Screening gene expression libraries for epitopes recognized in Mycobacterium leprae by mouse T cells

Eur J Immunol. 1991 Oct;21(10):2621-4. doi: 10.1002/eji.1830211047.

Abstract

Parasite expression libraries have so far been screened with antibodies, DNA probes or T cell clones. Immunity to many parasites, such as Mycobacterium leprae, is largely mediated by T cells, and so the screening of such libraries for T cell epitopes is an important step toward the development of effective vaccines and diagnostic reagents. A new method for screening of lambda gt11 libraries with uncloned T cell populations is presented here, which takes advantage of the fact that the recombinant proteins contain beta-galactosidase as their leader peptide; this allows them to be semipurified by means of anti-beta-galactosidase antibodies coated on the bottom of microtiter plate wells, within which a proliferation assay can then be carried out. Optimum conditions for the assay were determined, using the M. leprae 18-kDa antigen as a test antigen.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology*
  • CD4-Positive T-Lymphocytes / immunology*
  • Cloning, Molecular
  • Gene Library
  • Immunosorbent Techniques
  • In Vitro Techniques
  • Lymphocyte Activation
  • Mice
  • Mycobacterium leprae / genetics
  • Mycobacterium leprae / immunology*
  • Recombinant Fusion Proteins / immunology
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • Antigens, Bacterial
  • Recombinant Fusion Proteins
  • beta-Galactosidase