Trachomatis Antigen Research Articles

Correlation of Chlamydia and Chlamydophila spp. IgG and IgM antibodies by microimmunofluorescence with antigen detection methods

Correlation of serologic titers for Chlamydia trachomatis with other tests has been based on direct fluorescence antibody (DFA) testing and culture, but not on nucleic acid-based tests that are used for screening. We retrospectively reviewed the specificity of antibodies against C. trachomatis, Chlamydia psittaci, and Chlamydophila pneumoniae by microimmunofluorescence (MIF) when compared with DFA, culture, nucleic acid probe, and transcription-mediated amplification. Over a 6-year period, 226 cases had both MIF and one of these other methods performed for comparison. Agreement between C. trachomatis antigen or nucleic acid detection and MIF results was 87% (197/226). C. trachomatis serology had a negative predictive value of 98%, and 10.6% of cases were positive by serology and negative by antigen testing. Of the 13 patients who had a positive C. trachomatis antigen or nucleic acid test result, 9 had IgG and/or IgM titers highest against C. trachomatis, 3 had IgG titers highest against C. pneumoniae, and 1 had undetectable titers for the three chlamydial species. Twenty-five patients had positive IgG and/or IgM titers to C. trachomatis but negative antigen test results. Serologic testing can increase the sensitivity of detecting C. trachomatis infections.

Lower genital tract infections in HIV-seropositive women in India

The presence of STD facilitates shedding of HIV and increases HIV-1 disease progression, possibly by increasing plasma viremia. Our aim was to study the presence of various associated Sexually transmitted disease/Reproductory tract infections in HIV-seropositive women in India. The study included 40 HIV-seropositive women attending the antiretroviral therapy (ART) clinic at Lok Nayak Hospital. An informed consent was taken from all subjects. All cases were subjected to detailed gynecological examination and two types of swabs, i.e., a vaginal swab and a cervical swab were taken for STD/RTIs evaluation. The vaginal swabs were used for preparation of wet mount and KOH mount for diagnosis of trichomoniasis and candidiasis; to make a vaginal smear for Gram staining to diagnose bacterial vaginosis (BV) as per Nugent's criteria; for culture of aerobic bacteria and Candida spp. The cervical swab was used for isolation of Neisseria gonorrhoeae by culture and for detection of Chlamydia trachomatis antigen by Chlamydia microplate enzyme immunoassay kit (BIORAD). All data were analyzed using appropriate statistical tests. All 40 cases were evaluated for the presence of STD/RTIs associated with HIV infection. The women belonged to the reproductive age group (15-45 years) and majority (40%) of them were para 2. Most of the women (14, 35%) were in World Health Organization (WHO) stage I and maximum number (28, 70%) had their CD4 cell count more than 200 cells/ml. There was no significant correlation between WHO stage of HIV-seropositive women and their CD4 cell count (P=0.092). Out of 40 cases, 15 (37.5%) were on ART with maximum cases (53.3%) in WHO stage III. The duration of ART was more than 6 months in 9 (60%) cases. The most common presenting complaint was vaginal discharge in women with WHO stage II and III and 27.5% women showed vaginitis on per speculum examination. Laboratory tests showed high prevalence of BV (30%), mixed infection (30%), and candidiasis (10%) among HIV-seropositive women (P<0.001 in both). Women with BV were mostly in WHO stage I (38.4%) and stage II (36.3%), while those with mixed infection were mainly in WHO stage III (36.3%) and stage IV (40%).Women with candidiasis were mainly in WHO stage III. C. trachomatis antigen was found only in one subject (prevalence 2.5%). Both WHO stage and CD4 cell count had no significant correlation with presence of BV (P=0.056 and 0.063, respectively) and candidiasis (P=0.492 and 0.530, respectively). Maximum number of patients on ART had mixed infection (53.3%), while most of the patients (36%) not on ART had BV. There was no significant association between duration of ART and the presence of vaginal infections. The prevalence of gynecological symptoms and RTIs in HIV-seropositive women is high enough to warrant routine gynecologic evaluation and RTI screening in these patients. However, larger studies and trials are needed to evaluate the effects of ART on these abnormalities as well as to choose the best screening tool in HIV-seropositive women.

