| 1. |
Toyoda M, Fukuda T, Fujimoto R, Kawakami K, Hayashi C, Nakao Y, Watanabe Y, Aoki T, Shida M, Sanui T, Taguchi M, Yamamichi K, Okabe A, Okada T, Oka K, Nakayama K, Nishimura F, Kajioka S., Scaffold-free bone-like 3D structure established through osteogenic differentiation from human gingiva-derived stem cells., Biomed Res Int, 10.1016/j.bbrep.2024.101656, 15, 38, 101656-101656, 2024.02. |
| 2. |
Kawakami K, Fukuda T, Toyoda M, Nakao Y, Hayashi C, Watanabe Y, Aoki T, Shinjo T, Iwashita M, Yamashita A, Shida M, Sanui T, Uchiumi T, Nishimura F., Luteolin Is a Potential Immunomodulating Natural Compound against Pulpal Inflammation., Biomed Res Int, 10.1155/2024/8864513, 2024, 8864513-8864513, 2024.01. |
| 3. |
Taketomi T, Fukuda T, Takeshita G, Sanui T., A Case of a Dentigerous Cyst in the Maxillary Sinus Treated Preoperatively with Vascular Embolization to Avoid Intraoperative Abnormal Bleeding., Cureus, 10.7759/cureus.50228, 15, 12, e50228-e50228, 2023.12. |
| 4. |
Imagawa M, Shinjo T, Sato K, Kawakami K, Zeze T, Nishimura Y, Toyoda M, Chen S, Ryo N, Ahmed AK, Iwashita M, Yamashita A, Fukuda T, Sanui T, Nishimura F., Epithelial-to-mesenchymal transition, inflammation, subsequent collagen production, and reduced proteinase expression cooperatively contribute to cyclosporin-A-induced gingival overgrowth development., Front Physiol, 10.3389/fphys.2023.1298813, 13, 14, 1298813-1298813, 2023.12. |
| 5. |
Zeze T, Shinjo T, Sato K, Nishimura Y, Imagawa M, Chen S, Ahmed A-K, Iwashita M, Yamashita A, Fukuda T, Sanui T, Park K, King GL, Nishimura F., Endothelial Insulin Resistance Exacerbates Experimental Periodontitis., J Dent Res, 10.1177/00220345231181539, 102, 10, 1152-1161, 2023.09. |
| 6. |
Li R, Sano T, Mizokami A, Fukuda T, Shinjo T, Iwashita M, Yamashita A, Sanui T, Nakatsu Y, Sotomaru Y, Asano T, Kanematsu T, Nishimura F., miR-582-5p targets Skp1 and regulates NF-κB signaling-mediated inflammation., Arch Biochem Biophys, 10.1016/j.abb.2022.109501, 15, 734, 109501-109501, 2023.01. |
| 7. |
Hayashi C, Fukuda T, Kawakami K, Toyoda M, Nakao Y, Watanabe Y, Shinjo T, Sano T, Iwashita M, Yotsumoto K, Shida M, Taketomi T, Sanui T, Uchiumi T, Kanematsu T, Nishimura F., miR-1260b inhibits periodontal bone loss by targeting ATF6β mediated regulation of ER stress., Front Cell Dev Biol, doi: 10.3389/fcell.2022.1061216. eCollection 2022., 30, 10, 1061216.-1061216., 2022.11. |
| 8. |
Nishimura Y, Iwashita M, Hayashi M, Shinjo T, Watanabe Y, Zeze T, Yamashita A, Fukuda T, Sanui T, Sano T, Asano T, Nishimura F., XAF1 overexpression exacerbates diabetes by promoting pancreatic β-cell apoptosis., Acta Diabetol, doi: 10.1007/s00592-022-01930-y., 59, 10, 1275-1286, 2022.10. |
| 9. |
Watanabe Y, Fukuda T, Hayashi C, Nakao Y, Toyoda M, Kawakami K, Shinjo T, Iwashita M, Yamato H, Yotsumoto K, Taketomi T, Uchiumi T, Sanui T, Nishimura F., Extracellular vesicles derived from GMSCs stimulated with TNF-α and IFN-α promote M2 macrophage polarization via enhanced CD73 and CD5L expression., Sci Rep, doi: 10.1038/s41598-022-17692-0., 12, 1, 13344.-13344., 2022.08. |
| 10. |
Hayashi M, Iwashita M, Nishimura Y, Shinjo T, Sano T, Yamashita A, Fukuda T, Sanui T, Asano T, Nishimura F., Adipose-specific C-C motif chemokine ligand (CCL) 19 overexpression drives the mice to both insulin resistance and weight gain., BMJ Open Diabetes Res Care, doi: 10.1136/bmjdrc-2020-001871., 9, 1, e001871.-e001871., 2021.05. |
| 11. |
Nakao Y, Fukuda T, Zhang Q, Sanui T, Shinjo T, Kou X, Chen C, Liu D, Watanabe Y, Hayashi C, Yamato H, Yotsumoto K, Tanaka U, Taketomi T, Uchiumi T, Le AD, Shi S, Nishimura F., Exosomes from TNF-α-treated human gingiva-drived MSCs enhance M2 macrophage polarization and inhibit periodontal bone loss., Acta Biomaterials, doi: 10.1016/j.actbio.2020.12.046., 122, 306-324, 2021.03. |
| 12. |
Yamato H, Sanui T, Yotsumoto K, Nakao Y, Watanabe Y, Hayashi C, Aihara R, Iwashita M, Tanaka U, Taketomi T, Fukuda T, Nishimura F., Combined application of geranylgeranylacetone and amelogenin promotes angiogenesis and wound healing in human periodontal ligament cells., Journal of Cellular Biochemistry, doi: 10.1002/jcb.29903., 2021.02. |
| 13. |
Alshargabi R, Shinjo T, Iwashita M, Yamashita A, Sano T, Nishimura Y, Hayashi M, Zeze T, Fukuda T, Sanui T, Nishimura F., SPOCK1 induces adipose tissue maturation: New insights into the function of SPOCK1 in metabolism., Biochemical and Biophysical Research Communications, doi: 10.1016/j.bbrc.2020.09.129., 533, 4, 1076-1082, 2020.12. |
| 14. |
Alshargabi R, Sano T, Yamashita A, Takano A, Sanada T, Iwashita M, Shinjo T, Fukuda T, Sanui T, Kishida S, Nishimura F., SPOCK1 is a novel inducer of epithelial to mesenchymal transition in drug-induced gingival overgrowth., Scientific Reports, doi: 10.1038/s41598-020-66660-z., 10, 1, 9785-9785, 2020.06. |
| 15. |
Yotsumoto K, Sanui T, Tanaka U, Yamato H, Alshargabi R, Shinjo T, Nakao Y, Watanabe Y, Hayashi C, Taketomi T, Fukuda T, Nishimura F, Amelogenin downregulates interferon gamma-induced major histocompatibility complex class II expression through suppression of euchromatin formation in the class II transactivator promoter IV region in macrophages., Frontiers in Immunology, doi.org/10.3389/fimmu.2020.00709, 11, 709-709, 2020.03. |
| 16. |
Sano T, Sanada T, Sotomaru Y, Shinjo T, Iwashita M, Yamashita A, Fukuda T, Sanui T, Asano T, Kanematsu T, Nishimura F, Ccr7 null mice are protected against diet-induced obesity via Ucp1 upregulation and enhanced energy expenditure., Nutrition & Metabolism, 4, 16, 43-43, 2019.07. |
| 17. |
Taketomi T, Onimura T, Yoshiga D, Muratsu D, Sanui T, Fukuda T, Kusukawa J, Nakamura S, Sprouty2 is involved in the control of osteoblast proliferation and differentiation through the FGF and BMP signaling pathways., Cell Biology International, 10.1002/cbin.10876., 42, 9, 1106-1114, 2018.09. |
| 18. |
Sanui T, Fukuda T, Tanaka U, Toyoda K, Yotsumoto K, Nakao Y, Yamato H, Taketomi T, Nishimura F, Sprouty2 Inhibition Resolves Inflammation in Periodontal Disease and Creates a Suitable Environment for Periodontal Tissue Regeneration., Journal of Cell Biology & Immunology, 1, 101-101, 2017.11. |
| 19. |
