Messick J B, Rikihisa Y
Department of Veterinary Pathobiology, College of Veterinary Medicine, Ohio State University, Columbus 43210.
Infect Immun. 1992 Aug;60(8):3079-86. doi: 10.1128/iai.60.8.3079-3086.1992.
Indirect immunofluorescence staining of macrophages infected with Ehrlichia risticii by anti-E. risticii serum revealed a punctate staining pattern on the surface of the host cell. This pattern was distinguishable by fluorescence microscopy from E. risticii bound to the surface of the macrophage and from intracellular E. risticii. The surface localization of ehrlichial antigen on infected macrophages was confirmed by electron microscopy with immunoferritin labeling. As the intracellular ehrlichial burden increased, the amount of ehrlichial antigen on the host cell surface increased. Prokaryotic protein synthesis was necessary for the maintenance of ehrlichial antigen on the host cell surface, as demonstrated by disappearance of the surface antigen following treatment with oxytetracycline. However, host cell protein synthesis was not required, as demonstrated by the continued presence of ehrlichial antigen on the surface of host cells after cycloheximide treatment. Pronase treatment abolished the ehrlichial antigen present on the cell surface, indicating that this antigen is a protein. Anti-E. risticii serum or immunoglobulin G-mediated antibody-dependent cellular cytotoxicity of infected cells was demonstrated in a chromium release assay. These results imply that the parasite antigen on the host cell surface has a role in the pathogenesis of ehrlichiosis.
用抗瑞氏埃立克体血清对感染瑞氏埃立克体的巨噬细胞进行间接免疫荧光染色,结果显示在宿主细胞表面呈现点状染色模式。通过荧光显微镜观察,这种模式与结合在巨噬细胞表面的瑞氏埃立克体以及细胞内的瑞氏埃立克体是可区分的。通过免疫铁蛋白标记的电子显微镜技术证实了感染巨噬细胞上埃立克体抗原的表面定位。随着细胞内埃立克体载量的增加,宿主细胞表面的埃立克体抗原量也增加。如用土霉素处理后表面抗原消失所表明的,原核生物蛋白质合成对于维持宿主细胞表面的埃立克体抗原是必需的。然而,如用环己酰亚胺处理后宿主细胞表面仍持续存在埃立克体抗原所表明的,宿主细胞蛋白质合成并非必需。链霉蛋白酶处理消除了细胞表面存在的埃立克体抗原,表明该抗原是一种蛋白质。在铬释放试验中证实了抗瑞氏埃立克体血清或免疫球蛋白G介导的感染细胞的抗体依赖性细胞毒性。这些结果表明宿主细胞表面的寄生虫抗原在埃立克体病的发病机制中起作用。