Antibody response to outer-membrane antigens of Pseudomonas aeruginosa in human burn wound infection

Abstract

There is little information about the local and systemic antibody response to surface antigens of bacteria growing in situ in infected lesions in man. In this study, Pseudomonas aeruginosa was obtained directly from the infected wounds of two patients with burns and studied without subculture. Outer-membrane proteins (OMPs) were investigated and compared with those of cells cultivated in the laboratory, with the aim of selecting defined growth conditions to give surface antigens more closely resembling those found in vivo. Several high-mol. wt (77,000-101,000) proteins were expressed in the outer membranes of the bacteria from the patients and could be phenotypically induced by cultivating the same isolate in iron-depleted conditions in vitro. Other major OMPs (D, E, F, G and H) were also observed in cells taken from the lesions. Immunoblotting demonstrated that proteins D and E were recognised by different classes of immunoglobulins in the sera of both patients as was flagellar antigen present in the outer-membrane preparation of the P. aeruginosa from patient 1. Iron-regulated membrane proteins (IRMPs) were similarly detected, but more strongly by IgM from patient 1. Furthermore, a marked antibody response to IRMPs was noted at the site of infection. Bands of a similar intensity were seen after absorption of the sera with lipopolysaccharide (LPS) purified from the infecting strain. This indicated that the response observed was directed against OMPs (including IRMPs) and not against contaminating LPS.