Trichomoniasis is a common sexually transmitted infection caused by Trichomonas vaginalis, which actually does not exist a vaccine for control or prevention. Thus, the identification of new and potent immunogens in T. vaginalis, which can contribute to the development of a vaccine against this parasite, is necessary. Therefore, the aim of this work was to evaluate the potential of a recombinant Transient Receptor Potential-like channel of T. vaginalis (TvTRPV), as a promising immunogen in BALB/c mice. First, TvTRPV was cloned and expressed as a recombinant protein in Escherichia coli BL21 cells and purified by nickel affinity. Next, BALB/c mice were immunized and the antibody levels in mice serum and cytokines from the supernatant of macrophages and from co-culture systems were evaluated. Recombinant TvTRPV triggered high levels of specific total IgG in sera from the immunized mice. Also, a statistically significant increase of cytokines: IL-1β, IL-6, and TNF-α after stimulation with the corresponding antigens in vitro, was identified. Moreover, co-cultures using CD4⁺ T cells from immunized mice were able to identify higher levels of IL-10 and IFN-γ. These results were useful to validate the immunogenicity of TvTRPV in BALB/c mice, where IL-10-IFN-γ-secreting cells could play a role in infection control, supporting the potential of TvTRPV as a promising target for vaccine against T. vaginalis.
Animals ; Calcium ; Clone Cells ; Coculture Techniques ; Cytokines ; Escherichia coli ; Immunoglobulin G ; In Vitro Techniques ; Infection Control ; Interleukin-10 ; Interleukin-6 ; Macrophages ; Mice ; Nickel ; Parasites ; Sexually Transmitted Diseases ; T-Lymphocytes ; Trichomonas vaginalis ; Trichomonas

PCR is known to be the most sensitive method for diagnosing Trichomonas vaginalis infections. This study aimed to compare the sensitivity of a PCR assay for trichomoniasis (HY-PCR) developed in Hanyang University with the use of a Seeplex Ace Detection Kit®, using urine collected from four Korean men with prostatic disease. Overall, HY-PCR was more sensitive than the Seeplex Kit. The use of Chelex 100 is recommended for DNA isolation in order to increase the sensitivity of the PCR test.
DNA ; Humans ; Male ; Methods ; Polymerase Chain Reaction ; Prostatic Diseases ; Trichomonas vaginalis ; Trichomonas

Trichomoniasis is the most common curable sexually-transmitted infection. Most Trichomonas vaginalis-infected men are asymptomatic and can remain undiagnosed and untreated, and this has been thought to result in chronic persistent prostatic infection. Chronic inflammation is regarded as the major factor in the pathogenesis and progression of benign prostatic hyperplasia (BPH) and prostatic cancer (PCa). The aim of this study is to identify seropositivity to T. vaginalis in men with prostate tumors (BPH or PCa) visited to Hanyang University Hospital. A total of 183 men were enrolled between October 2013 and November 2014. They consisted of 139 with BPH (mean age: 64.0±0.07) and 44 with prostate cancer (mean age: 73.3±0.18). We carried out ELISA to identify the seropositivity to T. vaginalis. Mixed lysate antigen extracted from 8 strains of T. vaginalis was used in the ELISA. Also 58 male outpatients visited to Health Promotion Center in Hanyang University Hospital were evaluated for comparing group. As a results, seropositivity to T. vaginalis in patients with prostatic diseases was 19.7% (BPH: 18.7%, PCa: 22.7%) and it was significantly higher than the 1.7% of the comparing healthy group (P=0.001). Therefore, prostatic tumor showed higher seropositivity against T. vaginalis than normal men. As far as we know, this is the first report about seroprevalence in prostatic tumor in Korea.
Enzyme-Linked Immunosorbent Assay ; Health Promotion ; Humans ; Inflammation ; Korea ; Male ; Outpatients ; Passive Cutaneous Anaphylaxis ; Prostate ; Prostatic Diseases ; Prostatic Hyperplasia ; Prostatic Neoplasms ; Seroepidemiologic Studies ; Trichomonas vaginalis ; Trichomonas

BACKGROUND: Metronidazole susceptibility and the presence of Trichomonas vaginalis virus (TVV) are the phenotypes found to be significantly correlated with the microsatellite-based genotypes of T. vaginalis. These phenotypes were assessed in T. vaginalis isolates from select urban areas to determine preliminary "type" of Philippine T. vaginalis.

