Design, Engineering, and Production of Human Recombinant T Cell Receptor Ligands Derived from Human Leukocyte Antigen DR2

Justin W. Chang, Diane E. Mechling, Hans Peter Bächinger, Gregory G. Burrows

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Major histocompatibility complex (MHC) class II molecules are membrane-anchored heterodimers on the surface of antigen-presenting cells that bind the T cell receptor, initiating a cascade of interactions that results in antigen-specific activation of clonal populations of T cells. Susceptibility to multiple sclerosis is associated with certain MHC class II haplotypes, including human leukocyte antigen (HLA) DR2. Two DRB chains, DRB5*0101 and DRB1*1501, are co-expressed in the HLA-DR2 haplotype, resulting in the formation of two functional cell surface heterodimers, HLA-DR2a (DRA*0101, DRB5*0101) and HLA-DR2b (DRA*0101, DRB1*1501). Both isotypes can present an immunodominant peptide of myelin basic protein (MBP-(84-102)) to MBP-specific T cells from multiple sclerosis patients. We have previously demonstrated that the peptide binding/T cell recognition domains of rat MHC class II (α1 and β1 domains) could be expressed as a single exon for structural and functional characterization; Burrows, G. G., Chang, J. W., Bächinger, H.-P., Bourdette, D. N., Wegmann, K. W., Offner, H., and Vandenbark A. A. (1999) Protein Eng. 12, 771-778; Burrows, G. G., Adlard, K. L., Bebo, B. F., Jr., Chang, J. W., Tenditnyy, K., Vandenbark, A. A., and Offner, H. (2000) J. Immunol. 164, 6366-6371). Single-chain human recombinant T cell receptor ligands (RTLs) of ∼200 amino acid residues derived from HLA-DR2b were designed using the same principles and have been produced in Escherichia coli with and without amino-terminal extensions containing antigenic peptides. Structural characterization using circular dichroism predicted that these molecules retained the antiparallel β-sheet platform and antiparallel α-helices observed in the native HLA-DR2 heterodimer. The proteins exhibited a cooperative two-state thermal unfolding transition, and DR2-derived RTLs with a covalently linked MBP peptide (MBP-(85-99)) showed increased stability to thermal unfolding relative to the empty DR2-derived RTLs. These novel molecules represent a new class of small soluble ligands for modulating the behavior of T cells and provide a platform technology for developing potent and selective human diagnostic and therapeutic agents for treatment of autoimmune disease.

Original languageEnglish (US)
Pages (from-to)24170-24176
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number26
DOIs
StatePublished - Jun 29 2001

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Human Engineering
HLA Antigens
T-Cell Antigen Receptor
T-cells
Ligands
Major Histocompatibility Complex
T-Lymphocytes
Peptides
Haplotypes
Molecules
Multiple Sclerosis
Hot Temperature
Dichlororibofuranosylbenzimidazole
Peptide T
Myelin Basic Protein
Dichroism
Antigen-Presenting Cells
Surface Antigens
Circular Dichroism
Escherichia coli

ASJC Scopus subject areas

  • Biochemistry

Cite this

Design, Engineering, and Production of Human Recombinant T Cell Receptor Ligands Derived from Human Leukocyte Antigen DR2. / Chang, Justin W.; Mechling, Diane E.; Bächinger, Hans Peter; Burrows, Gregory G.

In: Journal of Biological Chemistry, Vol. 276, No. 26, 29.06.2001, p. 24170-24176.

Research output: Contribution to journalArticle

Chang, Justin W. ; Mechling, Diane E. ; Bächinger, Hans Peter ; Burrows, Gregory G. / Design, Engineering, and Production of Human Recombinant T Cell Receptor Ligands Derived from Human Leukocyte Antigen DR2. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 26. pp. 24170-24176.
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