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.
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Related Subject Headings
- Spleen
- Nitrohydroxyiodophenylacetate
- Mitogens
- Mice
- Lipopolysaccharides
- Immunology
- Immunoglobulin Light Chains
- Hemolytic Plaque Technique
- Epitopes
- Cells, Cultured
Citation
Published In
DOI
ISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- Spleen
- Nitrohydroxyiodophenylacetate
- Mitogens
- Mice
- Lipopolysaccharides
- Immunology
- Immunoglobulin Light Chains
- Hemolytic Plaque Technique
- Epitopes
- Cells, Cultured