Kensuke Yamamichi, Takao Fukuda, Terukazu Sanui, Kyousuke Toyoda, Urara Tanaka, Yuki Nakao, Karen Yotsumoto, Hiroaki Yamato, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura, Amelogenin induces M2 macrophage polarisation via PGE2/cAMP signalling pathway, Archives of Oral Biology, 10.1016/j.archoralbio.2017.08.005, 83, 241-251, 2017.11, Objectives Amelogenin, the major component of the enamel matrix derivative (EMD), has been suggested as a bioactive candidate for periodontal regeneration. Apart from producing a regenerative effect on periodontal tissues, amelogenin has also been reported to have an anti-inflammatory effect. However, the precise molecular mechanisms underlying these effects remain unclear. In the present study, we examined the immunomodulatory effects of amelogenin on macrophages. Design Human phorbol 12-myristate 13-acetate (PMA)-differentiated U937 macrophages and CD14+ peripheral blood-derived monocytes (PBMC)-derived macrophages were stimulated with recombinant amelogenin (rM180). After performing a detailed microarray analysis, the effects of rM180 on macrophage phenotype and signal transduction pathways were evaluated by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, confocal microscopy and flow cytometry. Results The microarray analysis demonstrated that rM180 increased the expression of anti-inflammatory genes in lipopolysaccharide (LPS)-challenged macrophages after 24 h, while it temporarily up-regulated inflammatory responses at 4 h. rM180 significantly enhanced the expression of M2 macrophage markers (CD163 and CD206). rM180-induced M2 macrophage polarisation was associated with morphological changes as well as vascular endothelial growth factor (VEGF) production. rM180 enhanced prostaglandin E2 (PGE2) expression, and the activation of the cAMP/cAMP-responsive element binding (CREB) signaling pathway was involved in amelogenin-induced M2 macrophage polarisation. Blocking of PGE2 signaling by indomethacin specifically abrogated rM180 with or without LPS-induced M2 shift in PBMC-derived macrophages. Conclusion Amelogenin could reprogram macrophages into the anti-inflammatory M2 phenotype. It could therefore contribute to the early resolution of inflammation in periodontal lesions and provide a suitable environment for remodeling-periodontal tissues.. |
| 20. |
Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Urara Tanaka, Rehab Alshargabi, Yoshitomi Aida, Fusanori Nishimura, Roles of serum in innate immune responses of human leukocytes to synthetic lipopeptide, International Immunopharmacology, 10.1016/j.intimp.2017.06.006, 50, 61-68, 2017.09, Tripalmitoyl-S-glyceryl-L-Cys-Ser-(Lys)4 (Pam3CSK4) is a highly conserved molecular motif found in various classes of lipoproteins. The requirement for leukocyte to respond to synthetic Pam3CSK4 were studied. Pam3CSK4 primed neutrophils for a respiratory burst in a serum-dependent manner. Pam3CSK4 upregulated CD11b, CD14, and cytochrome b558, and downregulated Leu-8. Treatment of neutrophils with anti-CD14 antibodies and treatment of serum with anti-LPS binding protein (LBP) antibodies resulted in the inhibition of priming for respiratory burst by Pam3CSK4. It should be noted that LBP could not replicate the effects of serum in priming of neutrophils for respiratory burst by Pam3CSK4. Serum LBP bound to immobilized Pam3CSK4. Pam3CSK4 induced the interleukin-8 (IL-8) production by leukocytes in a serum-dependent manner. Further, Pam3CSK4-induced priming of neutrophils for respiratory burst was not inhibited by the LPS antagonists LA-14-PP, Rhodobacter sphaeroides LPS, or E5531, and Pam3CSK4-induced IL-8 production by leukocytes was not affected by LPS antagonist, E5531, indicating that Pam3CSK4 was recognized by a different receptor than LPS. Thus, Pam3CSK4 and LPS had similar biological activities and similar requirement to act on leukocytes, but were recognized by different receptors. Serum in the action of Pam3CSK4 on leukocytes was not replicated by LBP, suggesting that Pam3CSK4 might be disaggregated by serum to result in the activation of leukocytes.. |
| 21. |
Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Urara Tanaka, Rehab Alshargabi, Yoshitomi Aida, Fusanori Nishimura, Adhesion attenuates respiratory burst induced by different modes of triggering in resting or LPS-primed neutrophils, Immunobiology, 10.1016/j.imbio.2017.05.001, 222, 8-9, 865-871, 2017.08, The effects of adherence on neutrophil superoxide anion (O2 −) generation triggered by surface, soluble ligand, or adherence were studied. Resting-neutrophils adhered to the uncoated tubes resulting in O2 − generation, but not on plasma-, fibrinogen-, vitronectin-, fibronectin-, laminin-, collagen-, or poly HEMA-coated surfaces. Enhanced N-formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated O2 − generation by LPS-primed-neutrophils was induced by the incubation on plasma, fibrinogen, vitronectin, fibronectin, or laminin in the absence of Mg2+. In the presence of Mg2+, this response was observed in cells on collagen or poly HEMA. LPS-primed-neutrophils adhered to uncoated, BSA- or IgG-coated tubes and did not respond to fMLP, indicating that the fMLP-response of LPS-primed-neutrophils was suppressed by adherence. Upon incubation on plasma, fibrinogen, vitronectin, fibronectin in the presence of Mg2+, LPS-primed-neutrophils showed O2 − generation. Upon incubation on collagen or poly HEMA, the primed-neutrophils neither generated O2 − nor adhered. We found that O2 − response of LPS-primed-neutrophils was attenuated depending on the time of exposure to plasma-coated surface. This attenuation was evident on plasma or fibrinogen, but not on collagen in the presence of Mg2+, indicating that O2 − generation by LPS-primed-neutrophils was attenuated dependent on adherence but not on Mg2+. Thus, adhesion attenuated the O2 − generation triggered by both soluble (fMLP) and insoluble (surface) stimuli.. |
| 22. |
Terukazu Sanui, 福田 隆男, 山道 研介, Kyosuke Toyoda, Urara Tanaka, 四本 かれん, 武富 孝治, 西村 英紀, Microarray analysis of the effects of amelogenin on U937 monocytic cells., American Journal of Molecular Biology, in press, 2017.04. |
| 23. |
Aiko Takano, Takao Fukuda, Takanori Shinjo, Misaki Iwashita, Etsuko Matsuzaki, Kensuke Yamamichi, Masaaki Takeshita, Terukazu Sanui, Fusanori Nishimura, Angiopoietin-like protein 2 is a positive regulator of osteoblast differentiation, Metabolism: Clinical and Experimental, 10.1016/j.metabol.2017.01.006, 69, 157-170, 2017.04, Introduction and Aims Several studies have reported that angiopoietin-like protein 2 (Angptl2) is expressed abundantly in adipocytes and is associated with adipose tissue inflammation. In the present study, we found that osteoblasts and mesenchymal stem cells also expressed Angptl2 at high levels. The aim of this study was to understand the role of Angptl2 in osteoblastic cell differentiation. Methods Angptl2 expression was examined during osteoblast and adipocyte differentiation. The role of Angptl2 on cell differentiation and associated signaling was analyzed by gene knockdown using Angptl2 small interfering ribonucleic acid (siRNA). Results Angptl2 was highly expressed in MC3T3-E1 cells, ST2 cells and primary osteoblasts, but not in RAW264 cells. Inhibition of Angptl2 expression using siRNA markedly inhibited alkaline phosphatase (ALP) expression and osteoblastic differentiation in MC3T3-E1, ST2 cells and primary osteoblasts. Angptl2 siRNA also inhibited adipocyte differentiation in ST2 cells. Treatment of MC3T3-E1 cells with Angptl2 siRNA led to the down-regulation of the activities of several cell signaling pathways, including extracellular signal-regulated kinase (ERK), Jun amino-terminal kinase (JNK), Akt, and nuclear factor kappa B (NF-κB) signals. It also down-regulated the expression of Osterix, but not that of runt-related transcription factor 2 (Runx2), suggesting that Angptl2 is a positive activator of Osterix and its down-stream signals. Treatment of MC3T3-E1 cells with anti-Angptl2 antibodies suppressed ALP gene expression. In addition, treatment of Angptl2 siRNA-treated cells with culture supernatants of normal MC3T3-E1 cells restored ALP gene expression, indicating that Angptl2 acts in an autocrine manner. Conclusions The results suggest that Angptl2 is an autocrine positive regulator of cell differentiation. Thus, it is suggested that Angptl2 regulates not only adipose tissue metabolism but also bone metabolism.. |
| 24. |
Tomomi Sano, S. Nagayasu, S. Suzuki, Misaki Iwashita, Akiko Yamashita, T. Shinjo, Terukazu Sanui, A. Kushiyama, T. Kanematsu, T. Asano, Fusanori Nishimura, Epicatechin downregulates adipose tissue CCL19 expression and thereby ameliorates diet-induced obesity and insulin resistance, Nutrition, Metabolism and Cardiovascular Diseases, 10.1016/j.numecd.2016.11.008, 27, 3, 249-259, 2017.03, Background and aims Epicatechin (EC) intake has been suggested to be beneficial for the prevention of cardiovascular disorders, and it is well known that adipose tissue inflammation is one of the major risk factors for coronary heart diseases. The purpose of the present study was to determine the in vitro and in vivo effects of EC on adipose tissue inflammation and obesity. Methods and results DNA microarray analysis was performed to evaluate the effects of EC on gene expression in adipocytes co-cultured with bacterial endotoxin-stimulated macrophages. To determine the in vivo effects of the catechin, C57BL/6 mice were fed either a high-fat diet (HFD) or HFD combined with EC, and metabolic changes were observed EC suppressed the expression of many inflammatory genes in the adipocytes co-cultured with endotoxin-stimulated macrophages. Specifically, EC markedly suppressed chemokine (C–C motif) ligand 19 (CCL19) expression. The target cell of EC appeared to macrophages. The in vivo study indicated that mice fed the EC-supplemented HFD were protected from diet-induced obesity and insulin resistance. Accordingly, the expression levels of genes associated with inflammation in adipose tissue and in the liver were downregulated in this group of mice. Conclusions EC exerts beneficial effects for the prevention of adipose tissue inflammation and insulin resistance. Since we previously reported that mice deficient in the CCL19 receptor were protected from diet-induced obesity and insulin resistance, it can be concluded that the beneficial effects of EC could be mediated, at least in part, by marked suppression of CCL19 expression.. |
| 25. |
Fusanori Nishimura, Tomomi Sano, Terukazu Sanui,, The influence of periodontal burden on metabolic control of diabetes - Myth or reality? From a nutritional perspective -, Current Oral Health Reports, doi:10.1007/s.40496-017-0136-0, ４, 59-63, 2017.03. |
| 26. |
Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Akira Haraguchi, Yoshitomi Aida, Fusanori Nishimura, Anti-CD14 Antibody-treated Neutrophils Respond to LPS
Possible Involvement of CD14 Upregulated by Anti-CD14 Antibody Binding, Immunological Investigations, 10.1080/08820139.2016.1238925, 46, 2, 190-200, 2017.02, CD14 and Toll-like receptor 4/MD2 (TLR4/MD2) mediate the action of LPS on neutrophils. The anti-CD14 antibody and the TLR4/MD2-antagonist, synthetic lipid IVa (LA-14-PP), are known to inhibit the response of neutrophils to LPS. We studied the role of CD14 in LPS-induced priming of neutrophils for enhanced release of the superoxide anion. The anti-CD14 antibody at much higher concentrations than required to saturate CD14 was required to inhibit priming by LPS. The inhibitory effect of the anti-CD14 antibody was overcome by LPS. After washing, anti-CD14-treated neutrophils showed upregulated CD14 upon incubation at 37°C and responded to LPS with a delayed time-course. Thus, CD14-blocked neutrophils gained responsiveness to LPS through newly upregulated CD14. These results suggested that the unbound/free anti-CD14 antibody was essential to inhibit LPS-induced priming by blocking CD14 that were newly expressed during incubation at 37°C. LA-14-PP inhibited the response of neutrophils to LPS in an anti-CD14 antibody sensitive manner. When neutrophils were treated with LA-14-PP followed by treatment with the anti-CD14 antibody, CD14 was upregulated upon warming, but priming was blocked, suggesting that TLR4/MD2 was not newly expressed by warming in association with CD14 molecules. Thus, in addition to blocking CD14, the anti-CD14 antibody was found to induce the expression of new CD14.. |
| 27. |
Masaaki Takeshita, Akira Haraguchi, Mayumi Miura, Takafumi Hamachi, Takao Fukuda, Terukazu Sanui, Aiko Takano, Fusanori Nishimura, Antibiotic effects against periodontal bacteria in organ cultured tissue, Clinical and Experimental Dental Research, 10.1002/cre2.48, 3, 1, 5-12, 2017.02, Mechanical reduction of infectious bacteria by using physical instruments is considered the principal therapeutic strategy for periodontal disease; addition of antibiotics is adjunctive. However, local antibiotic treatment, combined with conventional mechanical debridement, has recently been shown to be more effective in periodontitis subjects with type 2 diabetes. This suggests that some bacteria may invade the inflamed inner gingival epithelium, and mechanical debridement alone will be unable to reduce these bacteria completely. Therefore, we tried to establish infected organ culture models that mimic the inner gingival epithelium and aimed to see the effects of antibiotics in these established models. Mouse dorsal skin epithelia were isolated, and periodontal bacteria were injected into the epithelia. Infected epithelia were incubated with test antibiotics, and colony-forming ability was evaluated. Results indicated that effective antibiotics differed according to injected bacteria and the bacterial combinations tested. Overall, in organ culture model, the combination of amoxicillin or cefdinir and metronidazole compensate for the effects of less effective bacterial combinations on each other. This in vitro study would suggest effective periodontal treatment regimens, especially for severe periodontitis.. |
