Characterization of kinetics and target proteins for binding of human complement component C3 to the surface-exposed outer membrane of Chlamydia trachomatis serovar L2.
In order to characterize the interaction of human complement with Chlamydia trachomatis, flow cytometry was used to quantitate binding of complement component C3 to elementary bodies of C. trachomatis serovar L2 preincubated in fresh serum in the presence or absence of human polyclonal chlamydial antibody. Isolation of each of the complement activation pathways revealed that C3 was activated most effectively by the alternative pathway. The degree of binding by the classical pathway was proportional to the concentration of antibody, but dual-pathway-mediated binding was not greater than antibody-independent alternative pathway binding. Electrophoresis and immunoblotting of detergent-extracted outer membrane protein-C3b complexes indicated that the chlamydial major outer membrane protein was the primary cell surface moiety binding C3b in both the presence and absence of specific antibody. Hydroxylamine cleavage of outer membrane protein-C3b complexes provided evidence that the majority of C3b is bound to the major outer membrane protein by hydroxyl ester bonds. This result was also unchanged by the presence of specific antibody. An unexpected finding was the apparent binding of anti-C3 antibody to a 40-kDa protein of the chlamydial outer membrane complex, perhaps indicating C3 mimicry on the part of the chlamydial major outer membrane protein.
Hall
R T
RT
Department of Medicine, University of British Columbia, Vancouver, Canada.
Strugnell
T
T
Wu
X
X
Devine
D V
DV
Stiver
H G
HG
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology; D000937:Antigen-Antibody Reactions; D000954:Antigens, Surface / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002692:Chlamydia trachomatis / Q000145:classification / Q000276:immunology*; D003167:Complement Activation; D003176:Complement C3 / Q000378:metabolism*; D003179:Complement C3b / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000737:chemistry; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D011485:Protein Binding; D012703:Serotyping
D000907:Antibodies, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D003176:Complement C3; D006898:Hydroxylamines; D046911:Macromolecular Substances; D019811:Hydroxylamine; D003179:Complement C3b