.1007/s00251-002-0528-7, 54, 11, 807-815, 54: 807-815., 2003.01.
44. Nakao M, Hisamatsu S, Nakahara M, Kato Y, Smith SL, Yano T, Molecular cloning of the complement regulatory factor I isotypes from the common carp (Cyprinus carpio)., Immunogenetics, 10.1007/s00251-002-0518-9, 54, 11, 801-806, 54: 801-806., 2003.01.
45. Fujiki K, Nakao M, Dixon B, Molecular cloning and characterization of a carp (Cyprinus carpio) cytokine-like cDNA that shares sequence similarity with IL-6 subfamily cytokines CNTF, OSM and LIF., Developmental and Comparative Immunology, 10.1016/S0145-305X(02)00074-5, 27, 2, 127-136, 27: 127-136., 2003.01.
46. Nakao M, Fujiki K, Kondo M, Yano T, Detection of complement receptors on head kidney phagocytes of the common carp (Cyprinus carpio)., Fisheries Science, 10.1046/j.1444-2906.2003.00709.x, 69, 5, 929-935, 69: 927-933., 2003.01.
47. Nakao M, Uemura T, Yano T, Characterization of the soluble membrane attack complex (SMAC) of carp (Cyprinus carpio) complement., Journal of Faculty of Agriculture, Kyushu University, 48, 1-2, 127-134, 48: 127-134., 2003.01.
48. Kono T, Fujiki K, Nakao M, Yano T, Endo M, Sakai M, The immune responses of common carp, Cyprinus carpio L., injected with carp interleukin-1b gene., Journal of Interferon and Cytokine Research, 10.1089/10799900252952190, 22, 4, 413-419, 22: 413-419., 2002.01.
49. Shimasaki Y, Oshima Y, Yokota Y, Kitano T, Nakao M, Kawabata S, Imada N and Honjo T, Purification and identification of a tributyltin-binding protein from serum of Japanese flounder, Paralichthys olivaceus., Environmental Toxicology and Chemistry, 10.1897/1551-5028(2002)021<1229:PAIOAT>2.0.CO;2, 21, 6, 1229-1235, 21: 1229-1235., 2002.01.
50. Nakao M, Matsumoto M, Nakazawa M, Fujiki K and Yano T, Diversity of complement factor B/C2 in the common carp (Cyprinus carpio): Three isotypes of B/C2-A expressed in different tissues., Developmental and Comparative Immunology, 10.1016/S0145-305X(01)00083-0, 26, 6, 533-541, 26: 533-541., 2002.01.
51. Tomana M, Ishikawa J, Imai E, Moritomo T, Nakao M, Yano T, Characterization of immunoglobulin light chain isotypes in the common carp., Immunogenetics, 10.1007/s00251-002-0447-7, 54, 2, 120-129, 54: 120-129., 2002.01.
52. Shin DH, Fujiki K, Nakao M, Yano T, Organization of the NKEF gene and its expression in the common carp (Cyprinus carpio)., Developmental and Comparative Immunology, 10.1016/S0145-305X(01)00021-0, 25, 7, 597-606, 25: 597-606., 2001.01.
53. Nakao M, Osaka K, Kato Y, Fujiki K, Yano T, Molecular cloning of the complement C1r/C1s/MASP2-like serine proteases from the common carp (Cyprinus carpio)., Immunogenetics, 52, 3-4, 255-263, 52: 255-263., 2001.01.
54. Fujiki K, Bayne CJ, Shin DH, Nakao M, Yano T, Molecular cloning of carp (Cyprinus carpio) C-type lectin and pentraxin by use of suppression subtractive hybridisation., Fish & Shellfish Immunology, 10.1006/fsim.2000.0331, 11, 3, 275-279, 11:275-279., 2001.01.
55. Bayne CJ, Gerwick L., Fujiki K., Nakao M, Yano T, Immune relevant genes identified in the liver of rainbow trout, Oncorhynchus mykiss, by means of suppression subtractive hybridization., Developmental and Comparative Immunology, 10.1016/S0145-305X(00)00057-4, 25, 3, 205-217, 24: 205-217., 2001.01.
