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Review
. 2000 Nov;7(6):859-64.
doi: 10.1128/CDLI.7.6.859-864.2000.

New methods for assessing T-cell responses

N Bercovici  1 , M T Duffour, S Agrawal, M Salcedo, J P Abastado
Affiliations
Review

New methods for assessing T-cell responses

N Bercovici et al. Clin Diagn Lab Immunol. 2000 Nov.
No abstract available

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Figures

FIG. 1
FIG. 1
Melan-A/MART-1-specific IFN-γ-producing T cells detected by ELISPOT assay. A T-cell population containing Melan-A-MART-1-specific CTL was incubated with unpulsed or Melan-A/MART-1-pulsed dendritic cells (DC) in a 96-well plate precoated with anti-IFN-γ antibody. IFN-γ-secreting cells were revealed after 20 h of culture. Of 1,000 T cells, 280 are specific for HLA-A2/Melan-A.
FIG. 2
FIG. 2
Quantification of functional epitope-specific T cells using MHC tetramers and IFN-γ intracellular staining. T cells were incubated for 6 h with dendritic cells (DC) pulsed with the Melan-A/MART-1 peptide or an irrelevant (MAFlu) peptide. Brefeldin A was added for the last 3 h of incubation. IFN-γ produced by Melan-A-MART-1-specific T cells was quantified by flow cytometry after gating on CD8+ cells.
FIG. 3
FIG. 3
Correlation between the cytotoxic activity and the frequency of epitope-specific T cells detected by MHC tetramers and ELISPOT assay. The cytotoxic activity of two T-cell lines, B2 and F9, against autologous B-EBV cells pulsed or not pulsed with Melan-A/MART-1 peptide was measured in a 4-h 51Cr release assay. The frequency of Melan-A/MART-1-specific CTL in B2 and F9 cells was determined by ELISPOT assay after stimulation with peptide-pulsed dendritic cells and by HLA-A2/Melan-A tetramer staining after gating on CD8+ cells. SFC, spot-forming cell; PBMC, peripheral blood mononuclear cells.
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