Immunological reactions involving leukocytes: I. Detection of antibodies to human granulocytes by measurement of the metabolic events associated with phagocytosis.
During phagocytosis, glucose oxidation via the hexose monophosphate shunt (HMS) is markedly stimulated in human polymorphonuclear leukocytes. A method is described for the detection of antibodies to human granulocytes which exploits this fact. The source of antibodies was rabbit antihuman leukocyte serum (RAHLS) obtained by repeated immunization of normal rabbits with human granulocyte homogenates. HMS activity was quantitated by measuring the release of 14CO2 from glucose-1-14C. Under precisely defined experimental conditions, RAHLS and its IgG fraction inhibited 14CO2 release. The inhibitory effect of RAHLS on human granulocyte phagocytosis-associated HMS activity could be reduced by prior absorption of RAHLS with human granulocytes. Compared to the cytotoxic and leukoagglutination tests, this method appears more sensitive with respect to detection of xenogeneic antibodies to human granulocytes.
D000042:Absorption; D000906:Antibodies; D000961:Antilymphocyte Serum / Q000494:pharmacology; D002245:Carbon Dioxide / Q000378:metabolism; D003601:Cytotoxicity Tests, Immunologic; D003864:Depression, Chemical; D005947:Glucose / Q000378:metabolism; D006098:Granulocytes / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D006650:Histocompatibility Testing; D006801:Humans; D007962:Leukocytes / Q000276:immunology*; D010587:Phagocytosis
D000906:Antibodies; D000961:Antilymphocyte Serum; D002245:Carbon Dioxide; D005947:Glucose