Systemic effector and regulatory immune responses to chlamydial antigens in trachomatous trichiasis

Trachomatous trichiasis (TT) caused by repeated or chronic ocular infection with Chlamydia trachomatis is the result of a pro-fibrotic ocular immune response. At the conjunctiva, the increased expression of both inflammatory (IL1B, TNF) and regulatory cytokines (IL10) have been associated with adverse clinical outcomes. We measured in vitro immune responses of peripheral blood to a number of chlamydial antigens. Peripheral blood effector cells (CD4, CD69, IFNγ, IL-10) and regulatory cells (CD4, CD25, FOXP3, CTLA4/GITR) were readily stimulated by C. trachomatis antigens but neither the magnitude (frequency or stimulation index) or the breadth and amount of cytokines produced in vitro [IL-5, IL-10, IL-12 (p70), IL-13, IFNγ, and TNFα] were significantly different between TT cases and their non-diseased controls. Interestingly we observed that CD4+ T cells account for <50% of the IFNγ positive cells induced following stimulation. Further investigation in individuals selected from communities where exposure to ocular infection with C. trachomatis is endemic indicated that CD3−CD56+ (classical natural killer cells) were a major early source of IFNγ production in response to C. trachomatis elementary body stimulation and that the magnitude of this response increased with age. Future efforts to unravel the contribution of the adaptive immune response to conjunctival fibrosis should focus on the early events following infection and the interaction with innate immune mediated mechanisms of inflammation in the conjunctiva.

Lower genital tract infections in HIV-seropositive women in India

Objectives:The presence of STD facilitates shedding of HIV and increases HIV-1 disease progression, possibly by increasing plasma viremia. Our aim was to study the presence of various associated Sexually transmitted disease/Reproductory tract infections in HIV-seropositive women in India.Materials and Methods:The study included 40 HIV-seropositive women attending the antiretroviral therapy (ART) clinic at Lok Nayak Hospital. An informed consent was taken from all subjects. All cases were subjected to detailed gynecological examination and two types of swabs, i.e., a vaginal swab and a cervical swab were taken for STD/RTIs evaluation. The vaginal swabs were used for preparation of wet mount and KOH mount for diagnosis of trichomoniasis and candidiasis; to make a vaginal smear for Gram staining to diagnose bacterial vaginosis (BV) as per Nugent's criteria; for culture of aerobic bacteria and Candida spp. The cervical swab was used for isolation of Neisseria gonorrhoeae by culture and for detection of Chlamydia trachomatis antigen by Chlamydia microplate enzyme immunoassay kit (BIORAD). All data were analyzed using appropriate statistical tests.Results:All 40 cases were evaluated for the presence of STD/RTIs associated with HIV infection. The women belonged to the reproductive age group (15-45 years) and majority (40%) of them were para 2. Most of the women (14, 35%) were in World Health Organization (WHO) stage I and maximum number (28, 70%) had their CD4 cell count more than 200 cells/ml. There was no significant correlation between WHO stage of HIV-seropositive women and their CD4 cell count (P=0.092). Out of 40 cases, 15 (37.5%) were on ART with maximum cases (53.3%) in WHO stage III. The duration of ART was more than 6 months in 9 (60%) cases. The most common presenting complaint was vaginal discharge in women with WHO stage II and III and 27.5% women showed vaginitis on per speculum examination. Laboratory tests showed high prevalence of BV (30%), mixed infection (30%), and candidiasis (10%) among HIV-seropositive women (P<0.001 in both). Women with BV were mostly in WHO stage I (38.4%) and stage II (36.3%), while those with mixed infection were mainly in WHO stage III (36.3%) and stage IV (40%).Women with candidiasis were mainly in WHO stage III. C. trachomatis antigen was found only in one subject (prevalence 2.5%). Both WHO stage and CD4 cell count had no significant correlation with presence of BV (P=0.056 and 0.063, respectively) and candidiasis (P=0.492 and 0.530, respectively). Maximum number of patients on ART had mixed infection (53.3%), while most of the patients (36%) not on ART had BV. There was no significant association between duration of ART and the presence of vaginal infections.Conclusions:The prevalence of gynecological symptoms and RTIs in HIV-seropositive women is high enough to warrant routine gynecologic evaluation and RTI screening in these patients. However, larger studies and trials are needed to evaluate the effects of ART on these abnormalities as well as to choose the best screening tool in HIV-seropositive women.

