Fractionation of mycobacterial integral membrane proteins by continuous elution SDS-PAGE reveals the immunodominance of low molecular weight subunits for human T cells.
Integral membrane proteins (IMP) represent a serologically distinct class of mycobacterial antigens which are potent stimulators of human T cells (Mehrotra et al., Clin Exp Immunol 1995; 102:626). The range of IMP from Mycobacterium fortuitum was resolved by continuous elution SDS-PAGE to recover 31 discrete fractions covering bands up to approximately 58 kD. The fractions, after removal of SDS, were subjected to human T cell proliferation assays for the identification of immunodominant molecule(s). A low molecular weight (<20kD) fraction was able to stimulate T cells from 11 out of 12 donors comprising mainly tuberculoid leprosy patients. The described protocol is well suited to situations where large quantities of antigenic protein mixtures must be processed in order to get the purified molecules/fractions in amounts required for immunoepidemiological studies.
Mehrotra
J
J
Division of Membrane Biology, Central Drug Research Institute, Lucknow, India.
Mittal
A
A
Dhindsa
M S
MS
Sinha
S
S
D001426:Bacterial Proteins / Q000276:immunology* / Q000302:isolation & purification*; D002455:Cell Division / Q000276:immunology; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D016056:Immunodominant Epitopes; D015441:Leprosy, Tuberculoid / Q000276:immunology; D008565:Membrane Proteins / Q000276:immunology* / Q000302:isolation & purification*; D009170:Nontuberculous Mycobacteria / Q000276:immunology*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
D001426:Bacterial Proteins; D016056:Immunodominant Epitopes; D008565:Membrane Proteins