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Cell type-intrinsic differences in the host transcriptome of Chlamydia trachomatis-infected primary human endocervical epithelial cells and vaginal epithelial cells [HVE_seq]

ABSTRACT: To investigate the role of cell type-intrinsic gene expression to fibrotic sequelae of Chlamydia trachomatis (Ct) infection of the upper female genital tract, we compared the transcriptomic response of primary human endocervical epithelial cells (HCECs, see GSE198272) to that in vaginal epithelial cells (HVEs).

ORGANISM(S): Homo sapiens

PROVIDER: GSE228774 | GEO | 2023/09/27

REPOSITORIES: GEO

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Project description:Introduction Chlamydia trachomatis (C. trachomatis) is a Gram-negative bacterium and a common human pathogen. The World Health Organization (WHO) estimates that over 130 million people are infected with C. trachomatis globally each year and with increasing incidence. C. trachomatis causes long-lasting and recurrent infections that over time induce severe tissue damage in the female genital tract that can lead to ectopic pregnancy and infertility. Thus, the human immune system fails to control and eradicate C. trachomatis during primary infection and fails to develop protective immunity against secondary infections. In vivo infection models, using complement knock out mice, suggest that the complement system is critically involved in both anti-chlamydial immunity and infection-induced pathology. To increase our understanding of complement-mediated immunity against C. trachomatis we analyzed global complement deposition on serum-incubated C. trachomatis by mass spectrometry. Methods Purified C. trachomatis was incubated in seronegative normal human serum (NHS) or heat-inactivated normal human serum (HI-NHS) for 30 min, thoroughly washed, and processed for mass spectrometry. All samples were lysed, reduced and alkylated and digested with trypsin. Some samples were chemically modified to acetylate free amino groups (N-terminal and lysine amino groups) before trypsin digestion. Peptides were analyzed on a UltimateTM 3500 RSLCnano coupled to a Q Exactive HF-X mass spectrometer. Raw data files were searched against the Uniprot human reference proteome using MaxQuant. Results We demonstrate that C. trachomatis elicits potent complement activation demonstrated by deposition of both early and late complement factors together with several complement regulators. We further demonstrate proteolytically processing of complement C3b to “inactive” C3 cleavage fragments. Conclusion We demonstrate the deposition of several novel complement-associated proteins and -cleavage fragments on the surface of C. trachomatis.

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Cell type-intrinsic differences in the host transcriptome of Chlamydia trachomatis-infected primary human endocervical epithelial cells and vaginal epithelial cells [HVE_seq]

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