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A peptide of Chlamydia trachomatis shown to be a primary T-cell epitope in vitro induces cell-mediated immunity in vivo.

Chlamydiae are a major cause of infertility and preventable blindness and there is currently no effective vaccine in humans or rodents against these organisms. We have previously shown that a peptide of 12 amino acids (termed TINKP) from a conserved region of the major outer membrane protein (MOMP) of Chlamydia trachomatis (C. trachomatis) is a primary T-cell epitope in humans. Here we showed that when dendritic cells (DC) from C3H or BALB/c mice were pulsed in vitro with the peptide they stimulated proliferation of syngeneic T cells in vitro indicating that the peptide is also a primary T-cell epitope in mice. Since the skin is a rich source of DC, we immunized mice from each strain with an intradermal injection of the peptide. Humoral and cell-mediated immunity to peptide, MOMP or whole elementary bodies (EB) of C. trachomatis (F/NI1/GU) were assessed. No antibody response to TINKP was observed. However, immunized mice showed recall responses to all three chlamydial antigens. T-cell-mediated immunity in the absence of antibody was induced by a single injection of the peptide intradermally. C. trachomatis isolated from the human genital tract causes salpingitis in mice. Preliminary studies in susceptible C3H mice indicated that intradermal injection of peptide conferred some protection against the development of salpingitis. Thus, a primary T-cell epitope identified by in vitro stimulation using DC can also initiate cell-mediated immunity in vivo and this approach may be useful in the development of vaccines.

Animals↗

Chlamydia trachomatis - the agent.

Chlamydiae are obligate intracellular bacteria, parasitizing eukaryotic cells. Chlamydia trachomatis, C. psittaci and C. pneumoniae are the three species of chlamydiae pathogenic to humans. C. trachomatis shows a tropism for the genital and conjunctival epithelia and consists of 19 different serovars which are pathogenic predominantly for the urogenital tract.A distinguishing feature of chlamydiae is their transition between the infectious elementary body that enters the host cell and the non-infectious reticulate body that replicates intracellularly within an inclusion that does not fuse with lysosomes. Chlamydiae depend for some functions upon the host cell; in particular, chlamydiae have little capacity for generating energy. The complete sequence of the 1000-kb chromosome of C. trachomatis is known, as are most of the genes located on the 7.5-kb cryptic plasmid. Recently, several concepts about the biology and the metabolic pathways of C. trachomatis have been revised in relation to the genome sequence, and different novel proteins have been described.

Chlamydia Infections↗

The immunology of Chlamydia trachomatis.

Chlamydia trachomatis (C. trachomatis) is one of the most common sexually transmitted bacterial agents. What distinguishes it from other organisms is its intracellular reproductive cycle. Up to now, four antigens have been identified in the Chlamydia genus: genus-specific antigen as well as species-specific, type-specific and subspecies-specific. C. trachomatis is a powerful immunogen which stimulates the host's immunological processes. The intracellular parasitism of the bacteria is the basis for both symptomatic or asymptomatic infection as well as for chronic ones. The primary infection leads to a local inflammatory reaction due to penetration and reproduction of the bacteria in the epithelial cells and to IgA secretory antibody production. In most cases the host's reaction to the primary infection is transient and does not cause tissue damage. In the course of chronic infection or reinfection, the most important processes are those of delayed hypersensitivity, which lead to a fast and intense immunological reaction of specifically sensitized Th1 lymphocytes. This reaction leads to progressive damage of the epithelial cells and to cicatrization and fibrosis, which means irreversible complications. Interferon gamma is of special importance in the process of C. trachomatis infection. High concentrations of it inhibit the bacteria's reproductive cycle, while lower concentrations promote the development of atypical, non-contagious forms of Chlamydia of diminished metabolic activity and altered antigenicity. The chlamydial heat shock proteins are considered to be of great importance lately. Their molecular weights of 60 and 10 kDa are a powerful stimulant of immunological reactions and show significant homology (40-90%) to human and other bacterial heat shock proteins.

Animals↗

[Detection of Chlamydia trachomatis antigen for diagnosis of respiratory infection I. Cross reaction with bacteriae consisting normal flora respiratory tract].

