HUMAN MONOCLONAL ANTIBODIES TO HIV

抗HIV人源IgG单克隆抗体的研制 这是非常困难的，在这方面的报道很少。 文学 我们已经开发了方法，并验证了我们的 定期生产人IgG抗HIV MoAb的能力 来自HIV感染者淋巴结和脾脏的杂交瘤 科目 8个稳定的杂交瘤产生2- 5 ug/ml （106cells）/d。 它们与GP 160/120反应， GP 160/41、P55/24和推测的构象表位。 之一 这些中和HTLV-IIIB，另一种增强感染性。 我们 假设人抗HIV单克隆抗体可以中和HIV， 介导ADCC或C '裂解或对HIV感染的细胞具有细胞毒性 当与毒素偶联时可用于艾滋病预防， 疗法 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，这些也可能是潜在的临床有用。 此外，我们还发现淋巴结和脾脏 淋巴细胞可以储存在液氮中， 一年后产生的细胞可以很容易地融合和制造 杂交瘤中产生抗HIV单克隆抗体。 他们和其他人类 抗HIV单克隆抗体，可用于表征抗原， 在患者的免疫应答期间识别的表位 包括与抗体介导的增强 感染 淋巴结和脾脏，在临床上获得 相关手术将作为单细胞悬液处理， 与我们独特的融合伴侣（P3 U1）融合，通过ELISA筛选 针对HIV感染和未感染的细胞和细胞裂解物， 以及确定的相关合成GP 120或GP 41表位， 然后克隆并扩增以产生特异性单克隆抗体。 这些将 细胞和抗原反应性的特征在于 免疫荧光、流式细胞术、Western印迹和 中和/增强测定。 与我们的合作伙伴，我们 将制备合成肽， 鉴定相关表位或GP 120 a和GP 41，并鉴定 每个人MoAb的反应性表位。 根据我们的记录， 完成后，可以开发和表征10-20种MoAb 每一年。 各种临床相关实验，如ADCC， 通过MoAb或MoAb+患者血清的混合物进行中和， 将进行毒素-MoAb缀合物的细胞毒性试验，以选择 用于临床试验的有前途的单克隆抗体以及潜在的 诊断用途。 最后，我们已经开始并将进一步 利用这一独特的资源，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.