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PubMed1998

Lack of correlation between the detection of Chlamydia trachomatis DNA in synovial fluid from patients with a range of rheumatic diseases and the presence of an antichlamydial immune response.

OBJECTIVE: To resolve how frequently Chlamydia trachomatis and Chlamydia pneumoniae DNA are present in the joints of unselected patients with reactive arthritis (ReA) and undifferentiated oligoarthritis, and to determine if there is an accompanying serologic or cellular antichlamydial immune response. METHODS: Two polymerase chain reaction (PCR) protocols to detect the plasmid of C. trachomatis and the outer membrane protein 1 gene of C. pneumoniae were developed for specific use with synovial fluid (SF). Subsequently, the assays were used to detect DNA from the 2 organisms in SF from 54 adult patients with rheumatic diseases, including 4 with sexually acquired ReA and 31 with undifferentiated oligoarthritis. The presence of chlamydial antibodies and SF lymphocyte proliferation responses were determined in parallel. RESULTS: The PCR protocols were species-specific and highly sensitive. SF samples from 15 patients (8 with undifferentiated oligoarthritis, 3 with ReA, 1 with rheumatoid arthritis, and 1 with psoriatic arthritis) were positive for C. trachomatis. There was no significant correlation between the presence of C. trachomatis DNA in the joint and a Chlamydia-specific synovial T cell response or a serologic response. C. pneumoniae was not detected in any of the 54 patients, although it was identified in the SF from a rheumatoid arthritis patient outside this study, demonstrating that the assay was capable of detecting the organism in the joint. CONCLUSION: C. trachomatis DNA was present in ReA patients and in nearly one-third of unselected patients with undifferentiated oligoarthritis, which further supports the hypothesis that it plays an important role in disease pathogenesis. However, its presence did not correlate with evidence of an antichlamydial immune response. Despite previous anecdotal reports, C. pneumoniae does not appear to be a major cause of undifferentiated oligoarthritis or ReA.

Adult↗
PubMed1999

Chlamydia trachomatis nucleic acids can be found in the synovium of some asymptomatic subjects.

OBJECTIVE: The recent identification of antigens or nucleic acids of infectious agents in the joints of patients with reactive arthritis has raised questions about whether chlamydial or other infectious agent nucleic acids are also present in normal joints. We had the opportunity to study synovium from 30 asymptomatic volunteer subjects by use of polymerase chain reaction (PCR) for attempted identification of Chlamydia and other infectious agents. METHODS: All subjects had blind needle synovial biopsies with the Parker-Pearson needle. DNA was extracted and PCR performed using primers for Chlamydia trachomatis, Chlamydia pneumoniae, Borrelia burgdorferi, and pan bacterial 16S ribosomal RNA (rRNA). RESULTS: Two subjects were identified with nucleic acid for the 16S rRNA gene of C trachomatis. All other PCR reactions were negative except for the pan bacterial 16S rRNA in the C trachomatis-positive subjects. Both subjects, although symptom free, had some evidence of synovial reaction. CONCLUSION: C trachomatis appears to occasionally be disseminated to joints without producing overt disease.

Adult↗
PubMed1988

Cytopathologic detection of Chlamydia trachomatis in vaginopancervical (Fast) smears.

The value of the Papanicolaou-stained vaginopancervical (Fast) smear in the detection of chlamydial infection has been disputed. We examined 116 satisfactory Fast smears from 203 women enrolled in the Johns Hopkins Fertility Control Clinic and compared tissue-culture results with cytopathologic detection using various published morphologic criteria. All Chlamydia culture-positive cases were reviewed, and certain cytologic features considered helpful in the detection of chlamydial infection in cervical smears obtained from this selected high-risk population were identified. The changes that had the highest correlation with tissue culture included fine vacuolation of metaplastic endocervical cells, giving their cytoplasm a rarefied "moth-eaten" appearance. Using these criteria, cytopathologic changes of chlamydial infection were observed in 24 of 28 cases of tissue-culture-positive cases and in 8 of 88 tissue-culture-negative cases. The sensitivity and specificity of the Fast-smear cytodiagnosis of Chlamydia infection utilizing these morphologic changes and compared with tissue culture were 86% and 91%, respectively. Other cytologic features, including inflammatory background and intracytoplasmic structures consistent with initial and intermediate chlamydial bodies within the metaplastic cells, were found to be useful although less specific and less sensitive. The implications of these diagnostic features, the conditions to be considered in their differential diagnosis, and the pitfalls of chlamydial cytodiagnosis and the chlamydia culture studies have been critically reviewed. Study design and the high unsatisfactory cervical smear rate are discussed.