We examined the cross reaction of Chlamydia trachomatis (C. trachomatis) by antigen detection method using EIA against the aerobic bacteria which colonize in the respiratory tract of children. Chlamydiazyme showed a cross reaction among 5 out of 7 species of Gram negative bacteria (= 71.4%) and Neisseria and Branhamella catarrhalis showed a cross reaction even in low concentrations of 1 X 10(3-4) CFU/ml. There were no Gram positive bacteria which showed cross reaction. IDEIA Mark III did not react to Gram negative bacteria even in high concentrations of 1 X 10(7-10) CFU/ml. One strain of coagulase positive staphylococcus (CPS) showed positive at a concentration of 1 X 10(8): however, other Gram positive bacteria including two other strains of CPS exhibited no cross reaction. The detectable concentration of C. trachomatis (D/UW 3 strain) using IDEIA Mark III was 1 X 10(3) IFU/ml. Low cross reaction rate and sensitivity suggest IDEIA Mark III is preferable for diagnosis of respiratory infection of C. trachomatis though further clinical studies are necessary.

Antigens, Bacterial↗

Phylogenetic relationship of Chlamydia pneumoniae to Chlamydia psittaci and Chlamydia trachomatis as determined by analysis of 16S ribosomal DNA sequences.

The 16S ribosomal DNA sequence of Chlamydia pneumoniae was determined and compared with the corresponding gene sequences of Chlamydia psittaci and Chlamydia trachomatis. C. pneumoniae has been reported to exhibit little chromosomal DNA homology with the other chlamydial species, and its phylogenetic relationships within the genus Chlamydia have not been described. A polymerase chain reaction was employed to determine the 16S rRNA gene sequence of C. pneumoniae. Ten primers from the C. psittaci sequences were used to amplify a C. pneumoniae template in overlapping segments of the gene. Sequence data for 1,554 bases indicated that the levels of homology of C. pneumoniae with C. psittaci and C. trachomatis were 96.19 and 94.07%, respectively. These data support the results of previous biochemical and developmental studies indicating that C. pneumoniae is more closely related to C. psittaci than to C. trachomatis.

Base Sequence↗

Chlamydia trachomatis IncA is localized to the inclusion membrane and is recognized by antisera from infected humans and primates.

Chlamydia psittaci produces a collection of proteins, termed IncA, IncB, and IncC, that are localized to the chlamydial inclusion membrane. In this report we demonstrate that IncA is also produced by Chlamydia trachomatis. C. trachomatis IncA is structurally similar to C. psittaci IncA and is also localized to the inclusion membrane. Immunoblot analysis demonstrated that sera from C. trachomatis-infected patients and from experimentally infected monkeys both recognized C. trachomatis IncA.

Animals↗

Detection of HIV-1 and HIV-2 antibodies among Chlamydia trachomatis infected pregnant women in Japan.

An anonymous unlinked HIV antibody test was conducted on 1632 Chlamydia trachomatis (C. trachomatis) antibody positive women from 10 institutes of 7 prefectures in Japan. All the sera were negative for both HIV-1 and HIV-2 antibodies. The result may support the suggestion that HIV prevalence is low among general population in Japan. Such a test as this study will be useful not only for developing a reliable HIV surveillance system but also for the study of sexual behavior of general population, since C. trachomatis infection is sensitive to reflect sexual contact.

Adult↗

Chlamydial rRNA in the joints of patients with Chlamydia-induced arthritis and undifferentiated arthritis.

Synovial fluid and synovial membrane specimens of 11 patients with Chlamydia-induced arthritis (CIA), 24 patients with undifferentiated arthritis (UndA), 4 patients with post-enteritic reactive arthritis, 3 patients with Lyme arthritis and 9 patients with rheumatoid arthritis were investigated for the presence of Chlamydia trachomatis (C. trachomatis). A single stranded DNA-probe was used for nucleic acid hybridization with ribosomal RNA (rRNA) from C. trachomatis. In 4 patients (CIA = 1, UndA = 3) chlamydial rRNA was found in the synovial fluid. In one additional patient (CIA) the specimen of a synovial membrane biopsy was positive for chlamydial rRNA. The detection of intra-articular chlamydial rRNA is discussed as an indicator for the presence of viable Chlamydiae in inflamed joints.

