The enumeration of mouse IgE-secreting cells using plaque-forming cell assays.
With the aid of a specific rabbit antibody preparation to purified monoclonal murine IgE, two plaque-forming cell (PFC) assays have been developed for the detection and enumeration of mouse IgE-secreting cells. The first assay, utilizing protein A-coated sheep red cells (protein A-SRC), detected antibody-secreting cells  on the basis of the class  of the secreted Ig irrespective of antigen specificity. With this assay, 30% of the class of the viable cells of two distinct IgE-secreting hybridoma cell lines were scored as PFC. Under these conditions, plaques were not obtained with IgG1 or IgG2a-secreting hybridoma cells. The second PFC assay, which utilized SRC coated with ovalbumin (OA-SRC), enumerated cells secreting anti-OA IgE antibodies. Similar kinetic patterns were observed for the cellular (IgE PFC/spleen) and humoral (IgE serum levels) responses of (C57BL/6 x DBA/2)F1 mice following immunization with 10 micrograms of OA adsorbed to 1 mg of A1(OH)3. Thus, it is concluded that the reverse plaque assay detecting all IgE-secreting cells, as well as the antigen-specific IgE PFC assay, can be used for the quantitation of IgE responses at the cellular level.
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D000921:Antibody-Producing Cells / Q000276:immunology*; D002452:Cell Count; D002470:Cell Survival; D000939:Epitopes; D006462:Hemolytic Plaque Technique; D007073:Immunoglobulin E / Q000096:biosynthesis*; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D010047:Ovalbumin / Q000276:immunology; D051381:Rats
D000939:Epitopes; D007073:Immunoglobulin E; D010047:Ovalbumin