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Chlamydia trachomatis , a leading infectious cause of tubal infertility, induces upper genital tract pathology, such as hydrosalpinx, which can be modeled with Chlamydia muridarum infection in mice. Following C. muridarum inoculation, wild-type mice develop robust hydrosalpinx, but OT1 mice fail to do so because their T cell receptors are engineered to recognize a single ovalbumin epitope (OVA 457-462 ). These observations have demonstrated a critical role of Chlamydia -specific T cells in chlamydial pathogenicity. In the current study, we have also found that OT1 mice can actively inhibit chlamydial pathogenicity. First, depletion of CD8 + T cells from OT1 mice led to the induction of significant hydrosalpinx by Chlamydia , indicating that CD8 + T cells are necessary to inhibit chlamydial pathogenicity. Second, adoptive transfer of CD8 + T cells from OT1 mice to CD8 knockout mice significantly reduced chlamydial induction of hydrosalpinx, demonstrating that OT1 CD8 + T cells are sufficient for attenuating chlamydial pathogenicity in CD8 knockout mice. Finally, CD8 + T cells from OT1 mice also significantly inhibited hydrosalpinx development in wild-type mice following an intravaginal inoculation with Chlamydia Since T cells in OT1 mice are engineered to recognize only the OVA 457-462 epitope, the above observations have demonstrated a chlamydial antigen-independent immune mechanism for regulating chlamydial pathogenicity. Further characterization of this mechanism may provide information for developing strategies to reduce infertility-causing pathology induced by infections., (Copyright © 2020 American Society for Microbiology.)

Ascension to the oviduct is necessary for Chlamydia to induce tubal infertility. Using the Chlamydia muridarum induction of hydrosalpinx mouse model, we have demonstrated a significant role of the uterotubal junction in preventing chlamydial ascending infection. First, delivery of C. muridarum to either side of the uterotubal junction resulted in significant reduction in live organisms from the tissues on the opposite sides. However, the recovery yields remained similar among different sections of the uterine horn. These observations suggest that the uterotubal junction may function as a barrier between the uterine horn and oviduct. Second, deficiency in innate immunity signaling pathways mediated by either MyD88 or STING significantly compromised the uterotubal junction barrier function, permitting C. muridarum to spread freely between uterine horn and oviduct. Finally, transcervical inoculation of C. muridarum led to significantly higher incidence of bilateral hydrosalpinges in the STING-deficient mice while the same inoculation mainly induced unilateral hydrosalpinx in the wild type mice, suggesting that the STING pathway-dependent uterotubal junction plays a significant role in preventing tubal pathology. Thus, we have demonstrated for the first time that the uterotubal junction is a functional barrier for preventing tubal infection by a sexually transmitted agent, providing the first in vivo evidence for detecting chlamydial infection by the STING pathway.

Chlamydia trachomatis is the leading cause of bacterial sexually transmitted disease worldwide resulting in 4-5 million new cases of Chlamydia annually and an estimated 100 million cases per annum. Infections of the lower female genital tract (FGT) frequently are asymptomatic; thus, they often remain undiagnosed or untreated. If infections are either not resolved or left untreated, chlamydia can ascend to the upper FGT and infect the fallopian tubes (FTs) causing salpingitis that may lead to functional damage of the FTs and tubal factor infertility (TFI). Clinical observations and experimental data have indicated a role for antibodies against C. trachomatis proteins such as the 60-kDa heat shock protein 60 (cHSP60) in the immunopathogenesis of TFI. When released from infected cells, cHSP60 can induce proinflammatory immune responses that may functionally impair the FTs leading to fibrosis and luminal occlusion. Chlamydial pathogenesis of irreversible and permanent tubal damage is a consequence of innate and adaptive host immune responses to ongoing or repeated infections. The extracellular matrix that is regulated by metalloproteinases may also be modified by chlamydial infections of the FGT. This review will highlight protective and pathogenic immune responses to ongoing and repeated chlamydial infections of the FGT. It will also present two recent hypotheses to explain mechanisms that may contribute to FT damage during a C. trachomatis infection. If Chlamydia immunopathology can be controlled, it might yield a method of inducing fibrosis and thus provide a means of nonsurgical permanent contraception for women., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objective: Our long-term goal is to develop a nonsurgical method of fallopian tubal occlusion for the purpose of permanent contraception. We have previously demonstrated that transcervical administration of 5% polidocanol foam (PF) can create tubal occlusion in macaques but that multiple treatments are required. In this study, we assessed the efficacy of various regimens of PF with and without depomedroxyprogesterone acetate (DMPA) (to control ovarian cycle phase) in the baboon., Study Design: Adult cycling female baboons were evaluated for tubal patency by hysterosalpingography and then received a transcervical infusion of PF with (+) or without (-) an intramuscular injection of DMPA (3.5 mg/kg). Two concentrations of PF were compared: 1% [(+) DMPA, n=5; (-) DMPA, n=3] and 5% [(+) DMPA, n=4; (-) DMPA, n=3]. Controls received (+) DMPA (n=2) or (-) DMPA, (n=3) only. The reproductive tracts were removed 1-3 months after treatment for examination., Results: No fallopian tubal occlusion was observed in negative controls (±DMPA). Histologic complete tubal occlusion was observed in 3/8 of females treated with 1% PF and in 6/7 treated with 5% PF. Histologic evaluation suggested that 1% PF is associated with prolonged chronic inflammation (more than 2-3 months), while 5% treatment eliminates the epithelial lining, at least focally, and resolves into complete occlusion within 1-2 months. This pattern of complete occlusion was seen in all 4 females that received 5% PF (+DMPA) and in 2/3 that received 5% PF (-DMPA)., Conclusion: In a baboon model of transcervical permanent contraception, a single treatment with 5% PF resulted in complete tubal occlusion more reliably (85%) than 1% PF (38%). Cotreatment with DMPA may improve treatment results with 5% PF but requires additional study., Implications: A finding that a single transcervical treatment with 5% PF can occlude the fallopian tubes of baboon supports further study of this approach as a novel strategy for permanent contraception for women., (Copyright © 2015 Elsevier Inc. All rights reserved.)

