Scheider T, Beck A, Röpke C, Ullrich R, Harthus H P, Bröker M, Pauli G
AIDS-Zentrum des Bundesgesundheitsamtes, Robert Koch-Institut, Berlin, Germany.
AIDS. 1993 May;7(5):647-54. doi: 10.1097/00002030-199305000-00006.
To characterize antigens on uninfected T lymphocytes reactive with monoclonal antibodies (MAb) directed against the HIV-1 Nef protein, and to search for antibodies directed against this epitope in HIV-1-infected individuals.
Murine MAb directed against an epitope of Nef defined by amino acids 60-73 reacted with cell surface antigens of normal peripheral blood lymphocytes and permanent human T-cell lines.
The specificity of the MAb reaction was investigated by flow cytometry and immunofluorescence. The antigen was precipitated from lysates or uninfected cells using MAb or sera from HIV-1-infected individuals and analysed by Western blot and isoelectrofocusing.
An antigen with an apparent relative molecular mass of 137,000 and an isoelectric point of 8.45 was immunoprecipitated with the cross-reactive MAb from uninfected human T cells. Sera from HIV-positive individuals recognizing a Nef epitope partially overlapping with the binding site of the cross-reactive MAb stained the 137 kD protein precipitated with the MAb in Western blot analysis, while HIV-positive sera without antibodies to this Nef region and sera from uninfected individuals were negative.
The induction of autoantibodies cross-reactive with cellular surface proteins may play a role in the pathogenesis of AIDS.
鉴定未感染的T淋巴细胞上与针对HIV-1 Nef蛋白的单克隆抗体(MAb)发生反应的抗原,并在HIV-1感染个体中寻找针对该表位的抗体。
针对由氨基酸60 - 73定义的Nef表位的鼠源MAb与正常外周血淋巴细胞和永久性人T细胞系的细胞表面抗原发生反应。
通过流式细胞术和免疫荧光研究MAb反应的特异性。使用来自HIV-1感染个体的MAb或血清从未感染细胞的裂解物中沉淀抗原,并通过蛋白质印迹法和等电聚焦进行分析。
用来自未感染人T细胞的交叉反应性MAb免疫沉淀出一种表观相对分子质量为137,000且等电点为8.45的抗原。在蛋白质印迹分析中,识别与交叉反应性MAb结合位点部分重叠的Nef表位的HIV阳性个体的血清,可对用MAb沉淀出的137 kD蛋白进行染色,而没有针对该Nef区域抗体的HIV阳性血清和未感染个体的血清均为阴性。
与细胞表面蛋白发生交叉反应的自身抗体的诱导可能在艾滋病发病机制中起作用。