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PubMed2004

Cell death, BAX activation, and HMGB1 release during infection with Chlamydia.

Infection by a number of Chlamydia species leads to resistance of the host cell to apoptosis, followed by induction of host-cell death. In a population of infected cells that displays protection against staurosporine-induced apoptosis among the adherent cells, we find that cells that had been recovered from the supernatant share characteristics of both apoptosis and necrosis, as assayed by the propidium iodide (PI)-annexin V double-labeling technique. Cell death was observed in both an epithelial cell line and primary fibroblasts, although the primary cells had a higher propensity to die through apoptosis than the immortalized cell line. Staurosporine-mediated activation of the pro-apoptotic BCL-2 family member, BAX, was inhibited in the epithelial cell line infected for 32 h with the lymphogranuloma venereum (LGV/L2) but not the murine pneumonitis (MoPn) strain of C. trachomatis, but inhibition of staurosporine-mediated BAX activation disappeared after 48 h of infection with the LGV/L2 strain. Conversely, infection with MoPn (C. muridarum) but not LGV/L2 led to BAX activation after 72 h, as previously reported for shorter (48 h) infection with the guinea pig inclusion conjunctivitis (GPIC) serovar of C. psittaci (C. caviae). These results suggest that the ability to inhibit staurosporine-mediated BAX activation or to activate BAX due to the infection itself may vary as a function of the chlamydial strain. Interestingly, both the epithelial cells and the fibroblasts also released high mobility group box 1 protein (HMGB1) during infection, although much less HMGB1 was released from fibroblasts, consistent with the higher level of apoptosis observed in the primary cells. HMGB1 is released preferentially by necrotic or permeabilized viable cells, but not apoptotic cells. In the extracellular space, HMGB1 promotes inflammation through interaction with specific cell-surface receptors. Higher levels of HMGB1 were also measured in the genital-tract secretions of mice infected vaginally with C. trachomatis, compared to uninfected controls. These results suggest that cells infected with Chlamydia release intracellular factors that may contribute to the inflammatory response observed in vivo.

Animals↗
PubMed1995

Incidence of adenoviral and chlamydial coinfection in acute follicular conjunctivitis.

PURPOSE/METHODS: After we studied a case of chlamydial and adenoviral coinfection in a 20-year-old woman, we determined the incidence of chlamydial infection in patients with acute adenoviral conjunctivitis. In a randomized retrospective study, we evaluated 100 specimens of patients with culture-proven adenoviral conjunctivitis. RESULTS/CONCLUSIONS: Three of 100 (3%) specimens tested positive for chlamydial DNA using polymerase chain reaction. Adenoviral and chlamydial coinfection is rare, yet should be considered in patients with prolonged follicular keratoconjunctivitis.

Acute Disease↗
PubMed1981

Genital tract infection with Chlamydia trachomatis in prepubertal children.

Oculogenital strains of Chlamydia trachomatis are important causes of a wide range of genital tract infections in adults, but their recognized role in pediatric infections has been limited to neonatal inclusion conjunctivitis and interstitial pneumonia. We examined 23 prepubertal children with nongonococcal urethritis and vaginitis, and 31 with gonococcal anogenital infection. C. trachomatis was not found in 12 males and 11 females with nongonococcal urethritis or vaginitis. Nine of 33 episodes (27%) of gonorrhea were complicated by concurrent or subsequent chlamydial infection, including postgonococcal urethritis, vulvovaginitis, and asymptomatic anogenital colonization. C. trachomatis complicates gonococcal genital infections in prepubertal children at an incidence comparable to that seen in adults. The diagnostic and therapeutic approach to a prepubertal child with postgonococcal genital discharge should include consideration of this pathogen.

Age Factors↗
PubMed1987

Adult follicular conjunctivitis and neonatal ophthalmia in a Liverpool eye hospital, 1980-1984.

In the five year period between 1980 and 1984, 2146 adults and 172 neonates suffering from acute conjunctivitis underwent laboratory investigation for Chlamydia trachomatis (CT), Adenoviruses (AV), Herpes Simplex Virus (HSV) and pathogenic bacteria. Epidemiology and clinical features are presented and discussed. CT was detected in 29 per cent of neonates with conjunctivitis. 5.6 per cent of adults and older children investigated for follicular conjunctivitis were CT positive. There was a significant female preponderance among CT positive neonates of 1.9:1 (p less than 0.02). 91 per cent of neonates and 62 per cent of adults in whom CT was detected were receiving some sort of treatment. Serotypes 7, 3, 10, 4 and 8 were responsible in decreasing order of frequency for 96 per cent of AV infections. Serotype 7 was seen for the first time in an adult age distribution. HSV was isolated in 1.3 per cent of cases in the absence of typical lid or corneal lesions. Viral infection was not detected in any neonate. Bacterial infection was a more likely cause than CT in neonates if infection had persisted longer than 5 weeks (p much less than 0.001). Neonates with Staph aureus infection tended to present earlier in the course of disease than those with Haemophilus sp or Pneumococcus (p less than 0.05).

