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The effect of co-expression costimulatory molecule CD80 on uptake of antigen peptide-MHC class I-GFP complex by specific T cells

The effect of co-expression costimulatory molecule CD80 on uptake of antigen peptide-MHC class I-GFP complex by specific T cells

International Journal of Oncology 30(6): 1389-1396

ISSN/ISBN: 1019-6439

PMID: 17487359

CD80, a costimulatory molecule, plays an important role in eliciting antitumor immunity. Without costimulation, recognition of antigens by T cells may not cause a response, even if tumor cells express MHC class I molecules and specific antigens. On the basis of the recombinant GFP-tagged K-b molecule, we constructed a co-expression vector of CD80 and GFP-tagged Kb molecules. The recombinant fusion was transfected into mouse melanoma B16 cells by electroporation; positive cells were obtained by G418 screening. Highly expressing monoclonal cells, irradiated by Cs-137, were used to immunize mice to obtain specific T cells, which were then cultivated with tumor cells in vitro and examined with a laser confocal microscope. The evident and intense uptake of the antigen peptide-MHC class I-GFP complex by specific T cells was visualized from the culture of B16/CD80-K-b-GFP and T cells. However, little uptake was observed from the culture of B16/K-b-GFP and T cells. These results show that co-expression of CD80 molecules with K-b, an MHC class I molecule, on the surface of B16 tumor cells can enhance the response of specific T cells and thus increase the uptake of the antigen peptide-MHC class I-GFP complex. The absorbed green fluorescence was concentrated mainly on the T cell surface, and this result might pave the way to eluting specific antigen peptides directly from T cells to find and isolate novel tumor-specific antigen peptides.

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Accession: 013870232

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