Establishment of a highly reproducible model of immunological male infertility with analyses of related testicular autoantigens

• Project/Area Number: 23500504
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: Tokyo Medical University
• Principal Investigator: ITOH Masahiro 東京医科大学, 医学部, 教授 (00232471)
• Co-Investigator(Kenkyū-buntansha): KAWAMOTO Eiichi 東京医科大学, 医学部, 准教授 (20074718) ; SASAKI Hiraku 順天堂大学, スポーツ健康科学部, 助教 (20384969) ; NAITO Munekazu 愛知医科大学, 医学部, 准教授 (10384984) ; QU Ning 東京医科大学, 医学部, 講師 (70527952) ; HIRAI Shuichi 東京医科大学, 医学部, 講師 (70516054) ; TERAYAMA Hayato 東海大学, 医学部, 講師 (00384983) ; HAYASHI Shogo 東京医科大学, 医学部, 准教授 (60349496) ; IKEDA Ayumi 東京医科大学, 医学部, 助教 (90617515) ; HATAYAMA Naoyuki 東京医科大学, 医学部, 助教 (80534792)
• Research Collaborator: OGAWA Yuki ; KURAMASU Miyuki ; HIRAYANAGI Yoshie ; KUERBAN Maimaiti ; MUSHA Muhetaerjiang
• Project Period (FY): 2011-04-28 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 精子形成 / 自己免疫性精巣炎 / 自己抗原 / タンパク質同定 / マウス / 精子抗原 / 精巣 / 自己免疫疾患 / 抗原解析 / 精巣上体

Outline of Final Research Achievements

Experimental autoimmune orchitis (EAO) is one of the models of immunological male infertility. To establish a highly reproducible model of EAO, in 2011, we found that only one placement of a syngeneic donor’s testes, epididymides and vasa deferentia into the abdominal cavity was sufficient to induce EAO on the recipient’s testes in mice. It was also noted that the placement of testes, epididymides and vasa deferentia induced only orchitis without epididymo-vasitis. At the next three years, the testicular autoantigens were identified using sera obtained from EAO-affected mice. The testicular proteins were separated using two-dimensional gel electrophoresis, and the autoantigens proteins were identified by Western blotting and Mass spectrometry. Some identified testicular specific autoantigens have not been reported previously, and the clarification of these proteins will facilitate new approaches for understanding EAO.

