Abstract
This paper describes the optimal culture and assay conditions for the polyclonal activation of canine lymphocytes with pokeweed mitogen and the quantitation of immunoglobulin secreting plaque-forming cells (PFC) using a staphylococcal protein A—reverse hemolytic plaque assay. The assay permits the quantitation of total immunoglobulin secreting PFC as well as class-specific immunoglobulin secreting PFC. On the optimal day of culture, a mean of 176 IgA PFC/10 6, 575 IgM PFC/10 6, 1276 IgG PFC/10 6, and 2158 total PFC/10 6 cells were generated following polyclonal activation. This study provides a simple and reproducible assay for the delineation of the immunoregulatory mechanisms involved in the differentiation of canine B lymphocytes.
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