Inflammatory mediators of prostate epithelial cells stimulated with Trichomonas vaginalis promote proliferative and invasive properties of prostate cancer cells.

BACKGROUND: Trichomonas vaginalis (Tv) is the most common sexually transmitted parasite. It is detected in prostatic tissue of benign prostatic hyperplasia, prostatitis, and prostate cancer (PCa) and has been suggested to cause chronic prostatitis. Moreover, up to 20% of all cancers worldwide are associated with chronic inflammation. Here, we investigated whether inflammatory mediators produced by normal human prostate epithelial cells (RWPE-1) stimulated with Tv could promote growth and invasiveness of PCa cells. METHODS: Conditioned medium of RWPE-1 cells was prepared by stimulating them with Tv (trichomonad-conditioned medium [TCM]) and without Tv (conditioned medium [CM]). Promotion of PCa cells (PC3, DU145, and LNCaP) was assessed by wound healing, proliferation, and invasion assays. RESULTS: We observed that the production of interleukin (IL)-1ß, IL-6, CCL2, CXCL8, prostaglandin-E2 (PGE2 ), and COX2 by RWPE-1 cells was increased by stimulating them with Tv. When PCa cells were incubated with TCM, their proliferation, invasion, and migration increased. Moreover, they showed increased epithelial-mesenchymal transition (EMT)-related markers by a reduction in epithelial markers and an increase in mesenchymal markers. In vivo, xenograft tumor tissues injected with TCM also showed increased expression of cyclin D1 and proliferating cell nuclear antigen, as well as induction of EMT. Receptors and signal molecules of PCa cells increased in response to exposure to TCM, and blocking receptors (CXCR1, CXCR2, C-C chemokine receptor 2, glycoprotein 130, EP2, and EP4) reduced the proliferation of PCa cells with decreased production of cytokines (CCL2, IL-6, and CXCL8) and PGE2 , and expression of NF-κB and Snail1. CONCLUSIONS: Our results suggest that Tv infection may be one of the factors creating the supportive microenvironment to promote proliferation and invasiveness of PCa cells.

Experimental rat prostatitis caused by Trichomonas vaginalis infection.

BACKGROUND: Trichomonas vaginalis (T. vaginalis) is the most common sexually transmitted parasite. It has been detected in prostatic tissue of patients with prostatitis and reported to be associated with chronic prostatitis and benign prostatic hyperplasia as well as prostate cancer. Recently, experimental rodent models of prostatitis induced by pathogen infection have been developed. However, there have so far been no reports of prostatitis caused by T. vaginalis infection in animals. Here, we investigated whether infection with T. vaginalis via the rat urethra could cause prostatitis. METHODS: T. vaginalis was injected into prostate through urethra of rat (Wistar rats), and the rats were killed 1, 2, or 4 weeks later. The presence of T. vaginalis trophozoites in the rat prostates was examined by immunohistochemistry, and pathological changes of the prostate were observed by hematoxylin-eosin staining and evaluated by grading from 0 to 5 for inflammatory cell infiltration, acinar changes, and interstitial fibrosis. Infiltrated mast cells were observed by toluidine blue staining of rat prostate tissue. Chemokine C-C motif ligand 2 (CCL2) levels of the rat prostates were measured by ELISA. RESULTS: T. vaginalis trophozoites were observed in acini in the prostates of the injected rats. The prostate tissues had higher pathological scores, and 83% (5/6) and 100% (6/6) of the ventral and dorsolateral lobes (n = 6), respectively, were inflamed. Infiltration and degranulation of mast cells were observed at higher rates in prostate sections of the T. vaginalis-infected rats. Also, prostate tissues of the injected rats had increased CCL2 levels. CONCLUSIONS: Injection of T. vaginalis in rats caused prostatitis as revealed by pathologic changes, mast cell infiltration and increased CCL2 production. Therefore, this study provides the first evidence that T. vaginalis infection in rats causes prostatitis.

Interaction between Trichomonas vaginalis and the Prostate Epithelium.

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-1ß, 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.

Signalling pathways associated with IL-6 production and epithelial-mesenchymal transition induction in prostate epithelial cells stimulated with Trichomonas vaginalis.

