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针对重组人内源性逆转录病毒-K(HERV-K)包膜蛋白制备的单克隆抗体的反应性。

Reactivity of monoclonal antibodies established against a recombinant human endogenous retrovirus-K (HERV-K) envelope protein.

作者信息

Vogetseder W, Feng J, Dierich M P

机构信息

Institut für Hygiene, Leopold Franzens Universität, Innsbruck, Austria.

出版信息

Immunol Lett. 1995 May;46(1-2):129-34. doi: 10.1016/0165-2478(95)00033-2.

Abstract

Human endogenous retroviruses (HERVs) are widely believed to represent possible pathogenic agents in autoimmune disorders and carcinogenesis. We generated 2 monoclonal antibodies (mAbs) against a recombinant HERV-K outer-membrane envelope protein. These mAbs were capable of immunoprecipitating protein bands of 55, 51 and 45 kDa, respectively, from [35S]methionine-labeled lysates of T47D (human breast carcinoma), Raji, Bjab (both human beta-lymphoblastoid) and H9 (human T-cell lymphoma) cells. Furthermore, the 55 kDa band could be precipitated from cell-free supernatant of biosynthetically labeled T47D cells, indicating that this molecule may be secreted. Indirect immunofluorescence and subsequent FACS analysis demonstrated the presence of these molecules on T47D, HeLa, Raji, Bjab and H9 cells as well as on lymphocytes isolated from tonsils and stimulated with poke weed mitogen (PWM). Due to the broad distribution and lack of detection of HERV-K-env mRNA in these cells, we believe that these antigens are of cellular origin detected by crossreactivity of mAbs against HERV-K-env protein with cellular molecules.

摘要

人们普遍认为,人类内源性逆转录病毒(HERVs)可能是自身免疫性疾病和癌症发生过程中的致病因子。我们制备了两种针对重组HERV-K外膜包膜蛋白的单克隆抗体(mAb)。这些单克隆抗体能够分别从用[35S]甲硫氨酸标记的T47D(人乳腺癌)、Raji、Bjab(均为人β淋巴母细胞样)和H9(人T细胞淋巴瘤)细胞裂解物中免疫沉淀出55 kDa、51 kDa和45 kDa的蛋白条带。此外,55 kDa的条带可以从生物合成标记的T47D细胞的无细胞上清液中沉淀出来,这表明该分子可能会被分泌。间接免疫荧光和随后的流式细胞术分析表明,这些分子存在于T47D、HeLa、Raji、Bjab和H9细胞以及从扁桃体分离并用商陆有丝分裂原(PWM)刺激的淋巴细胞上。由于这些细胞中HERV-K-env mRNA分布广泛且未检测到,我们认为这些抗原是通过抗HERV-K-env蛋白的单克隆抗体与细胞分子的交叉反应而检测到的细胞来源抗原。

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