【摘要】 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。更多还原

【Abstract】 Objective To isolate and characterize monoclonal neutralizing antibodies from a HIV-1-infected Chinese donor. Methods Monoclonal antibodies were isolated from a donor by antigen specific single B cell sorting and monoclonal antibody expression.The program IMGT was used to analyze genes encoding the antibody variable region.The specific binding capacity was detected by ELISA and the neutralizing activity was measured by TZM-bl/pseudovirus assay. Results Two monoclonal antibodies(A1and A6) were isolated from a clade CRF07_BC HIV-1-infected Chinese donor.Heavy chains of A1 and A6 were derived from the germline IGHV5-51*01 and IGHV4-59*01.Light chains of A1 and A6utilized the lambda chain and kappa chain.Low levels of somatic hypermutations were found in the variable region of both antibodies.A1 and A6showed good binding capacity with clade CRF01_AE,B and CRF07_BC virus.A1 and A6could neutralize two Tier 1virus-SF162(clade B)and MW965(clade C),but could not neutralize twelve Tier 2virus(Global Panel).Conclusions In this study,two cross-reactive monoclonal neutralizing antibodies were identified from a HIV-1-infected donor.更多还原