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

Bernardo Villarreal‐Ramos, Javier Sanchez‐Garcia, Neil Stoker, Emma Timms, Deborah Chomer, Kim Raff, N. Avrion Mitchison*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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 λgt11 libraries with uncloned T cell populations is presented here, which takes advantage of the fact that the recombinant proteins contain β‐galactosidase as their leader peptide; this allows them to be semipurified by means of anti‐β‐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.

Original languageEnglish
Pages (from-to)2621-2624
Number of pages4
JournalEuropean Journal of Immunology
Volume21
Issue number10
DOIs
Publication statusPublished - 01 Oct 1991

Keywords

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

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