The production and characterization of murine monoclonal antibodies to a DNA receptor on human leukocytes

R. M. Bennet, S. H. Hefeneider, A. Bakke, M. Merritt, C. A. Smith, D. Mourich, M. C. Heinrich

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Abstract

Two murine mAb have been generated with a reactivity toward a 30,000 m.w. DNA binding protein found on the cell surface of human leukocytes; mAb 12A has an IgG1/k isotype, and mAb 24T has an IgG2b/k isotype. Both react with the DNA binding domain or adjacent region of the putative DNA receptor and inhibit the binding of [3H]DNA to PBMC at concentrations as low as 100 ng/ml. Stoichiometric studies indicate that both mAb react with monocytes and T cells with a kDa of 10-7 M; about 0.5 x 106 molecules bind per cell at saturation. Flow cytometry indicated that 67% of lymphocytes and 98% of monocytes bore the DNA receptor. Dual labeling studies showed that 90% of B cells and 50% of T cells express the receptor; 50% of CD4+ T cells are receptor positive. Immunomatrices constructed with both mAb 12A and 24T allowed the receptor to be purified to a high degree of purity. A single protein of M(r) 30,000 was readily observed after SDS-PAGE and silver staining of the gel; after electropheretic transfer of nitrocellulose this protein was shown to be a DNA binding molecule by use of a probe of biotin labeled DNA. These experiments provide further evidence to support the existence of a specific DNA receptor on human leukocytes; the availability of mAb to the receptor should be useful in its further characterization.

Original languageEnglish (US)
Pages (from-to)2937-2942
Number of pages6
JournalJournal of Immunology
Volume140
Issue number9
StatePublished - Jan 1 1988

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ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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