Salari S H, Ward M E
J Gen Microbiol. 1981 Apr;123(2):197-207. doi: 10.1099/00221287-123-2-197.
Strains of Chlamydia trachomatis representative of 14 serotypes were grown in HeLa 229 cells. HeLa cell susceptibility to chlamydial infection was increased by treating the host cells with DEAE-dextran. Optimal conditions of DEAE-dextran treatment were determined for each serotype of C. trachomatis to maximize chlamydial yields. Chlamydial polypeptides were selectively radiolabelled with 3H-labelled amino acids in the presence of emetine, an inhibitor of HeLa cell protein synthesis. The radiolabelled chlamydiae were purified from host cell components by density gradient centrifugation and their polypeptide composition was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The distribution of the major chlamydial polypeptide of 38 000 to 42 000 daltons amongst the different serotypes correlated closely with the predominant human infections caused by each serotype. Lymphogranuloma venereum agents possessed a unique polypeptide of 118 000 daltons not found amongst trachoma inclusion conjunctivitis strains of chlamydiae. Chlamydial surface polypeptides were selectively radiolabelled with 125I by lactoperoxidase-catalysed oxidation. The major chlamydial polypeptide and polypeptides of 155 000 and 29 000 daltons were thus identified as surface polypeptides of the chlamydial elementary body. It is suggested that the 155 000 dalton polypeptide is a species-specific antigen, the major polypeptide is the principal outer membrane protein, and the 29 000 dalton polypeptide is the type-specific antigen.
沙眼衣原体14种血清型的代表菌株在HeLa 229细胞中培养。用二乙氨基乙基葡聚糖(DEAE-葡聚糖)处理宿主细胞可增加HeLa细胞对衣原体感染的易感性。针对每种沙眼衣原体血清型确定了DEAE-葡聚糖处理的最佳条件,以使衣原体产量最大化。在放线菌酮(一种HeLa细胞蛋白质合成抑制剂)存在的情况下,用3H标记的氨基酸对衣原体多肽进行选择性放射性标记。通过密度梯度离心从宿主细胞成分中纯化放射性标记的衣原体,并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳确定其多肽组成。38000至42000道尔顿的主要衣原体多肽在不同血清型中的分布与每种血清型引起的主要人类感染密切相关。性病性淋巴肉芽肿病原体拥有一种118000道尔顿的独特多肽,在沙眼包涵体结膜炎衣原体菌株中未发现。通过乳过氧化物酶催化氧化用125I对衣原体表面多肽进行选择性放射性标记。因此,主要衣原体多肽以及155000和29000道尔顿的多肽被鉴定为衣原体原体的表面多肽。有人提出,155000道尔顿的多肽是种特异性抗原,主要多肽是主要外膜蛋白,29000道尔顿的多肽是型特异性抗原。