| 28. |
Kohji Nozoe, Terukazu Sanui, Masaaki Takeshita, Takao Fukuda, Akira Haraguchi, Yoshitomi Aida, Fusanori Nishimura, Innate immune-stimulatory activity of Porphyromonas gingivalis fimbriae is eliminated by phase separation using Triton X-114, Journal of Immunological Methods, 10.1016/j.jim.2016.11.012, 441, 31-38, 2017.02, Fimbriae are virulence factors of Porphyromonas gingivalis (P. gingivalis). In this study, the action of fimbriae on neutrophil respiratory burst and cytokine production by mononuclear cells (MNC) were investigated. Native or denatured form of purified P. gingivalis fimbriae contained endotoxin at an equivalence of 1– 3 μg lipopolysaccharides (LPS)/mg protein. The endotoxin could be reduced to the equivalent of 1 ng-LPS/mg protein by phase separation using Triton X-114. Unfractionated fimbriae caused serum-dependent priming of neutrophils for enhanced respiratory burst, but both native and denatured forms of Triton X-114-fractionated fimbriae were not active at 100 μg/mL. Unfractionated fimbriae induced serum-dependent production of IL-1β by MNC. Triton X-114-fractionated fimbriae (10 μg/mL)-induced production of IL-1β, IL-8 or TNF-α was much lower than that induced by unfractionated fimbriae or 10 ng/mL P. gingivalis-LPS preparation. Triton X-114-fractionated fimbriae immobilized on polystyrene tubes induced adhesion-stimulated superoxide release by LPS-primed neutrophils in a β2 integrin-dependent manner. P. gingivalis cells caused priming of neutrophils; however, Toll-like receptor (TLR) 4 antagonists did not affect this response. Thus, P. gingivalis fimbriae were ineffective in inducing innate immune response in leukocytes; however, they induced β2 integrin-mediated response by neutrophils. Immune-stimulatory components of P. gingivalis might be recognized by receptors other than TLR4.. |
| 29. |
Takaharu Taketomi, Tomohiro Onimura, Daigo Yoshiga, Daichi Muratsu, Terukazu Sanui, Takao Fukuda, Jingo Kusukawa, Seiji Nakamura, Sprouty2 is involved in the control of osteoblast proliferation and differentiation through the FGF and BMP signaling pathways, Cell Biology International, 10.1002/cbin.10876, 2017.01, Fibroblast growth factor (FGF) and bone morphogenetic protein (BMP) play essential roles in bone formation and osteoblast activity through the extracellular signal-regulated kinase 1/2 (ERK1/2) and Smad pathways. Sprouty family members are intracellular inhibitors of the FGF signaling pathway, and four orthologs of Sprouty have been identified in mammals. In vivo analyses have revealed that Sprouty2 is associated with bone formation. However, the mechanism by which the Sprouty family controls bone formation has not been clarified. In this study, we investigated the involvement of Sprouty2 in osteoblast proliferation and differentiation. We examined Sprouty2 expression in MC3T3-E1 cells, and found that high levels of Sprouty2 expression were induced by basic FGF stimulation. Overexpression of Sprouty2 in MC3T3-E1 cells resulted in suppressed proliferation compared with control cells. Sprouty2 negatively regulated the phosphorylation of ERK1/2 after basic FGF stimulation, and of Smad1/5/8 after BMP stimulation. Furthermore, Sprouty2 suppressed the expression of osterix, alkaline phosphatase, and osteocalcin mRNA, which are markers of osteoblast differentiation. Additionally, Sprouty2 inhibited osteoblast matrix mineralization. These results suggest that Sprouty2 is involved in the control of osteoblast proliferation and differentiation by downregulating the FGF-ERK1/2 and BMP-Smad pathways, and suppresses the induction of markers of osteoblast differentiation.. |
| 30. |
Takanori Shinjo, Misaki Iwashita, Akiko Yamashita, Tomomi Sano, Mitsudai Tsuruta, Hiroaki Matsunaga, Terukazu Sanui, Tomoichiro Asano, Fusanori Nishimura, IL-17A synergistically enhances TNFα-induced IL-6 and CCL20 production in 3T3-L1 adipocytes, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2016.06.049, 477, 2, 241-246, 2016.08, Interleukin-17A (IL-17A) is known to induce inflammatory responses and to be involved in the pathogenesis of not only autoimmune diseases, but also several metabolic and infectious diseases. In this study, IL-17A is shown to induce IL-6 expression in 3T3-L1 mature adipocytes. Interestingly, we found that IL-17A synergistically amplified TNFα-induced secretion of IL-6 and upregulation of IL-17RA expression in 3T3-L1 adipocytes. Its synergistic effects on IL-6 production were inhibited by pre-treatment with inhibitors of IκBα and JNK. Furthermore, IL-17A cooperatively enhanced LPS-mediated IL-6 production in 3T3-L1 adipocytes co-cultured with RAW264.7 macrophages. In addition, IL-17A also enhanced CCL20 production in 3T3-L1 adipocytes stimulated with TNFα or co-cultured with LPS-stimulated RAW macrophages. In high-fat diet-fed mouse epididymal adipose tissues, IL-17RA and RORγt mRNA levels were significantly increased and the serum level of CCL20 was also upregulated. Taken together, these data show that, in adipose tissues, IL-17A contributes to exacerbating insulin resistance-enhancing IL-6 production and promotes the infiltration of Th17 cells in cooperation with TNFα; these findings represent a novel hypothesis for the association between IL-17A-producing cells and type 2 diabetes.. |
| 31. |
Terukazu Sanui, 福田 隆男, Urara Tanaka, Kyosuke Toyoda, 山道 研介, 張 祥翼, 武富 孝治, 西村 英紀, Biological effects of Sprouty2 inhibition in periodontal ligament cells., Journal of Cell Signaling, 10.4172/jcs.1000117, 1, 117, 2016.07. |
| 32. |
Kohji Nozoe, Yoshitomi Aida, Takao Fukuda, Terukazu Sanui, Fusanori Nishimura, Mechanisms of the Macrolide-Induced Inhibition of Superoxide Generation by Neutrophils, Inflammation, 10.1007/s10753-016-0333-3, 39, 3, 1039-1048, 2016.06, The effect of macrolides on the superoxide (O2 −) production by neutrophils was studied. Resting neutrophils become primed by lipopolysaccharide (LPS) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), and primed neutrophils generate O2 − in response to fMLP or adhesion, respectively. Both LPS-primed fMLP-stimulated O2 − generation by macrolide-treated neutrophils and adhesion-stimulated O2 − generation by macrolide-treated fMLP-primed neutrophils were inhibited. Macrolide inhibition of O2 − generation was dependent on serum or pH. Serum could be substituted by NaHCO3. The intensity of inhibition was azithromycin = roxithromycin > clarithromycin > erythromycin, in that order. Non-antimicrobial derivatives of erythromycin, that is, EM703 and EM900, inhibited O2 − generation at pH 7.4. NH4Cl abolished the activity of azithromycin (AZ) only when added to neutrophils with AZ but not after incubation with AZ, suggesting that NH4Cl prevented the influx of AZ. AZ did not affect the expression of alkaline phosphatase, CD11b, and cytochrome b558 in both resting and LPS-primed neutrophils. These results suggested that macrolides did not affect granule mobilization but inhibited O2 − generation selectively.. |
