Cloning of mitogen- and antigen-reactive B lymphocytes on filter paper discs. I. A description of the technique and of methods for the analysis of colonies.
A novel technique for establishing short term clones of antigen- or mitogen-activated splenic B lymphocytes is described. Spleen cells are plated onto the surface of filter paper discs and subsequently stimulated by antigen or mitogen in situ; activated B cells proliferate and differentiate into pure colonies of cells analogous to bacterial colonies growing on agar. These colonies of lymphocytes may be characterized in a series of replica hemolytic-plaque, autoradiographic, or immunoenzyme assays making possible a full characterization of the frequency of secreted idiotopes and paratopes and of the cells that produce them. Colony induction by either antigen or mitogen occurs under identical conditions, thus a rigorous comparison between the mitogen-selected and antigen-selected antibody repertoires may be made.
Kelsoe
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D000818:Animals; D000916:Antibody Diversity; D000918:Antibody Specificity; D000941:Antigens; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology*; D002478:Cells, Cultured; D000939:Epitopes; D006462:Hemolytic Plaque Technique; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000276:immunology; D051379:Mice; D008934:Mitogens; D009592:Nitrohydroxyiodophenylacetate / Q000276:immunology; D013154:Spleen / Q000166:cytology
D000941:Antigens; D000939:Epitopes; D007147:Immunoglobulin Light Chains; D008070:Lipopolysaccharides; D008934:Mitogens; D009592:Nitrohydroxyiodophenylacetate