Morphology of human Fallopian tubes after infection with Mycoplasma genitalium and Mycoplasma hominis—in vitro organ culture study (original) (raw)
Related papers
2016
BACKGROUND: Female infertility can be caused by scarring and occlusion of the Fallopian tubes. Sexually transmitted bacteria can damage the delicate epithelial layer of human Fallopian tubes (HFT). Genital mycoplasmas are associated with human reproductive failure. Yet, there is not enough evidence that mycoplasmas can cause tubal factor infertility. We analysed the effects of infections with Mycoplasma hominis and Mycoplasma genitalium on the HFT epithelium and compared them with the effects of infections with genital pathogens: Chlamydia trachomatis and Neisseria gonorrhoeae. METHODS: We used an in vitro model in which pieces of normal HFT were infected with different bacteria, and the outcome of the infections was analysed by scanning electron microscopy (SEM) and confocal microscopy. RESULTS: The presence of M. hominis did not cause any morphological changes of the epithelium of HFT. Noticeable changes in the morphology of the ciliated cells were observed in M. genitalium-infected tissue. Five days post-infection, the cilia were abnormally swollen and some of the ciliated cells fell off the epithelium. These effects could be inhibited by pre-incubation of M. genitalium with antibody directed against the C-terminal part of the adhesion protein MgPa before infection of HFT organ culture. CONCLUSION: We have shown that the presence of M. genitalium, but not M. hominis, in the HFT organ culture affected the epithelium and resulted in cilia damage. The effect of infection with M. genitalium on the HFT was, however, very moderate when compared with the extensive damage of the epithelium caused by N. gonorrhoeae or C. trachomatis.
Serological investigation of Mycoplasma genitalium in infertile women
Human Reproduction, 2001
BACKGROUND: The role of Mycoplasma genitalium in the pathogenesis of pelvic inflammatory disease has not been characterized. METHODS: Sera from 308 infertile women were investigated for antibodies to M. genitalium by immunoblotting. Women with tubal factor infertility (TFI) made up 132 of the patients, 67 of the women had an infertile male partner and 109 were infertile for unknown reasons. RESULTS: Of the TFI patients 29 (22.0%) were seropositive to the major adhesin, MgPa, of M. genitalium versus 11 (6.3%) in the group of women with normal tubes. No cross-reactions between MgPa and P1 of the related Mycoplasma pneumoniae were found. Besides, MgPa positive sera were confirmed by immunoblotting using a cloned fragment of the C-terminal part of MgPa specific to M. genitalium. Chlamydia trachomatis is known to be able to cause infertility as a result of salpingitis. Therefore, the sera were tested against C. trachomatis using a commercial ELISA test. Seventy-five (56.8%) of the TFI patients were seropositive to C. trachomatis. Eight (27.6%) TFI patients seropositive to MgPa were negative to C. trachomatis. CONCLUSIONS: This study indicates that M. genitalium may be an independent risk factor in the development of an inflammatory process leading to scarring of the uterine tubes in women and thereby causing infertility.
Sexually Transmitted Infections, 1979
Quantatitive determinations of the mucociliary activity of human Fallopian tube epithelium maintained as organ cultures were performed using a light beam reflex method. In noninfected organ cultures the mucociliary wave (MCW) frequency slowly decreased during the first 54 hours of culture maintenance. In organ cultures experimentally infected with fresh isolates of Neisseria gonorrhoeae producing T1/T2 colonies the MCW frequency either decreased to subnormal values or completely ceased whereas in organ cultures infected with a laboratoryadapted gonococcal strain the MCW frequencies remained within normal range. In organ cultures exposed to gonococcal endotoxin prepared from the laboratory-adapted strain, as well as in cultures in which cell-free filtrates of medium from organ cultures infected with N. gonorrhoeae (producing T1/T2 colonies) were added to the culture medium, the ciliary activity decreased and subsequently ceased. The same phenomenon occurred when organ cultures were challenged with Escherichia coli endotoxin. The ciliostatic effect appeared before any morphological changes in the surface epithelium, including the cilia, were demonstrable by scanning electron microscopy.
