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Susceptibility to reinfection after a primary chlamydial genital infection is associated with a decrease of antigen-specific T cells in the genital tract.

We tested the hypothesis that the intensity of specific antichlamydial T cell-mediated immunity in the genital tract of female guinea pigs infected intravaginally with the chlamydial agent of guinea pig inclusion conjunctivitis would determine the resistance or susceptibility to reinfection after a primary chlamydial infection. T cell-enriched lymphocytes were isolated by collagenase treatment of genital tract tissues from either infected or control uninfected female guinea pigs at various times after infection. The nylon wool-enriched T lymphocytes were evaluated for expression of antigen-specific T cell-mediated immunity in vitro by using a blast transformation assay. Both uninfected and infected genital tracts contained T cells, as evidenced by reactivity to concanavalin A, although a greater number of T lymphocytes was detected in the genital tracts of infected animals compared with that in controls. Significant antigen-specific T-cell activity could be detected in the genital tract tissue by 7 days after a primary genital tract infection with the chlamydial agent of guinea pig inclusion conjunctivitis. When antigen-specific activity was assessed at different times after infection, the intensity of the response of genital tract-associated T lymphocytes was directly proportional to the degree of resistance of the animals to genital challenge. Thus, susceptibility of animals to reinfection by chlamydiae appears to be associated with the intensity of the local T cell-mediated immune responses in the genital tract of infected animals.

Animals↗

Chlamydia trachomatis infections in neonates and young children.

In 1911, Lindner and colleagues identified intracytoplasmic inclusions in infants with a nongonococcal form of ophthalmia neonatorum called inclusion conjunctivitis of the newborn (ICN). Mothers of affected infants were found to have inclusions in their cervical epithelial cells, fathers of such infants had inclusions in their urethral cells, and the epidemiology of sexually transmitted chlamydial infections was revealed. Fifty years later, chlamydial isolation procedures were developed, and studies again demonstrated Chlamydia trachomatis as an etiology of ICN and the female birth canal as the reservoir. In the late 1970s, a report by Beem and Saxon described respiratory tract colonization and a distinct pneumonia syndrome in infected infants. Genital chlamydial infection is recognized as the world's most common sexually transmitted disease, with estimates of greater than 4 million new infections occurring annually in the United States. Although most C. trachomatis infections in men and women are asymptomatic, infection can lead to severe reproductive complications in women. The high prevalence in women of child-bearing age results in exposure of an estimated 100,000 neonates in the United States annually. Many of these infants develop conjunctivitis, pneumonia, or both in the first few months of life. Clinical features, diagnosis, treatment, and approaches to prevention of conjunctivitis and pneumonia in the newborn and young infant are reviewed here. Appropriate testing for chlamydial infection in a pediatric victim of sexual assault and the implications of identifying C. trachomatis in suspected cases of childhood sexual abuse also are reviewed.

Adult↗

Chlamydial salpingitis in female guinea pigs receiving oral contraceptives.

Female guinea pigs were given daily doses of a combination of oral contraceptive (OC) agents, consisting of mestranol and norethynodrel suspended in sesame oil or distilled H2O, and were infected in the genital tract with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC). Counts of chlamydial inclusions in cells of vaginal smears collected during infection, showed prolongation and enhancement of infection in OC-treated animals as compared with controls. Appearance of IgG and IgA antibodies to GPIC in genital secretions, as determined by enzyme-linked immunosorbent assay (ELISA), was also delayed in OC-treated animals as compared with controls. OC-treated infected animals were killed on days 15 and 43, and gross pathological evidence for ascending infection culminating in salpingitis was found in all of five and four of five animals, respectively. On the other hand, among untreated infected controls on each sacrifice day, only one of five animals had any evidence for ascending infection. Chlamydiae were detected by light and electron microscopy in fallopian tube tissue collected on day 15 following OC-treatment but not in tissue from control animals.

Animals↗

Ocular delayed hypersensitivity: a pathogenetic mechanism of chlamydial-conjunctivitis in guinea pigs.

