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Immunity to vaginal reinfection in female guinea pigs infected sexually with Chlamydia of guinea pig inclusion conjunctivitis.

Guinea pig boars were inoculated intraurethrally with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC). At the heights of their urethral infections, they were caged with sows in estrus. Whereas some of the sows had not been previously exposed to GPIC agent, others had received an intravaginal inoculation 5 to 8 weeks earlier. Those sows for which infected boars provided the first exposure were challenged by intravaginal inoculation 5 to 8 weeks later. Vaginal and conjunctival scrapings were taken regularly and stained for chlamydial inclusions. Titers of serum anti-GPIC antibodies and of vaginal secretory IgA anti-GPIC antibodies were determined by immunofluorescence. Our results show for the first time that a sexually acquired vaginal GPIC infection induces immunity to manual reinfection of the vagina. Because of the high incidence of secondary conjunctival infections among the vaginally infected sows, we could not provide a sound statistical basis for our tentative conclusion that manual infection of the vagina induces immunity to sexual reinfection. The results of our antibody titrations confirm previous work showing that vaginal GPIC infection induces formation of both serum antibody and vaginal secretory immunoglobulin A antibody.

Animals↗

Cell-mediated and humoral immune responses to chlamydial antigens in guinea pigs infected ocularly with the agent of guinea pig inclusion conjunctivitis.

Cell-mediated immune response and humoral response to chlamydial antigens were investigated in guinea pigs infected with the agent of guinea pig inclusion conjunctivitis (GPIC). Pronounced cell-mediated immune response to the homologous antigen, as well as to two other chlamydial antigens, 6BC (Chlamydia psittaci) and LB-1 (C. trachomatis), occurred in all infected animals. Cell-mediated immune response to GPIC, and to a lesser extent to 6BC and LB-1 as well, was enhanced with time after infection even without the re-inoculation of the infectious agent. Extensive cross-reactions among the three chlamydial antigens during the cell-mediated immune response appeared to be due to shared species-specific and group-reactive antigens. Serum antibody response was pronounced and uniform to GPIC; it was less marked to 6BC and LB-1, with fewer cross-reactions than seen in tests for cell-mediated immunity.

Animals↗

Infection of genital tract and transmission of ocular infection to newborns by the agent of guinea pig inclusion conjunctivitis.

Female guinea pigs were inoculated intravaginally with the agent of guinea pig inclusion conjunctivitis (Gp-ic). Evidence for infection was obtained by demonstration of Gp-ic inclusions in epithelial cells of genital tract smears, histopathology, recovery of Gp-ic during infection, and antibody response. Infection of the genital tract was produced with low doses (20-200 median egg lethal doses) of the agent. The infection lasted about 3 weeks, and there was no marked clinical response. Ocular infection that could be detected as early as 3 days after birth in offspring of infected mothers was also demonstrated.

Animals↗

Tear and serum antibody response to Chlamydia trachomatis antigens during acute chlamydial conjunctivitis in monkeys as determined by immunoblotting.

In this study, we examined the temporal antibody response by immunoblotting analysis in tears and sera of three cynomolgus monkeys (Macaca fasicularis) with primary acute Chlamydia trachomatis serovar B conjunctivitis. The objective was to identify chlamydial antigens stimulating antibody during the host responses in the course of this self-limiting infection with the rationale that they may be protective antigens. The major outer membrane protein (MOMP), lipopolysaccharide (LPS), and polypeptides of 60 and 68 kilodaltons (kDa) were the predominant antigens recognized by immunoglobulin A (IgA) in monkey tears. Tear IgA antibody specific for the MOMP was first detected 14 days postinfection, whereas tear IgA reactive with LPS or the 68- and 60-kDa polypeptides was first detectable on day 21. Tear IgA antibodies specific for each of these antigens persisted in tears through day 56, 4 weeks after both peak clinical disease and recovery of the organism from the conjunctivae. In contrast, tear IgG antibodies peaked at approximately 28 days postinfection, the time of maximal inflammatory response. The IgG response in monkey sera was similar to that observed for tear antibodies, in that the MOMP, 60-, and 68-kDa polypeptides were the primary immunogens. The exception was that IgG antibody against these antigens was detected 1 week later than that observed for tear IgA antibodies. Of three monkeys that responded with tear IgA antibody against LPS, one did not have detectable serum IgG LPS antibody. The specificity of the tear IgA antibody response of monkeys was determined by immunoblotting nine other C. trachomatis serovars in addition to the homologous B serovar. The tear IgA response to the MOMP was predominantly B complex subspecies-specific (serovars B, Ba, D, and E), whereas the response to chlamydial LPS was found to be species-specific. The significance of these observations in relation to previous vaccine studies in nonhuman primates is discussed.

