A mutant human beta2-microglobulin can be used to generate diverse multimeric class I peptide complexes as specific probes for T cell receptors

J Immunol Methods. 1998 May 1;214(1-2):41-50. doi: 10.1016/s0022-1759(98)00035-0.

Abstract

Antigen-specific receptors (TCR) on CD8 T lymphocytes form relatively short-lived complexes with their natural ligands: peptides in association with major histocompatibility complex (MHC) class I molecules, which consist of a polymorphic heavy chain and a conserved light chain, beta2-microglobulin (beta2-M). To produce soluble MHC-peptide complexes in a form that would bind more stably and could be used to identify, count, and isolate CD8 T cells having the appropriate TCR, we prepared multimeric MHC-peptide complexes. Our work builds on the assembly of recombinant MHC class I peptide complexes using a mutant human beta2-M chain (Tyr 67 > Cys) which can form stable heterodimers with diverse MHC heavy chains. With biotin added to the SH group, the assembled MHC-peptide monomers formed multimers with avidin linked to a fluorochrome. The specific reactivity of the multimeric reagents with human and mouse cytotoxic T cells (CTL) is described. The present approach permits the production of class I multimers, without the necessity of genetic engineering each heavy chain, a significant advantage in view of the enormous polymorphism of MHC heavy chains. Because human beta2-M forms stable heterodimers with diverse class I heavy chains from various species (human and non human primates, mouse, etc.), this procedure is a general method for producing multimers of MHC-peptide complexes as T cell receptor-specific probes.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Binding Sites
  • Biotin / chemistry
  • Fluorescent Dyes / chemistry
  • Histocompatibility Antigens Class I / chemistry*
  • Humans
  • Immunoglobulin Heavy Chains / chemistry
  • Mice
  • Mutation
  • Peptides / chemical synthesis*
  • Receptors, Antigen, T-Cell / analysis*
  • Receptors, Antigen, T-Cell / chemistry
  • Substrate Specificity
  • beta 2-Microglobulin / chemistry*
  • beta 2-Microglobulin / genetics

Substances

  • Fluorescent Dyes
  • Histocompatibility Antigens Class I
  • Immunoglobulin Heavy Chains
  • Peptides
  • Receptors, Antigen, T-Cell
  • beta 2-Microglobulin
  • Biotin