We used a naturally occurring, Chlamydia psittaci-caused eye disease in guinea pigs, guinea pig inclusion conjunctivitis, as an animal model to understand both the immune response and the pathogenesis of chlamydial eye infections. When instilled into the conjunctival sac of guinea pigs that had been previously infected and were immune, viable chlamydiae or a Triton X-100-soluble extract of them produced a short-lived (12-48 hr) eye disease indistinguishable clinically and histologically from that observed during primary chlamydial eye infection. The clinical and histologic findings were consistent with those of ocular delayed hypersensitivity. Ocular delayed hypersensitivity was induced by primary chlamydial infection at mucosal sites other than conjunctival, such as vaginal and intestinal. Preliminary characterization of the hypersensitivity allergen shows that it is heat sensitive and common to the genus Chlamydia. The allergen is apparently not surface-exposed on chlamydiae and requires viable but not replicating organisms for activity. Our observation should be useful in understanding pathogenetic mechanisms of Chlamydia trachomatis-caused infections in humans, in particular those that produce chronic inflammatory diseases, such as blinding trachoma and urogenital diseases.

Animals↗

Effects of various doses of estradiol on chlamydial genital infection in ovariectomized guinea pigs.

The effect of various doses of estradiol on genital tract infection by the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC) was investigated in ovariectomized guinea pigs. Prolongation of infection, as determined by chlamydial inclusion counts of cells in Giemsa-stained smears of vaginal scrapings, was observed in animals receiving daily doses of 1.0, 10.0, 100.0, or 1000 micrograms of estradiol. In contrast to controls, ascending infection resulting in endometritis was found in animals receiving doses of greater than or equal to 1.0 microgram of estradiol per day. Response to estradiol treatment was reflected in an increase in cervical-uterine wet weight and uterine wall thickness. No differences were observed in time of appearance of antibody titers to GPIC in serum, but a delay in appearance of IgA antibody to GPIC in genital secretions was found in estradiol-treated animals receiving doses of greater than or equal to 1.0 microgram per day.

Animals↗

Prospective study of chlamydial infection in neonates.

Chlamydia trachomatis was recovered from the cervices of 4% (36/900) of pregnant women tested. 20 infants born through chlamydia-infected cervices were followed up for a year, as were 18 infants born to chlamydia-negative mothers. A statistically significant excess of conjunctivitis and pneumonia was found in infants exposed to chlamydia. The attack-rate for inclusion conjunctivitis was 35% (7/20) and for chlamydial pneumonia it was 20% (4/20). Chlamydiae were recovered from 10 of the 20 (50%) exposed infants, and seroconversion was demonstrated in 14 (70%). None of the 18 unexposed infants showed evidence of chlamydial infection. Thus in our clinic 2.8% of all newborns acquired chlamydial infection, with incidence-rates of 14 cases of conjunctivitis and 8 cases of pneumonia per 1000 live births. Neonatal chlamydial infection is thus a major public-health problem warranting a preventive programme based on the fuller provision of diagnostic services and the treatment of infected pregnant women.

Cervix Uteri↗

Absence of progesterone effects on chlamydial genital infection in female guinea pigs.

The effect of progesterone alone and in combination with estradiol was investigated in ovariectomized and gonadally intact female guinea pigs infected with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC). The course of the infection, as determined by the percentage of cells with GPIC (chlamydia) inclusions in Giemsa-stained vaginal scrapings, was not affected in animals receiving 5.0 mg of progesterone daily. Progesterone had no influence on the enhancement of infection by estradiol. In comparison with sesame oil-treated controls, infection was prolonged by four to six days (P less than .05) in animals receiving a combination of 5.0 mg of progesterone plus 1.0 microgram of estradiol or 1.0 microgram of estradiol alone each day. In ovariectomized animals, estradiol delayed the appearance of IgA antibody in genital secretions, whereas progesterone alone had no effect. Guinea pigs treated with estradiol or progesterone plus estradiol manifested an acute endometritis not observed in animals treated with progesterone alone or in controls receiving sesame oil. Although cervical ectopy, analogous to that seen in women with high levels of progesterone, was identified by histopathology in animals treated with progesterone, no enhancement of the chlamydial infection was observed.

Animals↗

[Follicular conjunctivitis due to Chlamydia trachomatis].

