Lee C K
Infect Immun. 1981 Feb;31(2):584-91. doi: 10.1128/iai.31.2.584-591.1981.
A system was devised for studying the interaction of a trachoma strain of Chlamydia trachomatis (G17) and mouse fibroblasts (McCoy cells) in the absence of centrifugation, which is usually employed to enhance the infection of cell cultures with non-lymphogranuloma venereum human strains of C. trachomatis. In this system, the conditions of infection more closely approached those encountered in natural infections, and the entry of G17 into host cells could be compared with the previously described entry of C. psittaci 6BC and a lymphogranuloma venereum strain (440L) of C. trachomatis. McCoy cells were infected by shaking at 37 degrees C with inocula suspended in 0.01 M phosphate buffer, pH 7.2, containing 0.2 M sucrose. The efficiency of infection (inclusion counts without centrifugation/inclusion counts with centrifugation) was 1.5% for monolayers and 7.5% for suspensions. When measured either by inclusion counts or by host cell-associated 14C-amino acid-labeled G17, association was proportional to G17 concentration and increased linearly for 60 min. Pretreatment of host cells with diethylaminoethyl-dextran (30 micrograms/ml, 30 min) raised the efficiency of infection to about 13% for both monolayers and suspensions. Host cells treated with cytochalasin B (2 x 10(-5) M, 90 min) or trypsin (50 micrograms/ml, 60 min) associated with G17 at undiminished rates. 14C-labeled G17 inactivated by heat (60 degrees C, 3 min) or ultraviolet light (1,800 ergs per mm2) associated with McCoy cells at the same rate as live G17. Comparison of these results with those previously reported for strains 6BC and 440L showed that strain G17 exhibited some, but not all, of the host cell association properties of the other two chlamydial strains.
设计了一种系统,用于研究沙眼衣原体(G17)的沙眼菌株与小鼠成纤维细胞(McCoy细胞)在无离心情况下的相互作用,离心通常用于增强细胞培养物被非淋巴肉芽肿性尿道炎人型沙眼衣原体菌株感染的能力。在该系统中,感染条件更接近自然感染中遇到的情况,并且可以将G17进入宿主细胞的情况与先前描述的鹦鹉热衣原体6BC和沙眼衣原体的淋巴肉芽肿性尿道炎菌株(440L)进入宿主细胞的情况进行比较。将McCoy细胞在37℃下与悬浮于含0.2M蔗糖的pH7.2的0.01M磷酸盐缓冲液中的接种物一起振荡感染。单层细胞的感染效率(无离心时的包涵体计数/有离心时的包涵体计数)为1.5%,悬浮液的感染效率为7.5%。当通过包涵体计数或通过宿主细胞相关的14C-氨基酸标记的G17进行测量时,结合与G17浓度成正比,并在60分钟内呈线性增加。用二乙氨基乙基葡聚糖(30微克/毫升,30分钟)预处理宿主细胞可使单层细胞和悬浮液的感染效率提高到约13%。用细胞松弛素B(2×10-5M,90分钟)或胰蛋白酶(50微克/毫升,60分钟)处理的宿主细胞与G17的结合速率未降低。经热(60℃,3分钟)或紫外线(每平方毫米1800尔格)灭活的14C标记的G17与McCoy细胞的结合速率与活G17相同。将这些结果与先前报道的6BC和440L菌株的结果进行比较表明,G17菌株表现出其他两种衣原体菌株的一些但不是全部宿主细胞结合特性。