Recombinant C. trachomatis MOMP (strain B/Tw-5/OT Serovar B) [His]

Trachoma and sexually transmitted diseases caused by Chlamydia trachomatis are major health problems worldwide. Epitopes on the major outer membrane protein (MOMP) of C. trachomatis have been identified as important targets for the development of vaccines. In order to examine the immunogenicity of a recombinant vector expressing a chlamydial epitope, a poliovirus hybrid was constructed in which part of neutralization antigenic site I of poliovirus type 1 Mahoney (PV1-M) was replaced by a sequence from variable domain I of the MOMP of C. trachomatis serovar A. The chlamydial sequence included the neutralization epitope VAGLEK. This hybrid was viable, grew very well compared with PV1-M, and expressed both poliovirus and chlamydial antigenic determinants. When inoculated into rabbits, this hybrid was highly immunogenic, inducing a strong response against both PV1-M and C. trachomatis serovar A. Antichlamydia titers were 10- to 100-fold higher than the titers induced by equimolar amounts of either purified MOMP or a synthetic peptide expressing the VAGLEK epitope. Furthermore, rabbit antisera raised against this hybrid neutralized chlamydial infectivity both in vitro, for hamster kidney cells, and passively in vivo, for conjunctival epithelia of cynomolgus monkeys. Because poliovirus infection induces a strong mucosal immune response in primates and humans, these results indicate that poliovirus-chlamydia hybrids could become powerful tools for the study of mucosal immunity to chlamydial infection and for the development of recombinant chlamydial vaccines.

Monoclonal antibodies (MAbs) were generated against an ovine abortive strain of Chlamydia psittaci. A plaque reduction assay was used to select 19 neutralizing antibodies which appeared to be heterogeneous in isotype, specificity, and recognized proteins. Different neutralizing MAbs were tested for their protective abilities against abortion in a pregnant-mouse model, All of the protective MAbs selected had the same isotype, were serotype 1 specific, and recognized a protein of about 110 kDa by immunoblotting, The recognized epitopes were resistant to sodium dodecyl sulfate and reducing agents, but all of them were heat sensitive, The protein was able to form disulfide-linked polymers. Immunological cross-reaction studies with rabbit sera showed a link between the 110-kDa protein and the major outer membrane protein (MOMP). The 110-kDa protein was purified by immunoaffinity and shown to be dissociated after heating into MOMP by silver staining and immunoblotting, These results show homogeneity among protective MAbs directed to heat-sensitive epitopes Located on an oligomer of the MOMP of C. psittaci.