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KMID : 0923620060060040179
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Immune Network
2006 Volume.6 No. 4 p.179 ~ p.184
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HLA-restricted and Antigen-specific CD8? T Cell Responses by K562 Cells Expressing HLA-A*0201
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Yun Sun-Ok
Yoon Sung-Hee
Sohn Hyun-Jung
Choi Hee-Baeg
Kim Tai-Gyu
Sohn Hyun-Jung
Yoon Sung-Hee
Choi Hee-Baeg
Kim Tai-Gyu
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Abstract
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Background: Identification of antigen-specific T cells has yielded valuable information on pathologic process and the disease state. Assays for quantification of inflammatory cytokines or lytic-granule molecules have been generally used to evaluate antigen specific T cell response, however their applicability have been hampered due to the limited source of autologous antigen-presenting target cells (APC).
Methods: K562, a leukemic cell line deficient of human leukocyte antigen (HLA), was transfected with a gene encoding HLA-A*02 (K562/A*02) and its function as stimulator cells in inducing activation of HLA-matched T cells was evaluated by IFN-γ enzyme linked immunospot (ELISPOT) assay.
Results: The stable transfectant K562/A*02 pulsed with HLA- A*02 restricted peptide could specifically induce IFN-γ secretion by CD8? T cells compared to no detectable secretion by CD4? T cells. However, CD56? NK cells secreted IFN-γ in both K562/A*02 with peptide and without peptide. The number of IFN-γ secreted CD8? T cells was increased according to the ratio of T cells to K562 and peptide concentration. Formalin-fixed K562/A*02 showed similar antigen presenting function to live K562/A*02. Moreover, K562/A*02 could present antigenic- peptide to not only A*0201 restricted CD8? T cells but also CD8? T cells from A*0206 donor.
Conclusion: These results suggest that K562/A*02 could be generally used as target having specificity and negligible background for measuring CD8? T cell responses and selective use of K562 with responsder matched HLA molecules on its surface as APC may circumvent the limitation of providing HLA-matched autologous target cells. (Immune Network 2006;6(4):179-184)
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KEYWORD
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K562, HLA-A*0201, peptides, CD8? T cells, IFN-γ ELISPOT assay
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