ANTIGENIC MAPPING OF CHLAMYDIAL PROTEINS

沙眼衣原体的主要外膜蛋白（MOMP）已被发现， 被鉴定为疫苗开发的可能免疫原。目标 本项目的目的是鉴定MOMP的氨基酸序列， 免疫球蛋白可及的表面，并确定这些序列是否 参与衣原体与宿主细胞的粘附。含肽 四个可变结构域（VD I、II、III和IV）的序列已经被 合成了这些肽已被用于定义表面可及性 表位B血清群在VD II和VD IV上具有可及表位; 相比之下，中间型和C血清群在 VD I和VD IV。血清型MOMP的VD II和VD IV的胰蛋白酶裂解 存活EB表面的B导致衣原体与 HeLa 229细胞抗血清型B MOMP的VD II和VD IV单克隆抗体 通过防止HAK细胞感染衣原体， 附件.这些数据表明，MOMP在附着中起作用。 衣原体感染宿主细胞MOMP似乎通过以下途径介导依恋： 亲水表面可及变量的静电相互作用 结构域，并通过结合到保守区中的疏水口袋 关于VD IV与疏水口袋的结合取决于疏水性。 MOMP的构象。目前的研究集中在使用重叠 合成肽（8个氨基酸）连接到引脚，以确定 免疫优势区的VD。使用豚鼠血清的研究 感染C.鹦鹉热GPIC株表明VD Ⅰ和VD Ⅳ是 免疫显性感染。通过这些研究，我们希望能够确定 如果在感染期间表面可及表位也是免疫显性的。

The major outer membrane protein (MOMP) of Chlamydia trachomatis has been identified as a possible immunogen for vaccine development. The objectives of this project are to identify amino acid sequences of MOMP which are surface accessible to immunoglobulin and to determine if these sequences are involved in chlamydia-host cell attachment. Peptides containing sequences of the four variable domains (VD I, II, III, and IV) have been synthesized. These peptides have been used to define surface accessible epitopes. The B serogroup have accessible epitopes on VD II and VD IV; in contrast, the intermediate and the C serogroup have accessible epitopes on VD I and VD IV. Trypsin cleavage of VD II and VD IV of the MOMP of serotype B on the surface of viable EBs results in loss of binding of chlamydiae to HeLa 229 cells. Monoclonal antibodies to VD II and VD IV of serotype B MOMP neutralize infectivity of chlamydiae for HAK cells by preventing attachment. These data indicate that MOMP has a role in attachment of chlamydiae to host cells. MOMP appears to mediate attachment through electrostatic interactions of the hydrophilic surface accessible variable domains and through binding to a hydrophobic pocket in a conserved region of VD IV. The binding to the hydrophobic pocket is dependent upon the conformation of MOMP. Current research is focused on using overlapping synthetic peptides (8 amino acids) attached to pins to identify immunodominant regions of the VDs. Studies using sera from guinea pigs infected with C. psittaci strain GPIC indicate that VD I and VD IV are immunodominant during infection. From these studies, we hope to determine if surface accessible epitopes are also immunodominant during infection.