Research Products

• [Journal Article] Busulfan pretreatment for transplantation of rat spermatogonia differentially affects immune and reproductive systems in male recipient mice. (2015)
  • Author(s): Hirayanagi Y, Qu N, Hirai S, Naito M, Terayama H, Hayashi S, Hatayama N, Kuramasu M, Ogawa Y, Itoh M.
  • Journal Title: Anat Sci Int. Volume: in press
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Contribution of IL-12/IL-35 common subunit p35 to maintaining the testicular immune privilege (2014)
  • Author(s): Terayama H, Yoshimoto T, Hirai S, Naito M, Qu N, Hatayama N, Hayashi S, Mitobe K, Furusawa J, Mizoguchi I, Kezuka T, Goto H, Suyama K, Moriyama H, Sakabe K, Itoh M.
  • Journal Title: PLoS One Volume: 9 Issue: 4 Pages: e96120-e96120
  • DOI: 10.1371/journal.pone.0096120
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Chronological changes of delayed-type hypersensitivity in mice immunised with testicular germ cells alone. (2014)
  • Author(s): Qu N, Naito M, Terayama H, Hirai S, Musha M, Itoh M.
  • Journal Title: Andrologia Volume: 46 Issue: 5 Pages: 556-563
  • DOI: 10.1111/and.12116
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Neonatal estrogen treatment with β-estradiol 17-cypionate induces in post-pubertal mice inflammation in the ductuli efferentes, epididymis, and vas deferens, but not in the testis, provoking obstructive azoospermia. (2014)
  • Author(s): Naito M, Hirai S, Terayama H, Qu N, Hayashi S, Hatayama N, Kawamura H, Nakano T, Itoh M.
  • Journal Title: Med Mol Morphol. Volume: 47 Issue: 1 Pages: 21-30
  • DOI: 10.1007/s00795-013-0034-z
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cadmium exposure increases susceptibility to testicular autoimmunity in mice. (2014)
  • Author(s): Ogawa Y, Itoh M, Hirai S, Suna S, Naito M, Qu N, Terayama H, Ikeda A, Miyaso H, Matsuno Y, Komiyama M, Mori C.
  • Journal Title: Journal of Applied Toxicology Volume: 33(7) Issue: 7 Pages: 652-660
  • DOI: 10.1002/jat.2713
  • Peer Reviewed
• [Journal Article] Lymphangiogenesis in chronic inflammation in the testis. (2013)
  • Author(s): Hirai S, Naito M, Terayama H, Qu N, Kuerban M, Musha M, Itoh M.
  • Journal Title: Andrology Volume: 1(1) Issue: 1 Pages: 147-154
  • DOI: 10.1111/j.2047-2927.2012.00015.x
  • Peer Reviewed
• [Journal Article] Effects on the Local Immunity in the Testis by Exposure to Di-(2-ethylhexyl) Phthalate (DEHP) in Mice (2013)
  • Author(s): Kitaoka M, Hirai S, Terayama H, Naito M, Qu N, Hatayama N, Miyaso H, Matsuno Y, Komiyama M, Itoh M, Mori C.
  • Journal Title: Journal of Reproduction and Development Volume: 59 Issue: 5 Pages: 485-490
  • DOI: 10.1262/jrd.2012-180
  • NAID: 130003361051
  • ISSN: 0916-8818, 1348-4400
  • Peer Reviewed
• [Journal Article] The Effects of Adjuvants on Autoimmune Responses Against Testicular Antigens in Mice (2013)
  • Author(s): Musha M, Hirai S, Naito M, Terayama H, Qu N, Hatayama N, Itoh M.
  • Journal Title: Journal of Reproduction and Development Volume: 59 Issue: 2 Pages: 139-144
  • DOI: 10.1262/jrd.2012-121
  • NAID: 10031170771
  • ISSN: 0916-8818, 1348-4400
  • Peer Reviewed
• [Journal Article] Xenogeneic and endogenous spermatogenesis following transplantation of rat germ cells into testes of immunocompetent mice. (2012)
  • Author(s): Qu N, Naito M, Li J, Terayama H, Hirai S, Itoh M
  • Journal Title: Reproduction, Fertility, and Development Volume: 24 Issue: 2 Pages: 337-343
  • DOI: 10.1071/rd10349
  • Peer Reviewed
• [Journal Article] Involvement of Fas/Fas-L and Bax/Bcl-2 systems in germ cell death following immunization with syngeneic testicular germ cells in mice. (2012)
  • Author(s): Kuerban M, Naito M, Hirai S, Terayama H, Qu N, Ikeda A, Musha M, Koji T, Itoh M
  • Journal Title: Journal of Andrology Volume: 33(5) Pages: 824-831
  • Peer Reviewed
• [Journal Article] Postinflammation stage of autoimmune orchitis induced by immunization with syngeneic testicular germ cells alone in mice. (2012)
  • Author(s): Naito M, Hirai S, Terayama H, Qu N, Kuerban M, Musha M, Kitaoka M, Ogawa Y, Itoh M
  • Journal Title: Medical Molecular Morphology Volume: 45(1) Issue: 1 Pages: 35-44
  • DOI: 10.1007/s00795-011-0539-2
  • NAID: 10030573640
  • Peer Reviewed
• [Journal Article] The effects of adjuvants on autoimmune responses against testicular antigens in mice (2012)