We compared the effectiveness of immunomodulators used in the treatment of patients with chronic salpingitis and oophoritis with or without changes in succinate dehydrogenase (SDH) activity in blood lymphocytes at incubation with the drug. Diurnal variations in individual reaction of SDH in blood lymphocytes to thymalin or ridostin were revealed. In the groups of women receiving ridostin or thymalin during the reaction of lymphocyte SDH to it, improvement of clinical laboratory and immunological parameters was observed in the majority of the patients and no effect was found in a lesser group of patients than in the groups treated with drugs during the absence of lymphocyte SDH reaction thereto. The timing of the presence of SDH reaction to drugs in the immunocompetent cells makes it possible to set the optimal daily regime of their application and to select a drug that would be most effective in each particular case.

Chlamydia trachomatis is the most common cause of acute salpingitis worldwide. The socioeconomic impact of sexually transmitted infections (STI) caused by C. trachomatis is considerable. The purpose of this study was to investigate secretion of a unique chemokine, CXCL13, during the inflammatory process in human fallopian tube tissue in response to infection with C. trachomatis. We employed two models for our experiments: archived fallopian tube paraffin sections from known cases of salpingitis of unknown etiology and human fallopian tube organ culture established from fresh fallopian tube biopsies subsequently infected in vitro with C. trachomatis serovar E. We used immunohistochemistry, microarray analysis and cytometric bead array to study these specimens. In both models, we found that the fallopian tissue infected with C. trachomatis expressed CXCL13 and other characteristics of tertiary lymphoid tissue. In addition, we found that CXCL13 was expressed in multiple cell types, including endothelial cells, demonstrating a mechanism for the lymphoid aggregation seen in fallopian tube tissue during salpingitis and infection with C. trachomatis., (Copyright 2009 Prous Science, S.A.U. or its licensors. All rights reserved.)

Sexually transmitted infections, and their associated sequelae, such as tubal infertility, ectopic pregnancy and preterm labour, are a major worldwide health problem. Chlamydia trachomatis infection is thought to be the leading global cause of tubal infertility and tubal ectopic pregnancy. Preterm birth occurs in around 10% of all deliveries, and nearly 30% of preterm deliveries are associated with intrauterine infection. The mucosal innate immune system of the female reproductive tract has evolved to eliminate such sexually transmitted pathogens whilst maintaining its ability to accommodate specialized physiological functions that include menstruation, fertilization, implantation, pregnancy and parturition. The aim of this review was to describe the role and distribution of key mediators of the innate immune system, the natural antimicrobial peptides (secretory leukocyte protease inhibitor, elafin and the defensins) and the pattern recognition toll-like receptors in the normal female reproductive tract and in the context of these pathological processes.

Objective: To evaluate and compare the inflammatory response and mediators in the endometrium of patients with hydrosalpinges compared with normal controls., Design: Retrospective case-control study., Setting: Urban medical center., Patient(s) and Intervention(s): Hysterectomy samples were identified as being affected by hydrosalpinx or salpingitis (n = 30) and were age-matched with control samples (n = 30)., Interventions: Fallopian tube and endometrial slides were analyzed for leukocytes and immunohistochemical techniques performed for cytokines (interleukin-2 [IL-2])., Main Outcome Measure(s): Evaluate and compare the endometrial inflammatory response (leukocytes and cytokines) from samples affected and non-affected by hydrosalpinx and salpingitis., Result(s): Examination of tubal and endometrial slides with hydrosalpinx demonstrated a statistically significant increase in the number of overall inflammatory cells. High-intensity immunohistochemical staining for IL-2 was demonstrated in 7.4% of controls versus 65% of cases., Conclusion(s): A defined, identifiable, local response to hydrosalpingeal fluid has been demonstrated in the endometrium. This response consists of statistically significant elevations of leukocytes and IL-2. An inflammatory endometrial response may be an independent contributor to the decreased reproductive outcome observed in patients with hydrosalpinges.

Following infection with Neisseria gonorrhoeae, bacteria may ascend into the Fallopian tubes (FT) and induce salpingitis, a major cause of infertility. In the FT, interactions between mucosal epithelial cells and gonococci are pivotal events in the pathogen's infection cycle and the inflammatory response. In the current study, primary FT epithelial cells were infected in vitro with different multiplicities of infection (MOI) of Pil+ Opa+ gonococci. Bacteria showed a dose-dependent association with cells and induced the secretion of tumor necrosis factor alpha (TNF-alpha). A significant finding was that gonococcal infection (MOI = 1) induced apoptosis in approximately 30% of cells, whereas increasing numbers of bacteria (MOI = 10 to 100) did not induce apoptosis. Apoptosis was observed in only 11% of cells with associated bacteria, whereas >84% of cells with no adherent bacteria were apoptotic. TNF-alpha was a key contributor to apoptosis, since (i) culture supernatants from cells infected with gonococci (MOI = 1) induced apoptosis in naïve cultures, suggesting that a soluble factor was responsible; (ii) gonococcal infection-induced apoptosis was inhibited with anti-TNF-alpha antibodies; and (iii) the addition of exogenous TNF-alpha induced apoptosis, which was inhibited by the presence of increasing numbers of bacteria (MOI = 10 to 100). These data suggest that TNF-alpha-mediated apoptosis of FT epithelial cells is likely a primary host defense mechanism to prevent pathogen colonization. However, epithelial cell-associated gonococci have evolved a mechanism to protect the cells from undergoing TNF-alpha-mediated apoptosis, and this modulation of the host innate response may contribute to establishment of infection. Understanding the antiapoptotic mechanisms used by Neisseria gonorrhoeae will inform the pathogenesis of salpingitis and could suggest new intervention strategies for prevention and treatment of the disease.