Cervix Uteri↗
PubMed1998

Cardiolipin remodeling in eukaryotic cells infected with Chlamydia trachomatis is linked to elevated mitochondrial metabolism.

Cardiolipin remodeling in mammalian eukaryotic cells was examined subsequent to infection with Chlamydia trachomatis, an intracellular parasite of eukaryotic cells. HeLa cells were labeled for 6 h with [1-14C]myristate or [1-14C]palmitate or [1-14C]oleate 20 h post infection with C. trachomatis and the radioactivity incorporated into glycerophospholipids examined. Chlamydia infection resulted in a 2-4 fold elevation of radioactive myristate, palmitate or oleate incorporation into phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine and phosphatidylinositiol compared to mock-infected cells. However, a 4-10 fold elevation in radioactivity incorporated into the mitochondrial glycerophospholipids phosphatidylglycerol and cardiolipin was observed in chlamydia-infected cells compared to mock-infected controls. Glycerophospholipid remodeling in CCL16-B2 cells, a mitochondrial respiration mutant with elevated glycerophospholipid metabolism, was compared to its parental cell line CCL16-B1 infected with C. trachomatis. Infection of the wild type CCL16-B1 cells with C. trachomatis resulted in an almost identical pattern of [1-14C]-palmitate labeling of glycerophospholipids compared to the uninfected mitochondrial mutant CCL16-B2 cells. The results suggest that cardiolipin, and glycerophospholipid, fatty acid molecular remodeling in eukaryotic cells infected with C. trachomatis may be linked to an elevation in mitochondrial metabolism.

Animals↗
PubMed1997

Does Chlamydia trachomatis MoPn enter a microbiologically-inapparent state during experimental infection of the mouse genital tract?

Microbiologically-inapparent chlamydial infection may contribute towards the immunopathogenesis of these diseases. Although morphologically and physiologically aberrant non-cultivable chlamydiae can be induced reversibly in cell culture, evidence for these forms in infections of animals and humans is indirect. A mouse model of salpingitis caused by the mouse pneumonitis biovar of Chlamydia trachomatis (MoPn) was used to determine the existence of non-cultivable organisms in vivo. Following intravaginal inoculation, mice yielded high chlamydial counts for 7-14 dyas, with a decline in culture-positivity by 21-28 days. A significant elevation of IFN gamma production in infected tissues was measured for 21 days and, from 28-70 days, all mice were culture-negative and developed characteristic hydrosalpinges. MoPn was detected by PCR in vaginal swabs of 80% and 69% respectively of culture-negative animals at 21 and 28 days. In a second study, 100%, 63% and 50% of culture-negative genital tissue homogenates were PCR-positive at 21, 28 and 42 days. Immunosuppression with either cyclophosphamide or hydrocortisone failed to regenerate cultivable chlamydiae. Tissues were disrupted by homogenization and inoculated intranasally to MF1 mice which are extremely susceptible to MoPn, but all culture-negative specimens were non-infectious. The significance of the PCR-positive culture-negative specimens requires further investigation, since these may represent a non-cultivable state in the deeper tissues of the mouse genital tract which may be beyond the reach of reactivating triggers.

Animals↗
PubMed1997

Ultrastructural and molecular analyses of the persistence of Chlamydia trachomatis (serovar K) in human monocytes.

Previous studies have suggested that monocytes may play a role in the dissemination of Chlamydia trachomatis, and in establishment of persistent infection with this bacterium. Infection of cultured human peripheral blood monocytes with C. trachomatis serovar K produced persistent, nonproductive infection. Transmission electron microscopy of such infected cultures revealed single or multiple Chlamydia in monocyte inclusions over a culture period of 10 days. Those inclusions were aberrant, and normal reticulate bodies within the inclusions were not observed. Immunoelectron microscopy showed the chlamydial major outer membrane protein and lipopolysaccharide to be associated with the bacterial plasma membrane. Lipopolysaccharide was also identified in the monocyte cytoplasm. Molecular analyses of primary chlamydial rRNA transcripts demonstrated that the organism is viable and metabolically active within monocyte inclusions. However, attempts to overcome chlamydial growth arrest by incubation of Chlamydia-infected monocytes with tryptophan, and antibodies against alpha interferon, gamma interferon, or tumor necrosis factor, were all ineffective, suggesting that known mechanisms of growth inhibition do not hold in human monocytes. These observations indicate that infection of human peripheral blood monocytes with C. trachomatis may be involved in the genesis/maintenance of extra-urogenital inflammation, since non-culturable, metabolically active bacteria persist in those cells.