Animals↗

Lack of allelic polymorphism for the major outer membrane protein gene of the agent of guinea pig inclusion conjunctivitis (Chlamydia psittaci).

The major outer membrane protein gene (omp1) was sequenced for each of six Chlamydia psittaci (guinea pig inclusion conjunctivitis [GPIC]) strains isolated from guinea pigs. Five of the isolates were obtained in the United States during the 1960s and 1970s, including the prototype strain isolated by Murray in 1962. The other isolate was obtained from a guinea pig in England. The nucleotide sequence of the omp1 gene for each strain was identical. The lack of omp1 allelic polymorphism among GPIC isolates suggests that, unlike C. trachomatis, the GPIC agent lacks antigenic variation in the major outer membrane protein.

Alleles↗

Immunity to chlamydial infections of the eye. II. Studies of passively transferred serum antibody in resistance to infection with guinea pig inclusion conjunctivitis.

Conjunctival infection of guinea pigs by the chlamydial agent of guinea pig inclusion conjunctivitis confers immunity. However, the mechanism of resistance to this intracellular pathogen is not yet defined. In the study reported here, serum immunoglobulin was passively transferred with resultant titers in excess of those known to be associated with immunity. Nonetheless, when the passive transfer recipients were challenged, they acquired infection which was neither delayed nor attenuated. Eye secretion antibody titers appeared and increased only after 11 days of infection in both passive transfer recipients and control groups, suggesting but not proving de novo local synthesis of secretory antibody. This study suggests that cellular or secretory immune mechanisms may predominate in resistance to this infection.

Animals↗

Experimental genital infection of male guinea pigs with the agent of guinea pig inclusion conjunctivitis and transmission to females.

Male guinea pigs were inoculated intraurethrally with the agent of guinea pig inclusion conjunctivitis (Gp-ic). Cytoplasmic inclusions were found in superficial epithelial cells of the urethra in smears and stained sections. Gp-ic antigen(s) was detected by immunofluorescent staining of sections. There was no marked urethral exudate, but many animals developed bullous lesions on the glans and the body of the penis and a severe inflammatory lesion of the hind leg. All males demonstrated an antibody response and most of them showed a positive skin test reaction. Venereal transmission to females of Gp-ic infection was shown to occur as determined by detection of inclusions in vaginal smears, antibody response, and positive skin tests.

Animals↗

Characterization of nontypable Streptococcus pneumoniae-like organisms isolated from outbreaks of conjunctivitis.

From January through June 1980 seven colleges and universities in various parts of New York State (NYS) reported outbreaks of conjunctivitis affecting at least 1,500 students of both sexes. Of the 125 conjunctival swabs tested in our laboratory, organisms identified as nontypable Streptococcus pneumoniae were isolated in pure culture from 24% and in combination with other organisms from 22%. Although bile-soluble and susceptible to optochin, the isolates had a dry-colony appearance and no typable capsule with the Neufeld capsular-swelling test. Mouse passage of four representative NYS isolates did not stimulate production of a typable capsule. We subsequently chose to refer to these isolates as S. pneumoniae-like organisms. Of primary importance to our study, all NYS isolates tested were similar in biochemical and immunological reactions, antibiotic susceptibility, and virulence in mice. Of 18 strains referred to us from three other outbreaks (California, 1980; NYS, 1981; Illinois, 1981), four of the six tested biochemically gave the same biochemical reactions as the four NYS isolates, and 16 of the 18 tested immunologically reacted strongly with antisera produced against those four isolates, showing line(s) of identity with each other and with the NYS isolates.

Animals↗

Comparison of lipopolysaccharides from Brazilian purpuric fever isolates and conjunctivitis isolates of Haemophilus influenzae biogroup aegyptius. Brazilian Purpuric Fever Study Group.

Haemophilus influenzae biogroup aegyptius (H. aegyptius) has been identified as the etiologic agent of the recently described disease Brazilian purpuric fever (BPF). Although there is heterogeneity among the strains associated with conjunctivitis, isolates from patients with BPF appear to be derived from a single clone. The clinical presentation of BPF suggests that bacterial lipopolysaccharides (LPS) are involved in its pathogenesis. We prepared LPS from H. influenzae biogroup aegyptius and found them to be similar to H. influenzae type b LPS in apparent size (by sodium dodecyl sulfate-polyacrylamide gel electrophoresis), biological activities, and fatty acid composition. We compared LPS from BPF clone isolates with LPS from non-BPF clone isolates in tests of Limulus lysate activation, spleen cell mitogenesis, promotion of neutrophil adherence to LPS-treated endothelial cells, and the dermal Shwartzman reaction. In none of these activities were LPS from the BPF clone isolates more potent. Because LPS shed from growing bacteria may be involved in the pathogenesis of purpura, we also measured the rate at which LPS were released into culture medium during bacterial growth and found no significant difference between BPF clone and non-BPF clone isolates.

