Abstract
Microbial adhesion is critical for parasitic infection and colonization of host cells. To study the host–parasite interaction in vitro, we established a flow cytometry-based assay to measure the adherence of Trichomonas vaginalis to epithelial cell line SiHa. SiHa cells and T. vaginalis were detected as clearly separated, quantifiable populations by flow cytometry. We found that T. vaginalis attached to SiHa cells as early as 30 min after infection and the binding remained stable up to several hours, allowing for analysis of drug treatment efficacy. Importantly, NADPH oxidase inhibitor DPI treatment induced the detachment of T. vaginalis from SiHa cells in a dose-dependent manner without affecting host cell viability. Thus, this study may provide an understanding for the potential development of therapies against T. vaginalis and other parasite infections.
Highlights
Trichomoniasis is a sexually transmitted disease (STD) caused by infection of the urogenital tract by the flagellate protozoan parasite Trichomonas vaginalis [1,2,3]
DPI, but not NAC, was involved in blockade of parasite adherence to host cells, suggesting that DPI-targetable reactive oxygen species (ROS) generation in T. vaginalis contributes to the pathogenic adherence process
Fluorescent dye-loaded T. vaginalis had been incubated with host cells and monitored by microscopy or flow cytometry [28,29]
Summary
Trichomoniasis is a sexually transmitted disease (STD) caused by infection of the urogenital tract by the flagellate protozoan parasite Trichomonas vaginalis [1,2,3]. Trichomoniasis in pregnant women may lead to premature membrane rupture, preterm delivery, and low birth weight [4] It has been associated with atypical pelvic inflammatory disease [5], infertility [6], predisposition to invasive cervical cancer [7], and increased susceptibility to HIV infection [8]. We established a flow cytometry-based method to examine the properties of T. vaginalis adhesion on the cervix carcinoma cell line, SiHa cells. Using antioxidant compounds such as N-acetyl-L-cysteine (NAC) and NADPH oxidase inhibitor diphenyleneiodonium (DPI), we determined whether ROS production was involved in the adhesion process during T. vaginalis infection. DPI, but not NAC, was involved in blockade of parasite adherence to host cells, suggesting that DPI-targetable ROS generation in T. vaginalis contributes to the pathogenic adherence process
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