HUMAN MONOCLONAL ANTIBODIES TO HIV

抗HIV人免疫球蛋白单抗的研制 一直非常困难，关于这一点的报道很少 文学。我们已经开发了方法，并验证了我们的 有规律地使人免疫球蛋白抗体产生抗HIV单抗的能力 HIV感染者的淋巴和脾杂交瘤 研究对象。8株稳定产生2-5ug/ml的杂交瘤细胞 (106个细胞)/d已研制成功。它们与GP160/120发生反应， Gp160/41、p55/24和推测的构象表位。其中之一 这些药物中和了HTLV-IIIB，另一种则增强了传染性。我们 假设可以中和HIV的人类抗HIV MoAbs， 介导ADCC或C‘lysis或对HIV感染细胞具有细胞毒作用 当与毒素结合在一起时，可能对预防艾滋病或 心理治疗。此外，如果我们的抗独特型抗体 利用我们的杂交瘤可以开发出增强抗体 方法学方面，这些也可能在临床上有用。 此外，我们还发现了淋巴和脾 淋巴细胞可以储存在液氮中，最多可以解冻一次 一年后生产出容易融合和制造的细胞 生产抗HIV MoAbs的杂交瘤。他们和其他人类 抗HIV单抗，可用于表征抗原和 在患者的免疫反应中识别的表位 包括与抗体介导的增强 感染。临床上获取的淋巴结和脾 相关手术将作为单细胞悬液进行处理， 与我们独特的融合伙伴(P3U1)融合，经ELISA筛选 抗HIV感染和未感染的细胞和细胞裂解物 以及定义了相关的合成GP120或GP41表位以及 然后进行克隆和扩增，以产生特异性MoAbs。这些遗嘱 以细胞和抗原反应性为特征 免疫荧光、流式细胞术、Western blotting和 中和/增强分析。与我们的合作者一起，我们 将准备代表已经存在的 确定了相关的主题或GP120a和GP41，并确定了 每个人摩押的反应性表位。根据我们的记录 完成后，可以开发出10-20个MoAbs并对其进行表征 每年。各种临床相关实验，如ADCC， 用MoAbs或MoAbbs+患者血清的混合物中和， 毒素-单抗结合物的细胞毒性将进行筛选 有望用于临床试验的MoAbs及其潜力 诊断用途。最后，我们已经并将进一步 开发这一独特的资源，并进行上述 工作。

The development of human IgG monoclonal antibodies (MoAbs) to HIV has been very difficult and there are few reports on this in the literature. We have developed the methodology and verified our ability to regularly make human IgG anti-HIV MoAb-producing hybridomas from the lymph nodes and spleens of HIV-infected subjects. Eight stable hybridomas producing 2-5ug/ml (106cells)/d have been developed. These react with GP160/120, GP160/41, P55/24 and presumed conformational epitopes. One of these neutralizes HTLV-IIIB and another enhances infectivity. We hypothesize that human anti-HIV MoAbs which can neutralize HIV, mediate ADCC or C'lysis or can be cytotoxic to HIV-infected cells when coupled to toxins may be useful in AIDS prophylaxis or therapy. Furthermore, if anti-idiotype antibodies to our enhancing antibody could be developed using our hybridoma methodology, these might also potentially be useful clinically. In addition, we have also found that lymph node and spleen lymphocytes can be stored in liquid nitrogen and thawed up to one year later to yield cells which can readily be fused and made into hybridomas producing anti-HIV MoAbs. They, and other human anti-HIV MoAbs, may be useful in characterizing the antigens and epitopes recognized during the patient's immune response including those related to antibody mediated enhancement of infection. Lymph nodes and spleens, obtained at clinically relevant surgery will be processed as single cell suspensions, fused with our unique fusion partner (P3U1), screened by ELISA against HIV-infected and non-infected cells and cell lysates as well as defined relevant synthetic GP120 or GP41 epitopes and then cloned and expanded to produce specific MoAbs. These will be characterized for cellular and antigenic reactivity by immunofluorescence, flow cytometry, Western blotting and neutralization/enhancement assays. With our collaborators, we will prepare synthetic peptides which represent the already identified relevant eptitopes or GP120a and GP41 and identify the reactive epitope of each human MoAb. Based on our record of accomplishment, 10-20 MoAbs can be developed and characterized per year. Various clinically relevant experiments such as ADCC, neutralization by mixtures of MoAbs or MoAbs + patient sera, cytotoxicity of toxin-MoAb conjugates will be done to select promising MoAbs for clinical trials as well as potential diagnostic uses. Finally, we have initiated and will further exploit this unique resource and carry on the above described work.