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Autophagy promotes citrullination of VIM (vimentin) and its interaction with major histocompatibility complex class II in synovial fibroblasts

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Title: Autophagy promotes citrullination of VIM (vimentin) and its interaction with major histocompatibility complex class II in synovial fibroblasts
Authors: Sugawara, Eri Browse this author
Kato, Masaru Browse this author →KAKEN DB
Kudo, Yuki Browse this author
Lee, Wenshi Browse this author
Hisada, Ryo Browse this author
Fujieda, Yuichiro Browse this author →KAKEN DB
Oku, Kenji Browse this author →KAKEN DB
Bohgaki, Toshiyuki Browse this author →KAKEN DB
Amengual, Olga Browse this author →KAKEN DB
Yasuda, Shinsuke Browse this author →KAKEN DB
Onodera, Tomohiro Browse this author →KAKEN DB
Hatakeyama, Shigetsugu Browse this author →KAKEN DB
Atsumi, Tatsuya Browse this author →KAKEN DB
Keywords: Autophagy
citrullination
MHC class II
rheumatoid arthritis
vimentin
Issue Date: 3-May-2020
Publisher: Taylor & Francis
Journal Title: Autophagy
Volume: 16
Issue: 5
Start Page: 946
End Page: 955
Publisher DOI: 10.1080/15548627.2019.1664144
Abstract: We aimed to investigate the involvement of macroautophagy/autophagy in autoimmunity in rheumatoid arthritis (RA) through citrullination of VIM (vimentin) and its interaction with MHC class II in synovial fibroblasts (SFs). The cell surface expression of MHC class II and B7 costimulatory molecules on SFs was analyzed by flow cytometry after treatment with IFNG/IFN-gamma (interferon gamma). Intracellular citrullinated autoantigens in SFs were analyzed by immunoblotting using serum from anti-citrullinated peptide antibodies (ACPA)-positive patient as a primary antibody. SFs were incubated in serum-free medium or treated with proteasome inhibitor MG132 to induce autophagy. An autophagy inhibitor 3-methyladenin (3-MA) was used. Intracellular citrullinated VIM (cVIM) was evaluated by immunoblotting and immunocytochemistry. The interaction between MHC class II and cVIM was evaluated with co-immunoprecipitation and proximity ligation assay (PLA). We demonstrated that MHC class II, CD274/B7-H1 and PDCD1LG2/B7-DC were expressed on SFs following treatment with IFNG whereas CD276/B7-H3 was detected on SFs regardless of the presence of IFNG. ACPA-positive sera recognized a 54 kDa protein in SFs. By immunoprecipitation, the 54 kDa protein recognized by RA sera was revealed to be cVIM. Following induction of autophagy, intracellular cVIM was increased in SFs but the effect was canceled by 3-MA. The interaction between MHC class II and cVIM was demonstrated by co-immunoprecipitation. Furthermore, PLA revealed the significant increase of MHC class II-cVIM interaction following induction of autophagy. Our findings suggest that SFs may contribute to the autoimmunity in RA through citrullination of VIM and its interaction with MHC class II promoted by autophagy.
Rights: This is an Accepted Manuscript of an article published by Taylor & Francis in Autophagy on 03 May 2020, available online: http://www.tandfonline.com/10.1080/15548627.2019.1664144.
Type: article (author version)
URI: http://hdl.handle.net/2115/81113
Appears in Collections:北海道大学病院 (Hokkaido University Hospital) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 加藤 将

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