Trichomonas vaginalis is a flagellated protozoan parasite that causes vaginitis and cervicitis in women and asymptomatic urethritis and prostatitis in men. Mast cells have been reported to be predominant in vaginal smears and vaginal walls of patients infected with T. vaginalis. Mitogen-activated protein kinase (MAPK), activated by various stimuli, have been shown to regulate the transcriptional activity of various cytokine genes in mast cells. In this study, we investigated whether MAPK is involved in ROS generation and exocytotic degranulation in HMC-1 cells induced by T. vaginalis-derived secretory products (TvSP). We found that TvSP induces the activation of MAPK and NADPH oxidase in HMC-1 cells. Stimulation with TvSP induced phosphorylation of MAPK and p47phox in HMC-1 cells. Stimulation with TvSP also induced up-regulation of CD63, a marker for exocytosis, along the surfaces of human mast cells. Pretreatment with MAPK inhibitors strongly inhibited TvSP-induced ROS generation and exocytotic degranulation. Finally, our results suggest that TvSP induces intracellular ROS generation and exocytotic degranulation in HMC-1 via MAPK signaling.
Cell Degranulation ; Cell Line ; *Exocytosis ; Humans ; Mast Cells/*drug effects/*metabolism ; Mitogen-Activated Protein Kinases/*metabolism ; Reactive Oxygen Species/*metabolism ; Trichomonas vaginalis/*metabolism ; Virulence Factors/*metabolism

Objectives: The aim of this study was to determine the usefulness of a diagnostic method for trichomoniasis that uses sequencing of the Tv-E650 gene. Methods: Totally 109 patients visited Ulaan Tuuz Hospital, National Center of Communicable Disease, Ulaanbaatar and were examined for trichomoniasis using wet mount examination, Papanicolaou (Pap) smear, culture and PCR. A family of 650 base pair (bp) long repeats from the T. vaginalis genome, designated as the Tv-E650 family, was sequenced for five Mongolian strains. Results: As a result, 21.1% (23/109) were positive by wet mount, 18.3% (20/109) were positive by Pap smear, 28.4% (31/109) were positive by culture and 36.6% (40/109) were positive by PCR. The differences among the strains are single-nucleotide and 2-nucleotide substitutions and insertions or deletions. The sequence uniformity of the strains as well as the presence of identical mutations in different isolates suggest efficient sequence homogenization mechanisms. In blast results, the Tv-E650 repeat family is conserved in all T. vaginalis strains examined, regardless of their diverse geographical origin. Conclusion: The Tv-E650 repeat family of T. vaginalis is a simple, rapid sensitive and specific accurate method for identification and diagnosis of trichomoniasis.