Differences in Rab protein expressions in McCoy cells with acute versus persistent Chlamydia trachomatis infection

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Objective To investigate differences in Rab protein expressions in McCoy cells with acute versus persistent Chlamydia trachomatis (Ct)infection. Methods Cultured McCoy cells were infected with different amounts(400, 500, 550 μl/well)of Ct strain D suspensions , then cultured with the medium containing 100 U/ml penicillin G(persistent Ct infection groups)or that without penicillin G (acute Ct infection groups). Ctuninfected McCoy cells receiving no penicillin G treatment served as the blank control group, and those receiving penicillin G treatment as the penicillin group. After 48-hour culture, McCoy cells were lysed, proteins were collected, and total RNA was extracted from the cells. Enzyme-linked immunosorbent assay (ELISA)was conducted to measure protein levels of Rab4A, Rab6A, Rab10, Rab11A and Rab14, and fluorescence-based quantitative PCR to quantify mRNA expressions of Rab4A and Rab14 (expressed as 2-ΔΔCt). Results Protein levels of Rab4A, Rab6A, Rab10, Rab11A and Rab14 were all significantly lower in the acute than in the persistent Ct infection groups (all Z = 3.621, P 0.05). In addition, the expressions of Rab4A and Rab14 mRNAs were consistent with those of their proteins in these groups. Conclusion The transcriptional and expression levels of Rab proteins are higher in McCoy cells persistently infected with Ct than in those acutely infected with Ct. Key words: Chlamydia trachomatis; Chlamydia infections; rab GTP-binding proteins; Acute infection; Persistent infection; McCoy cells