Antibodies, Blocking↗
PubMed1989

Prevalence of asymptomatic carriers of Chlamydia trachomatis among pregnant and non-pregnant women in south-western Greece.

One-hundred-ninety-three pregnant and seventy five non-pregnant women, aged 18-35 years, were examined for endocervical infection with Chlamydia trachomatis using an enzyme immunoassay technique on cervical specimens. Chlamydia trachomatis was discovered in 7.2% of the pregnant women and in 10.6% of the non-pregnant ones. There was no correlation between chlamydia infection and either previous vulvovaginitis, previous pregnancies or pregnancy outcome. However, younger, more highly educated women with high sexual promiscuity tended to be more frequently colonized.

Adolescent↗
PubMed1988

Chlamydia trachomatis and pregnancy: a cross-sectional study in pregnant Viennese women.

To identify the epidemiologic role of Chlamydia trachomatis in Vienna and its incidence in the first and third trimesters of pregnancy, 1116 pregnant women (age range 16-43 years, medium age 26) were screened. Of 459 women applying for legal abortion in the first trimester of pregnancy, Chlamydia trachomatis was detected from the cervices of 24 (5.2%). Chlamydial detection was also performed in 657 women during the thirty-fourth week of pregnancy. The infection rate was found to be very similar to that of the other group (5.9%). The diagnosis was established by the direct immunofluorescence test. The number of elementary bodies did not differ between the two groups, indicating that pregnancy itself may not influence shedding of Chlamydia trachomatis from the cervix.

Adolescent↗
PubMed1988

Diagnosis of Chlamydia trachomatis infection--culture versus serology.

The diagnostic value of different laboratory methods in detecting Chlamydia trachomatis infections in high risk groups was analysed. The efficiency of a direct specimen test was compared with serology (IgG and IgM ELISA) and culture in L929 cells, stained either with fluorescein conjugated monoclonal antibodies or with iodine. Patients (no. = 1041) with localized genital infections attending a STD clinic, sexual contacts and patients with ascending infections from urological and gynecological clinics were examined. Chlamydia trachomatis was detected in 225 patients: 210 (93.3%) were reactive in the direct test (smears stained with monoclonal antibodies), whereas culture missed only 5 (sensitivity 97.8%) when stained by the same method. Cultures stained with iodine produced the lowest recovery rate (73.8%), but this rate increased to 80.9% when a second passage was performed. In addition the prevalence of Neisseria gonorrhoeae, Mycoplasma hominis, Ureaplasma urealyticum, Candida albicans and Trichomonas vaginalis was investigated. In patients with non-gonococcal urethritis (no. = 331) and cervicitis (no. = 353), Chlamydia trachomatis was isolated in 32.3% and 12.8% respectively. However, this pathogen could be isolated in only 3 (15.8%) out of 19 patients with epididymitis and 15 (14%) out of 107 patients with adnexitis, although 66.7% and 93.3% respectively had specific IgG antibodies. Specific IgM could by detected with a sandwich ELISA in patients with adnexitis (46.7%), epididymitis (33.3%), cervicitis (22.2%), non-gonococcal urethritis (14%) and in the sexual partners of patients with genital infections (35.7%). The direct specimen test with monoclonal antibodies is the method of choice for the diagnosis of a C. trachomatis infection in patients with urethritis and cervicitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1985

Recent advances in Chlamydia trachomatis.

Chlamydia trachomatis is an obligate intracellular energy parasitic bacterium with a genome of 660 X 10(6) daltons, possessing a plasmid and unique life cycle which includes the differentiation of the infective elementary body to a replicative reticulate body. C. trachomatis is the etiological agent of trachoma, which affects approximately 500 million people in developing countries. Recently it became evident that in industrialised Western nations certain strains of C. trachomatis are the most common cause of sexually transmitted infections such as non-gonococcal urethritis, cervicitis, endometritis, salpingitis and subsequent ectopic pregnancies or infertility, perihepatitis, neonatal conjunctivitis and pneumonia, adult conjunctivitis and epididymitis. Since C. trachomatis infections are often asymptomatic, widespread screening of sexually active young people is needed in order to initiate early antibiotic treatment which may prevent serious complications such as ectopic pregnancies and infertility. Development of sensitive and simple techniques for mass screening for detection of Chlamydia in excretions as well as techniques for detection of specific markers of chronic internal infections (such as Chlamydia specific IgA antibodies) is of great importance.

Chlamydia Infections↗
PubMed1991

Chlamydiae as pathogens--an overview of diagnostic techniques, clinical features, and therapy of human infections.