| 33. |
Toshiya Komatsu, Yoshitomi Aida, Takao Fukuda, Terukazu Sanui, Shunji Hiratsuka, Michael J. Pabst, Fusanori Nishimura, Disaggregation of lipopolysaccharide by albumin, hemoglobin or high-density lipoprotein, forming complexes that prime neutrophils for enhanced release of superoxide, Pathogens and Disease, 10.1093/femspd/ftw003, 74, 3, 2016.04, We studied the interaction of LPS with albumin, hemoglobin or high-density lipoprotein (HDL), and whether the interaction affected the activity of LPS on neutrophils. These proteins disaggregated LPS, depending upon temperature and LPS:protein ratio. Albumin-treated LPS was absorbed by immobilized anti-albumin antibody and was eluted with Triton X-100, indicating that LPS formed a hydrophobic complex with albumin. Rd mutant LPS was not disaggregated by the proteins, and did not form a complex with the proteins. But triethylamine-treated Rd mutant LPS formed complexes. When LPS was incubated with an equal concentration of albumin and with polymyxin B (PMXB), PMXB-LPS-protein three-way complexes were formed. After removal of PMXB, the complexes consisted of 11-15 LPS monomers bound to one albumin or hemoglobin molecule. LPS primed neutrophils for enhanced release of formyl peptide-stimulated superoxide, in a serum- and LPS-binding protein (LBP)-dependent manner. Although LPS plus LBP alone did not prime neutrophils, albumin-, hemoglobin- or HDL-treated LPS primed neutrophils when added with LBP. Triethylamine-treated Rd mutant LPS primed neutrophils only when incubated with one of the proteins and with LBP. Thus, in addition to LBP, disaggregation and complex formation of LPS with one of these proteins is required for LPS to prime neutrophils.. |
| 34. |
後村 亮, 讃井 彰一, 福田 隆男, 田中 麗, 豊田 敬介, 山道 研介, 武富 孝治, 西村 英紀, Inhibition of Sprouty2 polarizes macrophages toward an M2 phenotype by stimulation with interferon γ and Porphyromonas gingivalis lipopolysaccharide., Immunity, Inflammation and Disease, 10.1002/iid3.99, 26, 4, 98-110, 2016.02. |
| 35. |
Kyousuke Toyoda, Takao Fukuda, Terukazu Sanui, Urara Tanaka, Kensuke Yamamichi, Ryo Atomura, Hidefumi Maeda, Atsushi Tomokiyo, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura, Grp78 Is Critical for Amelogenin-Induced Cell Migration in a Multipotent Clonal Human Periodontal Ligament Cell Line, Journal of Cellular Physiology, 10.1002/jcp.25087, 231, 2, 414-427, 2016.01, Periodontal ligament stem cells (PDLSCs) are known to play a pivotal role in regenerating the periodontium. Amelogenin, which belongs to a family of extracellular matrix (ECM) proteins, is a potential bioactive molecule for periodontal regenerative therapy. However, its downstream target molecules and/or signaling patterns are still unknown. Our recent proteomic study identified glucose-regulated protein 78 (Grp78) as a new amelogenin-binding protein. In this study, we demonstrate, for the first time, the cellular responses induced by the biological interaction between amelogenin and Grp78 in the human undifferentiated PDL cell line 1-17, which possesses the most typical characteristics of PDLSCs. Confocal co-localization experiments revealed the internalization of recombinant amelogenin (rM180) via binding to cell surface Grp78, and the endocytosis was inhibited by the silencing of Grp78 in 1-17 cells. Microarray analysis indicated that rM180 and Grp78 regulate the expression profiles of cell migration-associated genes in 1-17 cells. Moreover, Grp78 overexpression enhanced rM180-induced cell migration and adhesion without affecting cell proliferation, while silencing of Grp78 diminished these activities. Finally, binding of rM180 to Grp78 promoted the formation of lamellipodia, and the simultaneous activation of Rac1 was also demonstrated by NSC23766, a widely accepted Rac1 inhibitor. These results suggest that Grp78 is essential for enhancing amelogenin-induced migration in 1-17 cells. The biological interaction of amelogenin with Grp78 offers significant therapeutic potential for understanding the biological components and specific functions involved in the signal transduction of amelogenin-induced periodontal tissue regeneration. J. Cell. Physiol. 231: 414-427, 2016.. |
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Urara Tanaka, Terukazu Sanui, Takao Fukuda, Kyousuke Toyoda, T. Taketomi, R. Atomura, K. Yamamichi, Hidefumi Maeda, Fusanori Nishimura, Sprouty2 inhibition promotes proliferation and migration of periodontal ligament cells, Oral Diseases, 10.1111/odi.12369, 21, 8, 977-986, 2015.11, Objectives: We previously demonstrated that a dominant-negative Sprouty2 (Spry2) mutation promotes osteoblast proliferation and differentiation after basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) stimulation, whereas it diminishes proliferation of gingival epithelial cells, thereby inducing favourable conditions for periodontal tissue regeneration. In this study, we investigated how Spry2 inhibition affects the cellular physiology of periodontal ligament (PDL) cells. Methods: A total of 1-17 PDL cells (multipotent clonal human PDL cell line) were stimulated with bFGF and EGF after transfection of Spry2 siRNA. Cell proliferation, migration, ALP staining, real-time PCR, Western blot and immunofluorescence assays were performed. Results: ERK1/2 activation and proliferation of 1-17 PDL cells were significantly upregulated by the addition of Spry2 siRNA in the presence of bFGF and EGF. In addition, Spry2 siRNA reduced transcription of osteogenesis-related genes and ALP staining relative to control cells. Furthermore, it increased AKT/phosphatidylinositol 3-kinase (PI3K) phosphorylation; consequently, Rac1 but not Cdc42 was activated, thereby promoting lamellipodia formation, cell proliferation and migration after stimulation by bFGF and EGF. Conclusion: Spry2 combined with bFGF and EGF stimulation reduced PDL cell migration and proliferation with inducing osteoblastic differentiation. These in vitro findings may provide a molecular basis for novel therapeutic approaches for establishing periodontal tissue regeneration.. |
| 37. |
