Review: Evidence against the hypothesis that antibodies to the inner core of lipopolysaccharides in antisera raised by immunization with enterobacterial deep-rough mutants confer broad-spectrum protection during Gram-negative bacterial sepsis

Abstract

Antisera to rough enterobacterial mutants of chemotypes Ra, Rc, and Re have been reported to confer broad-spectrum protection against wild-type smooth strains. It has been hypothesized that binding and neutralization of lipopolysaccharides (LPS) by antibodies to common core epitopes underlies such protection. This review summarizes experiments by our laboratory and others that do not confirm this concept and proposes reasons for the divergent results. Studies indicating broad-spectrum protection by rough-mutant antisera often had defects in experimental design or methodology. These include the failure: (i) to use matched pre- and postimmune sera from the same donors to control for variable protective activity of normal sera; (ii) to exclude the role of natural and polyclonally stimulated antibodies with proven protective activity against the infecting bacterial strain (e.g. O-specific, capsular, Pseudomonas exotoxin A); (iii) to exclude protective effects of acute-phase serum factors; (iv) to exclude protective effects of endotoxin contamination after adsorption or fractionation of antibody preparations; (v) to use non-boiled bacteria and LPS not subjected to acid-hydrolysis or gel-fractionation, and to exclude nonspecific adsorption, to demonstrate physiologically meaningful binding of rough-mutant antibodies to smooth enterobacteria and their LPS.