Chlamydiae are Gram-negative bacteria with obligate intracellular reproduction and disability to synthesize high-energy compounds such as ATP. Their cycle of development is unique among the prokaryotes: the host cells, mainly epithelial cells, are infected by so-called elementary bodies (EB) which undergo reorganization to form metabolically active reticulate bodies (RB). These RB multiply by binary fission, and after transition into infectious EB they are released within 48-72 hours. Chlamydiae cause prolonged subclinical infections of the conjunctiva, lung, cervix, and urethra. Complications in newborns are inclusion conjunctivitis, nasopharyngitis and pneumonia; in females, salpingitis, infertility, and perihepatitis; in male patients, epididymitis and prostatitis; and in both sexes, Chlamydiae-induced arthritis. Identification of the pathogenic agent confirms clinical diagnosis; tissue culture identification remains the diagnostic method of choice. Therapeutical drugs are tetracycline, erythromycin, josamycin, and in certain cases quinolone derivatives.

Animals↗
PubMed1990

The correlation between Chlamydia antigen, antibody, vaginal colonization and contraceptive method in young unmarried women.

In a group of 55 unmarried women, mean age 25 years, attending a family planning clinic and having minor gynecological complaints, the correlation between Chlamydia trachomatis (CT) antigen, CT antibodies, vaginal colonization by Candida or bacteria and the method of contraception was investigated. The correlation between CT antigen and CT antibodies (IgG) was significant in oral contraceptive users (p = 0.003), as was the correlation with vaginal colonization by Candida and potential pathologic bacteria. In the group using the natural family planning method, a statistically significant correlation was found between CT antigen, IgG (p = 0.002), IgA (p = 0.02) antibodies, and vaginal candidiasis (p = 0.002), but not with bacterial colonization (p = 0.90). The discrepancy between CT antigen and antibodies is discussed. Differences in the prevalence of Chlamydia trachomatis infection were found among groups using different birth control methods, indicating an association between Chlamydia infection and the contraceptive method used.

Adolescent↗
PubMed1986

Quinolones in the treatment of gonorrhoea and Chlamydia trachomatis infections.

123 Female patients suffering from uncomplicated urogenital gonorrhoea were treated with either 200 mg or 400 mg enoxacin. The cure rate in the 400 mg group was 100%; the cure rate in the 200 mg group was 95.7% 212 Male patients suffering from urethral gonorrhoea were treated with either 250 mg or 500 mg ciprofloxacin (one tablet). Cure rates in both groups were 100%. Post-gonococcal urethritis was observed in 31 out of 85 (36%) patients in the first, and 21 out of 79 (27%) in the second group. In a pilot study 42 male patients suffering from non-gonococcal urethritis were treated during one week with I g ciprofloxacin daily. In 22 patients Chlamydia trachomatis was isolated from the urethra: in 20 of these 22 cases Chlamydia trachomatis could not be cultured after treatment (cure rate 91%), but in 4 of these 20 cases (20%) and in 8 of the 20 Chlamydia trachomatis negative cases (40%) urine-sediment abnormalities were present after treatment.

Adolescent↗
PubMed1993

Lack of evidence of Chlamydia pneumoniae infection in infants with acute lower respiratory tract disease.

In order to determine whether there is serologic evidence of Chlamydia pneumoniae infection in young infants with acute lower respiratory tract infection, serum samples from 86 subjects aged less than 6 months were assayed for IgG and IgM antibodies to Chlamydia pneumoniae using a microimmunofluorescence method. Infants hospitalized in Toronto, Canada, were enrolled between 15 March 1991 and 15 March 1992. No patient had infection determined by the serologic results. IgG antibody was detected at low concentrations in 32 patients, with an inverse relationship between titer and chronological age. In our setting, Chlamydia pneumoniae does not appear to be an important lower respiratory pathogen in young infants.

Acute Disease↗
PubMed1986

Diagnosis of genital Chlamydia trachomatis infection in males by cell culture and antigen detection test.

Urethral Chlamydia trachomatis infection was diagnosed in 204 of 1,011 (20.2%) male patients by cell culture, in 219 (21.7%) by an antigen detection test consisting of a solid phase immunoassay, and in 247 (24.4%) patients by both methods combined. The positive results of the two methods agreed for 176 patients, and both positive and negative results of the tests agreed for 940 patients (93%). With cell culture as the reference method, the antigen detection test had a sensitivity of 86.3%, a specificity of 94.7%, a positive predictive value of 80.4% and a negative predictive value of 96.5%. It gave false negative results in 28 patients. In 43 patients the antigen detection test gave a positive result, whereas culture was negative. Thirty-nine of these males were treated with antibiotics (tetracycline or erythromycin), 19 because their consorts had a proven Chlamydia trachomatis infection, and 20 for obvious clinical and/or microscopic findings of urethritis requiring treatment. According to this analysis there were 19 probable misses by cell culture test and four true false-positives by the antigen detection test, i.e. less than 0.4% of all patients examined. Since one-third of males with a final diagnosis of Chlamydia trachomatis infection were clinically asymptomatic efforts to control genital chlamydial infections must identify this reservoir. The antigen detection test provides an alternative diagnostic method to the more laborious and time-consuming cell culture procedure.

