Durda P J, Bacheler L, Clapham P, Jenoski A M, Leece B, Matthews T J, McKnight A, Pomerantz R, Rayner M, Weinhold K J
E.I. Du Pont de Nemours and Co., Inc., North Billerica, MA 01862.
AIDS Res Hum Retroviruses. 1990 Sep;6(9):1115-23. doi: 10.1089/aid.1990.6.1115.
We have developed a series of murine monoclonal antibodies to a region of the 120 kD envelope glycoprotein (gp120) of human immunodeficiency virus type 1 (HIV-1). This region has previously been implicated as a site for virus neutralization by antisera raised to recombinant proteins and by antibodies made to full-length gp120 purified from virus. The antigen employed was a synthetic peptide containing 15 amino acids, representing amino acid residues 308-322, RIQRGPGRAFVTIGK, of env gp120 (HTLV-IIIB isolate). Five of the monoclonal antibodies raised to this antigen have reactivity with gp120 from divergent strains of HIV-1 in Western blot assays. The two of these five which were tested with live cells infected with the divergent HIV-1 isolates IIIB, MN, and RF were specifically reactive by fluorescence analyses with cells infected with the MN and IIIB isolates. Four of the five monoclonal antibodies blocked the fusion of IIIB-infected cells with uninfected MOLT-4 target cells. The monoclonal antibody most reactive with MN-infected cells by fluorescence, #5025A, blocked the fusion of MN-infected cells with uninfected MOLT-4 cells. Four of the five monoclonal antibodies neutralized the IIIB isolate of HIV-1 in vitro, but none neutralized the MN or RF isolates at the levels of antibody tested (less than or equal to 50 micrograms/ml). Taken together these data indicate that monoclonal antibodies to the immunodominant neutralizing domain of HIV-1 gp120 display different levels of group reactivity depending on the assay system being examined.
我们已经针对人类免疫缺陷病毒1型（HIV-1）120 kD包膜糖蛋白（gp120）的一个区域开发了一系列鼠单克隆抗体。该区域先前已被认为是通过针对重组蛋白产生的抗血清以及针对从病毒中纯化的全长gp120制备的抗体进行病毒中和的位点。所使用的抗原是一种含有15个氨基酸的合成肽，代表env gp120（HTLV-IIIB分离株）的氨基酸残基308 - 322，即RIQRGPGRAFVTIGK。针对该抗原产生的五种单克隆抗体在蛋白质印迹分析中与来自不同HIV-1毒株的gp120有反应性。在这五种抗体中，对两种用感染了不同HIV-1分离株IIIB、MN和RF的活细胞进行测试时，通过荧光分析发现它们与感染MN和IIIB分离株的细胞有特异性反应。五种单克隆抗体中有四种阻断了感染IIIB的细胞与未感染的MOLT-4靶细胞的融合。在荧光分析中与感染MN的细胞反应性最强的单克隆抗体#5025A，阻断了感染MN的细胞与未感染的MOLT-4细胞的融合。五种单克隆抗体中有四种在体外中和了HIV-1的IIIB分离株，但在所测试的抗体水平（小于或等于50微克/毫升）下，没有一种能中和MN或RF分离株。综合这些数据表明，针对HIV-1 gp120免疫显性中和结构域的单克隆抗体根据所检测的分析系统显示出不同水平的群反应性。
