Generation of histamine-releasing activity from serum albumin by medium derived from stimulated neutrophils of rat.
1. Medium conditioned by rat neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine (FMLP) has been found to generate mast cell histamine-releasing activity (HRA) when incubated with bovine serum albumin (BSA). 2. Histamine release increased as the concentration of BSA used to generate HRA was increased from 0.25 to 10 mg ml-1, as the concentration of neurotrophil conditioned medium was increased and as the concentration of FMLP used to stimulate the neutrophils was increased. Histamine release was non-cytotoxic as it was inhibited by energy deprivation or by removal of calcium and it was accompanied by degranulation. 3. HRA was detectable after 30 min of incubation with BSA and its generation continued to increase over the 18 h of our measurements. 4. Generation of HRA was dependent upon the presence of medium from stimulated neutrophils and on the presence of BSA, although plasma could substitute for BSA. Likewise, HRA could be generated from gamma-globulin although to a lesser extent than with albumin. 5. Generation was optimum at acid pH and was inhibited by prior boiling of the neutrophil conditioned medium or by the addition of pepstatin. 6. It is suggested that an enzyme(s) released from the neutrophil during stimulation acts on an albumin-like substrate to generate HRA. It is proposed that HRA is peptide in nature and may be generated during an inflammatory response.
Cochrane
D E
DE
Department of Biology, Tufts University, Medford, Massachusetts 02155.
Boucher
W
W
Carraway
R E
RE
D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects; D002268:Carboxypeptidases / Q000378:metabolism; D006636:Histamine Release / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence
D012710:Serum Albumin, Bovine; D002268:Carboxypeptidases