Infertility is a condition that affects approximately 15-25% of couples with the desire to procreate. The integrity of the feminine reproductive tract is essential for this purpose, but the occlusion of the Fallopian tubes occurs in 12-33% of infertile women. The infection by Chlamydia trachomatis is one of the principle causes of tubal injury, which could finally lead to tubal occlusion. The tract infection has also been related to the use of intrauterine device, basically due to the fact that the insertion of the device could carry bacteria to the endometrial cavity. Keloid scars result from alterations in the normal process of wound healing, and it affects principally the population in reproductive age, maybe due to specific hormonal influence. These fibroproliferative alterations may produce significant deformations and alter organ function. The genetic factors have been studied in order to have a better understanding of the pathophysiology of keloid scarring. With these assessments, many other factors have been known to have a relationship with this abnormal healing process. This keloid scarring involves an excess in extracellular matrix production and inhibition of apoptosis, for which a several growth factors and interleukins are needed. One of the most important growth factors is IGF-1, which increases the expression of type I and III procollagen (found in the uterus); the IGF-1 receptor is overexpressed in the fibroblasts of keloids. Based on those previous observations a hypothesis that the chronic and repeated infection, and the use of IUD, generate an exaggerated inflammatory response in patients with a predisposition for keloid formation (which frequently form in childbearing age), in comparison to the patients that do not form this type of scarring, has been proposed. This makes a major frequency of adherences and finally tubal occlusion and infertility. The tendency of excessive scarring could not be exclusive of skin and generate abnormal scarring responses in feminine reproductive tract, leading to a major frequency of infertility. Thus, it could be suggested the use of other contraceptive methods and a more aggressive treatment against infections of the reproductive tract, taking in consideration the pathophysiology of keloid scar formation and its relationship with tubal occlusion.

The role of immune disturbances on the systemic and local levels is evaluated and the immunological mechanisms of tuboperitoneal sterility as the basis for the development of the methods of immunocorrection is grounded.

Chlamydial delayed-type hypersensitivity antigens were analyzed by using the subcutaneous salpingeal autotransplant model of Macaca nemestrina infected with Chlamydia trachomatis serovar E. Heat shock protein 60 was the only antigen shown to induce delayed-type hypersensitivity among other antigens tested, including UV-inactivated organisms, recombinant major outer membrane protein, purified outer membrane proteins, and heat shock protein 10.

Chlamydia trachomatis pgp3 DNA immunized (no. 300) and non-immunized (no. 300) C3H/HeN mice were infected by vaginal inoculation with infectious C. trachomatis serotype D elementary bodies (EBs) and the spread of infection to the salpinges was assessed by cell culture isolation from tissue homogenates 7, 14, 21, 28, 35 and 42 days post-infection (p.i.). Overall, the pgp3-DNA immunization prevented salpinx infection in 94 (56%) mice, if compared with the 168 positive animals found among the non-immunized animals (P < 0.001). A group of negative control animals (i.e. mice immunized with plasmid DNA containing an irrelevant insert) was not protected, whereas all the mice of a positive immune control group (mice that had resolved a primary genital C. trachomatis infection) were resistant to re-infection. Pgp3 DNA immunization induced both humoral and mucosal anti-pgp3 antibodies., (Copyright 2002 Elsevier Science Ltd.)

Perforin (pfp)/Fas ligand (FasL) double-deficient mice have previously been shown to be infertile, lose weight and die prematurely due to tissue destruction caused by a significant inflammatory infiltrate of monocytes/macrophages and T cells. Herein we have compared disease progression in mice additionally deficient in the inflammatory mediator TNF. Unlike pfp/FasL double-deficient mice (TNF+/+ pfp-/- gld), mice lacking functional TNF, FasL and pfp (TNF-/- pfp-/- gld) were comparatively fertile, with the majority of mice not suffering severe pancreatitis or hysterosalphingitis in the first 5 months of life. The mean lifespan of TNF-/- pfp-/- gld mice was 217 +/- 79 days compared with 69 +/- 10 days for TNF+/+ pfp-/- gld mice and the majority of moribund TNF-/- pfp-/- gld mice appeared to die as a result of severe pancreatitis, suggesting that loss of TNF was not completely protective. At 8 weeks of age, characteristics associated with the gld phenotype, such as expansion of B220+ CD4- CD8- T cells, lymphadenopathy and hypergammaglobulinemia were comparable between TNF+/+ pfp-/- gld and TNF-/- pfp-/- gld mice, although the lymphoid organs of TNF+/+ pfp-/- gld mice contained greater numbers of B220+ CD4- CD8- T cells, macrophages and T cells. We conclude that TNF is necessary for the full manifestation of immune dysregulation caused by pfp/FasL-deficiency, in particular in the early and overwhelming tissue infiltration and destruction caused by inflammatory cells.

