Barsoum I S, Goodman T A, Hardin L K, Colley D G
Department of Microbiology, Vanderbilt University School of Medicine, Nashville, TN.
Med Microbiol Immunol. 1989;178(2):113-20. doi: 10.1007/BF00203307.
A human epithelioid cell line (A-431) was tested in parallel with McCoy fibroblast cells for the growth of trachoma-related serovar A Chlamydia trachomatis without centrifugation or cycloheximide addition. A-431 cells were 4-7 times more susceptible to infection with serovar A than McCoy cells in such unmanipulated cultures. Murine monoclonal antibodies (MAbs) developed against serovar A were then evaluated for their ability to inhibit unmanipulated serovar A infectivity of A-431 cells. Two of seven MAbs tested neutralized infectivity by more than 50%. An IgG2a MAb (2C8) that is specific for serovar A, and another IgG2a MAb (4E3) that reacts equally with serovars A and L2 neutralized infectivity of serovar A by 72.2 +/- 3.7% and 56.0 +/- 5.8% (mean +/- SEM of 7 experiments) respectively. Mouse immune serum (MIS) raised against serovar A elementary bodies (EB) neutralized infectivity of serovar A by 76.0 +/- 4.9% (mean +/- SEM of 7 experiments). Immunoblot detection of serovar A EB polypeptides separated by SDS-PAGE indicated that 2C8 reacted with a 16 kD and 4E3 reacted with a 12 kD polypeptide while MIS reacted with several polypeptides including the major outer membrane protein (MOMP). These studies show that the human epithelioid cell line A-431 is a more susceptible host than McCoy cells in unmanipulated cultures, and that 2 MAbs neutralize serovar A infectivity of A-431 cells. Identification of antigenic moieties of importance in unmanipulated chlamydial infections may help in the development of potential vaccines against trachoma.
在不进行离心或不添加环己酰亚胺的情况下，将人上皮样细胞系（A-431）与 McCoy 成纤维细胞平行进行测试，以观察沙眼相关血清型 A 沙眼衣原体的生长情况。在这种未处理的培养物中，A-431 细胞对血清型 A 感染的敏感性比 McCoy 细胞高 4-7 倍。然后评估针对血清型 A 产生的鼠单克隆抗体（MAb）抑制 A-431 细胞未处理的血清型 A 感染性的能力。所测试的七种 MAb 中有两种中和感染性超过 50%。一种对血清型 A 特异的 IgG2a MAb（2C8）和另一种与血清型 A 和 L2 反应相同的 IgG2a MAb（4E3）分别将血清型 A 的感染性中和了 72.2±3.7%和 56.0±5.8%（7 次实验的平均值±标准误）。针对血清型 A 原体（EB）产生的小鼠免疫血清（MIS）将血清型 A 的感染性中和了 76.0±4.9%（7 次实验的平均值±标准误）。通过 SDS-PAGE 分离的血清型 A EB 多肽的免疫印迹检测表明，2C8 与一种 16 kD 的多肽反应，4E3 与一种 12 kD 的多肽反应，而 MIS 与包括主要外膜蛋白（MOMP）在内的几种多肽反应。这些研究表明，在未处理的培养物中，人上皮样细胞系 A-431 比 McCoy 细胞更易感染，并且两种 MAb 可中和 A-431 细胞的血清型 A 感染性。鉴定未处理的衣原体感染中重要的抗原部分可能有助于开发针对沙眼的潜在疫苗。
