Methotrexate Enhances Apoptosis of Transmembrane TNF-Expressing Cells Treated With Anti-TNF Agents
Methotrexate Enhances Apoptosis of Transmembrane TNF-Expressing Cells Treated With Anti-TNF Agents
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DOI：
10.3389/fimmu.2020.02042
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发表时间：
2020-08
影响因子：
7.3
通讯作者：
Qiaolei Wang;D. Oryoji;H. Mitoma;Y. Kimoto;M. Koyanagi;K. Yokoyama;M. Ayano;M. Akahoshi;Y. Arinobu;H. Niiro;K. Akashi;T. Horiuchi
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作者：
Qiaolei Wang;D. Oryoji;H. Mitoma;Y. Kimoto;M. Koyanagi;K. Yokoyama;M. Ayano;M. Akahoshi;Y. Arinobu;H. Niiro;K. Akashi;T. Horiuchi
背景甲氨蝶呤（MTX）合用可提高抗肿瘤坏死因子（tnf）治疗类风湿性关节炎（RA）的临床疗效。我们旨在阐明MTX对经抗TNF药物处理的跨膜TNF （tmTNF）表达细胞的细胞毒性作用。方法采用稳定表达tmTNF的Jurkat T细胞进行后续实验。用流式细胞术比较抗tnf药物（英夫利昔单抗、阿达木单抗或certolizumab pegol）联合MTX诱导的细胞毒性与单独使用MTX或单独使用抗tnf药物诱导的细胞毒性。研究了逆转录信号通过tmTNF介导的细胞凋亡诱导、补体依赖性细胞毒性（CDC）、抗体依赖性细胞毒性（ADCC）和抗体依赖性细胞吞噬（ADCP）。以叶酸和Rho激酶抑制剂Y-27632作为抑制剂，研究MTX和抗tnf诱导细胞凋亡的细胞内信号通路。结果与单用MTX或单用抗tnf相比，MTX和抗tnf合用可显著增加表达tmtnf的细胞凋亡。合并MTX诱导细胞凋亡在英夫利昔单抗治疗中最为明显。MTX和抗tnf药物对表达tmtnf的细胞的协同作用是由反向信号转导，而不是CDC或ADCC/ADCP。叶酸抑制mtx介导的细胞凋亡，而Y-27632通过tmTNF的反向信号抑制JNK激活和英夫利昔单抗诱导的细胞凋亡。结论MTX联合抗肿瘤坏死因子可增强表达tmtnf的细胞的凋亡作用。MTX和抗tnf诱导的细胞内通路似乎是独立的。这些发现可能至少在一定程度上解释了甲氨蝶呤和抗肿瘤坏死因子联合治疗RA后临床反应的改善。
Background Concomitant use of methotrexate (MTX) improves the clinical efficacy of anti-TNF agents in the treatment of rheumatoid arthritis (RA). We aimed to clarify the cytotoxic effect of MTX on transmembrane TNF (tmTNF)-expressing cells treated with anti-TNF agents. Methods Jurkat T cells stably expressing tmTNF were used for the following experiments. Cytotoxicity induced by an anti-TNF agent (infliximab, adalimumab, or certolizumab pegol) with concomitant MTX were compared with that by MTX alone or by an anti-TNF agent alone using flow cytometry. Apoptosis-induction mediated by reverse signal through tmTNF, complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), and antibody-dependent cellular phagocytosis (ADCP) were evaluated. Folic acid and Y-27632, a Rho kinase inhibitor, were used as inhibitors to study intracellular signaling pathway in apoptosis induced by MTX and anti-TNF agents. Results Apoptosis of tmTNF-expressing cells was significantly increased by the concomitant administration of MTX and an anti-TNF agent, compared with MTX alone or an anti-TNF agent alone. The apoptosis induction by concomitant MTX was most pronounced in infliximab-treatment. Reverse signal transduction, but not CDC or ADCC/ADCP, was responsible for the coordinate effect of MTX and an anti-TNF agent on tmTNF-expressing cells. Folic acid inhibited MTX-mediated apoptosis, while Y-27632 suppressed JNK activation and infliximab-induced apoptosis via revere signal through tmTNF. Conclusion The apoptotic effect was enhanced by combination of MTX and an anti-TNF agent in tmTNF-expressing cells. The intracellular pathways induced by MTX and anti-TNF agents seem to be independent. These findings might explain at least in part improved the clinical response upon co-therapy of MTX and an anti-TNF agent in RA.