Antibody- and complement-dependent cell injury assayed by 51Cr release from human peripheral blood mononuclear cells pretreated with lipopolysaccharide

Abstract

Exposure of human peripheral blood mononuclear (MN) cells to deesterified (alkali-treated) lipopolysaccharide (LPS-OH) and then to 51Cr rendered the cells susceptible to 51Cr release in the presence of specific antibody and complement. The assay was optimized by using rough (Rb2 or Re) LPS. 51Cr release did not occur from cells preexposed to untreated or electrodialyzed LPS. Studies of isolated monocytes and lymphocytes revealed that the majority of the 51Cr released was derived from monocytes. The optimum concentration of LPS-OH was 10 micrograms/ml. Antiyersinia agglutinin-positive serum, but not a negative serum, obtained from patients with reactive yersinia arthritis caused 51Cr release from MN cells pretreated with yersinia LPS-OH. This implies that during yersinia infection antibodies are generated that can attack the cell membrane--LPS-OH complex. We conclude that the method provides a tool to demonstrate binding of LPS to MN cells in a manner that leads to cell injury in an immune host.