Spontaneous secretion of interleukin-17 and -22 by human cervical cells in Chlamydia trachomatis infection

To investigate whether IL-17A (IL-17) and IL-22 are produced in response to Chlamydia trachomatis infection, the cervical washes of 27 women with C. trachomatis infection and 17 C. trachomatis negative controls were collected. The levels of cytokines were determined in the cervical wash and in the supernatant of cervical and systemic cell cultures upon C. trachomatis antigen stimulation. C. trachomatis infection appeared to activate local IL-17 and IL-22 production more efficiently than IFN-γ production. In the cervical wash of infected women, median concentrations of IL-17 and -22 were 5- and 3-fold higher, respectively, than in negative controls. The spontaneous intracellular expression of these cytokines was analysed by flow cytometry in blood and cervical cells and 26% of cervical mononuclear cells from infected women were shown to produce IL-22 and 12% to coproduce IL-17 and IL-22. In addition, it was demonstrated that 20–25% of IL-22 producing and IL-17-IL-22 coproducing cervical CD4+ T cells expressed the mucosal homing receptor CCR6. These results suggest that CCR6 is involved in the migration of these cells to the cervix and that IL-17 and IL-22 might play a role in the immune response at the site of C. trachomatis infection.

Prevalence of infection for HIV, HTLV, HBV and of syphilis and chlamydia in pregnant women in a tertiary health unit in the western Brazilian Amazon region

to estimate the prevalence infection of human immunodeficiency virus (HIV), human T-cell lymphotropic vírus (HTLV), hepatitis B virus (HBV), Chlamydia trachomatis (C. trachomatis) and syphilis in pregnant women, as well as risk factors associated with these infections, in Fundação de Medicina Tropical do Amazonas (FMTAM). a cross-sectional study was carried including 674 pregnant women consecutively attended of the spontaneous demand of FMTAM between March and September 2008. Demographic, epidemiologic, socioeconomic, clinical and obstetric information have been collected through specific questionnaires. Patients had blood sample collected by peripheral venous for accomplishment of serological tests of HIV, HTLV, HBV and syphilis. Cervical secretion sample has been collected for C. trachomatis antigens detection test. The Odds Ratio has been used to evaluate risk factors associated to infections. Statistical analysis has been done with the t-Student, chi2 and Fisher's exact tests. the average age was 23.9 years old (SD 6.3). The observed prevalence was 0.6% to infection by HIV; 0.7% by HBsAg; 1.0% of syphilis and 2.7% by C. trachomatis. All the samples went negatives to HTLV. There were no variables associated with infection by HIV, HBV and syphilis. Significative statistically association was observed between pregnant woman with age under 20 years and of first pregnancy with C. trachomatis infection. the study evidenced that the prevalence infection by HIV in pregnant women assisted in FMTAM is similar to the values described in the Brazilian literature, while the prevalence by HTLV, HBV, syphilis and C. trachomatis in the studied population are below found by other authors. The main risk factor for the infection by C. trachomatis was being under 20 years old.

Chlamydia trachomatis antigens in enteroendocrine cells and macrophages of the small bowel in patients with severe irritable bowel syndrome

BackgroundInflammation and immune activation have repeatedly been suggested as pathogentic factors in irritable bowel syndrome (IBS). The driving force for immune activation in IBS remains unknown. The aim of our study was to find out if the obligate intracellular pathogen Chlamydia could be involved in the pathogenesis of IBS.MethodsWe studied 65 patients (61 females) with IBS and 42 (29 females) healthy controls in which IBS had been excluded. Full thickness biopsies from the jejunum and mucosa biopsies from the duodenum and the jejunum were stained with a monoclonal antibody to Chlamydia lipopolysaccharide (LPS) and species-specific monoclonal antibodies to C. trachomatis and C. pneumoniae. We used polyclonal antibodies to chromogranin A, CD68, CD11c, and CD117 to identify enteroendocrine cells, macrophages, dendritic, and mast cells, respectively.ResultsChlamydia LPS was present in 89% of patients with IBS, but in only 14% of healthy controls (p < 0.001) and 79% of LPS-positive biopsies were also positive for C. trachomatis major outer membrane protein (MOMP). Staining for C. pneumoniae was negative in both patients and controls. Chlamydia LPS was detected in enteroendocrine cells of the mucosa in 90% of positive biopsies and in subepithelial macrophages in 69% of biopsies. Biopsies taken at different time points in 19 patients revealed persistence of Chlamydia LPS up to 11 years. The odds ratio for the association of Chlamydia LPS with presence of IBS (43.1; 95% CI: 13.2-140.7) is much higher than any previously described pathogenetic marker in IBS.ConclusionsWe found C. trachomatis antigens in enteroendocrine cells and macrophages in the small bowel mucosa of patients with IBS. Further studies are required to clarify if the presence of such antigens has a role in the pathogenesis of IBS.

