Differences in Chlamydia trachomatis serovar E growth rate in polarized endometrial and endocervical epithelial cells grown in three-dimensional culture.
In vitro studies of obligate intracellular chlamydia biology and pathogenesis are highly dependent on the use of experimental models and growth conditions that mimic the mucosal architecture and environment these pathogens encounter during natural infections. In this study, the growth of Chlamydia trachomatis genital serovar E was monitored in mouse fibroblast McCoy cells and compared to more relevant host human epithelial endometrium-derived HEC-1B and cervix-derived HeLa cells, seeded and polarized on collagen-coated microcarrier beads, using a three-dimensional culture system. Microscopy analysis of these cell lines prior to infection revealed morphological differences reminiscent of their in vivo architecture. Upon infection, early chlamydial inclusion distribution was uniform in McCoy cells but patchy in both epithelial cell lines. Although no difference in chlamydial attachment to or entry into the two genital epithelial cell lines was noted, active bacterial genome replication and transcription, as well as initial transformation of elementary bodies to reticulate bodies, were detected earlier in HEC-1B than in HeLa cells, suggesting a faster growth, which led to higher progeny counts and titers in HEC-1B cells upon completion of the developmental cycle. Chlamydial development in the less relevant McCoy cells was very similar to that in HeLa cells, although higher progeny counts were obtained. In conclusion, this three-dimensional bead culture system represents an improved model for harvesting large quantities of infectious chlamydia progeny from their more natural polarized epithelial host cells.
Guseva
Natalia V
NV
Department of Microbiology, East Tennessee State University, James H. Quillen College of Medicine, Johnson City, TN 37614, USA.
Dessus-Babus
Sophie
S
Moore
Cheryl G
CG
Whittimore
Judy D
JD
Wyrick
Priscilla B
PB
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D016764:Cell Polarity; D002692:Chlamydia trachomatis / Q000254:growth & development*; D003593:Cytoplasm / Q000382:microbiology; D004261:DNA Replication; D004269:DNA, Bacterial / Q000032:analysis; D004717:Endometrium / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology*; D005260:Female; D005347:Fibroblasts / Q000382:microbiology; D006367:HeLa Cells; D006801:Humans; D002479:Inclusion Bodies; D051379:Mice; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008863:Microspheres; D012329:RNA, Bacterial / Q000032:analysis; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D012329:RNA, Bacterial