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Identification of endogenous peptides recognized by in vivo or in vitro generated alloreactive cytotoxic T lymphocytes: distinct characteristics correlated with CD8 dependence

We studied the molecular basis for CD8 independence of in vivo generated (BM3.3) versus CD8 dependence of in vitro sensitized (KB5.C20/Des) alloreactive H‐2Kb‐specific cytotoxic T lymphocytes (CTL). Using microcapillary high‐performance liquid chromatography fractionation of H‐2Kb eluates, mass spec...

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Bibliographic Details
Published in:European journal of immunology 2001-02, Vol.31 (2), p.421-432
Main Authors: Guimezanes, Annick, Barrett‐Wilt, Gregory A., Gulden‐Thompson, Pamela, Shabanowitz, Jeffrey, Engelhardt, Victor H., Hunt, Donald F., Schmitt‐Verhulst, Anne‐Marie
Format: Article
Language:English
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Summary:We studied the molecular basis for CD8 independence of in vivo generated (BM3.3) versus CD8 dependence of in vitro sensitized (KB5.C20/Des) alloreactive H‐2Kb‐specific cytotoxic T lymphocytes (CTL). Using microcapillary high‐performance liquid chromatography fractionation of H‐2Kb eluates, mass spectrometry and CTL reconstitution assays, we determined that BM3.3 and KB5.C20 recognize, respectively, a single peptide (pBM1) expressed on 8,000 H‐2Kb molecules per allogeneic cell, and three distinct peptides (pKB1, 2, 3), each expressed on around 200 H‐2Kb molecules per allogeneic cell. CD8 (in)dependence was intrinsic to the respective TCR/H‐2Kb‐peptide interactions. KB5.C20 and BM3.3 TCR illustrate the correlation that appears to exist between CD8 dependence/low affinity and in vitro sensitization as opposed to low dependency on CD8 and high TCR affinity observed after in vivo sensitization. The results suggest that CD8‐dependent alloreactive CTL obtained in vitro with high frequency correspond to low‐affinity TCR from the MHC‐biased TCR repertoire unpurged by negative selection and have implications for cellular immunotherapeutic approaches.
ISSN:0014-2980
1521-4141
DOI:10.1002/1521-4141(200102)31:2<421::AID-IMMU421>3.0.CO;2-4