IDENTIFICATION AND CHARACTERIZATION OF A NEUTRALIZATION SITE WITHIN THE 2ND VARIABLE REGION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 GP120
IDENTIFICATION AND CHARACTERIZATION OF A NEUTRALIZATION SITE WITHIN THE 2ND VARIABLE REGION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 GP120
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DOI：
10.1128/jvi.66.2.848-856.1992
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发表时间：
1992-02-01
影响因子：
5.4
通讯作者：
HO, DD
中科院分区：
文献类型：
--
作者：
FUNG, MSC;SUN, CRY;HO, DD
用从人类免疫缺陷病毒1型（HIV-1）IIIB感染的H9细胞提取物中纯化的gp 120免疫BALB/c小鼠，制备了两种单克隆抗体，命名为BAT 085和G3-136。 在三种HIV-1实验室分离株（IIIB、MN和RF）中，BAT 085仅中和CEM-SS细胞的IIIB感染，而G3-136中和IIIB和RF。 这些抗体还中和了活化的人外周血单核细胞感染中的一些主要HIV-1分离株。 在间接免疫荧光试验中，BAT 085与感染IIIB或MN的H9细胞结合，而G3-136与感染IIIB或RF的H9细胞结合，但不与MN结合。 使用HIV-1 IIIB gp 120的序列重叠合成肽，将BAT 085和G3-136的结合位点定位于V2区的肽段（氨基酸残基169至183）。 这些抗体与固定化gp 120的结合不受针对V3区中的主要中和决定簇或gp 120的CD 4结合结构域的抗体的抑制。 在竞争性酶联免疫吸附试验中，可溶性CD 4抑制G3-136与gp 120的结合，但不抑制BAT 085与gp 120的结合。 通过内切-β-N-乙酰氨基葡萄糖苷酶H对gp 120进行去糖基化或通过二硫苏糖醇对gp 120进行还原，可降低其与G3-136的反应性，但不降低其与BAT 085的反应性。 这些结果表明，V2区的gp 120包含多个中和抗体识别的构象依赖性和非依赖性的方式决定簇。
Two monoclonal antibodies designated BAT085 and G3-136 were raised by immunizing BALB/c mice with gp120 purified from human immunodeficiency virus type 1 (HIV-1) IIIB-infected H9 cell extracts. Among three HIV-1 laboratory isolates (IIIB, MN, and RF), BAT085 neutralized only IIIB infection of CEM-SS cells, whereas G3-136 neutralized both IIIB and RF. These antibodies also neutralized a few primary HIV-1 isolates in the infection of activated human peripheral blood mononuclear cells. In indirect immunofluorescence assays, BAT085 bound to H9 cells infected with IIIB or MN, while G3-136 bound to H9 cells infected with IIIB or RF, but not MN. Using sequence-overlapping synthetic peptides of HIV-1 IIIB gp120, the binding site of BAT085 and G3-136 was mapped to a peptidic segment in the V2 region (amino acid residues 169 to 183). The binding of these antibodies to immobilized gp120 was not inhibited by the antibodies directed to the principal neutralization determinant in the V3 region or to the CD4-binding domain of gp120. In a competition enzyme-linked immunosorbent assay, soluble CD4 inhibited G3-136 but not BAT085 from binding to gp120. Deglycosylation of gp120 by endo-beta-N-acetylglucosaminidase H or reduction of gp120 by dithiothreitol diminished its reactivity with G3-136 but not with BAT085. These results indicate that the V2 region of gp120 contains multiple neutralization determinants recognized by antibodies in both a conformation-dependent and -independent manner.