Komura K, Fujimoto M, Yanaba K, Matsushita T, Matsushita Y, Horikawa M, Ogawa F, Shimizu K, Hasegawa M, Takehara K, Sato S
Department of Dermatology, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan.
Ann Rheum Dis. 2008 Jun;67(6):867-72. doi: 10.1136/ard.2007.073387. Epub 2007 Sep 6.
To assess the association of CD40/CD40 ligand (CD40L) interactions with the development of skin fibrosis and autoimmunity in tight-skin (TSK/+) mouse, which is a mouse model for human systemic sclerosis.
Newly born TSK/+ mice were treated with murine anti-CD40L monoclonal antibody (100 microg intraperitoneally weekly). Hypodermal thickness of 8-week-old female mice (defined as the thickness of a subcutaneous loose connective tissue layer beneath the panniculus carnosus) was measured under a light microscope. All skin sections were taken from the para-midline, upper back region. Serum anti-topoisomerase I autoantibody levels, serum immunoglobulin levels and plasma soluble CD40L levels were determined by enzyme-linked immunosorbent assay. For analysis of lymphocyte surface molecules, single cell suspensions of lymphocytes were stained by monoclonal antibodies. Proliferation of TSK/+ B cells and fibroblasts to anti-CD40 antibodies was assessed by the uptake of [3H]-labelled thymidine and bromodeoxyuridine, respectively.
The blockade of CD40/CD40L interactions by anti-CD40L monoclonal antibody significantly reduced cutaneous fibrosis (65%) and anti-topoisomerase I autoantibody in TSK/+ mice. Anti-CD40L monoclonal antibody also normalised B lymphocyte abnormal activation in TSK/+ mice, demonstrated by hyper-gamma-globulinaemia. Furthermore, augmented CD40/CD40L interactions in TSK/+ mice were suggested by upregulated expression of CD40L on CD4(+) T cells, elevated plasma soluble CD40L levels. The hyperresponsiveness to CD40 stimulation was also observed in TSK/+ B cells and fibroblasts.
Cutaneous fibrosis and autoimmunity in TSK/+ mice are closely correlated with CD40/CD40L interactions.
评估CD40/CD40配体（CD40L）相互作用与紧皮（TSK/+）小鼠皮肤纤维化和自身免疫发展的关联，该小鼠是人类系统性硬化症的模型。
新生TSK/+小鼠每周腹腔注射鼠抗CD40L单克隆抗体（100微克）。在光学显微镜下测量8周龄雌性小鼠的皮下厚度（定义为肉质膜下方皮下疏松结缔组织层的厚度）。所有皮肤切片取自背部中线上方区域。采用酶联免疫吸附测定法测定血清抗拓扑异构酶I自身抗体水平、血清免疫球蛋白水平和血浆可溶性CD40L水平。为分析淋巴细胞表面分子，用单克隆抗体对淋巴细胞单细胞悬液进行染色。分别通过[3H]标记胸腺嘧啶核苷和溴脱氧尿苷的摄取来评估TSK/+ B细胞和成纤维细胞对抗CD40抗体的增殖情况。
抗CD40L单克隆抗体阻断CD40/CD40L相互作用可显著减轻TSK/+小鼠的皮肤纤维化（65%）和抗拓扑异构酶I自身抗体。抗CD40L单克隆抗体还使TSK/+小鼠中B淋巴细胞的异常活化恢复正常，高丙种球蛋白血症证明了这一点。此外，TSK/+小鼠中CD40L在CD4(+) T细胞上的表达上调、血浆可溶性CD40L水平升高，提示CD40/CD40L相互作用增强。在TSK/+ B细胞和成纤维细胞中也观察到对CD40刺激的高反应性。
TSK/+小鼠的皮肤纤维化和自身免疫与CD40/CD40L相互作用密切相关。