Screening of pregnant women attending the antenatal care clinic of a tertiary hospital in eastern Saudi Arabia for Chlamydia trachomatis and Neisseria gonorrhoeae infections

Inroduction:Of the “top ten” sexually transmitted infections, Chlamydia trachomatis and Neisseria gonorrhoeae are ranked second and fifth, respectively, worldwide.Aim:The aim of this study was to screen the pregnant women for C. trachomatis and N. gonorrhoeae infections and to detect antimicrobial resistance pattern of N. gonorrhoeae.Materials and Methods:This study was a prospective, hospital-based analysis of a random sample of pregnant women visiting the antenatal clinic of a tertiary hospital in eastern Saudi Arabia. Endocervical and high vaginal swabs were collected both from pregnant women and female patients attending gynecology clinic with lower genital tract infection (control group). C. trachomatis antigen was detected using enzyme-linked immunosorbent assay (ELISA). N. gonorrhoeae was detected by culture and identification of isolates, and antimicrobial susceptibility testing was performed. Statistical Package for Social Sciences (SPSS) version 13.0 and Chi-square test were used for statistical analysis.Results:C. trachomatis antigen was detected in 10.5% (10/95) and 34.4% (35/102) of pregnant women and control group, respectively (P < 0.001). The isolation rate of N. gonorrhoeae among pregnant women was 0.0% compared to 7.8% (8/102) among the control group (P < 0.01). N. gonorrhoeae were resistant to penicillin (62.5%), tetracycline (50%), ampicillin (25%), amoxycillin–clavulinic acid (25%) and ciprofloxacin (37.5%), while they were susceptible to cefepime, ceftriaxone, ceftazidime, spectinomycin, and cefuroxime.Conclusion:Screening of pregnant women for C. trachomatis infection should be included in the antenatal care in this area. The detection rate of both organisms among the control group highlights the importance of preventive strategies. Certain antibiotics previously used in treating gonorrhea are no longer effective.

Prevalence of Chlamydia trachomatis Infection among Women attending infertility Clinics in Calabar, Nigeria.

The prevalence of Chlamydia trachomatis infection was investigated among 100 women attending infertility clinics and compared with another subset of 100 pregnant women who served as control. Chlamydia trachomatis antigen detection cassette supplied by Biomil Diagnostics was used to detect the presence of chlamydial antigen in endocervical swabs collected from each of the subjects. Questionnaire-based interview was used by the attending clinician to collect the demographic characteristics of the subjects for analysis. Twenty two 22 (22%) women with infertility and 2 (2%) of the control group tested positive for chlamydial antigen (p0.05). This study has demonstrated the involvement of Chlamydia trachomatis in female infertility and the need to put in place appropriate facilities to screen for Chlamydiatrachomatis among women with infertility problem in the study area.

Rapid Assessment of Trachoma among Children Living in Rural Northern India

ABSTRACTPurpose: Rapid assessment of active trachoma in children aged 1–9 years in a previously hyperendemic rural area in Haryana, India. Methods: Ten disadvantaged villages each with a population of 3000–5000 were chosen by cluster random sampling. One thousand children from 500 households in the most underdeveloped parts of the villages—identified by observation and consultation, between the ages of 1–9 years—were examined for signs of Trachomatous inflammation follicular (TF) and Trachomatous inflammation intense (TI). Assessment was done in a health care unit. Examination of both eyes for signs of trachoma and its complications was done with the aid of binocular loupe (2.5X magnification). Tarsal conjunctival swabs from patients of active trachoma were analyzed by direct immunoflourescence assay and polymerase chain reaction for Chlamydia trachomatis antigen. Results: Forty children (males 21, females 19) had signs of active trachoma that included TF (33) and TI (7). At least one ocular morbidity was present in 69% of all children that were examined. Unclean face carried a 2.70 (confidence interval [CI] = 1.30–5.37) times higher risk and poor ocular hygiene had 2.05 (CI = 1.02–4.11) times higher risk for trachoma infection. Among clinically positive cases, direct immunoflourescence assay and polymerase chain reaction assays were positive in 25% and 10%, respectively. Conclusions: Active trachoma is not a public health problem in previously hyperendemic areas of North India.