66 women of reproductive age with different course of the inflammatory process in the upper section of the reproductive tract (endometritis and salpingo-oophoritis) were examined. The cell composition, viability and functional activity of the phagocytizing cells of cervical and endometrial secretions, as well as peritoneal exudate, were studied. The study revealed that these characteristics of the phagocytizing cells of the reproductive tract in women with the inflammatory process differed from similar characteristics in healthy women. Different changes in the functional activity of neutrophils and macrophages in the biological fluids under study in different course of the inflammatory process were detected.

Plasma content of thymosin-alpha(1)and its circadian variations in patients with inflammatory gynecologic diseases differ from those in healthy donors and depend on the type of inflammation and efficacy of treatment. It is concluded that not only the absolute content of thymic hormones, but also their biorhythmic variations are important for immune regulation.

97 females at reproductive age with chronic nonspecific salpingo-oophoritis (CNSO) were examined and treated. The results of the treatment (vaginal and external impact) demonstrate positive effects of various cryotherapeutic techniques on CNSO clinical course, on hormonal and immune unbalance, functional activity of the uterine tubes, regional hemodynamics, psychoemotional status. Thus, cryotherapy is an effective adjuvant in combined therapy of CNSO.

We report that perforin/Fas-ligand double-deficient mice die early of severe pancreatitis. Female mice, in addition, are infertile and suffer from hysterosalpingitis. Tissue destruction is accompanied by infiltration with Mac-1 (CD11b)-positive monocytes/macrophages, Mac-1-positive T cells, and expansion of CD8+ T cells. In vivo inactivation of monocytes/macrophages by carrageenan reverses disease progression and restores fertility of female mice. Perforin/Fas-ligand double-deficient CD4+ or CD8+ CTL are unable to lyse cognate-activated macrophages, and therefore are unable to mediate negative feedback regulation by lysis of APCs, thereby preventing further T cell activation. These studies demonstrate a novel role for perforin in homeostatic regulation of the immune response.

The studies of 116 females with chronic nonspecific salpingo-oophoritis demonstrate that combination of low-frequency magnetic field with iodine-bromine water effectively corrects immunological indices, i.e. improves T- and B-cell immunity, production of IgM and IgA, proportion of immunoregulatory subpopulations of T-lymphocytes.

An evaluation was done of the immunity status in 100 women presenting with infertility of associated genesis. An immunologic investigation into blood serum, follicular fluid (FF) and peritoneal fluid (PF) was carried out at day 14-16 and 18-23 of the menstrual cycle. The functional status of spermatozoa was characterized with the aid of the index of their survival in FF and PF. Local humoral immunity of FF and PF differs from the indices in blood serum, which fact was manifested by a striking reduction of Igs G, A, M. Sperm-immobilizing antibodies (SIAB) have been identified in blood serum, FF, and PF in 14, 4%, 22.4%, and 25% of women. SIAB production is associated with inadequacy of local immunity, such as low levels of SIgA, IgA, and lysozyme. Occurrence of SIAB in PF and FF worsens general motility of spermatozoa by agglutination of the latter, which fact undoubtedly affects conception.

Clinical and histopathological correlations of immunoreactivity to Chlamydia trachomatis and to epitopes of the C. trachomatis 60 kDa heat shock protein (hsp60) among women with ectopic pregnancy were evaluated in a case-control study. Serological responses to 13 synthetic peptides corresponding to major epitopes of the chlamydial hsp60 were determined in 67 women treated for ectopic pregnancy and 45 women with uncomplicated pregnancy in utero. Plasma cell salpingitis was detected in 29 (43.3%) of the ectopic patients. Its presence correlated with antibodies to two hsp60 epitopes, encompassing amino acids 260-271 and 411-422 (P = 0.02). Antibodies to these two epitopes, along with five other epitopes, also correlated with peritubal adhesion formation in ectopic pregnant patients (P < 0.01). Antibodies to epitopes 260-271 and 188-199 also correlated with a history of pelvic inflammatory disease (PID; P = 0.05). Patients with ectopic pregnancy were also more likely than their intrauterine pregnant controls to have present anti-chlamydial immunoglobulin G (P < 0.005). Women positive for both C. trachomatis and hsp60 epitope antibodies had an increased prevalence over controls of salpingitis, pelvic adhesions or history of PID (P < 0.05). In contrast, patients who were positive for only C. trachomatis antibodies or only hsp60 epitope antibodies did not differ from antibody-negative patients in each of these categories.

Objective: Our purpose was to examine clinical, microbiologic, serologic, and laparoscopic findings associated with perihepatitis., Study Design: In a prospective study of 157 women with a clinical diagnosis of pelvic inflammatory disease, 27 women with laparoscopically confirmed perihepatitis and salpingitis were compared with 46 patients with salpingitis alone., Results: Both current use or a history of ever using oral contraceptives was negatively associated with perihepatitis (p = 0.05 and p = 0.008, respectively). Moderate-to-severe pelvic adhesions were present at laparoscopy significantly more often in the perihepatitis-salpingitis group (70%) than in the salpingitis alone group (35%, p = 0.003). Antibody to the chlamydial 60 kd heat-shock protein at > or =0.5 optical density was detected in 67% of the perihepatitis-salpingitis group and in 28% of the salpingitis alone group (p = 0.005), and the median titer was significantly higher in the former group (p = 0.02)., Conclusion: Compared with women with salpingitis alone, patients with perihepatitis-salpingitis do not have distinctive clinical or microbiologic findings but do manifest a higher prevalence of moderate-to-severe pelvic adhesions and both a higher prevalence and higher titers of antibody to the chlamydial heat-shock protein-60.

