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Demonstration and Characterization of Cyst-Like Structures in the Life Cycle of Trichomonas vaginalis

Beri, D and Yadav, P and Devi, HRN and Narayana, C and Gadara, D and Tatu, U (2020) Demonstration and Characterization of Cyst-Like Structures in the Life Cycle of Trichomonas vaginalis. In: Frontiers in Cellular and Infection Microbiology, 9 .

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Official URL: https://doi.org/10.3389/fcimb.2019.00430


Trichomonas vaginalis is the parasitic protozoan residing in human urogenital tract causing trichomoniasis, which is the leading non-viral sexually transmitted disease. It has cosmopolitan distribution throughout the globe and affects both men and women. Lifecycle of the parasite has been traditionally described as consisting of motile and symptom-causing trophozoites. Chemical and temperature perturbations in trophozoites have been shown to aid conversion to pseudocysts, which is poorly investigated. In the current study, we show the formation of viable cyst-like structures (CLS) in stationary phase of T. vaginalis axenic culture. We used a fluorescent stain called calcofluor white, which specifically binds to chitin and cellulose-containing structures, to score for T. vaginalis CLS. Using flow cytometry, we demonstrated and quantitated the processes of encystation as well as excystation; thus, completing the parasite's lifecycle in vitro without any chemical/temperature alterations. Like cysts from other protozoan parasites such as Entamoeba histolytica and Giardia lamblia, T. vaginalis CLS appeared spherical, immotile, and resistant to osmotic lysis and detergent treatments. Ultrastructure of CLS demonstrated by Transmission Electron Microscopy showed a thick electron-dense deposition along its outer membrane. To probe the physiological role of CLS, we exposed parasites to vaginal pH and observed that trophozoites took this as a cue to convert to CLS. Further, upon co- culturing with cells of cervical origin, CLS rapidly excysted to form trophozoites which abrogated the cervical cell monolayer in a dose-dependent manner. To further corroborate the presence of two distinct forms in T. vaginalis, we performed two-dimensional gel electrophoresis and global, untargeted mass spectrometry to highlight differences in the proteome with trophozoites. Interestingly, CLS remained viable in chlorinated swimming pool water implicating the possibility of its role as environmentally resistant structures involved in non-sexual mode of parasite transmission. Finally, we showed that symptomatic human patient vaginal swabs had both T. vaginalis trophozoites and CLS; thus, highlighting its importance in clinical infections. Overall, our study highlights the plasticity of the pathogen and its rapid adaption when subjected to stressful environmental cues and suggests an important role of CLS in the parasite's life cycle, pathogenesis and transmission.

Item Type: Journal Article
Publication: Frontiers in Cellular and Infection Microbiology
Publisher: Frontiers Media S.A.
Additional Information: The copyright for this article belongs to the Authors.
Keywords: cellulose; chitin; chlorine; fluorescent dye; pool water; proteome; unclassified drug; water; proteome; protozoal protein, adult; Article; chemical structure; clinical article; coculture; controlled study; cyst; cyst like structure; Entamoeba histolytica; female; flow cytometry; gel electrophoresis; Giardia intestinalis; human; in vitro study; life cycle; mass spectrometry; nonhuman; outer membrane; parasite transmission; pathogenesis; quantitative analysis; transmission electron microscopy; Trichomonas vaginalis; trophozoite; vagina pH; vagina smear; cell plasticity; cyst; HeLa cell line; life cycle stage; metabolism; parasite encystment; parasitology; physiological stress; physiology; proteomics; ultrastructure; vagina, Cell Plasticity; Cysts; Entamoeba histolytica; Female; Giardia lamblia; HeLa Cells; Humans; Life Cycle Stages; Microscopy, Electron, Transmission; Parasite Encystment; Proteome; Proteomics; Protozoan Proteins; Stress, Physiological; Trichomonas vaginalis; Trophozoites; Vagina
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 04 Jan 2023 10:17
Last Modified: 04 Jan 2023 10:17
URI: https://eprints.iisc.ac.in/id/eprint/79471

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