Moorman D R, Sixbey J W, Wyrick P B
J Gen Microbiol. 1986 Apr;132(4):1055-67. doi: 10.1099/00221287-132-4-1055.
Primary cultures of human endometrial and ectocervical epithelial cells were examined as a new model system to study genital infection by Chlamydia trachomatis. Initial studies demonstrated that these cells were indeed susceptible to chlamydial infection. Inocula, adjusted to produce inclusions in 50 to 80% of equivalent numbers of standard McCoy cells, resulted in infection rates of approximately 15 to 30% for the columnar cells of the endometrium and 5 to 10% for the squamous cells of the ectocervix. Exposure of cultures to DEAE-dextran and centrifugation-assisted inoculation, manipulations reported to enhance infection of HeLa and McCoy cells, did not alter the number of inclusion-positive genital cells. Addition of cycloheximide to the post-inoculation culture medium slightly increased numbers of inclusion-bearing cells while growth of genital cells in hormone-supplemented medium resulted in a variable effect on inclusion development and a significant reduction in the association of radiolabelled organisms with these cells. The basis for the different levels of infection in McCoy versus genital cell cultures was revealed by immunofluorescence analysis of chlamydial association with host cells immediately after inoculation. Chlamydiae failed to adhere to many cells in the genital cell cultures while adherence to McCoy cells was uniform. In addition, the association of radiolabelled C. trachomatis was significantly lower with genital cells than with McCoy cells. Finally, culture conditions were defined which markedly inhibited inclusion development without an immediate loss of chlamydial growth potential. This investigation indicates that primary genital cell cultures are susceptible to chlamydial infection and will be valuable for studies on the nature of C. trachomatis interactions with natural human target cells.
人子宫内膜和宫颈外膜上皮细胞的原代培养物被作为一种新的模型系统来研究沙眼衣原体引起的生殖道感染。初步研究表明,这些细胞确实易受衣原体感染。接种物经调整后可在50%至80%等量的标准McCoy细胞中产生包涵体,导致子宫内膜柱状细胞的感染率约为15%至30%,宫颈外鳞状细胞的感染率为5%至10%。将培养物暴露于二乙氨基乙基葡聚糖并进行离心辅助接种,据报道这些操作可增强对HeLa细胞和McCoy细胞的感染,但并未改变包涵体阳性生殖道细胞的数量。在接种后培养基中添加放线菌酮可略微增加含包涵体细胞的数量,而在添加激素的培养基中培养生殖道细胞对包涵体发育产生可变影响,并显著降低放射性标记生物体与这些细胞的结合。接种后立即通过免疫荧光分析衣原体与宿主细胞的结合情况,揭示了McCoy细胞培养物与生殖道细胞培养物中不同感染水平的基础。衣原体未能粘附于生殖道细胞培养物中的许多细胞,而对McCoy细胞的粘附则是均匀的。此外,放射性标记的沙眼衣原体与生殖道细胞的结合明显低于与McCoy细胞的结合。最后,确定了显著抑制包涵体发育而不立即丧失衣原体生长潜力的培养条件。这项研究表明,原代生殖道细胞培养物易受衣原体感染,对于研究沙眼衣原体与天然人类靶细胞相互作用的性质将具有重要价值。