Gorny M K, Moore J P, Conley A J, Karwowska S, Sodroski J, Williams C, Burda S, Boots L J, Zolla-Pazner S
New York University Medical Center, New York.
J Virol. 1994 Dec;68(12):8312-20. doi: 10.1128/JVI.68.12.8312-8320.1994.
A human immunoglobulin G1 lambda monoclonal antibody (MAb), 697-D, was developed that recognizes the V2 region of human immunodeficiency virus type 1 (HIV-1) gp120. Substitutions at amino acid positions 176/177, 179/180, 183/184, and 192 to 194 in the V2 loop of gp120 each completely abolished the binding capacity of 697-D in an enzyme-linked immunosorbent assay format. Competition analysis with three different neutralizing murine anti-V2 MAbs confirmed the specificity of 697-D. The 697-D epitope is primarily conformation dependent, although there was weak reactivity of the MAb with a V2 peptide spanning residues 161 to 180. Treatment of recombinant gp120 HIVIIIB with sodium metaperiodate, which oxidizes carbohydrates, abolished the binding of the MAb, showing the dependence of the epitope on intact carbohydrates. The broad reactivity of 697-D was displayed by its binding to the gp120 molecules from four of four laboratory isolates and five of five primary isolates. The MAb 697-D neutralized three out of four primary isolates but failed to neutralize any of four laboratory strains of HIV-1. 697-D and a human anti-V3 MAb, 447-52-D, displayed similar potency in neutralizing primary isolates, indicating that the V2 region of gp120, like the V3 region and the CD4-binding domain, can induce potent neutralizing antibodies against HIV-1 in humans.
开发了一种人免疫球蛋白G1λ单克隆抗体(MAb)697-D,它能识别1型人类免疫缺陷病毒(HIV-1)gp120的V2区域。在酶联免疫吸附测定中,gp120 V2环中第176/177、179/180、183/184位氨基酸以及第192至194位氨基酸的替换均完全消除了697-D的结合能力。与三种不同的中和性鼠抗V2单克隆抗体进行的竞争分析证实了697-D的特异性。697-D表位主要依赖构象,尽管该单克隆抗体与跨越161至180位残基的V2肽有弱反应性。用高碘酸钠处理重组gp120 HIVIIIB(高碘酸钠可氧化碳水化合物)可消除该单克隆抗体的结合,表明表位对完整碳水化合物的依赖性。697-D与来自四个实验室分离株中的四个以及五个原代分离株中的五个的gp120分子结合,显示出广泛的反应性。单克隆抗体697-D中和了五个原代分离株中的三个,但未能中和四个HIV-1实验室菌株中的任何一个。697-D和一种人抗V3单克隆抗体447-52-D在中和原代分离株方面表现出相似的效力,表明gp120的V2区域与V3区域和CD4结合域一样,可在人体内诱导针对HIV-1的强效中和抗体。