Sixty two and 59 patients with tuberculous and nontuberculous salpingo-oophoritis, respectively, and 30 healthy females were examined. It was found that indirect solid-phase enzyme immunoassay (EIA) using ultrasonography of Mycobacterium tuberculosis H37Rv could detect antituberculosis antibodies in 66.7% of patients with tuberculous salpingo-oophoritis, and in 10% of healthy females. That using the antigen isolated from the cell wall extract of M. tuberculosis (whose molecular weight was 38 - 42 kD) could reveal them in 70.2, 4.3%, and 6.7%, respectively. After dissociation of immune complexes, EIA inhibition using affinity-purified antimycobacterial antisera displayed mycobacterial antigens in 75.0 and 4.3% of patients with tuberculous and nontuberculous salpingo-oophoritis, respectively, and in none healthy female. Thus, the detection of mycobacterial antigens and antituberculosis antibody may be successfully used in the complex diagnosis of tuberculosis of the female genitals.

The aim of this study was to evaluate the prevalence of serum immunoglobulin (Ig)G and IgA antibodies to recombinant chlamydial 60 kDa heat shock protein (C-hsp60) and to assess the prevalence of serum IgG antibodies to recombinant chlamydial 70 kDa heat shock protein (C-hsp70) in Chlamydia-associated chronic salpingitis and/or salpingitis isthmica nodosa with tubal occlusion. Infertile patients (n =34) with Chlamydia-associated, histologically documented chronic salpingitis and/or salpingitis isthmica nodosa and bilateral tubal occlusions (group I) were compared with infertile patients (n = 19) without tubal occlusions (group II). The prevalence of chlamydial antigen in endocervical, urethral and urine samples was low in both groups. The median chlamydial serum IgG and IgA antibody titres were significantly higher in group I than in group II (P < 0.0001 and P = 0.0002 respectively). Serum IgG antibodies to C-hsp60 and C-hsp70 were detected in 24 out of 34 patients (71%) in group I compared with 10 out of 19 (53%) and nine out of 19 (47%) patients in group II (not significantly different). There was a significant difference (P = 0.035) between the prevalences of serum IgA antibodies to C-hsp60 in groups I (seven out of 34 patients; 21%) and II (none of the 19 patients). The association between the presence of serum IgA antibodies to C-hsp60 and Chlamydia-associated chronic salpingitis and/or salpingitis isthmica nodosa with tubal occlusion underlies the significance of chlamydial 60 kDa heat shock protein in the pathogenesis of tubal infertility.

Background and Objectives: Although it has been hypothesized that repeated infections with Chlamydia trachomatis result in an increased potential for the development of infertility, it is not know whether repeated chlamydial infection by the vaginal route will result in an increased incidence of upper tract pathology or enhanced pathology., Goal of This Study: To determine whether guinea pigs given two infections with the chlamydial agent of guinea pig inclusion conjunctivitis would experience an increased incidence of pathologic changes compared with animals having only a single infection., Study Design: Guinea pigs previously infected with guinea pig inclusion conjunctivitis were challenged with a fresh intravaginal inoculum 73-77 days after the primary infection. Oviducts were examined either nine or 30 to 37 days after the challenge infection for pathologic changes and compared with control unchallenged animals 75 to 85 days after a primary infection., Results: A significant increase in the number of animals with oviducts demonstrating marked tubal dilatation was observed in the challenged animals 30 to 37 days after the challenge infection. There was no association of increased antibody titer and chlamydial Hsp60 with the presence of tubal dilatation., Conclusion: These data strongly indicate that repeated infection via the natural vaginal route does increase the risk of tubal damage.

We identified, by two-dimensional electrophoretic analysis and microsequencing, a protein of Chlamydia trachomatis elementary bodies which corresponds to the polypeptide (pgp3) encoded by open reading frame 3 (ORF3). Amino acid analysis showed that the first residue (Gly) found in the native protein is the one encoded by the second ORF3 codon, implying a typical bacterial removal of the first Met residue. Relatively large amounts of recombinant pgp3 (r-pgp3) in a stable, water-soluble form were obtained by overexpressing ORF3 in Escherichia coli and purifying the product from periplasmic extracts under nondenaturing conditions. These r-pgp3 preparations allowed specific detection of anti-pgp3 antibodies by enzyme-linked immunosorbent assay. Analysis of a group of 170 sera from healthy blood donors and from patients who were seropositive or -negative for C. trachomatis and Chlamydia pneumoniae showed that an immune response to pgp3 occurs in the majority (ca. 81%) of patients with sexually transmitted diseases who are seropositive for C. trachomatis and generally correlates with the response to cell surface antigens. No reaction between r-pgp3 and C. pneumoniae-positive sera was detected.

Objective: Our objective was to determine whether an upper genital tract Chlamydia trachomatis infection sensitizes lymphocytes to heat-shock protein epitopes expressed in both the human and chlamydial 60 kd heat-shock protein., Study Design: Peripheral blood mononuclear cells were isolated from women with or without a prior documented salpingitis and tested for their ability to proliferate in response to the recombinant C. trachomatis heat-shock protein and to five synthetic peptides corresponding to conserved epitopes expressed in both the human and chlamydial heat-shock proteins., Results: Among 22 healthy women with no history of chlamydial infections or salpingitis and 10 women seen for complaints other than a C. trachomatis infection, none had positive lymphocyte responses to any of the peptides and only one responded to the chlamydial heat-shock protein. Among nine women with a single episode of salpingitis none responded to the chlamydial heat-shock protein and one exhibited a positive lymphocyte response to a single peptide. This woman was also positive for C. trachomatis in the cervix. In contrast, among the 10 women with two or more episodes of salpingitis four (40%) had proliferation in response to the chlamydial heat-shock protein and five (50%) had positive lymphocyte responses to one of the peptides; two of these women also had C. trachomatis detected in their cervices., Conclusion: In women with a history of C. trachomatis upper genital tract infections, infection with C. trachomatis or other microorganisms can induce a lymphocyte proliferative response to the chlamydial 60 kd heat-shock protein and to epitopes present in the human heat-shock protein.

