IMMUNOCHEMISTRY OF CHLAMYDIAL SURFACE ANTIGENS

该项目的主要重点是开发一种疫苗，以预防 或控制沙眼衣原体引起的感染。衣原体病 外膜蛋白(MOMP)被认为在血管紧张素转换酶活性中起重要作用。 衣原体感染保护性免疫的研究进展。因此，当前 疫苗战略的重点是开发重组或 可靶向诱导粘膜损伤的合成肽MOMP疫苗 豁免权。我们已经进行了一项广泛的努力，旨在描述 从分子水平鉴定MOMP的抗原性 这种蛋白的免疫相关结构，可能具有实用价值 对衣原体疫苗的合理设计。之前的努力是 集中于鉴定免疫优势的中和B细胞表位 最高统一部。血清型特异性和广泛交叉反应亚种特异性 中和位点被映射到位于 蛋白质的可变区(Vd)。在这一年里，我们承担了 MOMP辅助性T细胞抗原决定簇的研究 诱导B细胞产生特异性抗体的功能 中和B细胞表位。MOMP的两个区域被证明含有 能够指导B细胞克隆产生抗体的TH细胞表位 对免疫优势中和部位具有特异性。含有以下成分的嵌合肽 合成了Th细胞和B细胞表位，并显示出高度的 免疫原性。高表达嵌合多肽免疫小鼠的研究 与天然MOMP反应并中和的滴定抗体 在试管中。正在研究的Th细胞表位显然是“混杂的” 由于H-2基因不同的同源小鼠品系能够对 嵌合的多肽免疫原。免疫原性和免疫原性的评价 嵌合肽在亚人灵长类动物中的疫苗效力将是 在即将到来的一年中启动。 我们还致力于构建具有传染性的衣原体载体。 疫苗。为了达到这些目标，我们表达了完整的MOMP蛋白 并对重组疫苗的免疫原性进行了评价。 小鼠体内的病毒。结果令人振奋；感染重组病毒的小鼠 表达MOMP的痘苗病毒诱导特异性高滴度抗体 到蛋白质。我们计划评估该病毒的保护效果 重组病毒在衣原体感染动物模型中的应用 年。

The major focus of this project is the development of a vaccine to prevent or control infections caused by Chlamydia trachomatis. The chlamydial major outer membrane protein (MOMP) is considered to be important in the development of protective immunity to chlamydial infection. Thus, current vaccine strategies are focused on the development of recombinant or synthetic peptide MOMP vaccines that can be target to induce mucosal immunity. We have undertaken an extensive effort aimed at characterizing the antigenic properties of the MOMP at the molecular level to identify immunologically relevant structures of this protein that may have utility for the rational design of a chlamydial vaccine. Previous efforts were focused on characterizing immunodominant neutralizing B-cell epitopes of the MOMP. Serovar-specific and broadly cross reactive subspecies-specific neutralizing sites were mapped to contiguous epitopes located in within the variable domains (VD) of the protein. In this year, we have undertaken studies to characterize T-helper cell antigenic determinants of the MOMP that function in inducing B-cells to produce antibody specific to neutralizing B-cell epitopes. Two regions of the MOMP were shown to contain Th-cell epitopes capable of directing B-cell clones to produce antibody specific to immunodominant neutralizing sites. Chimeric peptides containing both Th-and B-cell epitopes were synthesized and shown to be highly immunogenic. Immunization of mice with the chimeric peptides induced high titered antibodies that reacted with the native MOMP and were neutralizing in vitro. The Th-cell epitopes under study are apparently "promiscuous" since strains of congeneic mice differing at H-2 are capable of responding to the chimeric peptide immunogens. Evaluation of the immunogenicity and vaccine efficacy of the chimeric peptides in sub-human primates will be initiated in the upcoming year. We are also working on the construction of infectious vectored chlamydial vaccines. Towards these goals we have expressed the complete MOMP protein in vaccinia virus and have evaluated the immunogenicity of the recombinant virus in mice. The results are promising; mice infected with recombinant vaccinia virus expressing the MOMP induced high titered antibodies specific to the protein. We plan to evaluate the protective efficacy of the recombinant virus in animal models of chlamydial infection in the upcoming year.