Terukazu Sanui, Urara Tanaka, Takao Fukuda, Kyousuke Toyoda, Takaharu Taketomi, Ryo Atomura, Kensuke Yamamichi, Fusanori Nishimura, Mutation of Spry2 Induces Proliferation and Differentiation of Osteoblasts but Inhibits Proliferation of Gingival Epithelial Cells, Journal of Cellular Biochemistry, 10.1002/jcb.25014, 116, 4, 628-639, 2015.04, Sprouty was identified as an inhibitor of the fibroblast growth factor (FGF) receptor, and Sprouty2 (Spry2) functions as a negative regulator of receptor tyrosine kinase signaling. In this study, we investigated how inhibition of Spry2 affects osteoblasts and gingival epithelial cells in periodontal tissue regeneration in vitro. Transduction of a dominant-negative mutant of Spry2 (Y55A-Spry2) enhanced basic fibroblast growth factor (bFGF)- and epidermal growth factor (EGF)-induced ERK activation in MC3T3-E1 osteoblastic cells. In contrast, it decreased their activation in GE1 cells. Consistent with these observations, Y55A-Spry2 increased osteoblast proliferation with bFGF and EGF stimulation, whereas the proliferation of Y55A-Spry2-introduced GE1 cells was decreased via the ubiquitination and degradation of EGF receptors (EGFRs). In addition, Y55A-Spry2 caused upregulation of Runx2 expression and downregulation of Twist, a negative regulator of Runx2, with treatment of bFGF and EGF, resulting in enhanced osteoblastogenesis accompanied by alkaline phosphatase activation and osteocalcin expression in MC3T3-E1 cells. These data suggest that suppression of Spry2 expression induces proliferation and differentiation of osteoblastic cells after the addition of a bFGF and EGF cocktail but inhibits proliferation in gingival epithelial cells. These in vitro experiments may provide a molecular basis for novel therapeutic approaches in periodontal tissue regeneration. Taken together, our study proposes that combined application of an inhibitor for tyrosine 55 of Spry2, bFGF, and EGF may effectively allow alveolar bone growth and block the ingrowth of gingival epithelial cells toward bony defects, biologically mimicking a barrier effect in guided tissue regeneration, with in vivo investigation in the future. J. Cell. Biochem. 116: 628-639, 2015.. |
| 38. |
讃井 彰一, 福田 隆男, 田中 麗, 豊田 敬介, 武富 孝治, 西村 英紀, Spry2 is a novel therapeutic target for periodontal tissue regeneration through fibroblast growth factor receptor signaling and epidermal growth factor signaling, Receptors & Clinical Investigation, 10.14800/rci.597, e597-e597, 2015.03. |
| 39. |
Takao Fukuda, Terukazu Sanui, Kyousuke Toyoda, Urara Tanaka, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura, Identification of Novel Amelogenin-Binding Proteins by Proteomics Analysis, PLoS One, 10.1371/journal.pone.0078129, 8, 10, 2013.10, Emdogain (enamel matrix derivative, EMD) is well recognized in periodontology. It is used in periodontal surgery to regenerate cementum, periodontal ligament, and alveolar bone. However, the precise molecular mechanisms underlying periodontal regeneration are still unclear. In this study, we investigated the proteins bound to amelogenin, which are suggested to play a pivotal role in promoting periodontal tissue regeneration. To identify new molecules that interact with amelogenin and are involved in osteoblast activation, we employed coupling affinity chromatography with proteomic analysis in fractionated SaOS-2 osteoblastic cell lysate. In SaOS-2 cells, many of the amelogenin-interacting proteins in the cytoplasm were mainly cytoskeletal proteins and several chaperone molecules of heat shock protein 70 (HSP70) family. On the other hand, the proteomic profiles of amelogenin-interacting proteins in the membrane fraction of the cell extracts were quite different from those of the cytosolic-fraction. They were mainly endoplasmic reticulum (ER)-associated proteins, with lesser quantities of mitochondrial proteins and nucleoprotein. Among the identified amelogenin-interacting proteins, we validated the biological interaction of amelogenin with glucose-regulated protein 78 (Grp78/Bip), which was identified in both cytosolic and membrane-enriched fractions. Confocal co-localization experiment strongly suggested that Grp78/Bip could be an amelogenin receptor candidate. Further biological evaluations were examined by Grp78/Bip knockdown analysis with and without amelogenin. Within the limits of the present study, the interaction of amelogenin with Grp78/Bip contributed to cell proliferation, rather than correlate with the osteogenic differentiation in SaOS-2 cells. Although the biological significance of other interactions are not yet explored, these findings suggest that the differential effects of amelogenin-derived osteoblast activation could be of potential clinical significance for understanding the cellular and molecular bases of amelogenin-induced periodontal tissue regeneration.. |
| 40. |
Kaori Matsumura, Takaharu Taketomi, Keigo Yoshizaki, Shinsaku Arai, Terukazu Sanui, Daigo Yoshiga, Akihiko Yoshimura, Seiji Nakamura, Sprouty2 controls proliferation of palate mesenchymal cells via fibroblast growth factor signaling, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2010.12.116, 404, 4, 1076-1082, 2011.01, Cleft palate is one of the most common craniofacial deformities. The fibroblast growth factor (FGF) plays a central role in reciprocal interactions between adjacent tissues during palatal development, and the FGF signaling pathway has been shown to be inhibited by members of the Sprouty protein family. In this study, we report the incidence of cleft palate, possibly caused by failure of palatal shelf elevation, in Sprouty2-deficient (KO) mice. Sprouty2-deficient palates fused completely in palatal organ culture. However, palate mesenchymal cell proliferation estimated by Ki-67 staining was increased in Sprouty2 KO mice compared with WT mice. Sprouty2-null palates expressed higher levels of FGF target genes, such as Msx1, Etv5, and Ptx1 than WT controls. Furthermore, proliferation and the extracellular signal-regulated kinase (Erk) activation in response to FGF was enhanced in palate mesenchymal cells transfected with Sprouty2 small interfering RNA. These results suggest that Sprouty2 regulates palate mesenchymal cell proliferation via FGF signaling and is involved in palatal shelf elevation.. |
| 41. |