Arthritis, Infectious↗

Unusual course of a Chlamydia pneumonia in an infant with IgG2/IgG4-deficiency.

An unusual case of Chlamydia trachomatis (C. trachomatis) pneumonia, complicated by the development of a pneumothorax, is reported in an IgG2/IgG4 deficient infant delivered by caesarean section. C. trachomatis was isolated initially from a throat smear and subsequently from pleural effusions. Serological examinations using the complement fixation test were negative in sera of both mother and child. However, using immunofluorescence the presence of an acute or recent infection was confirmed by IgM-antibodies in the serum of the infant and IgA-antibodies in the serum of the mother. At the age of 7 months the girl suffered from impetigo contagiosa which was partially resistant to antibiotic treatment. IgG-subclass deficiency was diagnosed after the onset of this disease and the girl was then treated by immunoglobulin transfusion.

Antibodies, Bacterial↗

[Chlamydia trachomatis in patients with genital contact infections in Austria--studies of 3,367 patients].

In order to evaluate the epidemiological importance of Chlamydia trachomatis (C. trachomatis) as a genital microorganism, data were obtained from 3,367 patients with sexually transmitted diseases in Vienna and analyzed by computer-assisted methods. C. trachomatis was cultured in 26.1% of 2,594 patients investigated for the first time. The microorganism was found more often in male patients (31.3%) than in female patients (21%). 32.2% of positive Chlamydia cultures were obtained from patients with non-gonococcal urethritis (NGU) and 64% from postgonococcal urethritis (PGU) patients. A high coincidence with Neisseria gonorrhoeae (N. gonorrhoeae) was detected in males (31.2%) and females (43.5%). Data on Mycoplasma hominis (M. hominis), and Ureaplasma urealyticum (U. urealyticum) show that, in contrast to the low incidence of M. hominis and U. urealyticum in males, the organisms were found predominantly in females.

Adult↗

[Study on the inhibitory effects of minocycline on genital Chlamydia trachomatis in McCoy cell culture].

In this study of Chlamydia trachomatis (C. trachomatis) from genital origin in McCoy cells, minocycline (MINO) was added to infected cell cultures at specific stages of their growth cycle. Sequential observations were made at each stage by electron microscope with the following results: 1) The minimal inhibitory concentration of MINO against C. trachomatis in McCoy cells was 0.025 micrograms/ml. 2) When MINO and chlamydial inoculation were simultaneously added (inoculum containing 1 microgram of MINO per ml), MINO did not inhibit attachment and invasion of elementary bodies (EBs) to McCoy cells in both cases of inoculation, with and without centrifugation. As 48 hours after inoculation 74% of the infectious EBs in McCoy cells were converted to early intermediate forms (IFs). This suggests that MINO did not inhibit the transformation of EBs to early IFs. Under the influence of MINO, even after 48 hours, reticulate bodies (RBs) and chlamydial inclusions were not recognized. At the stages of transformation of infectious EBs to early IFs, duplication of the cell wall and electron lucent areas in the cytoplasm were observed. 3) Meanwhile, in the case of replacing the media with one containing MINO (1 microgram/ml) at 12 hours after inoculation, the conversion of RBs to late IFs was blocked. 4) Electron microscopically, the average numbers of late IFs and EBs per cross section of the inclusion at 24 hours after inoculation, were 0.64 and 0.17, respectively. At 48 hours after inoculation without MINO (control), the average numbers of late IFs and EBs were 10.5 and 65.6, respectively. When MINO (1 microgram/ml) was added at 24 hours after inoculation and observed at 48 hours, the average numbers of late IFs and EBs numbered 0.10 and 0.67, respectively. This indicates A) statistically, a significant decrease (0.64----0.10) for late IFs and a significant increase (0.17----0.67) for EBs compared with the average number of those at 24 hours after inoculation, B) when compared with the average number of those at 24 hours after inoculation, the sum of late IFs and EBs were nearly the same (0.64 + 0.17 not equal to 0.10 + 0.67), C) somewhat similar ratios of late IFs to EBs (10.5:65.6 = 1:6.2 not equal to 0.10:0.67 = 1:6.7) compared to the average number of those at 48 hours after inoculation without MINO (control). B) suggests that the conversion of RBs to late IFs was blocked by MINO. A), B) and C) suggest that the conversion of late IFs to EBs was not inhibited by MINO.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

[Chlamydia trachomatis in the urogenital tract of pregnant women].