Adenoviridae Infections↗
PubMed1990

Per-nasal swabbing as an aid to the diagnosis of chlamydial and adenovirus conjunctivitis.

Two hundred and thirty four patients (adults and babies) with conjunctivitis were investigated by taking eye swabs and in addition by taking per-nasal swabs. Chlamydia trachomatis was isolated from 20 patients and adenovirus from 14 patients. Per-nasal swabbing led to a 53% increase in chlamydia diagnosis and a 27% increase in the diagnosis of adenovirus infection. It is suggested that per-nasal swabbing has an important role to play in detecting chlamydial conjunctivitis which itself may be an indicator for high morbidity in patients and their contacts.

Adenovirus Infections, Human↗
PubMed1991

The antigenic specificity of the humoral immune response to primary and repeated ocular infections of the guinea pig with the GPIC agent (Chlamydia psittaci).

The antigenic specificity of the humoral immune response in guinea pigs to primary and repeated ocular infections with the guinea pig inclusion conjunctivitis (GPIC) chlamydial agent was analysed using microbiological, serological and Western blotting techniques. The results indicate that although there was a response to many minor polypeptide antigens, there was a marked lack of reactivity to the major outer membrane protein (MOMP), particularly following reinfection of guinea pigs. It is suggested that, lack of a good antibody response to the MOMP, may be one of the reasons why guinea pigs are susceptible to repeated ocular infections with this chlamydial agent.

Animals↗
PubMed1997

Type III secretion genes identify a putative virulence locus of Chlamydia.

Four genes of Chlamydia psittaci strain guinea pig inclusion conjunctivitis (GPIC), whose predicted products are highly homologous to structural and regulatory components of a contact-dependent or type III secretion apparatus, were isolated. Related to genes present in several animal and plant bacterial pathogens, these genes may represent a section of a previously undetected chromosomal virulence locus analogous to several recently described virulence-associated type III secretion loci. The existence of contact-dependent secretion in Chlamydia strongly suggests that these bacteria use pathogenic mechanisms that are similar to those of other intracellular bacterial pathogens. Unlike other intracellular bacteria, however, chlamydiae are metabolically inactive extracellularly and only become capable of global protein synthesis several hours after infection. This implies that chlamydial contact-dependent secretion is only active from within, uniquely after the bacteria have been internalized by eukaryotic cells. The possible role(s) of this pathway in chlamydial pathogenesis are discussed.

Amino Acid Sequence↗
PubMed2001

Peptide immunization of guinea pigs against Chlamydia psittaci (GPIC agent) infection induces good vaginal secretion antibody response, in vitro neutralization and partial protection against live challenge.

Immunization of female guinea pigs with a chimeric peptide consisting of variable domain IV (VDIV) and a region known as GP8 from the major outer membrane protein of Chlamydophila caviae, formerly Chlamydia psittaci guinea pig inclusion conjunctivitis strain, was performed to assess whether humoral immune responses could be elicited in the reproductive tracts of immunized animals. The C. caviae strain is able to cause a sexually transmitted infection in the guinea pig that closely parallels C. trachomatis infections in humans. The best anti-VDIV antibody response in vaginal secretions was achieved by intraperitoneal priming with subsequent intravaginal boosting (P < 0.001). Dot-blot analyses of vaginal secretions confirmed that these anti-VDIV antibodies, produced against a linear peptide, were able to recognize and bind to whole conformational C. caviae elementary bodies. Following live intravaginal challenge with C. caviae, a significant reduction in the intensity (P = 0.01) and an apparent reduction in the duration of the infection was evident between the guinea pigs immunized with VDIV-GP8 and non-immunized controls.

Animals↗
PubMed1989

Chlamydial disease pathogenesis. Ocular hypersensitivity elicited by a genus-specific 57-kD protein.

Recurrent or persistent infections with Chlamydia trachomatis are thought to provide the antigenic stimulus for the chronic inflammation associated with blinding trachoma. We used the guinea pig model of inclusion conjunctivitis to identify chlamydial antigens that may be involved in this deleterious immune response. We purified from chlamydial elementary bodies a genus-specific 57-kD protein that elicited an ocular hypersensitivity response when placed topically onto the conjunctiva of ocular immune guinea pigs. This response was characterized by a predominantly mononuclear macrophage and lymphocyte cellular infiltrate of the submucosal epithelium. The clinical and histological findings were consistent with those of a delayed hypersensitivity response. These data demonstrated that the 57-kD chlamydial protein was a potent stimulator of ocular delayed hypersensitivity. Our findings may be critical to understanding the pathogenesis of the debilitating chlamydial diseases associated with chronic inflammation, such as trachoma and many urogenital syndromes.