During two years (1997-1999) an investigation of possible infections of chlamydial etiology in outpatients with follicular conjunctivitis was carried out, through the use of specific assays. Fifty-seven selected patients with presumptive inclusion conjunctivitis were diagnosed by means of ophthalmoscopic examination and bilateral tarsal-conjunctiva swabbing for microorganisms. The possible presence of Chlamydia trachomatis was tested by immunofluorescence microscopy and isolation in cell culture of McCoy line. Of the 57 conjunctivitis patients screened, 37 (65%) proved to be positive by cell culture (CC) and 27 (47%) by direct immunofluorescence (IFD). A good agreement between the two assays was observed, where the CC was more sensitive than IFD. Of these 37 patients with chlamydial conjunctivitis, 23 (62%) were women, with over one-third of them ranging in age from 45 to 65 years. Their clinical records revealed an evolution period of 1 to 12 months. Eighteen (78%) of these women reported previous genital pathology, while 4 (29%) of the 14 men had a history of urethritis by Chlamydia trachomatis. A high frequency of follicular conjunctivitis by Chlamydia (65%) in the screened patients was observed, without any evidence of urogenital signs and symptoms at the moment of the study.

Adolescent↗

Ocular chlamydial infection in a university eye clinic.

In the University Hospital, Kuala Lumpur, from 1984 to 1990, 184 patients with acute conjunctivitis were examined for chlamydial infection by direct immunofluorescence. Overall, 52 (28.3%) were found to be positive for chlamydial antigen. There was no significant difference in the detection rate between men and women and among the 3 major ethnic groups. The detection rate was highest among sexually active adults. Epidemiological and clinical features suggest that most of the chlamydial ocular infections seen were inclusion conjunctivitis and not classical trachoma.

Acute Disease↗

Stimulation of peroxidase by chlamydial infection: cytochemistry of guinea pig conjunctival epithelium.

Guinea pig inclusion conjunctivitis, a naturally occurring chlamydial disease of guinea pigs, resolves spontaneously after 3 to 4 weeks. The factors responsible for curbing the infection have not yet been specifically defined. Since Iwata (Invest. Ophthalmol. 15:297-301, 1976) reported cytochemical activity for peroxidase in the conjunctival epithelium of the normal rat, we undertook these studies to determine whether a similar activity exists in the guinea pig, and if so, whether it functions in the elimination of this Chlamydia psittaci infection. Tarsal conjunctivas of 14 normal guinea pigs, 34 infected ones, and 7 control guinea pigs (inoculated with yolk sac only) were excised and tested for peroxidase by the Graham and Karnovsky method (J. Histochem. Cytochem. 14:291-302, 1966). We found that peroxidase activity, virtually absent in normal animals, was intensely stimulated by the infection. This enzymatic activity appeared 2 days after inoculation of the conjunctiva with chlamydia and persisted for 6 to 7 weeks. The enzyme was localized in the rough endoplasmic reticulum and perinuclear cisternae of all layers of the conjunctival epithelium from the external surface to the basal lamina, including cells with no apparent inclusions as well as those heavily parasitized. Reaction in the Golgi complex was variable. No reaction, however, was ever evident in the chlamydial vacuoles or lysosomes, and the organisms continued to grow and multiply during peak enzymatic activity. We therefore concluded that the stimulated enzyme is apparently not directly responsible for the waning of the infection, but instead reflects an alteration of host metabolism that occurs as a consequence of the infection.

Animals↗

Development of chronic conjunctivitis with scarring and pannus, resembling trachoma, in guinea-pigs.

Guinea-pigs were repeatedly infected with guinea-pig inclusion conjunctivitis agent. Reinfection caused severe conjunctival inflammation, and repeated reinfection led to chronic inflammation lasting for many months. This was followed by the development of pannus, follicles on the palpebral conjunctivae, scarring of the lower palpebral conjunctiva, and deformities of the lower lid. Reinfection was accompanied by small numbers of inclusion-bearing cells, small numbers of polymorphonuclear cells, and high numbers of mononuclear cells. There was no increase in the level of serum antibodies. The chronic conjunctivitis was associated with high numbers of mononuclear cells and no inclusions or polymorphonuclear cells. The response to reinfection appears to be a delayed-type hypersensitivity reaction, and we suggest that the chronic inflammation, pannus, scarring, and lid deformities associated with hyperendemic trachoma may be due to repeated reinfection combined with delayed-type hypersensitivity.

Animals↗

Lymphogranuloma venereum. II. Characterization of some recently isolated strains.

Five Bedsonia (Chlamydia) isolates from lymphogranuloma venereum (LGV) patients were tested for inclusion type, sulfonamide sensitivity, and mouse virulence. Two matched the classical description of LGV agents. Two were not virulent for mice by the intracerebral route, therefore fitting the description for trachoma-inclusion conjunctivitis agents. One was highly virulent for mice and sulfonamide-resistant, and produced inclusions that did not stain with iodine, all characteristics generally associated with avian bedsoniae. A sixth isolate could not be adequately tested due to poor infective yields. Because of this variety of properties within the Bedsonia group, the term LGV might more appropriately be reserved for clinical disease rather than to describe a particular bedsonial agent.

