Screening gene expression libraries for epitopes recognized in Mycobacterium leprae by mouse T cells.
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.
Imperial Cancer Research Fund, Department of Biology, Medawar University College, London, GB.
B
B
Villarreal-Ramos
J
J
Sanchez-Garcia
N
N
Stoker
E
E
Timms
D
D
Chomer
K
K
Raff
N A
NA
Mitchison
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D003001:Cloning, Molecular; D015723:Gene Library; D007163:Immunosorbent Techniques; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D051379:Mice; D009166:Mycobacterium leprae / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D000942:Antigens, Bacterial; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase