Saw Valerie P J, Dart Robin J C, Galatowicz Grazyna, Daniels Julie T, Dart John K G, Calder Virginia L
Department of Ocular Biology and Therapeutics, University College London Institute of Ophthalmology, London, United Kingdom.
Invest Ophthalmol Vis Sci. 2009 Nov;50(11):5310-7. doi: 10.1167/iovs.08-3345. Epub 2009 Jun 3.
First, to determine whether tumor necrosis factor-(TNF)-alpha is expressed in the conjunctiva of ocular mucous membrane pemphigoid (MMP) and the consequences of systemic immunosuppressive treatment on this expression. Second, to investigate the in vitro effects of TNFalpha on human conjunctival fibroblasts.
The expression of TNFalpha in conjunctival tissues of patients with actively inflamed ocular MMP (n = 10), patients with clinically noninflamed ocular MMP after systemic immunosuppressive treatment (n = 10), and normal subjects (n = 10) was studied by immunohistochemistry. The effect of TNFalpha on functional assays of human conjunctival fibroblast activity were investigated, including migration, collagen lattice contraction, matrix metalloproteinase (mmp), and tissue inhibitor of matrix metalloproteinase (timp) secretion, proliferation, and surface expression of HLA-DR, ICAM, CD80, CD86, CD40, CD40-ligand.
In active ocular MMP, TNFalpha is expressed by a large number of stromal infiltrating cells (234 cells/mm(2)), and although the level of stromal TNFalpha expression is significantly reduced after immunosuppressive treatment (90 cells/mm(2)), these levels are still significantly elevated compared with normal conjunctiva (10 cells/mm(2), P < 0.05). TNFalpha stimulates increased migration by conjunctival fibroblasts (P < 0.001), increased production of mmp-9 (P = 0.01), decreased production of timp-2 (P = 0.01) and timp-4 (P = 0.04), and upregulated expression of CD40 and ICAM (P = 0.04). No significant effects of TNFalpha on fibroblast proliferation or collagen lattice contraction were detected.
Increased conjunctival expression of TNFalpha in ocular MMP suggests that systemic TNFalpha antagonists are likely to be effective in controlling severe disease unresponsive to conventional systemic immunosuppression. Residual TNFalpha expression persists in clinically noninflamed disease. TNFalpha appears to have profibrotic and proinflammatory effects on human conjunctival fibroblasts.
第一，确定肿瘤坏死因子-α（TNF-α）是否在眼黏膜类天疱疮（MMP）的结膜中表达，以及全身免疫抑制治疗对该表达的影响。第二，研究TNF-α对人结膜成纤维细胞的体外作用。
通过免疫组织化学研究活动性眼MMP患者（n = 10）、全身免疫抑制治疗后临床无炎症的眼MMP患者（n = 10）以及正常受试者（n = 10）结膜组织中TNF-α的表达。研究TNF-α对人结膜成纤维细胞活性功能测定的影响，包括迁移、胶原晶格收缩、基质金属蛋白酶（MMP）和基质金属蛋白酶组织抑制剂（TIMP）分泌、增殖以及HLA-DR、ICAM、CD80、CD86、CD40、CD40配体的表面表达。
在活动性眼MMP中，大量基质浸润细胞表达TNF-α（234个细胞/mm²），尽管免疫抑制治疗后基质TNF-α表达水平显著降低（90个细胞/mm²），但与正常结膜（10个细胞/mm²，P < 0.05）相比，这些水平仍显著升高。TNF-α刺激结膜成纤维细胞迁移增加（P < 0.001），MMP-9产生增加（P = 0.01），TIMP-2（P = 0.01）和TIMP-4（P = 0.04）产生减少，以及CD40和ICAM表达上调（P = 0.04）。未检测到TNF-α对成纤维细胞增殖或胶原晶格收缩有显著影响。
眼MMP中结膜TNF-α表达增加表明，全身TNF-α拮抗剂可能有效控制对传统全身免疫抑制无反应的严重疾病。在临床无炎症的疾病中仍存在残余的TNF-α表达。TNF-α似乎对人结膜成纤维细胞具有促纤维化和促炎作用。
