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- W3148340632 abstract "Objective To difine the factors which affect the specificity and sensitivity for detecting the antigen specific CTL immune response in ELISPOT assay, so as to optimize the ELISPOT assay. Methods PBMC-derived CD8~+ T cells specific to HLA-A * 0201 binding peptide influenza matrix peptide Flu p58-66 were induced in vitro 7 days^ stimulation with peptide-pulsed irradiated PBMCs or DCs from HLA-A2 positive healthy donors, the frequency of effector cells to secrete IFN-gamma in response to Flu p58-66 was detected with an improved ELISPOT assay. Results The frequency of effector cells to secrete IFN-gamma in response to Flu peptide can be increased when the CD8~+ T cells were separated from PBMC before antigen stimulation than the CD8~+ T cells were separated from PBMC after antigen stimulation (P < 0.05). For CTL activation, DCs was better than CD8~- PBMCs as APC (P < 0.05); the cytokine cocktail (IL-7 + IL-2 + IL-6) was more effective than the IL-2 alone for CTL proliferation. During the CTL functional monitoring in ELISPOT assay, T2-2 cells were more potent than that of T2-1 cells to stimulate effector cells, the former reflect to stimulate effector cells to generate more numbers of specific spots (P < 0.05). Conclusion When the critical factors are optimized, the specificity and sensitivity of IFN-gamma-producing ELISPOT assay can be substantially improved." @default.
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- W3148340632 date "2002-01-01" @default.
- W3148340632 modified "2023-09-26" @default.
- W3148340632 title "Technical optimization for detecting the antigen specific CTL immune response in ELISPOT assay" @default.
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