The Journal of Experimental Medicine
Avanti Polar Lipids, Inc.
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 287K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JEM
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Manning, T. C.
Right arrow Articles by Kranz, D. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Manning, T. C.
Right arrow Articles by Kranz, D. M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?
© The Rockefeller University Press, 0022-1007/1999/2/461/ $5.00
The Journal of Experimental Medicine, Volume 189, Number 3, February 1, 1999 461-470

Articles

Effects of Complementarity Determining Region Mutations on the Affinity of an {alpha}/β T Cell Receptor: Measuring the Energy Associated with CD4/CD8 Repertoire Skewing

Thomas C. Manning*, Evan A. Parke*, Luc Teyton{ddagger}, and David M. Kranz*

From the * Department of Biochemistry, University of Illinois, Urbana, Illinois 61801; and the {ddagger} Department of Immunology, The Scripps Research Institute, La Jolla, California 92037

It has been proposed that the generally low affinities of T cell receptors (TCRs) for their peptide–major histocompatibility complex (pMHC) ligands (Kd ~10–4 to 10–7 M) are the result of biological selection rather than an intrinsic affinity limitation imposed by the TCR framework. Using a soluble version of the 2C TCR, we have used complementarity determining region (CDR)-directed mutagenesis to investigate whether the affinity of this receptor for its allogeneic pMHC ligand can be improved upon. We report that several mutants at positions lying within CDR3{alpha} and CDR2β showed increased affinities for pMHC compared with the wild-type receptor. Additionally, we have investigated whether V{alpha} mutations that have been implicated in the phenomenon of CD8+ repertoire skewing achieve this skewing by means of generalized increases in affinity for MHC-I molecules. Two mutants (S27F and S51P), which each promote skewing toward a CD8+ phenotype, exhibited significantly reduced affinity for pMHC-I, consistent with a quantitative-instructional model of CD4/CD8 lineage commitment. This model predicts that CD8 is downregulated on thymocytes that have TCR–ligand interactions above a minimal energy threshold. Together, the results (a) demonstrate that engineering higher affinity TCRs is feasible, and (b) provide TCR–pMHC energy values associated with CD4/CD8 repertoire skewing.

Key Words: T cell receptor • peptide–major histocompatibility complex • affinity • CD4/ CD8 • complementarity determining region

Address correspondence to David M. Kranz, Department of Biochemistry, University of Illinois, 600 S. Mathews, Urbana, IL 61801. Phone: 217-244-2821; Fax: 217-244-5858; E-mail: d-kranz{at}uiuc.edu

Abbreviations used: {Delta}{Delta}G, free energy change; MCMV, murine CMV; pMHC, peptide–MHC complex; scTCR, single-chain TCR; SPR, surface plasmon resonance.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?




  Home | Help | Feedback | Subscriptions | Archive | Search
TABLE OF CONTENTS