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目的从中国人类免疫缺陷病毒1型(human immunodeficiency virus-1,HIV-1)感染者中分离单克隆中和抗体,并对其生物活性进行初步鉴定。方法采用抗原特异性单个B细胞分选和单克隆抗体表达技术从感染者中分离单克隆抗体,采用免疫遗传学数据库(immunogenetics database,IMGT)分析抗体的可变区基因,采用酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)检测抗体的结合能力,采用TZM-bl/假病毒中和实验检测抗体的中和能力。结果从一个HIV-1CRF07_BC病毒感染者中分离到了两个单克隆抗体(A1和A6);两个抗体的重链来源于不同家系(IGHV5-51*01和IGHV4-59*01),轻链分别利用lambda链和kappa链,可变区基因的突变率较低;两个抗体与CRF01_AE、B亚型和CRF07_BC病毒的结合能力较强,可以中和Tier 1的SF162病毒(B亚型)和MW965病毒(C亚型),不能中和12个Tier 2的病毒(Global Panel)。结论本研究成功分离到了两个具有交叉反应性的单克隆中和抗体,丰富了HIV-1中和抗体的研究,也为艾滋病的抗原检测和治疗等应用领域提供备选的免疫制剂。
OBJECTIVE: To isolate monoclonal neutralizing antibodies from human immunodeficiency virus-1 (HIV-1) infected individuals and to identify their biological activity. Methods Monoclonal antibodies (McAbs) were isolated from infected patients using antigen-specific single B cell sorting and monoclonal antibody expression. The variable region genes of the antibodies were analyzed by immunogenetics database (IMGT). The enzyme-linked immunosorbent assay The antibody binding ability was detected by enzyme linked immunosorbent assay (ELISA). The neutralizing ability of antibody was detected by TZM-bl / pseudovirus virus neutralization assay. Results Two monoclonal antibodies (A1 and A6) were isolated from one HIV-1 CRF07_BC virus-infected patient. The heavy chains of the two antibodies were derived from different families (IGHV5-51 * 01 and IGHV4-59 * 01) Mutations of lambda and kappa chains in the variable region genes were lower. The binding of the two antibodies to the CRF01_AE, B subtype and CRF07_BC virus was strong, which could neutralize the SF162 virus (subtype B) of Tier 1 and MW965 The virus (subtype C) can not neutralize 12 Tier 2 viruses (Global Panel). Conclusions This study successfully isolated two monoclonal neutralizing antibodies with cross-reactivity, enriched the study of neutralizing antibodies against HIV-1, and provided alternative immunological agents for the detection and treatment of AIDS.