Growth kinetics of Chlamydia trachomatis in primary human Sertoli cells
Scientific Reports, 2019
Chlamydia trachomatis (Ct) is the leading cause of bacterial sexually transmitted infections worldwide and has been associated with male infertility. Recently, it was hypothesized that Ct may infect the epithelium of the seminiferous tubule, formed by Sertoli cells, thus leading to impaired spermatogenesis. To date, there is a lack of data on Ct infection of the seminiferous epithelium; therefore, we aimed to characterize, for the first time, an in vitro infection model of primary human Sertoli cells. We compared Ct inclusion size, morphology and growth kinetics with those in McCoy cells and we studied F-actin fibres, Vimentin-based intermediate filaments and α-tubulin microtubules in Sertoli and McCoy cells. Our main finding highlighted the ability of Ct to infect Sertoli cells, although with a unique growth profile and the inability to exit host cells. Furthermore, we observed alterations in the cytoskeletal fibres of infected Sertoli cells. Our results suggest that Ct struggles to generate a productive infection in Sertoli cells, limiting its dissemination in the host. Nevertheless, the adverse effect on the cytoskeleton supports the notion that Ct may compromise the blood-testis barrier, impairing spermatogenesis. C. trachomatis is an obligate intracellular pathogen characterized by a distinctive developmental cycle, alternating between two morphologically and functionally distinct forms, the elementary body (EB), the extracellular infectious form, and the replicative body (RB), the intracellular replicative form 1,2. C. trachomatis is the leading cause of bacterial sexually transmitted infections worldwide, with more than 130 million new cases per year 3. In men, C. trachomatis is a major cause of urethritis, with up to 42% of all cases of non-gonococcal urethritis attributed to a chlamydial infection 4. However, its genuine prevalence remains unknown and probably underestimated, since 50% of chlamydial infections in men are asymptomatic and, hence, undetected and untreated, leading to potential complications 3. Indeed, it is now widely accepted that ascending chlamydial infections are involved in the etiopathogenesis of acute or chronic epididymitis 5,6 and chlamydial antigens have also been detected in men with epididymo-orchitis, suggesting a causative link between C. trachomatis and infection of the testis 5,6 that may contribute to the development of infertility. The etiopathogenesis of infertility in men is characterised by different causes, ranging from genetic disorders to testicular dysfunction, obstructive azoospermia, varicocele and hormonal imbalance 7,8. In more than half of all cases, the aetiology is unknown, and amongst all the potential risk factors, C. trachomatis has been the target of several studies that suggested an association between this pathogen and male infertility 8-15. To explain its involvement, several potential pathophysiological mechanisms have been investigated and, amongst them, one theory postulates that the infection of the seminiferous tubule epithelium by C. trachomatis might lead to inflammatory damage and, hence, result in impaired spermatogenesis 8,9. Interestingly, a report analysing a model of C. muridarum infection in male C57BL/6 mice demonstrated that this pathogen severely affects the seminiferous tubules, formed by Sertoli cells, leading to compromised spermatogenesis with reduced sperm count, motility and altered morphology of mature spermatozoa 16. Sertoli cells are key elements for the development of germ cells, since they finely regulate the spermatogenetic process through either the secretion of endocrine and paracrine mediators or by guiding germ cells from the basal to the adluminal compartment of the seminiferous tubule, that is physically divided by the blood-testis barrier
Mycoplasma genitalium, Chlamydia trachomatis, and tubal factor infertility—a prospective study
Fertility and Sterility, 2008
Objective: To determine the presence of M. genitalium and C. trachomatis in women attending fertility clinics and to follow these women for the effects of previous infections or tubal damage on pregnancy rate and outcome. Design: Prospective study. Setting: Fertility clinics and university. Patient(s): Two hundred twelve couples attending fertility clinics. Intervention(s): Blood and cervical swab samples from the women. Tubal status was assessed by culdoscopy and/ or laparoscopy. Main Outcome Measure(s): Presence of M. genitalium and C. trachomatis was determined by polymerase chain reaction. Serum samples were tested for antibodies against M. genitalium and C. trachomatis. Result(s): One swap sample was positive to C. trachomatis and none positive to M. genitalium. Thirty of the 194 women had tubal factor infertility (TFI); 23% and 17% of women with TFI had antibodies to C. trachomatis and M. genitalium, respectively, compared with 15% and 4%, respectively, of women with normal tubes; 36% and 14% of women with a self-reported history of pelvic inflammatory disease (PID) were seropositive to C. trachomatis and M. genitalium, respectively, compared with 10% and 6%, respectively, of women without past PID. Conclusion(s): A strong antibody response against M. genitalium or C. trachomatis but no sign of current or chronic infection was found in women with TFI, indicating that previous infections caused by these microorganisms may have resulted in permanent damage and occlusion of the fallopian tubes.