METHODS: Culture and microscopy were used to detect T. vaginalis in vaginal swab samples collected from women attending social hygiene clinics in Metro Manila and Angeles City, Philippines. Screening of TVV on T. vaginalis was performed using RNA gel electrophoresis and RT-PCR. A modified protocol for metronidazole susceptibility assay was used to determine the aerobic minimum lethal concentration (MLC) of metronidazole in axenized T. vaginalis isolates.

RESULTS: A total of 42 T. vaginalis were screened for the presence of TVV and assayed for metronidazole susceptibility. TVV was detected in 13 of the isolates. All but one of the samples was susceptible to metronidazole.

CONCLUSION: This is the first study to assess the in vitro metronidazole susceptibility of Philippine T. vaginalis isolates. The isolates are generally susceptible to metronidazole even with the presence of TVV. The metronidazole susceptibility and presence of TVV are not enough to classify the isolates into type 1 or type 2.

Adult ; Azoospermia/parasitology* ; Hormones/blood* ; Humans ; Infertility, Male/parasitology* ; Male ; Polymerase Chain Reaction ; Testicular Diseases/parasitology* ; Trichomonas Infections/parasitology* ; Trichomonas vaginalis

OBJECTIVE: Human papillomavirus (HPV) testing is widely incorporated into cervical cancer screening strategies. Current screening requires pelvic examination for cervical sampling, which may compromise participation. The acceptance could be raised by introducing testing on vaginal swabs. We explored the interchangeability of vaginal swabs and cervical smears for HPV testing, by means of a prospective study conducted in female sex workers (FSWs). Besides, we report on the occurrence of 32 different HPV genotypes in FSW with low-grade squamous intraepithelial lesion (LSIL) or high-grade squamous intraepithelial lesion (HSIL). METHODS: Paired physician-collected vaginal swabs and cervical smears from 303 FSW were tested for HPV using the Abbott RealTime High-Risk HPV assay. Cervical cytology was examined on cervical smears. In case of HSIL/LSIL cytological classification (n=52), both samples were genotyped using INNO-LiPa HPV Genotyping Extra II. RESULTS: The overall prevalence of high-risk (HR)-HPV was 51%. In FSW with HSIL/LSIL cervical cytology, the sensitivity and specificity of vaginal samples for the detection of HR-HPV was 100% and 70% and for probable HR-HPV 100% and 91%. The mean number of genotypes identified in vaginal samples (mean=3.5; 95% confidence interval [CI]=2.8–4.2) was significantly higher than in cervical smear samples (mean=2.6; 95% CI=2.1–3.0) (p=0.001). The most frequently encountered HR-HPV genotypes were HPV16, 31, 51, and 52. CONCLUSION: As our study shows that vaginal swabs are equivalent to cervical smears for the detection of (probable) HR-HPV, vaginal swabs can be used for HPV testing in cervical cancer screening strategies. Given the acceptance of vaginal sampling, this finding offers an opportunity to boost screening coverage.
Chlamydia trachomatis ; Classification ; DNA ; Female ; Genotype ; Gynecological Examination ; Humans ; Mass Screening ; Mycoplasma genitalium ; Neisseria gonorrhoeae ; Papillomaviridae ; Prevalence ; Prospective Studies ; Sensitivity and Specificity ; Sex Workers ; Sexually Transmitted Diseases ; Squamous Intraepithelial Lesions of the Cervix ; Trichomonas vaginalis ; Uterine Cervical Neoplasms ; Vaginal Smears

Most men infected with Trichomonas vaginalis are asymptomatic and can remain undiagnosed and untreated. This has been hypothesized to result in chronic persistent prostatic infection. Adhesion of the protozoan organisms to mucosal cells is considered a first and prerequisite step for T. vaginalis infection. Adhesion of T. vaginalis to prostate epithelial cells has not yet been observed; however, there are several reports about inflammation of prostate epithelial cells induced by T. vaginalis. The aim of this study was to investigate whether adhesion and cytotoxicity of T. vaginalis are involved in inflammation of prostate epithelial cells. When RWPE-1 cells were infected with T. vaginalis (1:0.4 or 1:4), adhesion of T. vaginalis continuously increased for 24 hr or 3 hr, respectively. The cytotoxicity of prostate epithelial cells infected with T. vaginalis (RWPE-1: T. vaginalis=1:0.4) increased at 9 hr; at an infection ratio of 1:4, cytotoxicity increased after 3 hr. When the RWPE-1 to T. vaginalis ratio was 1:0.4 or 1:4, production of IL-1β, IL-6, CCL2, and CXCL8 also increased. Epithelial-mesenchymal transition (EMT) was verified by measuring decreased E-cadherin and increased vimentin expression at 24 hr and 48 hr. Taken together, the results indicate that T. vaginalis adhered to prostate epithelial cells, causing cytotoxicity, pro-inflammatory cytokine production, and EMT. Our findings suggest for the first time that T. vaginalis may induce inflammation via adhesion to normal prostate epithelial cells.
Cadherins ; Cell Adhesion ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Epithelium* ; Humans ; Inflammation ; Interleukin-6 ; Male ; Prostate* ; Trichomonas vaginalis* ; Trichomonas* ; Vimentin