Animals↗
PubMedDate not supplied

Pathogenesis and immunology of chlamydial infections of the genital tract.

In recent years there has been a steady increase in our understanding of the pathogenesis and immunobiology of lower genital tract infections caused by Chlamydia trachomatis. Much of this increase in knowledge has stemmed from two areas of research: clinical, pathologic, and immunologic observations of patients and the development and study of animal models. Research on patients has included the use of techniques such as histologic examination of biopsy specimens and measurement of levels of antibody and/or leukocyte responses to chlamydial antigens. The animal models that have been developed include experimental genital tract infections not only with C. trachomatis but also with appropriate Chlamydia psittaci organisms, such as the agent of guinea pig inclusion conjunctivitis. Continued investigation of the pathophysiologic and immunologic bases of chlamydial genital tract disease should lead to the rational development of potential control measures, such as vaccines or other forms of prophylaxis or therapy.

Animals↗
PubMed1978

Use of enteric vaccines in protection against chlamydial infections of the genital tract and the eye of guinea pigs.

Guinea pigs in a test group were fed living guinea pig inclusion conjunctivitis (GPIC) organisms classified as Chlamydia psittaci in 60% yolk-sac suspensions as enteric vaccines, while animals in a control group received uninfected yolk sac. Seven test animals and 14 control animals were challenged 11 or 22 days later with 1,000 50% infectious doses of GPIC organisms in either the conjunctiva or the vagina. Evidence of protection from mucosal infection in both sites was noted in test animals. Clinically, the disease was less severe, and microbiologically, lower percentages of mucosal cells were infected. The results suggest that enteric vaccination against mucosal infections of the eye and the genital tract with chlamydial agents is possible.

Administration, Oral↗
PubMedDate not supplied

Cystitis associated with chlamydial infection of the genital tract in male guinea pigs.

Male guinea pigs were infected with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC) by intraurethral injection of chlamydiae or by placement of a drop of chlamydial suspension on the meatus of the extruded penis. Transient urethritis and cystitis were observed in animals infected by either method. The production of cystitis by the drop-on technique indicated that chlamydiae are able to ascend the urethra and that the bladder may be a target organ of chlamydial infection. When infected animals were immunosuppressed with cyclophosphamide, the number of guinea pigs with cystitis was increased, and chlamydiae could be detected in the bladder for as long as 50 days after infection. In contrast, GPIC was not detected in the bladders of untreated animals after day 18.

Animals↗
PubMed1981

Chlamydial genital infections: manifestations and management.

Infection with Chlamydia trachomatis is an important cause of nongonococcal urethritis and cervicitis, and may be the most common sexually transmitted disease in the United States. Associated complications include epididymitis, proctitis, salpingitis, bartholinitis, arthritis, perihepatitis, and endocarditis. Perinatal transmission of infection may result in neonatal inclusion conjunctivitis and/or pneumonia of infancy. Chlamydial genital infection should be suspected in a patient (male or female) who presents with a gonorrhea-like syndrome but whose laboratory studies fail to demonstrate Neisseria gonorrhoeae. Such patients, together with their sex partners, should receive antichlamydial therapy; the uncomplicated genital infections respond well to oral treatment with tetracycline, erythromycin, and sulfonamide. The most important cause of treatment failure in nongonococcal urethritis is lack of simultaneous treatment of both patient and partner.

Erythromycin↗
PubMed1981

Polypeptide composition of Chlamydia trachomatis.

Strains of Chlamydia trachomatis representative of 14 serotypes were grown in HeLa 229 cells. HeLa cell susceptibility to chlamydial infection was increased by treating the host cells with DEAE-dextran. Optimal conditions of DEAE-dextran treatment were determined for each serotype of C. trachomatis to maximize chlamydial yields. Chlamydial polypeptides were selectively radiolabelled with 3H-labelled amino acids in the presence of emetine, an inhibitor of HeLa cell protein synthesis. The radiolabelled chlamydiae were purified from host cell components by density gradient centrifugation and their polypeptide composition was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The distribution of the major chlamydial polypeptide of 38 000 to 42 000 daltons amongst the different serotypes correlated closely with the predominant human infections caused by each serotype. Lymphogranuloma venereum agents possessed a unique polypeptide of 118 000 daltons not found amongst trachoma inclusion conjunctivitis strains of chlamydiae. Chlamydial surface polypeptides were selectively radiolabelled with 125I by lactoperoxidase-catalysed oxidation. The major chlamydial polypeptide and polypeptides of 155 000 and 29 000 daltons were thus identified as surface polypeptides of the chlamydial elementary body. It is suggested that the 155 000 dalton polypeptide is a species-specific antigen, the major polypeptide is the principal outer membrane protein, and the 29 000 dalton polypeptide is the type-specific antigen.

