Expression and analysis of soluble MHC- and tcrimmunoglbulin super dimers

Scan M. O'Herrin, Jonathan P. Schneck

Research output: Contribution to journalArticlepeer-review

Abstract

Specificity in immune responses is in part controlled by the selective interaction of T cell receptors with their cognate ligands, peptide/MHC molecules. The discriminating nature of this interaction makes these molecules, in soluble form, good candidates for selectively regulating immune responses. Attempts to exploit soluble analogs of these proteins has been hampered by the intrinsic low avidity of these molecules for their ligands. To increase the avidity of soluble analogs for their cognates to biologically relevant levels, divalent peptide/MHC complexes or T cell receptors (superdimers) were constructed. Using a recombinant DNA strategy, DNA encoding either the MHC class 11/peptide or TCR heterodimers was ligated to DNA coding for murine Ig heavy and light chains. These constructs were subsequently expressed in a baculovirus expression system. Enzyme-linked immunosorbant assays specific for the Ig and polymorphic determinants of either the TCR or MHC fraction of the molecule indicated that infected insect cells secreted approximately 1 M-g/rol °f soluble, conformationally intact chimeric superdimers. SDS PAGE gel analysis of purified protein showed the expected molecular weight species. The results of flow cytometry demonstrated that the TCR and class II chimeras bound specifically with high avidity to cells bearing their cognate receptors. These superdimers will be useful reagents for studying TCRTMHC interactions, lymphocyte tracking and have possible use as specific regulators of immune responses. The method of construction and expression outlined here is useful as a general approach to express and study the function of soluble superdimers of heterodimeric integral membrane proteins.

Original languageEnglish (US)
Pages (from-to)A1473
JournalFASEB Journal
Volume10
Issue number6
StatePublished - Dec 1 1996

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Expression and analysis of soluble MHC- and tcrimmunoglbulin super dimers'. Together they form a unique fingerprint.

Cite this