  • Author(s): Musha M, Hirai S, Naito M, Terayama H, Qu N, Hatayama N, Itoh M
  • Journal Title: Journal of Reproduction and Development Volume: 59(2) Pages: 139-144
  • NAID: 10031170771
  • Peer Reviewed
• [Journal Article] Experimental model of autoimmune orchitis with abdominal placement of donor's testes, epididymes and vasa deferentia in recipient mice (2011)
  • Author(s): Hayato Terayama, Masahiro Itoh, Munekazu Naito, Shuichi Hirai, Ning Qu, Maimaiti Kuerban, Muhetaerjiang Musha
  • Journal Title: Journal of Reproductive Immunology Volume: 90(2) Issue: 2 Pages: 195-201
  • DOI: 10.1016/j.jri.2011.03.008
  • Peer Reviewed
• [Journal Article] Intratesticular Expression of mRNAs of Both Interferon γ and Tumor Necrosis Factor α is Significantly Increased in Experimental Autoimmune Orchitis in Mice (2011)
  • Author(s): Hayato Terayama, Munekazu Naito, Ning Qu, Shuichi Hirai, Miyuki Kitaoka, Yuki Ogawa, Masahiro Itoh
  • Journal Title: Journal of Reproduction and Development Volume: 57 Issue: 2 Pages: 296-302
  • DOI: 10.1262/jrd.10-121N
  • NAID: 10028275659
  • ISSN: 0916-8818, 1348-4400
  • Peer Reviewed
• [Presentation] 微量カドミウム曝露は自己免疫障害を増悪させる (2014)
  • Author(s): 小川夕輝, 倉升三幸, 曲 寧, 平井宗一, 林 省吾, 畑山直之, 伊藤正裕
  • Organizer: 第29回日本生殖免疫学会総会・学術集会
  • Place of Presentation: 東京
  • Year and Date: 2014-12-13
• [Presentation] ブスルファン処置マウスの精子形成障害における牛車腎気丸の治療効果 (2014)
  • Author(s): 曲 寧, 平栁淑恵, 林 省吾, 平井宗一, 畑山直之, 倉升三幸, 小川夕輝, 伊藤正裕
  • Organizer: 第29回日本生殖免疫学会総会・学術集会
  • Place of Presentation: 東京
  • Year and Date: 2014-12-12 – 2014-12-13
• [Presentation] 進化と発生からみた「ヒト精子の生きにくさ」 (2014)
  • Author(s): 伊藤 正裕
  • Organizer: 第32回日本受精着床学会総会・学術講演会
  • Place of Presentation: 東京
  • Year and Date: 2014-07-31 – 2014-08-01
  • Invited
• [Presentation] Contribution of IL-35 to maintaining the testicular immune privilege (2014)
  • Author(s): 寺山隼人, 内藤宗和, 善本隆之, 平井宗一, 曲 寧, 倉升三幸, 金沢輝久, 隅山香織, 坂部 貢, 伊藤正裕
  • Organizer: アンドロロジー学会第33回学術大会および第20回精子形成・精巣毒性研究会
  • Place of Presentation: 長野
  • Year and Date: 2014-06-12 – 2014-06-13
• [Presentation] Role of macrophages in newly formed lymphatic vessels in the testis. (2013)
  • Author(s): Hirai S, Naito M, Terayama H, Qu N, Hatayama N, Musha M, Itoh M.
  • Organizer: 12th International Congress of Reproductive Immunology
  • Place of Presentation: Boston, USA
• [Presentation] Testicular immune-microenvironment in the Epstein-Barr virus-induced gene-3 (EBI3) knock out mice. (2013)
  • Author(s): Terayama H, Hirai S, Yoshimoto T, Naito M, Qu N, Hatayama N, Sakabe K*, Itoh M.
  • Organizer: 12th International Congress of Reproductive Immunology
  • Place of Presentation: Boston, USA
• [Presentation] 免疫正常マウスにおけるブスルファンの免疫系と生殖系への影響 (2013)
  • Author(s): 平柳淑恵, 曲 寧, 平井宗一, 内藤宗和, 倉升三幸, 林 省吾, 畑山直之, 伊藤正裕
  • Organizer: 第20回日本免疫毒性学会学術大会
  • Place of Presentation: 東京
• [Presentation] 微量カドミウム曝露は自己免疫性精子形成障害を増悪させる (2013)
  • Author(s): 小川夕輝, 倉升三幸, 曲 寧, 平井宗一, 林 省吾, 畑山直之, 内藤宗和, 伊藤正裕
  • Organizer: 第20回日本免疫毒性学会学術大会
  • Place of Presentation: 東京
• [Presentation] 低用量フタル酸曝露による自己免疫性精巣炎誘導への影響 (2013)
  • Author(s): 内藤宗和, 平井宗一, 曲 寧, 倉升三幸, 小川夕輝, 畑山直之, 林 省吾, 伊藤正裕
  • Organizer: 第28回日本生殖免疫学会総会・学術集会
  • Place of Presentation: 兵庫
• [Presentation] 精巣とサイトカイン (2013)
  • Author(s): 寺山 隼人, 内藤 宗和, 曲 寧, 平井 宗一, 畑山 直之, 伊藤 正裕
  • Organizer: 第118回日本解剖学会総会・学術集会
  • Place of Presentation: 香川
  • Invited
• [Presentation] Autoimmune responses induced by immunization with xenogenic testicular germ cells alone (2013)
  • Author(s): 曲 寧, 寺山 隼人, 平井 宗一, 平栁 淑恵. 内藤 宗和, 伊藤 正裕
  • Organizer: 第118回日本解剖学会総会・学術集会
  • Place of Presentation: 香川
• [Presentation] 後天性精子障害の病態解析 異種生殖細胞に対する免疫正常マウスの精巣組織反応 (2012)
  • Author(s): 曲 寧, 寺山 隼人, 平井 宗一, 内藤 宗和, 伊藤 正裕
  • Organizer: 第27回日本生殖免疫学会総会・学術集会
  • Place of Presentation: 大阪
• [Presentation] Autoimmune responses induced by immunization with xenogenic testicular germ cells alone (2012)
  • Author(s): Ning Qu, Hayato Terayama, Shuichi Hirai, Yoshie Hirayanag, Munekazu Naito, Masahiro Itoh
  • Organizer: 日本アンドロロジー学会第31回学術大会ならびに総会および第18回精子形成・精巣毒性研究会 共同開催学会
  • Place of Presentation: 兵庫
• [Presentation] 異種生殖細胞に対する免疫正常マウスの精巣組織反応 (2011)
  • Author(s): 曲 寧、内藤宗和、寺山隼人、平井宗一、伊藤正裕
  • Organizer: 第67回日本顕微鏡学会学術講演会(招待講演)
  • Place of Presentation: 福岡
• [Presentation] 自己免疫性精巣炎における細胞死 (2011)
  • Author(s): 平井宗一、内藤宗和、寺山隼人、曲 寧、池田あゆみ、伊藤正裕
  • Organizer: 第43回日本臨床分子形態学会総会・学術集会(招待講演)
  • Place of Presentation: 大阪