Chlamydia trachomatis↗
PubMed1986

Antigenic specificity of human antibody to chlamydia in trachoma and lymphogranuloma venereum.

An understanding of the molecular basis of the humoral immune response to chlamydial infections in man requires the identification of target antigens to which antibodies are directed. The antigenic specificity of antibody from patients with lymphogranuloma venereum (LGV) or trachoma was therefore assessed by Western blotting. Surface polypeptides were first identified using purified chlamydial outer membrane complex as antigen. Antibodies in sera from patients with LGV but not from control negative sera reacted with a wide range of chlamydial surface polypeptides with molecular masses of 19, 29, 41, 58, 63 and 65 kDa. The major component of the antibody response detected by both immunoblotting and immunoprecipitation assay was directed against the major outer membrane protein (MOMP). Antibody to MOMP was species-specific on Western blotting, whereas antibody to several other polypeptides recognized common immunodeterminants on polypeptides of C. psittaci Cal-10 of equivalent molecular mass. Immunologically C. psittaci Cal-10 was more closely related to LGV strains of C. trachomatis than a guinea pig inclusion conjunctivitis strain of C. psittaci. Trachoma sera collected from a village in southern Iran showed predominantly type-specific antibody on micro-immunofluorescence to serotype A or B trachoma agents. These sera showed a weak immune response to MOMP, a pronounced response to a polypeptide of 36 kDa and much less widespread reactivity with other chlamydial polypeptides. The lack of an immune response to SDS-stable immunodeterminants on MOMP might contribute to the susceptibility of trachoma patients to repeated cycles of ocular infection with chlamydiae.

Antibodies↗
PubMed1988

Distribution of plasmid sequences in avian and mammalian strains of Chlamydia psittaci.

Plasmid DNA from an avian strain of Chlamydia psittaci was purified and estimated to be 7.9 kb in size using restriction endonuclease analysis. A 5.9 kb fragment of this plasmid was cloned, mapped and used to screen a range of chlamydial strains. Hybridizing DNA was absent from ovine abortion and arthritis isolates and also from the Cal 10 strain but related sequences were detected in C. psittaci strains of feline pneumonitis, guinea-pig inclusion conjunctivitis, ovine conjunctivitis and C. trachomatis serovar L2. The plasmid DNA from the feline strain was shown to have a distinct restriction endonuclease profile. Similar plasmid sequences were detected in all avian isolates tested: thus the clone may have a useful diagnostic role for the detection of the pathogen in its natural host and in zoonotic episodes.

Animals↗
PubMed2006

Chlamydiales in guinea-pigs and their zoonotic potential.

The aim was to detect and characterize chlamydial infections in guinea-pigs (GP) with ocular disease, study their pathogenicity and zoonotic potential and to test for the presence of Acanthamoebae spp. in GP eyes and to investigate whether they could act as vectors for Chlamydia-like organisms. Overall 126 GP, of which 77 were symptomatic, were screened by clinical examination, cytology, gross pathology, histology, immunohistochemistry, polymerase chain reaction (PCR) and bacteriology. A new Chlamydiaceae-specific intergenic spacer rRNA gene PCR, designed to amplify this segment linking the 16S and 23S regions, was performed. DNA samples were also received from one owner including samples of his cat and rabbit. Guinea-pigs: 48 of 75 symptomatic, but only 11 of 48 asymptomatic GP were positive by PCR for Chlamydophila caviae guinea-pig inclusion conjunctivitis (GPIC) (P < 0.0001). Eighteen of 75 or 15/48, respectively, were positive for DNA from Chlamydia-like organisms. Acanthamoebae-DNA could be found in two GP, of which one was symptomatic. Owner, cat and rabbit: Samples of all three species were positive by PCR for C. caviae GPIC and the owner's one-day disposable contact lenses showed a positive PCR result for the Chlamydia-like organism Parachlamydia acanthamoebae. No Acanthamoebae-DNA could be detected. This study is the first to describe Chlamydia-like organisms in GP and to detect C. caviae GPIC in human, cat and rabbit. Therefore, C. caviae GPIC could pose a zoonotic potential. We believe that the finding of C. caviae GPIC in species other than GP is probably not unique.

Animals↗