Trichomonas vaginalis (Tv) has been found in patient tissue of benign prostatic hyperplasia (BPH), and suggested to cause chronic prostatitis. IL-6 is known as one of the important factors of chronic inflammation in prostate cancer. Patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) had higher levels of IL-6 in seminal plasma. Furthermore, inflammatory conditions induced by pathogen infections have been shown to promote epithelial-mesenchymal transition (EMT). Here, we investigated the signals involved in IL-6 production by human prostate epithelial cells (PECs) stimulated with Tv and examined whether Tv induces EMT in PECs. We found that PECs stimulated with Tv increased the production of IL-6, as well as the expression of TLR2, TLR4, MAPKs (p38, JNK, ERK), NF-κB and JAK2/STAT3, and levels of ROS. Inhibition of TLR2 or TLR4 reduced IL-6 production as well as expression of these other factors, and agents inhibiting ROS, MAPKs, NF-κB and JAK reduced IL-6 production. However, when PECs were stimulated with Tv, transcripts of mesenchymal cell markers increased, and epithelial cell markers decreased. In addition, the induction of EMT was suppressed by inhibitors of JAK or NF-κB. These findings are the first evidence that Tv infection of prostate epithelial cells may induce EMT.

Trichomonas vaginalis induces IL-1ß production in a human prostate epithelial cell line by activating the NLRP3 inflammasome via reactive oxygen species and potassium ion efflux.

BACKGROUND: Trichomonas vaginalis is a sexually transmitted protozoan parasite that causes vaginitis in women, and urethritis and prostatitis in men. IL-1ß is synthesized as immature pro-IL-1ß, which is cleaved by activated caspase-1. Caspase-1 is, in turn, activated by a multi-protein complex known as an inflammasome. In this study, we investigated the inflammatory response of a prostate epithelial cell line (RWPE-1) to T. vaginalis and, specifically, the capacity of T. vaginalis to activate the NLRP3 inflammasome. METHODS: RWPE-1 cells were stimulated by live T. vaginalis, and subsequent expression of pro-IL-1ß, IL-1ß, NLRP3, ASC and caspase-1 was determined by real-time PCR and Western blotting. IL-1ß and caspase-1 production was also measured by ELISA. To evaluate the effects of NLRP3 and caspase-1 on IL-1ß production, the activated RWPE-1 cells were transfected with small interfering RNAs to silence the NLRP3 and caspase-1 genes. Activation of the NLRP3 inflammasome was observed by fluorescence microscopy. Intracellular reactive oxygen species (ROS) were evaluated by spectrofluorometry. RESULTS: When RWPE-1 cells were stimulated with live T. vaginalis, the mRNA and protein expression of IL-1ß, NLRP3, ASC, and caspase-1 increased. Moreover, silencing of NLRP3 and caspase-1 attenuated T. vaginalis-induced IL-1ß secretion. The NADPH oxidase inhibitor DPI and high extracellular potassium ion suppressed the production of IL-1ß, caspase-1, and the expression of NLRP3 and ASC proteins. The specific NF-κB inhibitor, Bay 11-7082, inhibited IL-1ß production, and also inhibited the production of caspase-1, ASC and NLRP3 proteins. CONCLUSIONS: T. vaginalis induces the formation of the NLRP3 inflammasome in human prostate epithelial cells via ROS and potassium ion efflux, and this results in IL-1ß production. This is the first evidence for activation of the NLRP3 inflammasome in the inflammatory response by prostate epithelial cells infected with T. vaginalis. Prostate 76:885-896, 2016. © 2016 Wiley Periodicals, Inc.

Prevalence of Trichomonas vaginalis by PCR in men attending a primary care urology clinic in South Korea.

Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.

PCR for diagnosis of male Trichomonas vaginalis infection with chronic prostatitis and urethritis.