Acute salpingitis complicating cervical gonococcal infection is a significant cause of infertility. Relatively little data are available concerning the pathophysiologic mechanisms of this disease. A cohort of 243 prostitutes residing in Nairobi were followed between March 1985 and April 1988. Gonococcal cultures were performed at each visit, and acute salpingitis was diagnosed clinically. Serum at enrollment was tested by immunoblot for antibody to gonococcal outer membrane proteins. 8.6% (146/1689) of gonococcal infections were complicated by salpingitis. Increased risk of salpingitis was associated with younger age, shorter duration of prostitution, HIV infection, number of gonococcal infections, and episodes of nongonococcal salpingitis. Rmp antibody increased the risk of salpingitis. Antibody to Opa decreased the risk of salpingitis. By logistic regression analysis, antibody to Opa was independently associated with decreased risk of gonococcal salpingitis (adjusted odds ratio [OR], 0.35; 95% confidence interval [95%CI], 0.17-0.76); HIV infection (adjusted OR, 3.5; 95% CI, 0.96-12.8) and episodes of nongonococcal salpingitis (adjusted OR, 3.4; 95% CI, 1.8-6.4) were independently associated with an increased risk of salpingitis. Antibody to Opa appears to protect against ascending gonococcal infection, perhaps by interfering with Opa mediated adherence and endocytosis. The demonstration of natural immunity that protects against upper genital tract infection in women suggests that a vaccine to prevent gonococcal salpingitis is possible.

The role of delayed hypersensitivity in the pathogenesis of Chlamydia t trachomatis salpingitis was studied in the monkey "pocket" model. Pigtailed monkeys (Macaca nemestrina) were sensitized by inoculation of live C. trachomatis organisms (E/UW-5/Cx) into subcutaneous pockets containing salpingeal autotransplants. At 21 days, affinity-purified recombinant C. trachomatis heat-shock protein (rhsp60) was injected into pockets either previously sensitized with C. trachomatis or not sensitized in the same monkey. Delayed-type hypersensitivity reaction was observed, characterized by mononuclear cell infiltration with peak reaction at 48 h. Injection of rhsp60 into the pockets of a naive animal did not induce inflammation. This study showed that C. trachomatis infection in monkeys induced delayed hypersensitivity, which is mediated by hsp60. Histologic findings of the salpinx were consistent with delayed hypersensitivity reaction observed in ocular C. trachomatis infection, further suggesting a similar pathogenesis for both salpingitis and trachoma.

Histological evaluation of sections from the human fallopian tube revealed an inflammatory reaction in 21 of 31 women using an intrauterine contraceptive device (IUCD) and in four of 29 controls (non-IUCD users). The inflammatory cells were mainly localized at the epithelium-lamina propria interface and at the center of the mucosal folds. The immunohistochemical study revealed leukocytes (CD45+), T lymphocytes (CD3+), T helper cells (CD4+), T suppressor/cytotoxic cells (CD8+), B lymphocytes (CD22+, CD19+), granulocytes, monocytes and null cells (CD11b+) mainly localized at the lamina propria in both groups. T lymphocytes were the predominant cell type, and the ratio between T helper and T suppressor/cytotoxic cells was fairly close to one both in IUCD-users and controls. B lymphocytes were the least frequent cell type identified. In IUCD users, the numbers of the different leukocytes were increased. In both groups, IgA-, IgG- and IgM-positive cells were demonstrated and were predominantly located at the lamina propria of the mucosal folds. The IgA-positive cells dominated in both groups, whereas IgG- and IgM-positive cells were less frequent. Cell positive for IgA, IgG or IgM were significantly increased in number in the IUCD users. The data confirm the presence of an immune system in the normal human fallopian tube and indicate that the IUCD can induce a prominent recruitment of inflammatory cells, with a tubal inflammation as the result. The IUCD may disturb the immunological function of the fallopian tube and its rôle in fertilization.

Peripheral blood mononuclear cells (PBMC) from 9 of 18 women with laparoscopy-verified salpingitis proliferated in response to recombinant Chlamydia trachomatis 57-kDa heat shock protein (hsp). In contrast, PBMC from 0 of 10 women with cervicitis, 1 of 5 women with recurrent abortions, and 3 (7.1%) of 42 healthy reproductive-age women were responsive to hsp (P < .001). After passage of the hsp through an endotoxin-removing column, PBMC from 6 of 14 additional women with salpingitis were responsive to hsp, while those from 10 controls, including the 3 previously positive women, were negative. PBMC from all patients responsive to the chlamydial hsp were unresponsive to Mycobacterium bovis 65-kDa hsp. PBMC from 6 of the 15 women with a positive hsp-induced lymphocyte response were unresponsive to C. trachomatis elementary bodies. Induction of a cell-mediated immune response to the chlamydial 57-kDa hsp is a common feature of an upper genital tract infection but does not appear to be limited to women with apparent chlamydial infections.

