title: An investigation of the optimal conditions for the in vivo production of immunologically sensitised rat mast cells.
abstract: Experiments were designed to develop an optimal method for inducing in vivo production of sensitised peritoneal mast cells. Rats of different strains were sensitised with whole egg-white and killed at suitable intervals to harvest the peritoneal mast cells. Release of histamine was induced in vitro by both whole egg-white and its major protein constituents, and assayed by a standard spectrofluorometric method. Wistar rats showed higher levels of sensitisation than black-hooded Lister rats; it was more convenient to harvest erythrocyte-free peritoneal mast cells from males than females. Very young (less than 150 g) and very old (greater than 300 g) rats showed sub-optimal sensitisation. Optimal sensitisation was obtained by simultaneous administration of antigen (doses of 50 micrograms whole egg-white and above) and adjuvant (1.0 ml pertussis vaccine); mast cells harvested between 20 and 50 days after the sensitising dose exhibited maximal histamine release upon in vitro challenge with 'whole' egg-white (100 micrograms). Routine use of plastic ware, and ice-cold phosphate-buffered saline (pH 7.2) for handling cells and avoidance of heparin and excessive centrifugation ensured optimal preservation of histamine-releasing capacity of the harvested peritoneal mast cells.
mesh_terms: D000375:Aging; D000818:Animals; D004306:Dose-Response Relationship, Immunologic; D004492:Edetic Acid / Q000494:pharmacology; D000939:Epitopes; D005260:Female; D006493:Heparin / Q000494:pharmacology; D006636:Histamine Release; D007158:Immunologic Techniques; D008297:Male; D008407:Mast Cells / Q000276:immunology*; D008722:Methods; D010047:Ovalbumin / Q000008:administration & dosage; D051381:Rats; D013997:Time Factors
chemical_list: D000939:Epitopes; D006493:Heparin; D010047:Ovalbumin; D004492:Edetic Acid