GENERATION OF HUMAN MONOCLONAL-ANTIBODIES AGAINST HIV-1 PROTEINS - ELECTROFUSION AND EPSTEIN-BARR-VIRUS TRANSFORMATION FOR PERIPHERAL-BLOOD LYMPHOCYTE IMMORTALIZATION
GENERATION OF HUMAN MONOCLONAL-ANTIBODIES AGAINST HIV-1 PROTEINS - ELECTROFUSION AND EPSTEIN-BARR-VIRUS TRANSFORMATION FOR PERIPHERAL-BLOOD LYMPHOCYTE IMMORTALIZATION
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DOI：
10.1089/aid.1994.10.359
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发表时间：
1994-04-01
影响因子：
1.5
通讯作者：
KATINGER, H
中科院分区：
文献类型：
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作者：
BUCHACHER, A;PREDL, R;KATINGER, H
通过永生化hiv -1阳性志愿者血液中的人pbl，研究电融合和EBV转化。通过细胞融合或EBV转化，建立了33株产生人类HIV-1单克隆抗体（hu - mab）的细胞系。在第一次融合实验中，B淋巴细胞的来源是CD4细胞计数高于500/mm的CDC II期或III期hiv -1感染供者的外周血(3)。后来，仅从这些患者中选择抗艾滋病毒滴度高的患者作为献血者。通过这种方法，稳定的特异性杂交瘤的产量增加了两倍。在我们的实验中，电熔合是一种比EBV转化更有效的永生化方法，因为电熔合的永生化率高且恒定。经过密集的亚克隆后，杂交瘤稳定，可以培养8个月而不损失单克隆抗体的产生。免疫球蛋白的类别、亚型、对HIV-1蛋白的反应性、Western blot模式、免疫荧光和表位进行了表征。所有抗体的亚型均为IgG(1)或IgG(3)。轻链主要是kappa。所有抗体对HIV-1包膜或核心蛋白均有反应性。所有杂交瘤稳定，适合批量生产。一些单克隆抗体正在成为诊断、研究和免疫治疗领域的重要工具。
Electrofusion and EBV transformation were studied by immortalizing human PBLs from blood of HIV-1-positive volunteers. A panel of 33 cell lines producing human monoclonal antibodies (Hu-MAbs) against HIV-1 was established by cell fusion or EBV transformation. For the first fusion experiments the source of B lymphocytes was peripheral blood of HIV-1-infected donors in CDC stages II or III with CD4 cell counts higher than 500/mm(3). Later on, from these patients only, those with high anti-HIV titers were chosen as blood donors. By that means the yield of stable specific hybridomas was increased twofold. In our experiments electrofusion turned out to be a more efficient immortalization method than EBV transformation, due to a high and constant immortalization rate. The hybridomas were stable after intensive subcloning and could be cultivated over a period of 8 months without loss in monoclonal antibody production. Immunoglobulin class, subtype, reactivity against HIV-1 proteins, Western blot patterns, immunofluorescence, and epitopes were characterized. The subtype of all antibodies was IgG(1) or IgG(3). The light chain was predominantly kappa. All antibodies showed reactivity against HIV-1 envelope or core protein. All hybridomas were stable and suited for mass production. Several Hu-MAbs are becoming an important tool in the field of diagnosis, research, and immunotherapy.