Background and Objectives: Infertility after murine chlamydial salpingitis can be prevented by tetracycline treatment given before or at the time of infection. This study evaluates the efficacy and timing of tetracycline treatment and anti-inflammatory agents in the prevention of inflammation and subsequent infertility., Study Design: The left ovarian bursae and uterine horns were inoculated with the mouse pneumonitis strain of Chlamydia trachomatis. Mice were mated 54 days after inoculation., Results: Infected mice treated with tetracycline beginning 2 days after infection showed mild inflammation and no significant reduction in fertility. However, when tetracycline treatment was delayed until 5 days after infection, a moderate degree of inflammation and a significant reduction in fertility (P < 0.01) was noted. Treatment with ibuprofen, prostaglandin E1 (PGE-1), or hydrocortisone beginning day 2 post-inoculation did not significantly alter the degree of inflammation or subsequent fertility. Mean anti-chlamydial IgG titers were significantly lower in mice treated with either PGE1 or hydrocortisone compared with infected, untreated mice., Conclusion: These results indicate that while early treatment of chlamydial salpingitis may prevent infertility, delay in treatment may result in significant inflammatory damage and infertility. There was no apparent benefit from the addition of anti-inflammatory agents.

The effect of treatment with doxycycline on serum IgG and IgA antichlamydial antibodies was evaluated in 33 women who had had acute salpingitis associated with high titers of serum IgG (> or = 1:128) and/or IgA (> or = 1:16) antichlamydial antibodies. Overall, 29 women (87.9%) remained with high titers of IgG and/or IgA antibodies. No change or insignificant change in IgG antibody titer was demonstrated in 21 women (63.6%) and in IgA antibody titer in 21 women (63.6%). Positive seroconversion or a significant increase (> or = 4-fold) in IgG antibody titer was demonstrated in eight women (24.2%) and in IgA antibody titer in six women (18.1%). Negative seroconversion or a significant decrease in IgG antibody titer was demonstrated in four women (12.1%) and in IgA antibody titer in six women (18.1%). It is concluded that in most patients who had acute salpingitis associated with pretreatment high titers of serum antichlamydial antibodies, posttreatment titers may remain high even if treatment with doxycycline results in complete resolution of clinical signs and symptoms of the disease.

A Swiss Webster white mouse model of salpingitis was used to characterize the immune response following an intrauterine infection with the Chlamydia trachomatis mouse pneumonitis biovar. Western blot (immunoblot) analyses of the serum samples showed that the immunodominant bands corresponded to molecular masses of 72, 60, 42, and 28 kDa and to the lipopolysaccharide. Antibodies to the 60-kDa heat shock protein and to the 60-kDa cysteine-rich protein were detected at 2 and 3 weeks postinfection, respectively. Neutralization was observed in an in vitro assay with serum samples as early as the 3rd day postinfection and remained high for the 7 weeks of observation. The mice were mated in the 7th week following infection. Of the infected experimental mice, 71.4% were found to be either unilaterally or bilaterally infertile, whereas only 27.4% of the noninfected control mice were found to be infertile.

Intrauterine infection of mice with a human genital tract isolate of Chlamydia trachomatis (serovar F) resulted in salpingitis. In some cases, oviduct damage was sufficient to cause infertility due to lumenal blockage. Parenteral immunization with a purified, heterologous, recombinant major outer-membrane (rMOMP) preparation reduced the proportion of animals developing severe salpingitis by 77% compared with mock-immunized controls, but failed to reduce chlamydial colonization of the lower genital tract. In contrast, mice immunized with rMOMP directly into the Peyer's patches to stimulate mucosal immunity shed fewer chlamydiae from the vagina than controls, but showed little reduction in oviduct damage. No consistent correlation was observed between antibody levels to rMOMP in immunized mice and reduced lower genital tract colonization. Immunization with rMOMP via the presacral space, a route previously shown to stimulate mucosal immunity in the genital tract, produced high levels of circulating anti-rMOMP IgG but only traces of anti-rMOMP IgA in vaginal secretions. There was no difference in the severity of salpingitis in these animals compared with mock-immunized controls. Immunization with rMOMP conferred no protection against infertility resulting from direct inoculation of chlamydiae into the oviducts.

Immune mediated mechanisms might contribute to damage of the fallopian tube in instances of salpingitis. Using a filter paper technique to obtain samples during the surgical procedure, we examined fluids from the reproductive tract organs of seven women with salpingitis and five controls, for evidence of tumor necrosis factor (TNF). TNF, produced principally by macrophages, is a substantial mediator of inflammatory responses. In three women culture-positive for Chlamydia trachomatis, TNF was identified only in those fallopian tubes with visual evidence of disease. Fluids obtained from morphologically normal tubes, as well as from the ovaries and uterus, were negative. In three women with negative fallopian tube cultures but visual evidence of salpingitis, TNF was also identified in fluids from damaged, but not from normal, tubes. Ovarian and uterine fluids of the women were also TNF positive. The last patient, also culture-negative, had TNF only in one affected tube. All five patients in the control group had negative findings at all genital tract sites. Only one patient had TNF in her serum. Thus, localized cell-mediated immune system activation, identified by TNF production, appears to be a typical component of salpingitis.

Serum antichlamydial antibodies were studied in three groups. Group A included 49 women with recent ectopic pregnancy, group B included 56 women with normal first trimester pregnancy, and group C included 20 women with a first episode of acute salpingitis. In the ectopic pregnancy group, a significantly higher percentage of women had IgG antichlamydial antibodies than in the normal pregnancy group (75.5% vs. 46.4%, P less than 0.001). Women with acute salpingitis had the same prevalence of IgG antibodies as patients who had an ectopic pregnancy (60% vs. 75.5%, P less than 0.10). In the ectopic pregnancy group, the women had a significantly higher geometric mean titer (GMT) of antichlamydial IgG antibodies than women with an intra-uterine pregnancy (70.4% vs. 27.3%, P less than 0.001). The results suggest that there is a positive relation between prior Chlamydia trachomatis infection and risk for tubal pregnancy.

