Silvestris F, Nagata S, Cafforio P, Silvestris N, Dammacco F
Department of Biomedical Sciences and Human Oncology, University of Bari, Italy.
J Exp Med. 1996 Dec 1;184(6):2287-300. doi: 10.1084/jem.184.6.2287.
Previous studies have demonstrated that T cell-reactive antibodies in HIV-1 infection contribute to lymphocyte depletion by cytotoxicity that involves differential membrane targets, such as the 43.5-kD receptor on CEM cells. Here, we show that these antibodies bind Fas as result of a molecular mimicry of the gp120. Both flow cytometry and immunoblotting using the human Fas-transfected mouse WC8 lymphoma revealed positive binding of immunoglobulin G from several patients to a 43.8-kD membrane receptor that also reacts with the CH11 anti-Fas monoclonal antibody. Specificity to Fas was further confirmed to chimeric recombinant human Fas-Fc by ELISA, whereas overlapping peptide mapping of a Fas domain (VEINCTR-N) shared by gp120 V3 loop demonstrated a predominant affinity to the full-length 10-mer peptide. Four anti-Fas affinity preparations greatly increased the subdiploid DNA peak of CEM cells similar to agonist ligands of Fas. In addition, anti-Fas immunoglobulin G strongly inhibited the [3H]thymidine uptake of CEM cells in proliferative assays, inducing a suppression as high as provoked by both CH11 mAb and recombinant human Fas ligand. Since anti-Fas were reactive to gp120, it is conceivable that antibodies binding that domain within the V3 region are effective cross-linkers of Fas and increase apoptosis in peripheral T cells. These results suggest that autologous stimulation of the Fas pathway, rather than of lymphocytotoxic antibodies, may aggravate lymphopenia in a number of HIV-1+ subjects.
先前的研究表明,HIV-1感染中的T细胞反应性抗体通过细胞毒性导致淋巴细胞耗竭,这种细胞毒性涉及不同的膜靶点,如CEM细胞上的43.5-kD受体。在此,我们表明这些抗体由于对gp120的分子模拟而结合Fas。使用人Fas转染的小鼠WC8淋巴瘤进行的流式细胞术和免疫印迹均显示,数名患者的免疫球蛋白G与一种43.8-kD膜受体呈阳性结合,该受体也与CH11抗Fas单克隆抗体发生反应。通过ELISA进一步证实了对Fas的特异性,而gp120 V3环共享的Fas结构域(VEINCTR-N)的重叠肽图谱显示对全长10聚体肽具有主要亲和力。四种抗Fas亲和制剂与Fas激动剂配体相似,大大增加了CEM细胞的亚二倍体DNA峰。此外,在增殖试验中,抗Fas免疫球蛋白G强烈抑制CEM细胞的[3H]胸苷摄取,诱导出与CH11单克隆抗体和重组人Fas配体所引发的抑制作用一样高的抑制作用。由于抗Fas对gp120有反应,因此可以想象,结合V3区域内该结构域的抗体是Fas的有效交联剂,并增加外周T细胞的凋亡。这些结果表明,Fas途径的自体刺激而非淋巴细胞毒性抗体可能会加重一些HIV-1阳性受试者的淋巴细胞减少症。