ANTIGENICITIES OF AIDS VIRUS RECOMBINANT PROTEINS

其目的是了解免疫系统如何试图防御 防治艾滋病，特别是艾滋病前期的体液反应 (ARC)和艾滋病患者对抗各种艾滋病病毒抗原。这个 两个最多样化的分离株HTLV-III和ARV-2的抗原性将 也可以通过比较来确定常见的抗原决定簇。我们建议 克隆和表达不同的HTLV-III和ARV-2抗原基因。 使用lac和pr启动子的细菌表达载体将被使用 用于抗原表达。蛋白质印迹分析和放射免疫分析将 用于监测表达和抗原性。克隆表达 抗原性蛋白将进一步鉴定。更短的衍生品 这些克隆将通过使用限制性内切酶去除 克隆插入片段的不同片段。对这些缺失的分析 然后，克隆人将定位编码该病毒基因的片段 抗原决定簇(表位)。在体外将被用来修饰 表位内的单个氨基酸使每个氨基酸的作用 将测定赋予抗原性的酸。病人的抗体 将从免疫化学的角度对单个表位进行研究 特异性、异质性和中和病毒感染性的能力。 细菌表达的抗原将通过亲和层析纯化 利用患者的抗体吸收重组抗原。小白鼠 对这些纯化抗原的抗体反应将与 人类对病毒抗原的典型免疫反应。这个 将研究应对措施的多样性以及特殊性。 产生单抗。它们中和病毒的能力 传染性也将被研究。 这项工作将使人们更好地了解艾滋病病毒的生物学 病毒基因产物及其主要和保守抗原的鉴定 决定因素在患者中表达。

The objective is to understand how the immune system attempts to defend against the disease AIDS, particularly the humoral responses of pre-AIDS (ARC) and AIDS patients against various AIDS viral antigens. The antigenicities of the two most diverse isolates, HTLV-III and ARV-2, will also be compared to determine common antigenic determinants. We propose to clone and express various HTLV-III and ARV-2 antigen genes in E. coli. Bacterial expression vectors using the lac and pR promoters will be used for antigen expressions. Western blot analysis and radioimmunoassays will be used to monitor expression and antigenities. Clones expressing antigenic proteins will be characterized further. Shorter derivatives of these clones will be generated by using restriction enzymes to remove different fragments of the cloned insert. Analysis of these deletion clones will then locate segments of the viral genes that encode the antigenic determinants (epitopes). In vitro will be used to modify individual amino acid within the epitope so that the role of each amino acid in conferring antigenicity will be determined. Patients' antibodies towards individual epitope will be studied immunochemically in terms of specificities, heterogeneity and ability to neutralize viral infectivity. Bacterial expressed antigens will be purified by affinity chromatography using patients' antibodies to absorb the recombinant antigens. Mouse antibody response towards these purified antigens will be compared to that of a typical immune reesponse to the viral antigens in humans. The diversity as well as specificities of the response will be studied by generating monoclonal antibodies. Their ability to neutralize viral infectivity will be studied as well. This work will lead to a better understanding of AIDS virus biology, the viral gene products and identification of the major and conserved antigenic determinants expressed in patients.