Terukazu Sanui, R. L. Gregory, Analysis of Streptococcus mutans biofilm proteins recognized by salivary immunoglobulin A, Molecular Oral Microbiology, 10.1111/j.1399-302X.2009.00523.x, 24, 5, 361-368, 2009.10, Introduction: The purpose of this study was to examine the Streptococcus mutans biofilm cellular proteins recognized by immunoglobulin A (IgA) in saliva from various caries-defined populations. Methods: Biofilm and planktonic S. mutans UA159 cells were prepared. The proteins were extracted, separated by two-dimensional gel electrophoresis, transferred to blotting membranes, and probed for IgA using individual saliva samples from three groups of subjects; those who developed 0 caries (no active caries), 5-9 caries (medium), or more than 10 caries (severe) over a 12-month interval. Results: Several proteins were recognized by salivary IgA in all groups of saliva but spot distribution and intensity varied greatly between the groups, and some proteins were recognized more strongly in biofilm cells than in planktonic culture, and vice versa. Furthermore, 15 proteins were only recognized by saliva from the 'no active caries' group, and four proteins were recognized by saliva samples from subjects in all three groups. Specifically, antigen I/II was recognized less in biofilm cells by caries-free saliva compared with planktonic cells. However, salivary IgA antibody to antigen I/II was absent in blots using saliva from the 'medium caries' and 'severe caries' groups. Conclusion: The bacterial molecules recognized by caries-free saliva are significant factors for S. mutans caries formation, and their inhibition could be a therapeutic target. In addition, saliva of caries-free subjects includes significant IgA antibody against antigen I/II of S. mutans, indicating a protective mechanism. However, microorganisms may protect themselves from host immune attack by forming biofilms and decreasing expression of antigen I/II.. |
| 42. |
Yuya Kunisaki, Yoshihiko Tanaka, Terukazu Sanui, Ayumi Inayoshi, Mayuko Noda, Toshinori Nakayama, Michishige Harada, Masaru Taniguchi, Takehiko Sasazuki, Yoshinori Fukui, DOCK2 is required in T cell precursors for development of Vα14 NK T cells, Journal of Immunology, 176, 8, 4640-4645, 2006.04, Mouse CD1d-restricted Vα14 NKT cells are a unique subset of lymphocytes, which play important roles in immune regulation, tumor surveillance and host defense against pathogens. DOCK2, a mammalian homolog of Caenorhabditis elegans CED-5 and Drosophila melanogaster myoblast city, is critical for lymphocyte migration and regulates T cell responsiveness through immunological synapse formation, yet its role in Vα14 NKT cells remains unknown. We found that DOCK2 deficiency causes marked reduction of Vα14 NKT cells in the thymus, liver, and spleen. When α-galactesylceramide (α-GalCer), a ligand for Vα14 NKT cells, was administrated, cytokine production was scarcely detected in DOCK2-deficient mice, suggesting that DOCK2 deficiency primarily affects generation of Vα14 NKT cells. Supporting this idea, staining with CD1d/α-GalCer tetramers revealed that CD44 -NK1.1- Vα14 NKT cell precursors are severely reduced in the thymuses of DOCK2-deficient mice. In addition, studies using bone marrow chimeras indicated that development of Vα14 NKT cells requires DOCK2 expression in T cell precursors, but not in APCs. These results indicate that DOCK2 is required for positive selection of Vα14 NKT cells in a cell-autonomous manner, thereby suggesting that avidity-based selection also governs development of this unique subset of lymphocytes in the thymus.. |
| 43. |
Hongsi Jiang, Fan Pan, Laurie M. Erickson, Mei Shiang Jang, Terukazu Sanui, Yuya Kunisaki, Takehiko Sasazuki, Masakazu Kobayashi, Yoshinori Fukui, Deletion of DOCK2, a regulator of the actin cytoskeleton in lymphocytes, suppresses cardiac allograft rejection, Journal of Experimental Medicine, 10.1084/jem.20050911, 202, 8, 1121-1130, 2005.10, Allograft rejection is induced by graft tissue infiltration of alloreactive T cells that are activated mainly in secondary lymphoid organs of the host. DOCK2 plays a critical role in lymphocyte homing and immunological synapse formation by regulating the actin cytoskeleton, yet its role in the in vivo immune response remains unknown. We show here that DOCK2 deficiency enables long-term survival of cardiac allografts across a complete mismatch of the major histocompatibility complex molecules. In DOCK2-deficient mice, alloreactivity and allocytotoxicity were suppressed significantly even after in vivo priming with alloantigens, which resulted in reduced intragraft expression of effector molecules, such as interferon-γ, granzyme B, and perforin. This is mediated, at least in part, by preventing potentially alloreactive T cells from recruiting into secondary lymphoid organs. In addition, we found that DOCK2 is critical for CD28-mediated Rac activation and is required for the full activation of alloreactive T cells. Although DOCK2-deficient, alloreactive T cells were activated in vitro in the presence of exogenous interleukin-2, these T cells, when transferred adoptively, failed to infiltrate into the allografts that were transplanted into RAG1-deficient mice. Thus, DOCK2 deficiency attenuates allograft rejection by simultaneously suppressing multiple and key processes. We propose that DOCK2 could be a novel molecular target for controlling transplant rejection. JEM. |
| 44. |
Yuya Kunisaki, Sadahiko Masuko, Mayuko Noda, Ayumi Inayoshi, Terukazu Sanui, Mine Harada, Takehiko Sasazuki, Yoshinori Fukui, Defective fetal liver erythropoiesis and T lymphopoiesis in mice lacking the phosphatidylserine receptor, Blood, 10.1182/blood-2003-09-3245, 103, 9, 3362-3364, 2004.05, Clearance of apoptotic cells by macrophages is considered important for prevention of inflammatory responses leading to tissue damage. The phosphatidylserine receptor (PSR), which specifically binds to phosphatidylserine (PS) exposed on the surface of apoptotic cells, mediates uptake of apoptotic cells in vitro, yet the physiologic relevance of PSR remains unknown. This issue was addressed by generating PSR-deficient (PSR -/-) mice. PSR-/- mice exhibited severe anemia and died during the perinatal period. In the PSR-/- fetal livers, erythroid differentiation was blocked at an early erythroblast stage. In addition, PSR-/- embryos exhibited thymus atrophy owing to a developmental defect of T-lymphoid cells. Clearance of apoptotic cells by macrophages was impaired in both liver and thymus of PSR-/- embryos. However, this did not induce up-regulation of inflammatory cytokines. These results indicate that during embryonic development, PSR-mediated apoptotic cell uptake is required for definitive erythropoiesis and T lymphopoiesis, independently of the prevention of inflammatory responses.. |
| 45. |
Terukazu Sanui, Ayumi Inayoshi, Mayuko Noda, Eiko Iwata, Jens V. Stein, Takehiko Sasazuki, Yoshinori Fukui, DOCK2 regulates Rac activation and cytoskeletal reorganization through interaction with ELMO1, Blood, 10.1182/blood-2003-01-0173, 102, 8, 2948-2950, 2003.10, Although the migratory property of lymphocytes is critical for protective immunity, tissue infiltration of lymphocytes sometimes causes harmful immune responses. DOCK2 plays a critical role in lymphocyte migration by regulating actin cytoskeleton through Rac activation, yet the mechanism by which DOCK2 activates Rac remains unknown. We found that DOCK2 associates with engulfment and cell motility (ELMO1) through its Src-homology 3 (SH3) domain. When DOCK2 was expressed in T-hybridoma cells lacking endogenous expression of DOCK2, Rac activation and actin polymerization were induced. However, such responses were not elicited by the DOCK2 mutant lacking the region required for ELMO1 binding. On the other hand, we found that the expression of ELMO1 induces Rac activation in the plasmacytoma cells expressing DOCK2 but not ELMO1. These results indicate that the association of DOCK2 with ELMO1 is critical for DOCK2-mediated Rac activation, thereby suggesting that their association might be a therapeutic target for immunologic disorders caused by lymphocyte infiltration.. |