Identification and characterization of novel recombinant vaccine antigens for immunization against genitalChlamydia trachomatis

Chlamydia trachomatis infection is the most common sexually transmitted bacterial infection worldwide, with over 91 million cases estimated annually. An effective subunit vaccine against Chlamydia may require a multivalent subunit cocktail of antigens in a single formulation for broad coverage of a heterogeneous major histocompatibility complex population. Herein, we describe the identification of novel C. trachomatis antigens by CD4+ and CD8+ T-cell expression cloning, serological expression cloning, and an in silico analysis of the C. trachomatis genome. These antigens elicited human CD4+ T-cell responses, and a subset proved to be immunogenic and protective when administered as immunoprophylactic vaccines against C. trachomatis challenge. Candidate vaccines consisting of the prioritized C. trachomatis antigens adjuvanted in a GlaxoSmithKline proprietary AS01B adjuvant were prioritized based on induction of solid protection against challenge in C57BL/6 and BALB/c mice with C. trachomatis. Some of the vaccines prevented bacterial shedding and colonization of the upper genital tract to varying degrees by mechanisms that may include CD4+ T cells.

Chlamydia trachomatis in women

The authors present results of a planned prospective study of infections with Chlamydia trachomatis in women. Direct immunofluorescence test (DIF) for proving Chlamydia trachomatis antigens in endocervical smear (by means of monoclonal antibodies), ELISA test for proving IgG antibodies against Chlamydia trachomatis in sera and RVK method for proving antibodies against group Chlamydia-antigen were applied to test 288 women (aged 19-67), previously thoroughly anamnestically and clinically examined Chlamydia trachomatis infection was found in 29.51% of the cases, equally distributed in all age groups. The authors analyzed frequency of Chlamydia positive findings according to clinical diagnoses, parity, relevant data from gynecological history, as well as results of vaginal secretion tests and Papanicolaou smear findings.

Prevalence and risk factors for trachoma in primary school children in Datong City in China

To assess the prevalence and risk factors for trachoma among primary school children in Datong City, China. An epidemiological survey was carried out in Datong City in 2006. Children aged 6 to 14 years old in primary school were selected by a cluster sampling in which class shift was the sampling unit. Trachoma was clinically assessed according to the simplified classification for trachoma proposed by the World Health Organization (WHO). If the child was clinically diagnosed as trachoma, a conjunctival swab would be taken for detecting C. trachomatis by antigen test and polymerase chain reaction (PCR). A total of 1236 children were examined and 135 cases of trachoma were found, the prevalence of trachoma was 10.9% (95% CI: 9.2% - 12.6%). Follicular trachoma was found in 86.7% of the cases and intense trachoma in 13.3%. The overall prevalence of trachoma was higher for females than for males (13.1% versus 8.8%, P = 0.016). Thirteen (9.6%) cases were positive in C. trachomatis antigen test, and 86 (63.7%) cases were positive by PCR. Poor ocular hygiene and sharing wash cloth and face basin with households were the main risk factors. Trachoma is endemic among primary school children in Datong City in China.

Antigenic Profiling of aChlamydia trachomatisGene‐Expression Library

The obligate intracellular bacterium Chlamydia trachomatis is the causative agent of sexually transmitted chlamydia infections. A panel of 116 recombinant C. trachomatis proteins was evaluated comparatively to characterize both cell-mediated and humoral immune responses in patients with confirmed C. trachomatis genital infection. The antigens identified were categorized as being recognized exclusively by T cells (CT004, CT043, CT184, CT509, and CT611), B cells (CT082, CT089, CT322, CT396, and CT681), or both T cells and B cells (CT110 and CT443). This grouping of C. trachomatis antigens was correlated to their predicted cellular localization. The comparative evaluation presented here indicates that T cell antigens are located in all bacterial compartments, whereas antibody targets are mainly localized to the outer membrane (P = .0013). Overall, we have identified 5 T cell antigens, 5 B cell antigens, and 2 T/B cell antigens that are potential components for a future chlamydia vaccine.

