Evaluation of the opsonic capacity of core lipopolysaccharide antiserum of equine origin against smooth Escherichia coli 0111:B4, using macrophage chemiluminescence.
A study was performed to determine whether equine antiserum to core lipopolysaccharide (LPS) would enhance phagocytosis of smooth gram-negative (GN) organisms by equine macrophages. Five healthy adult horses (group A) were immunized with a bacterin prepared from the J-5 mutant of Escherichia coli 0111:B4 and Salmonella minnesota R595 to produce antibodies to core LPS. Five horses (group B) served as nonimmunized controls and were given physiologic saline solution instead of the rough mutant bacterin. Serum antibody titers to core LPS and to smooth E coli 0111:B4 were determined by indirect ELISA. Four serum pools were prepared: pool 1 = sera from horses in group B prior to immunization; pool 2 = sera from horses in group A prior to immunization (preimmune serum); pool 3 = sera from horses in group B, 7 days after the last saline injection; pool 4 = sera from horses in group A, 7 days after the last immunization (core LPS antiserum). The serum pools, either unheated or heated 30 minutes at 56 C, in 3 dilutions (1/50, 1/100, 1/500) were used to opsonize smooth E coli 0111:B4 in an assay of equine peritoneal macrophage chemiluminescence (CL). Peritoneal fluid was collected from clinically normal horses and the macrophages were purified by adherence to borosilicate glass scintillation vials. Each serum type and dilution was added to triplicate vials containing 10(7) colony-forming units of E coli 0111:B4. Luminol-dependent CL was measured with a liquid scintillation counter in the out-of-coincidence mode. Each serum dilution was tested in duplicate vials without bacteria to asses serum-induced nonspecific CL.(ABSTRACT TRUNCATED AT 250 WORDS)
Morris
D D
DD
Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens 30602.
Harmon
B G
BG
Moore
J N
JN
D000818:Animals; D004926:Escherichia coli / Q000276:immunology*; D006736:Horses / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology*; D008163:Luminescent Measurements; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism; D009895:Opsonin Proteins / Q000276:immunology*; D010587:Phagocytosis; D012475:Salmonella / Q000276:immunology
D007106:Immune Sera; D008070:Lipopolysaccharides; D009895:Opsonin Proteins