| 46. |
Terukazu Sanui, Ayumi Inayoshi, Mayuko Noda, Eiko Iwata, Masahiro Oike, Takehiko Sasazuki, Yoshinori Fukui, DOCK2 is essential for antigen-induced translocation of TCR and lipid rafts, but not PKC-θ and LFA-1, in T cells, Immunity, 10.1016/S1074-7613(03)00169-9, 19, 1, 119-129, 2003.07, DOCK2 is a mammalian homolog of Caenorhabditis elegans CED-5 and Drosophila melanogaster Myoblast City which are known to regulate actin cytoskeleton. DOCK2 is critical for lymphocyte migration, yet the role of DOCK2 in TCR signaling remains unclear. We show here that DOCK2 is essential for TCR-mediated Rac activation and immunological synapse formation. In DOCK2-deficient T cells, antigen-induced translocation of TCR and lipid rafts, but not PKC-θ and LFA-1, to the APC interface was severely impaired, resulting in a significant reduction of antigen-specific T cell proliferation. In addition, we found that the efficacy of both positive and negative selection was reduced in DOCK2-deficient mice. These results suggest that DOCK2 regulates T cell responsiveness through remodeling of actin cytoskeleton via Rac activation.. |
| 47. |
T. Oono, Yoshinori Fukui, S. Masuko, O. Hashimoto, T. Ueno, Terukazu Sanui, A. Inayoshi, Mami Noda, M. Sata, T. Sasazuki, Organ-specific autoimmunity in mice whose T cell repertoire is shaped by a single antigenic peptide, Journal of Clinical Investigation, 10.1172/JCI200113256, 108, 11, 1589-1596, 2001.12, Organ-specific autoimmune diseases have been postulated to be the result of T cell response against organ-specific self-peptides bound to MHC molecules. Contrary to this paradigm, we report here that transgenic mice lacking MHC class I expression and expressing an MHC class II I-Ab molecule that presents only a single peptide (Eα52-68) spontaneously develops peripheral nervous system-specific autoimmune disease with many of the histopathological features found in experimental allergic neuritis. Reciprocal bone marrow chimeras produced using susceptible and resistant lines revealed that bone marrow-derived cells determined disease susceptibility. While the expression of the I-Ab-Eα52-68 complex in the periphery was readily detectable in both lines, its expression on thymic dendritic cells responsible for tolerance induction was markedly lower in the susceptible line than in the resistant line. Consistent with this, CD4+ T cells that can be activated by the I-Ab-Eα52-68 complex were found in the susceptible line, but not in the resistant line. Such CD4+ T cells conferred the disease to the resistant line by adoptive transfer, and administration of Ab specific for the I-Ab-Eα52-68 complex inhibited disease manifestation in the susceptible line. These results indicate that disease development involves systemic T cell reactivity to I-Ab-Eα52-68 complex, probably caused by incomplete negative thymocyte selection.. |
| 48. |
Yoshinori Fukui, Osamu Hashimoto, Terukazu Sanui, Takamasa Oono, Hironori Koga, Masaaki Abe, Ayumi Inayoshi, Mayuko Noda, Masahiro Oike, Toshikazu Shirai, Takehiko Sasazuki, Haematopoietic cell-specific CDM family protein DOCK2 is essential for lymphocyte migration, Nature, 10.1038/35090591, 412, 6849, 826-831, 2001.08, Cell migration is a fundamental biological process involving membrane polarization and cytoskeletal dynamics, both of which are regulated by Rho family GTPases. Among these molecules, Rac is crucial for generating the actin-rich lamellipodial protrusion, a principal part of the driving force for movement. The CDM family proteins, Caenorhabditis elegans CED-5, human DOCK180 and Drosophila melanogaster Myoblast City (MBC), are implicated to mediate membrane extension by functioning upstream of Rac. Although genetic analysis has shown that CED-5 and Myoblast City are crucial for migration of particular types of cells, physiological relevance of the CDM family proteins in mammals remains unknown. Here we show that DOCK2, a haematopoietic cell-specific CDM family protein, is indispensable for lymphocyte chemotaxis. DOCK2-deficient mice (DOCK2-/-) exhibited migration defects of T and B lymphocytes, but not of monocytes, in response to chemokines, resulting in several abnormalities including T lymphocytopenia, atrophy of lymphoid follicles and loss of marginal-zone B cells. In DOCK2-/- lymphocytes, chemokine-induced Rac activation and actin poly- merization were almost totally abolished. Thus, in lymphocyte migration DOCK2 functions as a central regulator that mediates cytoskeletal reorganization through Rac activation.. |
| 49. |
Yoshinori Fukui, Takamasa Oono, Jean Pierre Cabaniols, Kazuki Nakao, Katsuiku Hirokawa, Ayumi Inayoshi, Terukazu Sanui, Jean Kanellopoulos, Eiko Iwata, Mayuko Noda, Motoya Katsuki, Philippe Kourilsky, Takehiko Sasazuki, Diversity of T cell repertoire shaped by a single peptide ligand is critically affected by its acid residue at a T cell receptor contact, Proceedings of the National Academy of Sciences of the United States of America, 10.1073/pnas.250470797, 97, 25, 13760-13765, 2000.12, T cell differentiation in the thymus is driven by positive selection through the interaction of αβ T cell receptors (TCRs) with selfpeptides bound to self-major histocompatibility complex molecules, yet the influence of the peptide sequence on this process remains unknown. To address this issue, we have compared CD4+ T cell differentiation between two sets of mouse lines in which MHC class II I-Ab molecules are occupied with either Eα chain-derived peptide (pEα) or its variant, (p)60k, with one amino acid substitution from leucine to lysine at P5 residue of TCR contacts. Here, we show that despite the comparable expression of I-Ab-peptide complex in the thymus, this substitution from leucine to lysine affects efficiency of positive selection, resulting in extremely small numbers of CD4+ T cells to be selected to mature on I-Ab-(p)60k complex. Furthermore, we show that, although I-Ab-pEα complex selects diverse T cells, T cell repertoire shaped by I-Ab-(p)60k complex is markedly constrained. Our findings thus suggest that positive selection is both specific and degenerate, depending on the amino acid residues at TCR contacts of the selecting self-peptides.. |
