| Autor: |
I M, Belyakov, J, Wang, R, Koka, J D, Ahlers, J T, Snyder, R, Tse, J, Cox, J S, Gibbs, D H, Margulies, J A, Berzofsky |
| Rok vydání: |
2001 |
| Předmět: |
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| Zdroj: |
European journal of immunology. 31(12) |
| ISSN: |
0014-2980 |
| Popis: |
Detection of the functional CD8(+) CTL response usually requires in vitro restimulation. The differences between the CD8(+) CTL repertoire in freshly isolated precursor cells and CD8(+) CTL after short-term in vitro expansion have been generally assumed to be minimal, but have never been defined experimentally. Using staining with P18-I10/H-2D(d) tetramers and monoclonal antibodies (mAb) against Vbeta, we show the surprising result that there was significant skewing of the CD8(+) CTL repertoire after just 7 days of stimulation. In contrast, we found that overnight incubation of precursor cells with peptide allows the functional assessment of CD8(+) CTL (which cannot be detected ex vivo from freshly isolated cells) without changing the absolute number of antigen-specific CTL as measured by tetramer staining or the repertoire of TCR analyzed with mAb. This study affords a better understanding of the differences between the ex vivo and in vitro stimulated CTL repertoire, and provides an approach to reveal a more faithful representation of the functional in vivo CTL response without skewing of the repertoire of T cells detected. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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