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Peptide presentation by the MHC class Ib molecule, H2-M3

G P Smith1, V M Dabhi, E G Pamer

  • 1Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas 75235-9050.

International Immunology
|December 1, 1994
PubMed
Summary

The H2-M3 molecule can present unformylated peptides to cytotoxic T cells, contrary to previous beliefs. Specific mutations in H2-M3 affect T cell responses, with Q34 and L167 being crucial for M3-specific cytotoxic T lymphocyte responses.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Cellular Biology

Background:

  • The mouse class I MHC molecule, H2-M3, is known to present N-formylated peptides to cytotoxic T cells.
  • Previous research indicated H2-M3's inability to present unformylated peptides.

Purpose of the Study:

  • To investigate the presentation of unformylated peptides by H2-M3.
  • To determine the role of specific amino acid residues in the H2-M3 peptide-binding groove in N-formylated peptide presentation and T cell recognition.

Main Methods:

  • Utilized site-directed mutagenesis to alter specific residues (34, 167, 171) in the H2-M3 molecule.
  • Assessed the presentation of N-formylated and unformylated ND1 peptides using H2-M3-transfected fibroblasts.
  • Measured target cell killing by ND1-specific and alloreactive cytotoxic T cells.

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Main Results:

  • Demonstrated that H2-M3 can present unformylated ND1 peptide, sensitizing target cells for killing.
  • N-formylated and unformylated ND1 peptides induced equivalent killing levels at high concentrations, but differed significantly in potency (10^4-fold).
  • Mutations Q34V and L167W, individually or combined, impaired killing by alloreactive T cells, while F171Y had a lesser effect.

Conclusions:

  • H2-M3 can present both N-formylated and unformylated peptides, with significant differences in binding affinity.
  • Residues Q34 and L167 in H2-M3 are critical for the recognition of M3-specific alloreactive cytotoxic T cells.
  • The specific amino acid changes at positions 34, 167, and 171 are not essential for N-formylated peptide presentation but are important for alloreactive T cell responses.