Enrichment of IgG anti-DNA-producing lymphoblastoid cell lines by antigen-coated immunomagnetic beads

Clin Immunol Immunopathol. 1992 Oct;65(1):39-44. doi: 10.1016/0090-1229(92)90245-j.

Abstract

In this paper we describe the establishment of four anti-DNA-producing lymphoblastoid cell lines (LCL) by Epstein-Barr virus (EBV) infection of peripheral blood B-cells from a patient with systemic lupus erythematosus. The LCL showed a heterogeneous cell composition: the frequencies of cells in active proliferation, cells secreting IgG or IgM, and cells effectively producing IgG or IgM anti-DNA were estimated by limiting dilution analysis. The cells producing anti-DNA antibodies were a small fraction of the whole cell population constituting the LCL. In order to enrich the fraction of cells producing the desired antibody we performed a positive selection by DNA-coated immunomagnetic beads. Results show that in two out of three IgG anti-DNA-secreting LCL the frequency of DNA-producing cells increased after incubation with DNA-coated beads. At variance, IgM anti-DNA-secreting cells were completely lost after immunomagnetic separation. This approach could represent a further tool to obtain better fusion partners to construct stable hybrids secreting human monoclonal antibodies. The advantages of the presented technique would be: (a) removing of the fraction of low-affinity IgM-producing cells, which is often prevalent in EBV-induced LCL; and (b) the enrichment of IgG anti-DNA producing cells, which better represent the antibodies involved in the pathogenesis of the disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antibodies, Antinuclear / immunology*
  • Antibody Formation
  • Antibody-Producing Cells / cytology
  • Antigens
  • B-Lymphocytes / microbiology
  • Cell Line
  • Cell Separation / methods
  • Female
  • Herpesviridae Infections
  • Herpesvirus 4, Human
  • Humans
  • Magnetics*
  • Microspheres

Substances

  • Antibodies, Antinuclear
  • Antigens