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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.

Publication ,  Journal Article
Kelsoe, G
Published in: J Immunol Methods
February 11, 1985

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.

Duke Scholars

Published In

J Immunol Methods

DOI

ISSN

0022-1759

Publication Date

February 11, 1985

Volume

76

Issue

2

Start / End Page

345 / 363

Location

Netherlands

Related Subject Headings

  • Spleen
  • Nitrohydroxyiodophenylacetate
  • Mitogens
  • Mice
  • Lipopolysaccharides
  • Immunology
  • Immunoglobulin Light Chains
  • Hemolytic Plaque Technique
  • Epitopes
  • Cells, Cultured
 
Journal cover image

Published In

J Immunol Methods

DOI

ISSN

0022-1759

Publication Date

February 11, 1985

Volume

76

Issue

2

Start / End Page

345 / 363

Location

Netherlands

Related Subject Headings

  • Spleen
  • Nitrohydroxyiodophenylacetate
  • Mitogens
  • Mice
  • Lipopolysaccharides
  • Immunology
  • Immunoglobulin Light Chains
  • Hemolytic Plaque Technique
  • Epitopes
  • Cells, Cultured