Evaluation of a Developed Species-Specific Monoclonal Antibody for DetectingChlamydia trachomatisInfections in Endocervical Specimens from Female Patients

To increase the sensitivity and reliability of immunodiagnostic assay for direct detection of chlamydial genital infection, we developed monoclonal antibodies (MAbs) that recognize the major outer membrane protein (MOMP) of all serovars of Chlamydia trachomatis. ELISA and dot-ELISA were used to select hybridomas that produced anti-C. trachomatis MAbs. Four MAb clones--A12.6, B serogroup specific; C1.5, C serogroup specific; G10.2, species specific; and E9.1, genus specific--were characterized by SDS-PAGE and immunoblot and were reactive with MOMP of C. trachomatis. The species-specific MAb G10.2 had a molecular mass of 40 kDa. The reactivity of developed species-specific monoclonal antibody for detection of C. trachomatis antigen in endocervical specimens was compared with the commercially available DFA test. We found that developed antibody had high sensitivity (97.22%) compared to DFA for detection of chlamydial infection in endocervical samples. It can be used as a reliable method in laboratories for the diagnosis of chlamydial infections.

External quality control program: Experience in the department of Microbiology and Immunology, Muhimbili University College of Health Sciences, Dar es Salaam, Tanzania

Objective: To share experiences and lessons learned from 2002 National External Quality Assessment Schemes for Leukocyte Immunophenotyping (UK NEQAS) and College of American Pathologists (CAP) proficiency testing (PT) panels received and tested as part of External Quality Control Program Setting: Department of Microbiology and Immunology, Muhimbili University College of Health Sciences, Dar es Salaam. Methods: All PT panel records from January – December 2002 were reviewed and summarized to show the type of panel and date received, date results sent after analysis and our performances. Results: Six and fourteen panels were received from UKNEQAS and CAP, respectively. The mean time interval between receipt of panels and reporting back of the results was 9.4 days (range 2-17). UKNEQAS performance was good in 5/6 panels (83.3%) analyzed. Among CAP panels, satisfactory (100%) performances included two viral markers, two C. trachomatis antigen detection and two comprehensive hematology with automated differentials whereas unsatisfactory (79%, 66%, 71%) performances were all 3 general chemistry with an overall score of (6/9) 66.7%. Conclusion: To implement an effective and efficient quality assurance in an established laboratory requires committed long-term efforts that are potentially constrained with multiple obstacles. Continuing education and training of laboratory personnel is an integral part of quality control/quality assurance program. Keywords: Proficiency Testing, Quality Control, and Quality Assurance. > Tanzania Medical Journal Vol. 19 (1) 2004

Urogenitale Chlamydieninfektionen bei Frau und Mann

Genital infections with Chlamydia trachomatis occur in all social groups in Germany. About 100,000 German women are sterile because of tubal scarring due to chlamydiae. Genital chlamydial infections are asymptomatic in 70% of patients, even if salpingitis occurs. Typical symptoms of chlamydial infection are purulent cervicitis with vaginal discharge, painful cervical bleeding because of endometritis, lower abdominal pain with dyspareunia, and upper abdominal pain because of perihepatitis. DNA amplification tests on first voided urine or cervical swab are the most sensitive routine tests. Specific serum antibodies to C. trachomatis indicate a previous infection in sterile women. For treatment, a 10-14 day course of doxycycline 200 mg daily or a macrolide antibiotic in the patient as well as in the sexual partner is recommended. In the male, C. trachomatis causes urethritis and epididymitis. Opinions differ about involvement of the prostate gland and seminal vesicles. Identification of C. trachomatis antigen or DNA in the accessory gland secretions is not sufficiently reproducible. The two vectors are easily diagnosed in urethral swabs or in urine. The occurrence of chlamydial antibodies in serum or in seminal fluid is not a sign of current infection. Reliable studies which indicate a reduced fertility of men infected with C. trachomatis are not available.

