Differential effects of anti-TNF-α drugs on fibroblast-like synoviocyte apoptosis
Differential effects of anti-TNF-α drugs on fibroblast-like synoviocyte apoptosis
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DOI：
10.1093/rheumatology/kep358
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发表时间：
2010-03-01
期刊：
影响因子：
5.5
通讯作者：
Salvarani, Carlo
中科院分区：
文献类型：
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作者：
Pattacini, Laura;Boiardi, Luigi;Salvarani, Carlo
Objective.靶向TNF-α的新型药物可用于治疗RA。成纤维细胞样滑膜细胞（FLS）在RA进展中起着重要作用，通过其部分由细胞死亡诱导抗性引起的扩增。本研究的目的是确定不同的抗TNF-α药物对FLS增殖的影响。将来自RA或OA患者的FLS与外周血单核细胞（PBMC）共培养，并与各种药物孵育6天。随后，通过核小体ELISA和末端脱氧核苷酸转移酶dUTP缺口末端标记检测凋亡诱导。Western blot检测10号染色体上缺失的磷酸酶和张力蛋白同源物（PTEN）-粘着斑激酶（FAK）通路的激活以及Bax和Bcl-2的水平。免疫沉淀法用于研究跨膜TNF-α（tmTNF-α）的磷酸化。只有当两个细胞群通过激活PTEN-FAK途径和增加Bax水平直接接触时，所有测试的药物在从同一患者获得的PBMC存在下诱导FLS的凋亡。这种效应不是由于抗体依赖性细胞介导的细胞毒性。只有英夫利昔单抗和阿达木单抗能够上调Bcl-2。依那西普在诱导FLS凋亡方面比其他药物更有效。这种诱导依赖于PBMC的存在，并且涉及PTEN-FAK途径的激活。单克隆抗体英夫利昔单抗和阿达木单抗诱导的Bcl-2增加可能起保护作用，从而抵消其对FLS的促凋亡作用。
Objective. Novel drugs targeting TNF-alpha are available for treatment of RA. Fibroblast-like synoviocytes (FLSs) play a fundamental role in RA progression, through their expansion caused in part by resistance to cell death induction. The aim of our study was to determine the effects of different anti-TNF-alpha agents on FLS apoptosis.Methods. FLS from patients with either RA or OA were co-cultured with peripheral blood mononuclear cells (PBMCs), and incubated with various drugs for 6 days. Subsequently, apoptosis induction was detected by Nucleosome ELISA and terminal deoxynucleotidyl transferase dUTP nick end labeling. Western blot was used to determine the activation of the phosphatase and tensin homologue deleted on chromosome 10 (PTEN)-focal adhesion kinase (FAK) pathway as well as Bax and Bcl-2 levels. Immunoprecipitation was used for studying phosphorylation of transmembrane TNF-alpha (tmTNF-alpha).Results. All the tested drugs induced apoptosis of FLSs in the presence of PBMCs obtained from the same patient only when the two cell populations were in direct contact by activating the PTEN-FAK pathway and increasing Bax levels. This effect was not due to antibody-dependent cell-mediated cytotoxicity. Only the two antibodies infliximab and adalimumab were able to up-regulate Bcl-2.Conclusions. Etanercept is more effective in inducing FLS apoptosis compared with the other drugs tested. This induction is dependent on the presence of PBMCs, and involves the activation of PTEN-FAK pathway. Bcl-2 increase induced by the monoclonal antibodies infliximab and adalimumab may play a protective role and thus counteract their pro-apoptotic effect on FLSs.