Journal of Experimental Medicine, Vol 173, 561-568, Copyright © 1991 by Rockefeller University Press

ARTICLES

T cell receptor-mediated selection of functional rat CD8 T cells from defined immature thymocyte precursors in short-term suspension culture

T Hunig and R Mitnacht
Institute for Virology and Immunobiology, University of Wurzburg, Federal Republic of Germany.

Recent results have indicated that positive and negative repertoire selection act on the major population of CD4,8 double-positive (DP) thymocytes that express 5-10-fold less T cell receptor (TCR) than mature T cells (i.e., they are TCRlow). Since DP cells obtained ex vivo are heterogeneous with regard to their stage within thymic selection, a homogeneous population of virgin DP cells suitable for selection studies was generated in vitro from their immediate precursors, the CD8 single-positive (SP) immature blast cells. To mimic TCR-mediated selection signals, these virgin DP cells were then cultured for another 2 d in the presence of immobilized anti-TCR monoclonal antibodies with or without interleukin 2 (IL-2). Daily monitoring of recovery and phenotype showed that without TCR stimulation, the cells remained DP and became small, TCRlow cells that were lost with a half-life of 1 d, regardless of the presence of IL-2. TCR stimulation resulted in rapid downregulation of CD4 and CD8, maintenance of a larger cell size, and induction of the CD53 antigen that marks mature and CD4,8 double- negative rat thymocytes. In the absence of IL-2, viability decreased as rapidly as without TCR stimulation. Addition of IL-2 rescued TCR- stimulated virgin DP cells and prevented CD8 downregulation, so that 50- 80% of input DP cells were recovered after 2 d as CD4-8+53+ cells. After release from modulation, these in vitro generated CD8 SP cells quantitatively upregulated the TCR to the TCRhigh phenotype and were readily induced to proliferate and exhibit cytotoxic T lymphocyte (CTL) activity in a polyclonal readout. Evidence is presented implicating an IL-2 receptor (IL-2R) not containing the p55 chain (i.e., most likely the p70 intermediate affinity IL-2R) in the TCR plus IL-2-driven in vitro differentiation of virgin DP cells towards the mature CD8 SP phenotype.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• van den Brandt, J., Kwon, S.-H., Hunig, T., McPherson, K. G., Reichardt, H. M. (2005). Sustained Pre-TCR Expression in Notch1IC-Transgenic Rats Impairs T Cell Maturation and Selection. J. Immunol. 174: 7845-7852 [Abstract] [Full Text]  
• Jimenez, E., Sacedon, R., Vicente, A., Hernandez-Lopez, C., Zapata, A. G., Varas, A. (2002). Rat Peripheral CD4+CD8+ T Lymphocytes Are Partially Immunocompetent Thymus-Derived Cells That Undergo Post-Thymic Maturation to Become Functionally Mature CD4+ T Lymphocytes. J. Immunol. 168: 5005-5013 [Abstract] [Full Text]  
• Akashi, K., Kondo, M., Weissman, I. L. (1998). Two distinct pathways of positive selection for thymocytes. Proc. Natl. Acad. Sci. USA 95: 2486-2491 [Abstract] [Full Text]  
• Mitnacht, R., Bischof, A., Torres-Nagel, N., Hunig, T. (1998). Opposite CD4/CD8 Lineage Decisions of CD4+8+ Mouse and Rat Thymocytes to Equivalent Triggering Signals: Correlation with Thymic Expression of a Truncated CD8{alpha} Chain in Mice But Not Rats. J. Immunol. 160: 700-707 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search

TABLE OF CONTENTS