A role for both the cellular and humoral components of the immune response has been established for chlamydial infection. The significance of helper (L3T4) T cells was evaluated by using a Chlamydia trachomatis murine salpingitis model for upper genital tract chlamydial infection. Mouse oviducts were inoculated with C. trachomatis by using the mouse pneumonitis agent (MoPn) or control medium. Mice depleted of L3T4-bearing lymphocytes had significantly higher (P less than 0.05) numbers of organisms recovered at day 7 postinoculation. The rate of hydrosalpinx formation was significantly higher in the mice depleted of L3T4-bearing lymphocytes (27 of 31 [87%] ) than in the infected undepleted group (8 of 16 [50%] ) (P less than 0.01). The geometric mean antichlamydial immunoglobulin G titers at day 54 postinoculation were significantly higher in the L3T4-depleted mice (mean titer, 2,030) than in the undepleted group (mean titer, 776; P less than 0.05). The rate of fertility was lower in the L3T4-depleted group (2 of 31 [6%]) than in the infected, undepleted mice (2 of 16 [13%]), but this difference did not reach statistical significance. In conclusion, the greater persistence of organisms in the oviduct and higher rates of hydrosalpinx formation in mice depleted of L3T4-bearing cells suggests that these cells play a role in the clearing of organisms following infection and thus in reducing the degree of oviduct obstruction and damage.

The major outer-membrane protein (MOMP) of Chlamydia trachomatis is the focus of attention for chlamydial vaccine design, particularly those serovar- and subspecies-specific epitopes which provoke neutralizing immune responses. Selected surface-exposed B-cell epitopes of MOMP, incorporating B-subspecies specificities, were expressed as fusions with LamB, an inducible outer-membrane transport protein of Escherichia coli. These recombinant chlamydial-LamB proteins were correctly transported to the outer membrane of both E. coli and an aro A mutant of Salmonella typhimurium. The immunogenicity of the constructs was investigated in a mouse model of chlamydial salpingitis. After oral immunization, recombinant S. typhimurium were recovered from the livers of mice for up to two weeks, and a serum IgG response was induced both to the Salmonella and to the inserted chlamydial epitopes. By contrast, intravenous inoculation was ineffective. Although these LamB fusions proved only weakly immunogenic, this approach should be useful for investigating the ability of attenuated S. typhimurium vaccines incorporating chlamydial epitopes to stimulate protective mucosal immunity in the mouse model of chlamydial salpingitis.

Nonspecific and selected determinants of humoral immunity have been evaluated in 122 patients who clinically presented with "chronic salpingo-oophoritis". An objective confirmatory study is laparoscopy. The patients with clinical presentations of chronic salpingo-oophoritis displayed depressed phagocytic activity of neutrophils and function of complement in its both pathways. Alternatively, humoral immune parameters (serum levels of immunoglobulins A, M, G and circulating immune complexes) were normal. Functional evaluation of the classical and alternative complement activation pathways may provide a noninvasive objective test for differential diagnosis of chronic salpingo-oophoritis.

The existence of a secretory immune system in the female genital tract has been demonstrated by the predominance of immunoglobulin (Ig)A-producing plasma cells in human fallopian tube, uterine cervix, and vagina. Epithelium lining fallopian tubes expresses a receptor for IgA, secretory component (SC), and thus resembles other secretory tissues such as intestine, mammary, lacrimal, and salivary glands. The present study extends the characterization of the local immune system in the fallopian tube and assesses its response to infection. We examined normal and infected fallopian tubes from surgical specimens, obtained at tubal ligation and abdominal hysterectomy, for the presence of Ig-producing cells, T cells, and natural killer cells. All tubular segments contained a predominance of IgA plasma cells in the subepithelial lamina propria. The epithelial cells were strongly positive for SC. Luminal contents stained positively for IgA, SC, and J chain, suggesting that this material contained secretory IgA. Submucosal plasma cells of IgM and IgG classes were less frequent than IgA. T cells were present in numbers approximately twofold greater than plasma cells in normal fallopian tubes. T-suppressor (CD8+) cells, which may function in the induction of immune tolerance, were present in the intraepithelial spaces. Infected segments of fallopian tubes demonstrated six- to tenfold increased numbers of plasma cells of all classes. These data suggest that a local immune system is functioning in the human fallopian tube and may provide a first line of defense against tubal infection and the prevention of tubal factor infertility.

Groups of inbred female mice of strains CBA or C3H were infected genitally with a pathogenic human strain of Chlamydia trachomatis (N.I.1, serovar F) known to produce salpingitis and infertility in mice. Mice were inoculated under the ovarian bursa or directly into the uterine cavity with chlamydiae (test groups) or with sucrose-phosphate transport medium (control groups) before being challenged with chlamydiae by the same route 12-17 weeks later. Twenty-five pairs of test and control animals were killed from 7 to 77 days after challenge and oviductal inflammatory changes, recovery of organisms, and antibody responses were compared in the two groups. Salpingitis in the mice infected previously (tests groups) was more severe than in the controls in 56% of comparisons, the same in 24% and less severe in 20%. However, despite the increase in the severity of disease, shedding of C. trachomatis from the lower genital tract was less prolonged after rechallenge or did not occur. Salpingitis occurred in spite of the almost certain presence of pre-existing serum antibody, and accelerated and accentuated antibody response in the rechallenged mice. Furthermore, the continued existence of high titres of antibody was not associated with less severe disease. Thus, the results reveal that previous exposure to chlamydiae does not prevent salpingitis and suggest that its severity is influenced by cell-mediated immune mechanisms.