Campbell S, Richmond S J, Yates P S, Storey C C
Department of Obstetrics and Gynaecology, University Hospital of South Manchester, West Didsbury, UK.
Microbiology (Reading). 1994 Aug;140 ( Pt 8):1995-2002. doi: 10.1099/13500872-140-8-1995.
In view of the controversy concerning the expression of chlamydial lipopolysaccharide (LPS) in the eukaryotic host cell, the presence of this molecule was examined in three cell types which were experimentally infected with Chlamydia trachomatis serotype E. LPS was detected in the McCoy cell line, human endometrial epithelium and human amniotic epithelium with two monoclonal antibodies. The appearance and distribution of LPS at the host cell surface during the chlamydial developmental cycle and its transmission to neighbouring cells were examined by immunofluorescence microscopy after air drying of the host cells. LPS distribution was not uniform; it was first observed on regions of the cell surface in close proximity to the chlamydial endosome (inclusion). Soon after, the antigen was also detected at points of contact with neighbouring uninfected cells. Immunofluorescent plaques of host cells contaminated with LPS were thus formed in the vicinity of infected cells. These plaques increased in size over 2 d before becoming smaller as host cell lysis occurred. The major outer-membrane protein (MOMP) was not visualized on the host cell surface after air drying. No cell-surface LPS antigen was observed in live cells or those fixed in formaldehyde without air drying. Conventional methanol fixation and immunolocalization of LPS and MOMP in parallel infected cultures stained these antigens within inclusions in the expected fashion. Radio-immunoassays were used to quantify LPS in confluent McCoy cell monolayers during the chlamydial developmental cycle. Cell-surface-associated and inclusion-associated LPS, measured by direct binding of 125I-labelled anti-LPS monoclonal antibodies to air-dried or methanol-fixed monolayers respectively, increased for up to 3 d then declined.(ABSTRACT TRUNCATED AT 250 WORDS)
鉴于衣原体脂多糖(LPS)在真核宿主细胞中表达存在争议,我们使用沙眼衣原体E血清型对三种细胞类型进行实验性感染,检测该分子的存在情况。使用两种单克隆抗体在 McCoy 细胞系、人子宫内膜上皮细胞和人羊膜上皮细胞中检测到了 LPS。通过对宿主细胞进行空气干燥后,利用免疫荧光显微镜检查衣原体发育周期中宿主细胞表面 LPS 的外观和分布及其向邻近细胞的传递。LPS 的分布并不均匀;首先在细胞表面靠近衣原体包涵体的区域观察到。不久之后,在与未感染的邻近细胞接触点也检测到了该抗原。因此,在感染细胞附近形成了被 LPS 污染的宿主细胞免疫荧光斑块。这些斑块在 2 天内尺寸增大,随后随着宿主细胞裂解而变小。空气干燥后,宿主细胞表面未观察到主要外膜蛋白(MOMP)。在活细胞或未经空气干燥的甲醛固定细胞中未观察到细胞表面 LPS 抗原。在平行感染的培养物中,常规甲醇固定和 LPS 及 MOMP 的免疫定位以预期方式在包涵体内对这些抗原进行了染色。使用放射免疫测定法定量衣原体发育周期中汇合的 McCoy 细胞单层中的 LPS。通过分别将 125I 标记的抗 LPS 单克隆抗体直接与空气干燥或甲醇固定的单层结合来测量细胞表面相关和包涵体相关的 LPS,其含量在长达 3 天内增加,然后下降。(摘要截短至250字)