Chemical Phenomena↗

Chronic conjunctivitis caused by oral anaerobes and effectively treated with systemic metronidazole plus amoxicillin.

In this study, we report on a case of refractory, unilateral anaerobic conjunctivitis. The predominant anaerobic flora consisted of Prevotella intermedia (formerly Bacteroides intermedius) and Peptostreptococcus micros. By using the technique of restriction endonuclease fingerprinting of genomic DNA, it was shown that the P. intermedia likely originated from the oral cavity. Topically applied antibiotics had failed to suppress the infection in the past. Successful treatment was achieved after systemic administration of metronidazole plus amoxicillin.

Adult↗

Rapid diagnosis of adenoviral conjunctivitis by PCR and restriction fragment length polymorphism analysis.

To detect and identify adenovirus (Ad), we used a combination of PCR and restriction fragment length polymorphism (RFLP) analysis. Nested PCR with two primer sets that hybridize to the conserved region for hexon proteins of 14 prototypes of Ad, Ad serotype 1 (Ad1) to Ad8, -11, -14, -19, -37, -40, and -41, amplified a 956-bp DNA fragment. The amplified fragments from the 14 prototypes were completely differentiated with a combination of three restriction endonucleases, EcoT14I, HaeIII, and HintI. We applied this new method for 127 samples of conjunctival scrapings from patients with conjunctivitis and compared the results with those obtained with the combination of culture isolation and a neutralization test (NT). PCR gave a positive result in 69 of 127 cases (54.3%), while only 61 of the 127 samples (48.0%) tested positive by culture isolation. Compared with isolation, the PCR method had a sensitivity of 100% (61 of 61). Positive PCR samples were further classified as Ad37 (59.5%), -3(31.9%), -11 (4.3%), -8 (2.9%), and -4 (1.4%) by PCR-RFLP analysis. Of eight samples that were PCR positive and culture isolation negative, six were Ad37 and two were Ad8 by PCR-RFLP analysis. These differentiations of isolation-positive samples were identical to the results obtained by the NT. It took only 3 days to detect and identify Ad by PCR-RFLP analysis, whereas it took at least 3 weeks by culture isolation and NT. Our newly developed method of detecting and typing human Ad by PCR-RFLP analysis is more sensitive, accurate, and rapid than the conventional method of culture isolation and an NT.

Adenoviruses, Human↗

Identification of a new strain of fastidious enterovirus 70 as the causative agent of an outbreak of hemorrhagic conjunctivitis.

A 1994 outbreak of acute hemorrhagic conjunctivitis in Israel was caused by an enterovirus 70 strain that was distinct from previously reported strains. Characterization was by electron microscopy (eye washes), reverse transcription-PCR (RT-PCR; eyewash, specimens, eye swabs, and tears), and sequence analysis of RT-PCR-amplified fragments from the 5' noncoding region and VP1.

Amino Acid Sequence↗

Tsukamurella conjunctivitis: a novel clinical syndrome.

In this report, we describe the first three cases of Tsukamurella conjunctivitis in the literature. All three patients presented with congestion of one eye with small amounts of serous discharge for 1 to 2 days. All three recovered after 10 days of treatment with polymyxin B-neomycin or chloramphenicol eyedrops. Sequencing of the 16S rRNA genes of the three isolates recovered from the serous discharge of the three patients showed that they were all Tsukamurella species. The phenotypic characteristics of the isolate obtained from one patient best fit the phenotypic profile of Tsukamurella pulmonis, whereas those of the other two best fit that of Tsukamurella tyrosinosolvens.

Actinomycetales↗

Confirmation of nontypeable Streptococcus pneumoniae-like organisms isolated from outbreaks of epidemic conjunctivitis as Streptococcus pneumoniae.

Eleven isolates representing five distinct outbreaks of pneumococcal conjunctivitis were examined for phenotypic and genetic characteristics. None of the strains possessed capsules, and all strains were susceptible to optochin, bile soluble, and Gen-Probe AccuProbe test positive. All 11 isolates were confirmed as Streptococcus pneumoniae by DNA-DNA reassociation experiments.

Conjunctivitis↗

Purification and biophysical properties of acute haemorrhagic conjunctivitis virus.

The buoyant density of acute haemorrhagic conjunctivitis virions labeled with either [(3)H]uridine or [(3)H]leucine was 1.34 g/ml in CsCl and 1.25 g/ml in sucrose. RNA extracted from the virions gave a sedimentation coefficient of approximately 34S in sucrose, and was found to be sensitive to RNase. Molecular weight of RNA was calculated to be 2.5 x 10(6) using poliovirus RNA for reference.

Centrifugation, Density Gradient↗