Animals↗

Temporal analysis of the developing Chlamydia psittaci inclusion by use of fluorescence and electron microscopy.

The chlamydiae are obligate intracellular parasites that develop and multiply within a vacuole (termed an inclusion) that does not fuse with lysosomes. Inclusion morphology varies dramatically among the different chlamydiae, particularly within the species Chlamydia psittaci. Some strains develop within a single vacuole, while the mature inclusion of other strains consists of several distinct lobes, each filled with chlamydial developmental forms. The development of this lobed structure was investigated in HeLa cells infected with the guinea pig inclusion conjunctivitis (GPIC) strain of C. psittaci. We employed two recently described probes for the chlamydial inclusion to study the development of these unique lobed structures. The novel probes were an antiserum directed at a protein localized to the GPIC inclusion membrane (anti-IncA) and the fluorescent sphingolipid (N-[7-(4-nitrobenzo-2-oxa-1,3-)]) aminocaproyl sphingosine (NBD-ceramide). Lobed inclusions developed in cells infected at very low multiplicities of infection, suggesting that the structure is not a function of infection by more than one elementary body (EB). Double-label fluorescent-antibody analysis with anti-IncA and an antibody directed at a chlamydial outer membrane protein showed that, prior to 18 h postinfection (p.i.), the inclusion membrane and the chlamydial membrane were tightly associated. After 18 to 20 h p.i., the lobes began to expand and fill with developmental forms and the inclusion membrane and chlamydial membrane became distinct. At times from 8 to 48 h p.i., GPIC inclusions were shown to receive fluorescent derivatives of NBD-ceramide and to be localized to the perinuclear region of the host cell. Labeled lectins with affinity for carbohydrate moieties localized to the Golgi apparatus showed that the lobes of mature inclusions surround the Golgi apparatus. Labeling with NBD-ceramide and the Golgi apparatus-specific lectins therefore demonstrated a functional and physical association of the inclusion with the Golgi apparatus throughout the developmental cycle. Collectively, these results lead to a model for the development of the lobed chlamydial inclusion. We propose that the lobed structure is a result of division of inclusions occurring in parallel with the multiplication of reticulate bodies (RB) early in the developmental cycle. The division of inclusions slows or stops in mid-cycle, and dividing RB accumulate within the enlarging lobes. The RB then differentiate to EBs, the inclusion and cell are lysed, and EBs are freed to infect another cell.

Chlamydophila psittaci↗

Protective role of serum antibody in immunity to chlamydial genital infection.

Female guinea pigs were injected intraperitoneally with pooled immunoglobulin derived from animals immunized to the chlamydial agent of guinea pig inclusion conjunctivitis. Genital infections in animals receiving pooled immunoglobulin from immune animals were markedly decreased with regard to the number of inclusions detected compared with control animals. These data indicated that serum-derived antibody was able to provide a degree of protection against a chlamydial genital tract infection.

Animals↗

Antibiotic susceptibility of Chlamydia trachomatis.

The antibiotic susceptibility of Chlamydia trachomatis isolates was determined in a tissue culture system. Representatives of all currently recognized serotypes of trachoma-inclusion conjunctivitis agents were tested. Tetracycline and erythromycin yielded similar results, with 1.0 mug/ml preventing chlamydial replication. Rifampin was the most active antibiotic, with 0.25 mug/ml completely suppressing inclusion formation of all strains. Fifty percent end points were usually achieved at one-fourth to one-eighth the 100% suppression level. Penicillin was not as effective, and the assays were often irregular. Antibiotic susceptibility of these chlamydiae was essentially the same, regardless of serotype, anatomic site infected, geographic origin, or antibiotic use in the community.

Chlamydia trachomatis↗

Experimental chlamydial salpingitis in the guinea pig.

Acute self-limited salpingitis developed when guinea pigs' fallopian tubes were infected with the guinea pig inclusion conjunctivitis agent (Chlamydia psittaci). The disease peaked at approximately 8 days post infection and had diminished markedly by day 14. The inflammatory reaction was concentrated at the luminal surface. Chlamydial replication was demonstrated and inclusions were found in epithelial cells.

Animals↗