Chlamydia trachomatis infection of cultured motile cells after uptake of chlamydiae from the substratum

Abstract

The ability of motile cells to remove small inanimate particles from solid substrata is well documented. We show here that motile cells will pick up and internalize infectious particles of the obligate intracellular parasite Chlamydia trachomatis when they are adherent to the substratum over which the host cells move. Two cell types were used to assess chlamydial uptake; a feeder independent human squamous cell carcinoma variant (AC3A cells) and the McCoy cell line. Purified chlamydia elementary bodies were attached to glass or collagen-coated glass by centrifugation. Suspensions of cells were then allowed to sediment on to the substrata to which chlamydiae had attached. Both types of cell picked up chlamydiae and transported them over their surface during the course of attachment and spreading. Stereoscopic images obtained by confocal microscopy demonstrated that chlamydiae were found mainly on the surface of non-spread cells. After the cells had spread on the substratum they began to move around forming tracks where the chlamydiae had been removed. Some cell-surface-attached chlamydiae were endocytosed and a proportion of these proliferated during the 48 h after plating. However, chlamydiae attached to the substratum lost infectivity by a simple exponential decay process within a few hours of incubation in the extracellular environment. Therefore, increasing numbers of non-viable organisms were probably endocytosed as the time of extracellular incubation increased. This mode of infection may be relevant to in vivo situations where cell migration occurs after damage to mucosal surfaces.