ANTIGENICITIES OF AIDS VIRUS RECOMBINANT PROTEINS

目的是了解免疫系统如何试图防御 对抗艾滋病，特别是艾滋病前的体液反应 (ARC)和艾滋病患者对抗各种艾滋病病毒抗原。 的 两种最不同的分离株HTLV-III和ARV-2的抗原性将 也可以进行比较，以确定共同的抗原决定簇。 我们建议 在E.杆菌 将使用使用lac和pR启动子的细菌表达载体 用于抗原表达。 Western印迹分析和放射免疫测定将 用于监测表达和抗原性。 克隆表达 抗原性蛋白将被进一步表征。 的短导数 这些克隆将通过使用限制性内切酶除去 克隆插入片段的不同片段。 对这些删除的分析 然后克隆将定位病毒基因的编码片段， 抗原决定簇（表位）。 体外将用于修饰 表位内的单个氨基酸，使得每个氨基酸的作用 将测定赋予抗原性的酸。 患者的抗体 针对单个表位将进行免疫化学研究， 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 利用病人的抗体来吸收重组抗原。 鼠标 针对这些纯化抗原的抗体应答将与 对病毒抗原的典型免疫反应。 的 多样性以及具体的反应将进行研究， 产生单克隆抗体。 它们能够中和病毒 还将研究传染性。 这项工作将有助于更好地了解艾滋病病毒的生物学， 病毒基因产物及主要和保守抗原的鉴定 患者表达的决定因素。

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.