Production and Characterization of Monoclonal Antibodies toChlamydia trachomatis

In order to develop an indigenous and reliable immunodiagnostic assay for Chlamydia trachomatis in India, monoclonal antibodies (MAbs) were developed. Serovar D of C. trachomatis (which was previously isolated from the genital tract of infected women) was propagated, purified, and used for production of monoclonal antibody. A total of 12 murine hybrid clones producing immunoglobulin G (IgG) class monoclonal antibodies to C. trachomatis (species-specific, B serogroup-specific, and serovar-specific) were developed. Enzyme-linked immunosorbent assay (ELISA) was used to screen developed murine MAbs with C. trachomatis antigen. Dot-ELISA was used to check the specificity of clones and was used for selecting hybridomas that produced anti-C. trachomatis MAb. There was no cross-reactivity of species-specific, B serogroup-specific, and D serovar-specific anti-major outer membrane protein (MOMP) monoclonal antibodies with other species of Chlamydiae i.e., C. pneumoniae and C. psitacci. Immunoblotting was done for further characterization of six of these clones, i.e., B2.2 and D5.1 (B serogroup-specific), D10.4 and G1.5 (species-specific), and H5.6 and E4.2 (D serovar-specific). Three of these clones D10.4 (species-specific), B2.2 (B serogroup-specific), and H5.6 (D serovar-specific) which reacted with 40 kd MOMP protein in Immunoblotting were used for further screening to detect C. trachomatis in endocervical specimens. The percent positivity with these clones for detection of C. trachomatis antigen by enzyme immunoassay (EIA) was 45% with D10.4, 43% with H5.6, and 35% with B2.2, while 46% of the specimens were found positive by cell culture method. This indicates a high prevalence of C. trachomatis infection in the female genital tract. The sensitivity and specificity of developed anti- MOMP monoclonal antibody in EIA for chlamydial antigen detection was 91.3% and 94.4% for D10.4 clone (species-specific), 91.30% and 98.1% for H5.6 (D serovar-specific) and 75.00% and 99.07% for B2.2 (B serogroup-specific) compared to cell culture method.

Identification of CT521 as a Frequent Target of Th1 Cells in Patients with UrogenitalChlamydia trachomatisInfection

The human immune response to a Chlamydia trachomatis serovar D lysate was investigated in patients with urogenital C. trachomatis infection, to identify novel T cell targets. A C. trachomatis lysate was fractionated on the basis of molecular mass, and each fraction was used to stimulate peripheral-blood mononuclear cells from patients with C. trachomatis infection. In frequently recognized fractions, proteins were identified by mass spectrometry, recombinantly expressed, and tested for T cell recognition. T cell recognition of the fractions was highly heterogeneous in patients with C. trachomatis infection (n=16). Four patients exhibited responses that were strongly targeted to antigens of 16-20-kDa molecular mass. Three proteins were identified in this fraction: CT043, CT511, and CT521. The T cell response to the individual recombinant proteins were investigated, and CT521 was found to induce the highest level of interferon (IFN)- gamma. The recognition of CT521 was investigated in a larger study population (n=41), and a positive IFN-gamma response was measured in 83% of the patients. Several T cell epitopes were identified in CT521; in particular, peptide 5 in the central part of the protein was frequently recognized by T cells (63%). We have identified a novel C. trachomatis antigen, CT521, that is frequently recognized in patients with urogenital C. trachomatis infection.

Pathological study of Fitz–Hugh–Curtis syndrome evaluated from fallopian tube damage

To analyze the causative pathogen, surgical indication and fallopian tube damages in Fitz-Hugh-Curtis syndrome (FHCS) cases diagnosed by laparoscopy. Laparoscopic findings in 3568 cases at our hospital were reviewed retrospectively. Of these, 108 cases were diagnosed to have FHCS and examined for causative pathogens. Chlamydial infection was determined by both Chlamydia trachomatis (CT) antigens in the cervix and serum anti-CT antibodies. One hundred and four cases with chronic FHCS were divided into hydrosalpinx (H) and normal (N) groups based on the fallopian tube conditions, and were compared for patient profiles, tubal functions and perihepatic adhesion. Among surgical indications, tubal infertility and ectopic pregnancy were more often associated with FHCS than any other diseases. The primary causative pathogen of FHCS was CT. In 104 cases with chronic FHCS, there was no difference in CT antigen positivity between the H and N groups. The serum anti-CT IgG titer also did not differ between the two groups, although anti-CT IgA was slightly higher in the H group. The peritubal adhesion score was significantly higher in the H group, but it was not correlated with the degree of perihepatic adhesion. While some FHCS patients had severe fallopian tube dysfunctions, such as hydrosalpinx and tubal occlusion, 30% of FHCS cases did not show any abnormalities in the fallopian tube. Our study indicated that the severity of tubal dysfunctions associated with FHCS is determined by the host's reactivity to CT.