Antigens, Bacterial↗
PubMed1987

Chlamydia trachomatis serology in ankylosing spondylitis.

Demonstration of chlamydial antibodies in patients with ankylosing spondylitis (AS) could show an etiological role of Chlamydia trachomatis in this condition. We studied serum specimens from 50 HLA-B27 positive patients with AS (Group I), 34 HLA-B27 positive patients with other rheumatic diseases (Group II), 67 HLA-B27 positive healthy blood donors (Group III) and 37 healthy untyped blood donors. (Group IV). Measured by an immunoperoxidase assay (IPA) chlamydial IgA (titre greater than or equal to 1:20) was more prevalent in the HLA-B27 positive persons than in the healthy controls not selected for HLA-group (Groups I + II + III vs IV : p less than 0.02). Chlamydia trachomatis IgA-IPA containing sera also had specific IgG-IPA antibodies (greater than or equal to 1:80) in 29 (96%) out of 30 sera from HLA-B27 positive individuals and controls. Conversely, 45% of specific IgG-positive (greater than or equal to 1:80) AS sera, 27.7% sera in Group II, 39.4% Group III sera vs. 11.1% of sera in Group IV had concomitant chlamydial IgA (greater than or equal to 1:20). The differences in the prevalence of specific IgA were statistically significant: Group I vs. IV : p less than 0.01; Group III vs. IV :p less than 0.05 and Gr. I + II + III vs. IV: p less than 0.05. Our data suggest an enhanced antibody production against Chlamydia trachomatis among the HLA-B27 positive individuals whether they have AS or are healthy.

Antibodies, Bacterial↗
PubMed1994

Detection of amplified Chlamydia trachomatis DNA using a microtiter plate-based enzyme immunoassay.

A microtiter plate-based method to detect amplified DNA was developed. The method uses on biotin-labeled primer in the polymerase chain reaction (PCR) mixture. The labeled amplicon is bound to streptavidin-coated microtiter plates, denatured and hybridized to a digoxigenin-labeled probe. The specificity of the hybridization reaction was optimized by varying the temperature of the subsequent washing step and adding urea to the washing buffer. The digoxigenin label was detected using an enzyme immunoassay (EIA). This PCR-EIA was compared with a standard PCR assay that uses gel electrophoresis, blotting and hybridization to detect the amplicon, with isolation in cell culture, and with an antigen detection EIA (Chlamydiazyme) in the diagnosis of Chlamydia trachomatis infection in 309 female patients attending a sexually transmitted diseases outpatient clinic. The prevalence of Chlamydia trachomatis infection as determined by isolation in cell culture, EIA, PCR-EIA and standard PCR assay was 9.1%, 8.7%, 12.3%, and 12.9%, respectively. Compared with results of a reference set of confirmed-positive cases (defined by a positive result in two or more independent assay after analysis of discrepancies), the sensitivity and specificity was 71.1% and 99.6% for cell culture, 65.8% and 99.3% for the EIA, 92.1% and 98.9% for the PCR-EIA, and 97.4% and 98.9% for the standard PCR assay. It is concluded that the PCR-EIA described is a fast, sensitive and specific method for detecting Chlamydia trachomatis in clinical specimens.

Amino Acid Sequence↗
PubMed1994

Intracellular location of inapparently infecting Chlamydia in synovial tissue from patients with Reiter's syndrome.

Culture of Chlamydia trachomatis from synovial tissues/fluids from Reiter's syndrome (RS) patients frequently yields negative results. However, we have identified chlamydial RNA at that site in such patients, suggesting that viable organisms may be present. Here we define the cellular location of chlamydia within the synovium via in situ hybridization. Using a chlamydial ribosomal RNA-directed probe, we show that synovial tissue from culture-negative RS patients gives strong hybridization which is often localized to a subsynovial cell layer, rather than to the synovial lining; in some cases, hybridizing cells are dispersed through the synovium. All hybridization signal is located within host cells, indicating that infectious extracellular elementary bodies are rare or absent. These data confirm the extensive intracellular presence of inapparent chlamydia in the synovia of RS patients and provide some insight into the usual culture negativity of synovial tissues for the organism.

Adult↗