Differential sensitivity of T cell activation vs deletion

D. A. Peterson, R. J. DiPaolo, E. R. Unanue

Research output: Contribution to journalArticle

Abstract

It has been previously shown from studies using super-antigen or TCR transgenic mice that deletion of double positive (DP) thymocytes is more sensitive than activation of mature T cells. In this study we quantitate the magnitude of this difference in sensitivity in identical culture conditions. We used the 3A9 transgenic mouse which expresses a transgenic T cell receptor (TCR) specific for hen egg lysozyme (HEL) peptide 48-61 on I-Ak. The thymus of this mouse contains roughly 35% mature CD4 single positive(SP) and 45% immature DP. 1x106 thymocytes were placed into a well, with 1x105 B-lymphomas transfected with I-Ak which have been pulsed with HEL 48-61 overnight and washed. After 20 hours in culture the viable cells are counted, and then stained for CD4, CD8, and CD69. Deletion is then measured as % of control DP cells, and activation is measured as % of maximum mean fluorescence of CD69 on CD4 SP cells. A monoclonal antibody specific for I-Ak-48-61 peptide-MHC complex was used to estimate the number of complexes per antigen presenting cell. Our results indicate that negative selection of DP thymocytes is a highly sensitive process that requires 10-15 fold less antigen than activation of mature CD4 T cells.

Original languageEnglish (US)
JournalFASEB Journal
Volume12
Issue number5
StatePublished - Mar 20 1998
Externally publishedYes

Fingerprint

T-cells
Thymocytes
thymocytes
T-lymphocytes
Chemical activation
T-Cell Antigen Receptor
T-Lymphocytes
Transgenic Mice
genetically modified organisms
lysozyme
Antigens
hens
Thymus
mice
peptides
Antigen-Presenting Cells
antigens
Thymus Gland
receptors
eggs

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Peterson, D. A., DiPaolo, R. J., & Unanue, E. R. (1998). Differential sensitivity of T cell activation vs deletion. FASEB Journal, 12(5).

Differential sensitivity of T cell activation vs deletion. / Peterson, D. A.; DiPaolo, R. J.; Unanue, E. R.

In: FASEB Journal, Vol. 12, No. 5, 20.03.1998.

Research output: Contribution to journalArticle

Peterson, DA, DiPaolo, RJ & Unanue, ER 1998, 'Differential sensitivity of T cell activation vs deletion', FASEB Journal, vol. 12, no. 5.
Peterson DA, DiPaolo RJ, Unanue ER. Differential sensitivity of T cell activation vs deletion. FASEB Journal. 1998 Mar 20;12(5).
Peterson, D. A. ; DiPaolo, R. J. ; Unanue, E. R. / Differential sensitivity of T cell activation vs deletion. In: FASEB Journal. 1998 ; Vol. 12, No. 5.
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