High affinity xenoreactive TCR

MHC interaction recruits CD8 in absence of binding to MHC

Jennifer Buslepp, Samantha E. Kerry, Doug Loftus, Jeffrey A Frelinger, Ettore Appella, Edward J. Collins

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The TCR from a xenoreactive murine cytotoxic T lymphocyte clone, AHIII 12.2, recognizes murine H-2Db complexed with peptide p1058 (FAPGFFPYL) as well as human HLA-A2.1 complexed with human self-peptide p1049 (ALWGFFPVL). To understand more about T cell biology and cross-reactivity, the ectodomains of the AHIII 12.2 TCR have been produced in E. coli as inclusion bodies and the protein folded to its native conformation. Flow cytometric and surface plasmon resonance analyses indicate that human p1049/A2 has a significantly greater affinity for the murine AHIII 12.2 TCR than does murine p1058/Db. Yet, T cell binding and cytolytic activity are independent of CD8 when stimulated with human p1049/A2 as demonstrated with anti-CD8 Abs that block CD8 association with MHC. Even in the absence of direct CD8 binding, stimulation of AHIII 12.2 T cells with "CD8-independent" p1049/A2 produces p56lck activation and calcium flux. Confocal fluorescence microscopy and fluorescence resonance energy transfer flow cytometry demonstrate CD8 is recruited to the site of TCR:peptide MHC binding. Taken together, these results indicate that there exists another mechanism for recruitment of CD8 during high affinity TCR:peptide MHC engagement.

Original languageEnglish (US)
Pages (from-to)373-383
Number of pages11
JournalJournal of Immunology
Volume170
Issue number1
StatePublished - Jan 1 2003
Externally publishedYes

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varespladib methyl
T-Lymphocytes
Peptides
Fluorescence Resonance Energy Transfer
Surface Plasmon Resonance
Inclusion Bodies
Cytotoxic T-Lymphocytes
Fluorescence Microscopy
Confocal Microscopy
Cell Biology
Flow Cytometry
Clone Cells
Escherichia coli
Calcium
Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Buslepp, J., Kerry, S. E., Loftus, D., Frelinger, J. A., Appella, E., & Collins, E. J. (2003). High affinity xenoreactive TCR: MHC interaction recruits CD8 in absence of binding to MHC. Journal of Immunology, 170(1), 373-383.

High affinity xenoreactive TCR : MHC interaction recruits CD8 in absence of binding to MHC. / Buslepp, Jennifer; Kerry, Samantha E.; Loftus, Doug; Frelinger, Jeffrey A; Appella, Ettore; Collins, Edward J.

In: Journal of Immunology, Vol. 170, No. 1, 01.01.2003, p. 373-383.

Research output: Contribution to journalArticle

Buslepp, J, Kerry, SE, Loftus, D, Frelinger, JA, Appella, E & Collins, EJ 2003, 'High affinity xenoreactive TCR: MHC interaction recruits CD8 in absence of binding to MHC', Journal of Immunology, vol. 170, no. 1, pp. 373-383.
Buslepp J, Kerry SE, Loftus D, Frelinger JA, Appella E, Collins EJ. High affinity xenoreactive TCR: MHC interaction recruits CD8 in absence of binding to MHC. Journal of Immunology. 2003 Jan 1;170(1):373-383.
Buslepp, Jennifer ; Kerry, Samantha E. ; Loftus, Doug ; Frelinger, Jeffrey A ; Appella, Ettore ; Collins, Edward J. / High affinity xenoreactive TCR : MHC interaction recruits CD8 in absence of binding to MHC. In: Journal of Immunology. 2003 ; Vol. 170, No. 1. pp. 373-383.
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