<i>Chlamydia trachomatis</i> infection induces CiHHV-6 DNA replication.

<p>(<b>A</b>) qPCR analysis of viral DNA in HSB-ML cells after 2 days of <i>C. trachomatis</i> infection. HSB-ML cells were infected with <i>C. trachomatis</i> (Ctr) for 2 days. In parallel, the same number of HSB-ML cells was cultured in the presence of 1 µM hydrocortisone or 80 ng/ml of TSA for 2 days. As TSA was dissolved in DMSO, a DMSO control was also included in the study. Data represent the mean ± SEM of 5 independent experiments. (<b>B</b>) qPCR analysis of viral DNA in HeLa cells carrying latent HHV-6A after 2 days of <i>C. trachomatis</i> infection. Cells were infected with <i>C. trachomatis</i> or were treated with TSA or hydrocortisone as mentioned in (A). Data represent the mean ± SEM of 5 independent experiments. (<b>C, D, E</b>) qPCR analysis of viral DNA in HSB-ML (C), JL-LCL (D) and PL-LCL (E) cells after 9 days of <i>C. trachomatis</i> infection. Cells were infected with <i>C. trachomatis</i> at an MOI of 2–5 for 2 days after which cells were washed thoroughly and grown in presence of fresh media with 1 µg/ml of doxycycline for another 7 days. Total DNA was isolated and used for qPCR assay. Data represent the mean ± SEM of three independent experiments. (<b>F</b>) qPCR analysis of viral DNA in freshly isolated blood cells from CiHHV-6 individuals after 5 days of <i>C. trachomatis</i> infection. Fresh PBMCs were isolated from total blood and were infected with <i>C. trachomatis</i> for 2 days at an MOI of 5 after which cells were washed thoroughly and grown in presence of fresh media with 1 µg/ml of doxycycline for another 3 days. In parallel, the same numbers of PBMCs were cultured in presence of 80 ng/ml of TSA for 5 days. Total leukocytes were separated from monocyte-derived macrophages and were infected with <i>C. trachomatis</i> in parallel. Heat inactivated <i>C. trachomatis</i> (hiCtr) were added to total PBMCs in a separate well and were included as negative control. Data represent the mean ± SEM of results from blood samples of 5 different CiHHV-6 individuals. (<b>G</b>) qPCR analysis of viral DNA in total leukocytes and monocyte-derived macrophages from CiHHV-6 individuals after 5 days of <i>C. trachomatis</i> infection. Total leukocytes were separated from monocyte-derived macrophages and were infected with <i>C. trachomatis</i> for 2 days at an MOI of 5 after which cells were washed thoroughly and grown in presence of fresh media with 1 µg/ml of doxycycline for another 3 days. Data represent the mean ± SEM of three independent experiments performed at the same time from one CiHHV-6 individual.