Sexually Transmitted Diseases, 2005
Objectives: We sought to determine if intraluminal occluding fibrosis of the oviduct occurs after urogenital Chlamydia muridarum infection in mice. Study: Oviduct occlusion was assessed by infusing dye into the distal uterus and tracking the diffusion of the dye into the oviduct. We also conducted histologic assessment of the affected tissues using hematoxylin and eosin (H&E) and Masson trichrome stains. Results: All previously infected susceptible mice had occluded oviducts compared with 17.5% of previously uninfected mice. Oviduct occlusion correlated with hydrosalpinx formation and infertility. Intraluminal oviduct fibrosis was observed in several sections of tissue displaying hydrosalpinx but not in tissues without hydrosalpinx. Fibrosis was localized to the oviduct isthmus and oviduct proper, proximal to the uterus. Conclusion: Intralumenal occluding fibrosis of the oviduct is a sequela of infection with C. muridarum in this model. These observations support the use of the murine model to study pathogenesis of chlamydial upper genital tract infection.
Impaired Function of the Spermatozoa: Links with Mycoplasmal and Chlamydial Infections
Mycoplasmal and chlamydial infections of the male genital tract have been associated with impairment of the human sperm functionality. However this topic remains doubtful due to contradictory results obtained from several studies around the globe. In this review we will discuss recent evidence from in vitro studies that have confirmed that Mycoplasma hominis, Ureaplasma urealyticum, U. parvum, M. genitalium and/or Chlamydia trachomatis can attach to and invade intact spermatozoa. Moreover we will explore how these bacterial infections can cause impaired sperm function and male infertility. First, bacteria attached to sperm membrane may provoke oxidative stress through release of reactive oxygen species, resulting in lipid peroxidation and reduction of membrane fluidity. Second, chlamydial lipopolysacharide interaction with sperm membrane alters the phosphorylation pattern of citosolic proteins, triggering apoptotic pathways. Third, internalized mycoplasmal and chlamydial cells disru...
Journal of Comparative Pathology, 2008
An experimental murine model of bovine genital tritrichomonosis is described. Female mice were inoculated per vaginam with Tritrichomonas foetus and a sample of the study population was killed every 3 days up to 60 days post-infection. Microscopical changes in the reproductive organs were assessed and immunohistochemistry was used to detectT. foetus within these tissues. Lectin histochemistry was used to determine changes in the expression of carbohydrates within the reproductive mucosa. A range of microscopical changes were detected in the uterine endometrium by 10 days post-inoculation and these were associated with the presence of the protozoan. The endometrial changes included endometritis and ulceration, mucosal atrophy and glandular metaplasia, and were similar to those reported in naturally infected cows. Changes in lectin binding were recognized ¢rst in the vagina where there was increased binding of Ulex europaeus agglutinin-1 (UEA-1) which was maximal on day 16 postinoculation. Within the uterus, there was increased binding of soy bean agglutinin (SBA) which was maximal on day 19 post-inoculation, and of peanut agglutinin (PNA) which was maximal on day 16 post-inoculation. These changes in carbohydrate expression parallel the infection kinetics, since they appeared ¢rst in the vagina and later in the uterus. The changes may re£ect either a host reaction against the infection or the production of enzymes by T. foetus, which act to enhance adhesion and colonization of the genital organs by the organism. The kinetics and pathogenesis of this murine infection are similar to those of the natural bovine disease, suggesting that this model system may be valuable for further studies of this disease. r
Journal of Clinical Pathology, 2005
In vitro growth and elementary body (EB) associated cytotoxicity of two Chlamydia trachomatis strains belonging to serovars D and H and C muridarum were compared to identify difference(s) that correlate with virulence variations between these strains in the mouse model of human female genital tract infection, and phenotypic characteristics that could explain human epidemiological data on serovar prevalence and levels of shedding during serovar D and H infection. Methods: Replication cycle kinetics, inclusion characteristics, and EB associated cytotoxicity were assessed in McCoy cell monolayers using culture, light microscopy, and lactate dehydrogenase release. Results: Over 72 hours, more rapid production and release of inclusion forming units (ifu) allowed C muridarum to initiate two replication rounds, resulting in 4-8 times more ifu/input unit of infection than with serovars D and H. Although C muridarum EBs were significantly more cytotoxic to McCoy cell monolayers than serovar D at moderate and high multiplicity of infection ratios (MOI), serovar H EBs were significantly more cytotoxic than C muridarum, even at the lowest MOI tested. Conclusions: These phenotypic differences are consistent with the more invasive course and severe pathological outcome of infection in mice infected with C muridarum, providing an objective basis for questioning the appropriateness of C muridarum as a surrogate for the human biovar of C trachomatis in the murine model of female genital tract infection. The differences seen between the human strains could help explain human epidemiological data relating to differences in prevalence and level of shedding that occurs during infection with oculogenital serovars D and H.