abstract
Introduction: Trichomoniasis is the most common non-viral sexually transmitted infection that increases the risk of cervical cancer. Trichomonas vaginalis (T. vaginalis) can regulate the pro-inflammatory cytokine production in the host cells. Toll-like receptors (TLRs) are a family of the pattern recognition receptors (PRRs) of mammalian cells, expressed in various host cells and have an important role in recognizing pathogens, and pro-inflammatory responses. The aim of the present study is to investigate the role of TLR5 in cervical cancer cells (HeLa) and human vaginal epithelial cells (HVECs) exposed to T. vaginalis. Methodology: First, the cells and parasites were cultured in RPMI and trypticase yeast extract maltose (TYM), respectively. After adaption of parasite and epithelial cells by RPMI-TYM medium co-culture (9:1 vol/vol), HVECs and HeLa cells were stimulated with T. vaginalis trophozoites (24-hour incubation at 37 degrees C, 5% CO2). Following RNA extraction and cDNA synthesis, the gene expression levels of TLR5, IRAK1, and NF-kappa B were assessed using real-time PCR. Besides, the protein levels were measured using western blotting. All tests and controls were normalized using beta-actin as a housekeeping control. Results: Real-time PCR results showed an increased gene expression of TLR5, IRAK1, and NF-kappa B in T. vaginalis exposed HVECs and HeLa cells compared to the control group (p < 0.05). Additionally, western blot analysis showed a statistically significant increase in TLR5, and NF-kappa B proteins in both groups after exposure to the parasite (p < 0.05). Conclusions: These findings provide insight into the host-parasite interaction, and the results indicated that T. vaginalis could stimulate TLR5 and activate related pathways.
keywords
TOLL-LIKE RECEPTORS; GENE-EXPRESSION; UP-REGULATION; TRIGGERS; IMMUNITY; CYTOKINES
subject category
Infectious Diseases
authors
Esfanjani, SM; Maleki, LA; Nami, S; Ebrahimi, M; Baghbanzadeh, A; Perez-Cordon, G; Oliveira, SMR; Pereira, MD; Barac, A; Ahmadpour, E; Micic, J
our authors
Maria de Lourdes Gomes Pereira
Associate Professor with Aggregation
Sónia M. R. Oliveira
Junior Researcheracknowledgements
This work has been done as part of the M.Sc. thesis for Soraya Mohammadi Esfanjani. This study was funded by Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran (Grant No. 62666) . The sponsor or funding organization had no role in the design or conduct of this study. M.d.L.P. thanks to project CICECO-Aveiro Institute of Materials, UIDB/50011/2020 and UIDP/50011/2020, national funds by FCT/MCTES.