The aim of this study was to assess the usefulness of PCR for diagnosis of Trichomonas vaginalis infection among male patients with chronic recurrent prostatitis and urethritis. Between June 2001 and December 2003, a total of 33 patients visited the Department of Urology, Hanyang University Guri Hospital and were examined for T. vaginalis infection by PCR and culture in TYM medium. For the PCR, we used primers based on a repetitive sequence cloned from T. vaginalis (TV-E650). Voided bladder urine (VB1 and VB3) was sampled from 33 men with symptoms of lower urinary tract infection (urethral charge, residual urine sensation, and frequency). Culture failed to detect any T. vaginalis infection whereas PCR identified 7 cases of trichomoniasis (21.2%). Five of the 7 cases had been diagnosed with prostatitis and 2 with urethritis. PCR for the 5 prostatitis cases yielded a positive 330 bp band from bothVB1 and VB3, whereas positive results were only obtained from VB1 for the 2 urethritis patients. We showed that the PCR method could detect T. vaginalis when there was only 1 T. vaginalis cell per PCR mixture. Our results strongly support the usefulness of PCR on urine samples for detecting T. vaginalis in chronic prostatitis and urethritis patients.

Subclinical leishmaniasis associated with infertility and chronic prostatitis in a dog.

A stud dog was presented for acquired infertility. Haematospermia and teratozoospermia were found on two ejaculates 2 weeks apart. A presumptive diagnosis of prostatitis was made follo-wing ultrasound examination. An ultrasound-guided needle core biopsy was performed under general anaesthesia, revealing a mild chronic macrophagic and plasma cell prostatitis with intracytoplasmic amastigotes consistent with Leishmania spp. infection. Presence of Leishmania infantum, Leishmania donovani or Leishmania chagasi was confirmed by polymerase chain reaction in seminal plasma. Serology and serum protein electrophoresis confirmed the diagnosis of a subclinical active systemic leishmaniasis. A meglumine antimoniate and allopurinol treatment was given which clearly improved within 3 months both general condition and the quality of sperm. To the authors' knowledge, this is the first reported case of a prostatitis secondary to a Leishmania spp. infection. Subclinical systemic leishmaniasis should be considered in the differential diagnosis of infertility in dogs suffering from semen alterations.

[Combined physiotherapy of chronic infectious prostatitis].

Our experience with therapy of 259 outpatients with chronic infectious prostatitis (CIP) aged 16-55 years has demonstrated that combined treatment of CIP with rectal digital massage of the prostate, electrophoresis of chimotripsin solution with dimexide and local magnetotherapy (Intramag unit) significantly raises treatment efficacy, shortens treatment, prevents complications.

The role of unusual pathogens in prostatitis syndrome.

A total of 1442 patients with symptoms of chronic prostatitis were examined over a 4-year period at the Outpatient Department for Urogenital Infections, University Hospital for Infectious Diseases "Dr. Fran Mihaljevic", Zagreb, Croatia. An infectious aetiology was determined in 1070 (74.2%) patients. In 561 of 1070 (52.4%) patients the inflammatory finding (>10 WBC/hpf) was found in expressed prostatic secretions (EPS) or voided bladder urine (VB(3)). Normal, <10 WBCs/hpf was found in 362 of 536 (67.5%) patients with symptoms of chronic prostatitis in whom Chlamydia trachomatis was detected in EPS or VB(3), in 51 of 151 (33.8%) patients with isolated Trichomonas vaginalis and in 40 of 72 (55.6%) patients with isolated Ureaplasma urealyticum. Escherichia coli was the causative pathogen in 95, Enterococcus in 68, Proteus mirabilis in 37, Klebsiella pneumoniae in 16, Streptococcus agalactiae in 19, and Pseudomonas aeruginosa in 3 patients with chronic prostatitis. Other patients had a mixed infection. In patients with chronic bacterial prostatitis (CBP) caused by E. coli, P. mirabilis, K. pneumoniae, E. or S. agalactiae, an inflammatory finding was regularly found in EPS or VB(3).

[Parasitic diseases are a risk factor of inflammations of the small pelvis organs. Trichomoniasis]. / Parazitozy--faktor riska vospalitel'nykh zabolevanii organov malogo taza. Trikhomoniaz.

