Polyclonal activation of canine B lymphocytes evaluated by a protein A reverse hemolytic plaque assay.
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.
D000818:Animals; D000921:Antibody-Producing Cells / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D002469:Cell Separation; D004285:Dogs; D006462:Hemolytic Plaque Technique; D007070:Immunoglobulin A / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D066298:In Vitro Techniques; D007700:Kinetics; D008213:Lymphocyte Activation; D009000:Monocytes; D011043:Pokeweed Mitogens / Q000494:pharmacology; D013205:Staphylococcal Protein A
D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011043:Pokeweed Mitogens; D013205:Staphylococcal Protein A