In two hundred and thirty three prenatal patients from the Prenatal Clinic of the University of Würzburg cervical, vaginal and urethral swabs were taken for microscopic examination and culture for Chlamydia trachomatis (C. trachomatis), Neisseria gonorrhoeae (N. gonorrhoeae), Trichomonas vaginalis (T. vaginalis), Beta-hemolytic streptococci, Candida species, Ureaplasma urealyticum (U. urealyticum), Mycoplasma species and facultatively pathogenic and apathogenic aerobic bacteria. In ten cases (4%) C. trachomatis was found. In six cases hemolytic streptococci group B were detected. Fifty two cases showed a Candida species. Seventy nine cases presented U. urealyticum. One case showed a Mycoplasma species and one case showed Trichomonas vaginalis. Striking was the significantly increased rate of premature rupture of the membranes in the group with U. urealyticum compared to the group without U. urealyticum.

Adult↗

Evaluation of an enzyme immunoassay for detection of Chlamydia trachomatis in urine of asymptomatic men.

In a study of 1,486 men attending two sexually transmitted disease clinics, of whom 891 had no symptoms of urethritis, we compared an enzyme immunoassay (EIA) (Baxter-Bartels, formerly Northumbria AntigEnz) of urine sediment to urethral culture for the detection of Chlamydia trachomatis. C. trachomatis prevalence by culture alone was 7.7% in asymptomatic men and 10.9% in symptomatic men. Discrepant results between EIA of urine and urethral culture were evaluated by direct fluorescent-antibody staining (DFA) for elementary bodies in urine sediment or in culture transport media. When chlamydial infection was defined as either a positive urethral culture or positive EIA confirmed by DFA, chlamydia prevalence increased to 8.9% in asymptomatic men and 11.6% in symptomatic men. The urine EIA sensitivity, specificity, and positive and negative predictive values for chlamydia detection in asymptomatic men were 84.8, 99.3, 91.8, and 98.5%, respectively, with nearly identical results for symptomatic men. The sensitivities of urethral culture alone compared with the combination of urethral culture and urine EIA (with DFA confirmation) were 87.3 and 94.3% for asymptomatic and symptomatic men, respectively. The present EIA of urine sediment is both highly sensitive and specific for the detection of C. trachomatis in asymptomatic men, thus providing a noninvasive screening method for chlamydia infection in asymptomatic men attending sexually transmitted disease clinics.

Adolescent↗

[Prevalence of antibodies to Chlamydia pneumoniae (strain TWAR) and other Chlamydia in Japan].

Chlamydia pneumoniae (Strain TWAR), the new third species of genus Chlamydia, is emerging as an important human respiratory pathogen in Western countries. However, little is known about it in Japan. In this study, 1330 serum samples were tested for seroprevalence of three species of genus Chlamydia including C. pneumoniae in Hiroshima. Microimmunofluorescence test was used to measure serum IgG antibody, and formalin fixed elementary bodies of C. trachomatis, C. psittaci, and C. pneumoniae were used as antigen. Serum titer of 1:32 or greater was considered positive. Seropositive rate of C. pneumoniae began to increase around the 6 month to 3 year old age group and reached 44% in the 8 to 11 year olds. A peak of seroprevalence according to age was around 20 year old, and about half of the adults were seropositive. Outpatients and healthy persons held almost the same seropositive rate. The positive rate in childhood was higher and the peak appeared earlier than some former studies in Western countries. The seropositive rate of C. trachomatis was noted 1-2% in the 6 month-7 year old age group, but no seropositive child was found in the 8-15 year olds. The rate was increased after adolescence and reached 10% in adults. The seroprevalence of C. psittaci was 1%. These data suggest C. pneumoniae is the most prevalent Chlamydia in Japan and as prevalent as in the Western countries.

Adolescent↗

Detecting of Mycoplasma genitalium in male patients with urethritis symptoms in Turkey by polymerase chain reaction.