56. Muturo J, Nakao M, Fujiki K, Yano T, Multiple forms of a2-macroglobulin from a bony fish, the common carp (Cyprinus carpio): Striking sequence diversity in functional sites., Immunogenetics, 10.1007/s002510000216, 51, 10, 847-855, 51: 847-855., 2000.01.
57. Fujiki K, Shin DH, Nakao M, Yano T, Molecular cloning of carp (Cyprinus carpio) leukocyte cell-derived chemotaxin 2, glia maturation factor b, CD45 and lysozyme C by use of suppression subtractive hybridization., Fish & Shellfish Immunology, 10.1006/fsim.2000.0294, 10, 7, 643-650, 10: 642-650., 2000.01.
58. Nakao M, Mutsuro J, Obo R, Fujiki K, Nonaka M, Yano T, Molecular cloning and protein analysis of divergent forms of the complement component C3 from a bony fish, the common carp (Cyprinus carpio): presence of variants lacking the catalytic histidine., European Journal of Immunology, 10.1002/1521-4141(200003)30:3<858::AID-IMMU858>3.3.CO;2-D, 30, 3, 858-866, 30: 858-866., 2000.01.
59. Fujiki K, Shin DH, Nakao M, Yano T, Molecular cloning and expression analysis of the putative carp (Cyprinus carpio) pre-B cell enhancing factor., Fish & Shellfish Immunology, 10.1006/fsim.2000.0263, 10, 4, 383-385, 10: 383-385., 2000.01.
60. Fujiki K, Shin DH, Nakao M, Yano T, Molecular cloning and expression analysis of carp (Cyprinus carpio) interleukin-1b, high affinity immunoglobulin E Fc receptor g subunit and serum amyloid A., Fish & Shellfish Immunology, 10.1006/fsim.1999.0253, 10, 3, 229-242, 10: 229-241., 2000.01.
61. Nagai T, Mutsuro J, Kimura M, Kato Y, Fujiki K, Yano T, Nakao M, A novel truncated isoform of the mannose-binding lectin-associated serine protease (MASP) from the common carp (Cyprinus carpio)., Immunogenetics, 10.1007/s002510050031, 51, 3, 193-200, 51: 193-200., 2000.01.
62. Fujiki K, Shin DH, Nakao M, Yano T, Molecular cloning of carp (Cyprinus carpio) CC chemokine, CXC chemokine receptors, allograft inflammatory factor-1, and natural killer cell enhancing factor by use of suppression subtractive hybridization., Immunogenetics, 10.1007/s002510050573, 49, 10, 909-914, 49: 909-914., 1999.01.
63. Tomana M, Nakao M, Moritomo T, Fujiki K, Yano T, Isolation of cDNA encoding immunoglobulin light chain from common carp (Cyprinus carpio)., Fish & Shellfish Immunology, 9: 71-80., 1999.01.
64. Endo Y, Takahashi M, Nakao M, Saiga H, Sekine H, Matsushita M, Nonaka M, Fujita T, Two lineages of mannose-binding lectin-associated serine protease (MASP) in vertebrates., Journal of Immunology, 161, 9, 4924-4930, 161: 4924-4930., 1998.01.
65. Nakao M, Fushitani Y, Fujiki K, Nonaka M, Yano T, Two diverged complement factor B/C2-like cDNA sequences from a teleost, the common carp (Cyprinus carpio)., Journal of Immunology, 161, 9, 4811-4818, 161: 4811-4818., 1998.01.
66. Nakao M, Moritomo T., Tomana M, Fujiki K, Yano T, Isolation of cDNA encoding the constant region of the immunoglobulin heavy-chain from common carp (Cyprinus carpio L.)., Fish & Shellfish Immunology, 10.1006/fsim.1998.0149, 8, 6, 425-434, 8: 425-434., 1998.01.
67. Oshima Y, Nirmala K, Yokota Y, Go J, Shimasaki Y, Nakao M, Lee RF, Imada N, Kobayashi K, Accumulation of tributhyltin (TBT) in the blood of flounder and dab intraperitoneally administered with TBT., Marine Environment Research, 10.1016/S0141-1136(97)00082-2, 46, 1-5, 587-590, 46: 587-590., 1998.01.