Wyrick P B, Choong J, Davis C H, Knight S T, Royal M O, Maslow A S, Bagnell C R
Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill 27599.
Infect Immun. 1989 Aug;57(8):2378-89. doi: 10.1128/iai.57.8.2378-2389.1989.
To study the initial invasion process(es) of genital chlamydiae, a model system consisting of hormonally maintained primary cultures of human endometrial gland epithelial cells (HEGEC), grown in a polarized orientation on collagen-coated filters, was utilized. After Chlamydia trachomatis inoculation of the apical surface of polarized HEGEC, chlamydiae were readily visualized, by transmission electron microscopy, in coated pits and coated vesicles. This was true for HEGEC maintained in physiologic concentrations of estrogen (proliferative phase) and of estrogen plus progesterone (secretory phase), despite the finding that association of chlamydiae with secretory-phase HEGEC is significantly reduced (P = 0.025; A.S. Maslow, C.H. Davis, J. Choong, and P.B. Wyrick, Am. J. Obstet. Gynecol. 159:1006-1014, 1988). In contrast, chlamydiae were rarely observed in the clathrin-associated structures if the HEGEC were cultured on plastic surfaces. The same pattern of coated pit versus noncoated pit entry was reproducible in HeLa cells. The quantity of coated pits associated with isolated membrane sheets derived from HeLa cells, grown on poly-L-lysine-coated cover slips in medium containing the female hormones, was not significantly different as monitored by radiolabeling studies and by laser scanning microscopy. These data suggest that culture conditions which mimic in vivo cellular organization may enhance entry into coated pits for some obligate intracellular pathogens.
为了研究生殖道衣原体的初始侵入过程,我们利用了一个模型系统,该系统由在胶原包被的滤膜上以极化方向生长的人子宫内膜腺上皮细胞(HEGEC)的激素维持原代培养物组成。在用沙眼衣原体接种极化的HEGEC的顶表面后,通过透射电子显微镜很容易在有被小窝和有被小泡中观察到衣原体。对于在生理浓度雌激素(增殖期)以及雌激素加孕激素(分泌期)中维持的HEGEC来说都是如此,尽管发现衣原体与分泌期HEGEC的结合显著减少(P = 0.025;A.S. Maslow、C.H. Davis、J. Choong和P.B. Wyrick,《美国妇产科杂志》159:1006 - 1014,1988年)。相比之下,如果HEGEC在塑料表面培养,则很少在网格蛋白相关结构中观察到衣原体。在HeLa细胞中也可重现相同的有被小窝与无被小窝进入模式。通过放射性标记研究和激光扫描显微镜监测,在含有女性激素的培养基中在聚-L-赖氨酸包被的盖玻片上生长的HeLa细胞来源的分离膜片相关的有被小窝数量没有显著差异。这些数据表明,模拟体内细胞组织的培养条件可能会增强某些专性细胞内病原体进入有被小窝的能力。