The enumeration of mouse IgE-secreting cells using plaque-forming cell assays

✍ Scribed by Edward S. Rector; Glen M. Lang; Brian G. Carter; Ken A. Kelly; Peter G. Bundesen; Irmgard Böttcher; Alec H. Sehon

• Publisher: John Wiley and Sons
• Year: 1980
• Tongue: English
• Weight: 708 KB
• Volume: 10
• Category: Article
• ISSN: 0014-2980
• DOI: 10.1002/eji.1830101210

✦ Synopsis

Abstract

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 viable cells of two distinct IgE‐secreting hybridoma cell lines were scored as PFC. Under these conditions, plaques were not obtained with IgG~1~ or IgG~2a~‐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 (C 57 BL/6 × DBA/2)F~1~ mice following immunization with 10 μg of OA adsorbed to 1 mg of Al(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.