title: Specific autoantigens identified by sera obtained from mice that are immunized with testicular germ cells alone.
abstract: There are various autoimmunogenic antigens (AIs) in testicular germ cells (TGCs) recognized as foreign by the body's immune system. However, there is little information of TGC-specific AIs being available. The aim of this study is to identify TGC-specific AIs. We have previously established that immunization using viable syngeneic TGC can also induce murine experimental autoimmune orchitis (EAO) without using any adjuvant. This study is to identify TGC-specific AIs by TGC liquid chromatography-tandem mass spectrometry analysis, followed by two-dimensional gel electrophoresis that reacted with serum IgG from EAO mice. In this study, we identified 11 TGC-specific AIs that reacted with serum from EAO mice. Real-time RT-PCR analysis showed that the mRNA expressions of seven TGC-specific AIs were significantly higher in only mature testis compared to other organs. Moreover, the recombinant proteins of identified 10 (except unnamed protein) TGC-specific AIs were created by using human embryonic kidney 293 (HEK293) cells and these antigencities were reconfirmed by Western blot using EAO serum reaction. These results indicated Atp6v1a, Hsc70t, Fbp1 and Dazap1 were candidates for TGC-specific AIs. Identification of these AIs will facilitate new approaches for understanding infertility and cancer pathogenesis and may provide a basis for the development of novel therapies.
authors.affiliation: Department of Anatomy, Division of Basic Medical Science, Tokai University School of Medicine, Kanagawa, Japan.|Department of Anatomy, Tokyo Medical University, Tokyo, Japan.
authors.forename: Hayato
authors.initials: H
authors.lastname: Terayama
authors.affiliation: Department of Anatomy, Tokyo Medical University, Tokyo, Japan.|Department of Anatomy, Aichi Medical University, Aichi, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Hirai
authors.affiliation: Department of Anatomy, Tokyo Medical University, Tokyo, Japan.|Department of Anatomy, Aichi Medical University, Aichi, Japan.
authors.forename: Munekazu
authors.initials: M
authors.lastname: Naito
authors.affiliation: Department of Anatomy, Tokyo Medical University, Tokyo, Japan.
authors.forename: Ning
authors.initials: N
authors.lastname: Qu
authors.affiliation: Department of Molecular and Cellular Physiology, Graduate School of Medicine, University of the Ryukyus, Okinawa, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Katagiri
authors.affiliation: Department of Anatomy, Tokyo Medical University, Tokyo, Japan.
authors.forename: Kenta
authors.initials: K
authors.lastname: Nagahori
authors.affiliation: Department of Anatomy, Tokyo Medical University, Tokyo, Japan.
authors.forename: Shogo
authors.initials: S
authors.lastname: Hayashi
authors.affiliation: Department of Health Science, School of Health and Sports Science, Juntendo University, Chiba, Japan.
authors.forename: Hiraku
authors.initials: H
authors.lastname: Sasaki
authors.affiliation: Department of Biochemistry, Tokyo Medical University, Tokyo, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Moriya
authors.affiliation: Department of Biochemistry, Tokyo Medical University, Tokyo, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Hiramoto
authors.affiliation: Department of Biochemistry, Tokyo Medical University, Tokyo, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Miyazawa
authors.affiliation: Department of Anatomy, Tokyo Medical University, Tokyo, Japan.|Department of Anatomy, Aichi Medical University, Aichi, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Hatayama
authors.affiliation: Department of Anatomy, Tokyo Medical University, Tokyo, Japan.
authors.forename: Zhong-Lian
authors.initials: ZL
authors.lastname: Li
authors.affiliation: Department of Anatomy, Division of Basic Medical Science, Tokai University School of Medicine, Kanagawa, Japan.
authors.forename: Kou
authors.initials: K
authors.lastname: Sakabe
authors.affiliation: Department of Molecular and Cellular Physiology, Graduate School of Medicine, University of the Ryukyus, Okinawa, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Matsushita
authors.affiliation: Department of Anatomy, Tokyo Medical University, Tokyo, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Itoh
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000097:blood; D001324:Autoantigens / Q000097:blood*; D001327:Autoimmune Diseases / Q000097:blood / Q000276:immunology; D004822:Epididymis / Q000378:metabolism; D005786:Gene Expression Regulation; D005854:Germ Cells / Q000166:cytology*; D057809:HEK293 Cells; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000378:metabolism; D008297:Male; D051379:Mice; D009920:Orchitis / Q000097:blood / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D013094:Spermatozoa / Q000378:metabolism; D013737:Testis / Q000166:cytology*
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D007074:Immunoglobulin G; D012333:RNA, Messenger