VACCINE FOR EQUINE INFECTIOUS ANEMIA VIRUS VARIANTS

逆转录病毒会引起人类和动物的疾病，包括肿瘤。 与免疫抑制有关。慢病毒(一个非致癌亚家族)是 由HTLV-III/LAV/ARV人类病毒组组成；小型反刍动物 山羊关节炎病毒群-脑炎、Visna和进行性 肺炎病毒；马传染性贫血病毒(EIAV)。控制 慢病毒是困难的，因为前病毒的持久性和 引起表面抗原性变异的env基因序列 糖蛋白。这项建议的长期目标是理解 表面关键功能区的结构组织 研究抗原性的独特逆转录病毒EIAV的糖蛋白 变化性和持久性。这项调查将审查单一的 功能区--受体结合部位--可以用来诱导 与EIAV抗原变异体反应的中和抗体反应。 这些抗体单独和联合使用的保护作用 针对不变的EIAV的细胞毒性T淋巴细胞反应 然后将对受感染细胞上的抗原进行评估。这些方法将 包括从纯化病毒粒子表面分离受体结合部位 糖蛋白通过蛋白水解酶和高效液相色谱构建结合蛋白 通过多肽合成或克隆环境表达实现的受体类似物 载体系统中的基因，用受体结合部位免疫马匹 与刺激细胞毒性T细胞的EIAV抗原类似和不变 淋巴细胞和用EIAV抗原变异体免疫的马。 具体目标是： 1.确定EIAV抗原变异体是否有共同的受体 结合部位。 2.分离和鉴定病毒gp90片段 受体结合部位。 3.使合成或重组产品在免疫化学上类似于 受体结合部位。 4.检测受体结合位点类似物诱导 马对EIAV抗原变异体的交叉保护性免疫反应。 5.为了证明用不变的EIAV抗原免疫 刺激细胞毒性T淋巴细胞将通过一种 受体结合部位类似物。

Retroviruses cause diseases in humans and animals, ranging from neoplasia to immunosuppression. Lentiviruses (a non-oncogenic subfamily) are comprised of the HTLV-III/LAV/ARV human virus group; the small ruminant virus group of caprine arthritis-encephalitis, visna and progressive pneumonia viruses; and equine infectious anemia virus (EIAV). Control of lentiviruses is difficult because of provirus persistence and changes in the env gene sequence resulting in antigenic variation of surface glycoproteins. The long term objective of this proposal is to understand the structural organization of critical functional regions of surface glycoproteins of EIAV, a unique retrovirus for studies of antigenic variation and persistence. This investigation will examine how a single functional region--the receptor binding site--can be used to elicit a neutralizing antibody response that reacts with EIAV antigenic variants. The protective effects of these antibodies both alone and in conjunction with a cytotoxic T lymphocyte response directed against invariant EIAV antigens on infected cells will then be evaluated. The methods will include isolating a receptor binding site from purified virion surface glycoproteins by proteolytic digestion and HPLC, constructing a binding site receptor analog by peptide synthesis or by expression of cloned env gene in a vector system, immunizing horses with the receptor binding site analog and with invariant EIAV antigens that stimulate cytotoxic T lymphocytes and challenging immunized horses with EIAV antigenic variants. The specific aims are: 1. To determine whether antigenic variants of EIAV have a common receptor binding site. 2. To isolate and characterize a fragment of viral gp90 containing the receptor binding site. 3. To make a synthetic or recombinant product immunochemically analogous to the receptor binding site. 4. To test the capacity of the receptor binding site analog to elicit a cross protective immune response in horses against EIAV antigenic variants. 5. To demonstrate that immunization with an invariant EIAV antigen that stimulates cytotoxic T lymphocytes will augment immunication with a receptor binding site analog.