OBJECTIVE: The aim of this study was to investigate the incidence of Mycoplasma genitalium in the urine samples of 63 male patients who had urethritis symptoms. Along with Neisseria gonorrhoeae (N. gonorrhoeae) and Chlamydia trachomatis (C. trachomatis). We also investigated Mycoplasma hominis (M. hominis) and Ureaplasma urealyticum (U. urealyticum), both of which are known to cause urethritis. METHODS: Microorganisms were investigated in urine samples of the patients with polymerase chain reaction. The study was conducted between September 2003 - February 2004 at the Department of Microbiology and Clinical Microbiology Ankara University School of Medicine, Ankara, Turkey. RESULTS: A total of 63 urine samples were analyzed and 6 (9.52%) patients had N. gonorrhoeae, 4 (6.34%) had C. trachomatis, while 4 (6.34%) urines were positive in terms of M. genitalium. Nevertheless, 3 (4.76%) patients had U. urealyticum and 2 (3.17%) patients had M. hominis. One urine sample was positive in terms of both N. gonorrhoeae and U. urealyticum, and another urine sample was positive in terms of both M. hominis and U. urealyticum. The results were compared with the control group and found no statistically significant difference. CONCLUSION: Mycoplasma species are found in normal flora of urogenital system and also as an agent of urogenital infection. In our study, we found low microorganism rates when compared with Europe and America. This difference may be due to the conservative sexual behavior in Turkey.

Chlamydia trachomatis↗

[Genetic diagnoses of Chlamydia trachomatis--DNA probe and PCR method].

Chlamydial infections such as male urethritis or female cervicitis, although not receiving as much attention as the AIDS epidemic, are on the increase around the world as sexually-transmitted diseases (STD). A number of diagnostic methods i.e., direct fluorescence antibody method and enzyme immunoassay (EIA) methods, have been developed for detecting Chlamydia trachomatis (C. trachomatis) infection, but they each have a defect in terms of sensitivity or specificity. Then, to overcome these disadvantages, the genetic diagnostic method such as DNA-RNA hybridization (DNA probe) method, PCR method and LCR method have been clinically introduced. In in vitro studies, the PCR method has higher sensitivity than the EIA method or DNA probe method. All genetic diagnostic methods have specificity superior over the EIA method. In clinical studies, the PCR method was the most useful for detection of C. trachomatis from various infectious lesions.

Chlamydia Infections↗

Gynecological chlamydial infections.

A review is presented of infections in the female genital tract caused by Chlamydia trachomatis. C. trachomatis primarily infects the cuboidal epithelium of the cervix. Chlamydial cervicitis is four to six time more common in Europe than gonococcal cervicitis. The infection is sexually transmitted and can cause urethritis in the male, endometritis and salpingitis in the female as well as infections in the neonate. The risk of salpingitis in women with chlamydia cervicitis has been estimated to be about 1:10, i.e. the same as for gonorrhoea. Chlamydial cervicitis is asymptomatic in about half of the culture-positive cases. A correlation between colposcopic follicular cervicitis and the isolation of C. trachomatis has been reported. Examination of serum specimens for IgG antibody to the organism is of little use for the diagnosis of cervicitis. Treatment with macrolide antibiotics or tetracyclines eradicates the organism.

Adolescent↗

Diagnosis of Chlamydia trachomatis infection.

Important progress in the diagnosis of Chlamydia trachomatis (C. trachomatis) includes the development of nucleic acid amplification techniques such as polymerase chain reaction (PCR) and ligase chain reaction (LCR). Commercial kits are available, but they are costly, sporadic in availability, must be imported, and are economically beyond the reach of common people. To overcome this limitation, most research laboratories have standardized their in-house-developed PCR methods for diagnosing this infection. However, each laboratory has to spend a great deal of time and money to accomplish this. Published reports do not always elaborate the steps involved in standardizing a test so that it can immediately be reproduced in another setting. In the present study we attempted to elaborate the steps involved in standardizing a sensitive and specific PCR technique followed by hybridization with specific C. trachomatis probe to diagnose this infection in cervical, introital, and urine specimens, and used it to determine the infection rate in a clinical population.

Chlamydia Infections↗