Matsushita S, Maeda H, Kimachi K, Eda Y, Maeda Y, Murakami T, Tokiyoshi S, Takatsuki K
Transfusion Service Department, University Medical School, Kumamoto, Japan.
AIDS Res Hum Retroviruses. 1992 Jun;8(6):1107-15. doi: 10.1089/aid.1992.8.1107.
A chimeric mouse-human antibody (C beta 1) was constructed that recognized the principal neutralizing domain (PND) of human immunodeficiency virus type 1 (HIV-1) gp120. The constant (C) immunoglobulin regions (C gamma 1 and C kappa) of a mouse monoclonal antibody, 0.5 beta, were substituted for the human C gamma 1 and C kappa by recombining the DNA modules encoding variable or C regions. The DNA constructs were then transfected into X63 Ag8.653 myeloma cells. A clone with a high production of the chimeric antibody (C beta 1) was selected. This antibody was tested for its biological activity against HIV-1. It bound to the surface of HTLV-IIIB-infected cells and reacted with gp120/160 with equal affinity and specificity to that of the parental 0.5 beta murine monoclonal antibody in a Western blot assay. Neutralization and/or enhancement of HIV infection were evaluated with C beta 1 and 0.5 beta. Both C beta 1 and 0.5 beta neutralized cell-to-cell infection and cell-free virus infection by HTLV-IIIB. Antibody-dependence enhancement of HIV infection was not observed with either C beta 1 or 0.5 beta in the presence or absence of human complement. Antibody-dependent cell-mediated cytolysis (ADCC) and antibody-dependent complement-mediated cytolysis (ACC) were observed with C beta 1 but not with the parental 0.5 beta. These findings suggest that the neutralizing antibodies to PND may neutralize but not enhance HIV infection. Furthermore, the high levels of ACC and ADCC shown against HIV-infected cells by C beta 1 indicate that the clinical application of such monoclonal antibodies may be possible.
构建了一种嵌合型小鼠 - 人抗体（Cβ1），它能识别1型人类免疫缺陷病毒（HIV - 1）gp120的主要中和结构域（PND）。通过重组编码可变区或恒定区的DNA模块，将小鼠单克隆抗体0.5β的恒定（C）免疫球蛋白区域（Cγ1和Cκ）替换为人的Cγ1和Cκ。然后将DNA构建体转染到X63 Ag8.653骨髓瘤细胞中。筛选出一个高表达嵌合抗体（Cβ1）的克隆。对该抗体针对HIV - 1的生物学活性进行了检测。它能结合HTLV - IIIB感染细胞的表面，并且在蛋白质印迹分析中与gp120/160反应，其亲和力和特异性与亲本0.5β小鼠单克隆抗体相同。用Cβ1和0.5β评估HIV感染的中和和/或增强作用。Cβ1和0.5β都能中和HTLV - IIIB介导的细胞间感染和无细胞病毒感染。在有或无人类补体存在的情况下，Cβ1和0.5β均未观察到HIV感染的抗体依赖性增强。Cβ1可观察到抗体依赖性细胞介导的细胞毒作用（ADCC）和抗体依赖性补体介导的细胞毒作用（ACC），而亲本0.5β则未观察到。这些发现表明，针对PND的中和抗体可能会中和但不会增强HIV感染。此外，Cβ1对HIV感染细胞显示出高水平的ACC和ADCC，这表明此类单克隆抗体可能具有临床应用价值。
