Modification of T-cell antigenic properties of tetanus toxoid by SDS-PAGE separation. Implications for T-cell blotting.
Using Tetanus Toxoid (TT) as a model antigen the T-cell Blotting method was evaluated. Peripheral blood mononuclear cell (PBMC) cultures were stimulated by blotted nitrocellulose-bound TT or soluble TT. SDS-Poly-Acrylamide-Gel-Electrophoresis separated TT only induced proliferation in 20% of the PBMC cultures whereas proliferation was induced in 79% of the same cultures offered similar treated TT (except for the PAGE separation). When T-cell blotting was performed with TT separated in a SDS-agarose matrix, proliferation was induced in 80% of donors responding to soluble TT. The results show that SDS-PAGE alters the ability of TT to induce T-cell proliferation, possibly due to unpolymerized acrylamide binding to proteins during SDS-PAGE. The use of SDS-PAGE T-cell blotting in the screening for T-cell antigens must therefore be reconsidered. We suggest the use of SDS-Agarose Gel Electrophoresis as an alternative when doing T-cell blots.
Christensen
C B
CB
Centre for Medical Parasitology, University of Copenhagen, Denmark.
Theander
T G
TG
D020106:Acrylamide; D000178:Acrylamides / Q000494:pharmacology; D003101:Collodion; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D008213:Lymphocyte Activation / Q000187:drug effects; D012995:Solubility; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology*; D013745:Tetanus Toxoid / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology
D000178:Acrylamides; D018984:Epitopes, T-Lymphocyte; D013745:Tetanus Toxoid; D020106:Acrylamide; D003101:Collodion