Clinical and laboratory studies were made in 2259 persons by using serological assays, including cultural and bacterioscopic assays in 1824 and 363 persons, respectively, in order to indicate Trichomonas infection among men, women, and 2-15-year-old children who had chronic inflammation of the urogenital system, as well as among children aged 2 months to 16 years who had a severe somatic disease. Parasitic cenoses of the urogenital tract were studied in women with trichomoniasis and reproductive dysfunction. Trichomonas invasion is an etiological factor of inflammatory small pelvic diseases, chronic prostatitis, reproductive dysfunction in males and females. Furthermore, trichomoniasis is an essential factor, that predisposes to recurrent viral infections: genital herpes and pointed condyloma, and substantially increases the risk of birth of babies with clinical manifestations of intrauterine infection. According to the studies, a laboratory diagnosis of trichomoniasis should be made by cultural assay due to the low sensitivity of bacterioscopy by sampling a biological material from females in the periovulatory period that corresponds to the hyperestrogenicity, which substantially increases the number of isolated cultures and characterizes the depth of integration of metabolic processes of a parasite and its host.

Chlamydia trachomatis: probable cause of prostatitis.

Seventy-eight men with symptoms of chronic or subacute prostatitis were enrolled. Investigations for the presence of Chlamydia trachomatis in urethral swabs were carried out. The expressed prostatic secretions were additionally examined for Mycoplasma hominis, Ureaplasma urealyticum, Gardnerella vaginalis, other gram-negative and gram-positive bacteria, Trichomonas vaginalis, yeast-like fungi and leucocyte count. Furthermore, all patients were evaluated for the presence of serum anti-chlamydial IgG antibodies. Signs of inflammation on the basis of the count of leucocytes per hpf in the prostatic secretions were detected in 42 patients (group I). Prostatodynia was found in the remaining 36 men (group II). In group I, chlamydial antigen was detected in the urethra and expressed prostatic secretions (EPS) in 6 (14.3%) and 9 (21.4%) patients, respectively. No evidence of current chlamydial infection was found in group II. The presence of serum anti-chlamydial IgG antibodies was demonstrated in 13/42 (30.9%) patients with prostatitis and in 3/36 (8.3%) patients with prostatodynia (P < 0.01). The results suggest that chlamydia may be one of the causative agents of chronic prostatitis.

[Rare cases of prostatitis caused by invasion of Trichomonas vaginalis with Candida albicans]. / Rzadkie przypadki zapalenia stercza wywolane inwazja Trichomonas vaginalis z Candida albicans.

Three cases of prostatitis caused by the invasion of T. vaginalis and C. albicans which has been found in different biological materials were described. After per rectum examination perineum biopsy of prostata was performed in all patients; in histopathological preparations pointing at the inflammation reaction of that gland the fungi were detected. Also, the same microorganisms were proved in sexual partners of those patients but multifocal invasion of C. albicans--including genital and urinary organs, mouth and alimentary tract--also in members of their family.

Prostatitis and epididymitis.

Ascending spread of urethral pathogens may be the mechanisms of infection of the prostate and epididymis. Sexually transmitted organisms cause most epididymitis in young men. Although evidence is mounting to suggest that sexually transmitted organisms may cause prostatitis, data are insufficient at this time to base a therapeutic approach on this concept. The authors present approaches to the management of these two disorders.

[Prostatitis due to the circular form of trichomonas (author's transl)]. / Les prostatites à trichomonas formes rondes (T.F.R.).

The authors underline the frequency with which trichomoniasis is located in the prostate. In these cases the parasite is rarely in its trophozoite form but has a circular shape which resembles the resistant pseudocystic type. They are found by phase contrast microscopy after staining with 1% cresyl blue which makes the difference with white blood cells. Out of a total of 178 men presenting with urethroprostatic signs (urethral discharge and burning), sterility with oligospermia, or a history of dyspareunia, the circular type of trichomoniasis was found in 40 (22%) of the cases, with urethritis in 26 cases, and prostatitis in the other 14 patients. Two courses of treatment, at one month's interval, with Tinidazole (2 tablets daily for 6 days) or Nimorazole (2 tablets daily for 8 days) produced healing in 4 cases with urethroprostatic effections, disappearance of dyspareunia in 2 out of the 3 cases treated, and return of fertility in 3 out of 7 patients. An up-to-date list of references is given.