BACKGROUND: The multiplex real-time PCR assay is a sensitive test for simultaneous detection of various pathogens of sexually transmitted infections (STIs). We evaluated the performance of two multiplex real-time PCR assays for six STI pathogens. METHODS: DNA samples after being used to conduct PCR for STI pathogens were stored below −70℃. Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), and Trichomonas vaginalis (TV) were detected by multiplex real-time PCR with GeneFinder STD I (CT/NG/UU)/II (MG/MH/TV) Multiplex Real-time PCR Kits (Infopia, Korea; GeneFinder assay) and Real-Q CT&NG/MH&TV/MG&UU Kits (BioSewoom, Korea; Real-Q assay). Discrepant results were resolved by another multiplex real-time assay, Anyplex II STI-7 Detection (Seegene, Korea). Any two positive results for the assays were considered true positive. RESULTS: Among 81 samples, the GeneFinder assay detected 63 pathogens from 45 cases (16 CT, 2 NG, 6 MG, 20 MH, 18 UU, and 1 TV) and Real-Q assay detected 66 pathogens from 47 cases (16 CT, 2 NG, 8 MG, 20 MH, 19 UU, and 1 TV). For the results of positive cases and negative cases, the overall concordance rate between the two multiplex real-time assays was 93.8% (Kappa=0.87). For each pathogen, the agreement rates of the two assays ranged from 97.5 to 100% (Kappa>0.8). CONCLUSION: There was no significant difference between the results of GeneFinder assay and Real-Q assay. Both multiplex real-time PCR assays can be useful methods for the detection of STI pathogens in clinical laboratories.
Chlamydia trachomatis ; DNA ; Korea ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction* ; Sexually Transmitted Diseases* ; Trichomonas vaginalis ; Ureaplasma urealyticum

Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected (V⁺) and uninfected (V⁻) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in V⁺ compared with V⁻ isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in V⁺ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in V⁺ and V⁻ isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.
Glucose-6-Phosphate Isomerase ; Glycogen Phosphorylase ; Heat-Shock Proteins ; Host-Parasite Interactions ; Malate Dehydrogenase ; Metabolism ; Polymerase Chain Reaction ; Proteome ; Reticuloendotheliosis virus ; Ribosomal Proteins ; RNA, Double-Stranded ; RNA, Messenger ; Trichomonas vaginalis* ; Trichomonas* ; Triose-Phosphate Isomerase ; Virulence

Avian trichomoniasis caused by Trichomonas gallinae is a serious protozoan disease worldwide. The domestic pigeon (Columba livia domestica) is the main host for T. gallinae and plays an important role in the spread of the disease. Based on the internal transcribed spacers of nuclear ribosomal DNA of this parasite, a pair of primers (TgF2/TgR2) was designed and used to develop a PCR assay for the diagnosis of T. gallinae infection in domestic pigeons. This approach allowed the identification of T. gallinae, and no amplicons were produced when using DNA from other common avian pathogens. The minimum amount of DNA detectable by the specific PCR assay developed in this study was 15 pg. Clinical samples from Guangzhou, China, were examined using this PCR assay and a standard microscopy method, and their molecular characteristics were determined by phylogenetic analysis. All of the T. gallinae-positive samples detected by microscopic examination were also detected as positive by the PCR assay. Most of the samples identified as negative by microscopic examination were detected as T. gallinae positive by the PCR assay and were confirmed by sequencing. The positive samples of T. gallinae collected from Guangzhou, China, were identified as T. gallinae genotype B by sequencing and phylogenetic analyses, providing relevant data for studying the ecology and population genetic structures of trichomonads and for the prevention and control of the diseases they cause.
China* ; Columbidae* ; Diagnosis ; DNA ; DNA, Ribosomal ; Ecology ; Genetic Structures ; Genotype ; Methods ; Microscopy ; Parasites ; Polymerase Chain Reaction* ; Trichomonas*