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.     

The first case of lymphogranuloma venereum in a woman in East-Central Europe and its multiple co-infections

We are reporting the first case of lymphogranuloma venereum in women in East-Central Europe. A 22-year-old heterosexual woman attended our department of venereology. She complained about a burning sensation in the urethra and vaginal discharge. Many tests were performed, and lymphogranuloma venereum, syphilis, gonorrhea, chlamydial urethritis and cervicitis, genital herpes, genital warts, and hepatitis C were diagnosed. Lymphogranuloma venereum was originally endemic in tropical and subtropical areas, but since 2003, outbreaks of this infection have been reported in North America, Europe, and Australia in men who have sex with men (MSM) community. To date, all cases of lymphogranuloma venereum in the Czech Republic appeared in men, predominantly in HIV-positive MSM. There are not many evidences about lymphogranuloma venereum (LGV) in women in developed countries. This report underlines the need for awareness of lymphogranuloma venereum in women among gynecologists, venereologists, and other physicians not only in Western Europe, but across all European countries.     

Purification of Chlamydia trachomatis lymphogranuloma venereum elementary bodies and their interaction with HeLa cells

A procedure has been developed to yield infectious elementary bodies of the lymphogranuloma venereum strains LGV 434 and 404 of Chlamydia trachomatis, labelled during intracellular growth in HeLa 229 cells. The final preparation, obtained after velocity sedimentation of a polycarbonate membrane-filtered sample through a sucrose gradient, is free of host proteins and, more importantly, of chlamydial reticulate bodies. Using such purified preparations, it was found that the association of LGV 434 elementary bodies with HeLa 229 cultures was unaffected by the pretreatment of the host cells with a variety of lectins or with neuraminidases from Clostridium perfringens and Vibrio cholerae. The association was inhibited by dextran sulphate and by mild trypsin treatment of HeLa cultures. Treatment of purified elementary bodies with trypsin, chymotrypsin, neuraminidases and a variety of carbohydrates and lectins did not produce any change in the rate of association with HeLa cultures. Heat-inactivated elementary bodies were significantly less able to associate with the host cells.     

Association and uptake studies of a Chlamydia trachomatis lymphogranuloma venereum strain by eukaryotic cells

We have studied association and uptake of Chlamydia trachomatis serovar L1 under various infection conditions. Chlamydiae attached to a greater extent to McCoy cells than to HeLa cells, although the number of inclusions formed in the latter was the same or higher. The amount of internalised chlamydiae was similar in the 2 cell types. Centrifugation of McCoy cell-attached chlamydiae did not affect the uptake of this serovar. However, if the inoculum was centrifuged to the cell surface and then incubated at 37°C, there was a pronounced increase in the relative amount of ingested chlamydiae in comparison with stationary infection. Chlamydiae were attached to and internalised insubconfluent McCoy cell monolayers as efficiently as in confluent layers. If the monolayers were sparse or very sparse, there was a great reduction of associated chlamydiae. Our results indicate that the host cell binding for chlamydiae vary with cell type, cell density, and mode of infection.     

Detection of type-specific antibody to lymphogranuloma venereum serotypes ofChlamydia trachomatis with a microneutralization test

Ten-day monospecific mouse antisera prepared against serotypes L1, L2, and L3 ofChlamydia trachomatis were titrated against homologous and heterologous antigens in the microimmunofluorescence (micro-IF) test and by a new microneutralization method. While broad crossreactivity between serotypes was detected in the micro-IF test, the same antisera demonstrated serotype specific neutralizing activity. The microneutralization test could be used for serotyping new chlamydial isolates when equivocal results have been obtained by the microimmunofluorescence method.