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Immunogenetics

, Volume 63, Issue 12, pp 821–834 | Cite as

Porcine major histocompatibility complex (MHC) class I molecules and analysis of their peptide-binding specificities

  • Lasse Eggers Pedersen
  • Mikkel Harndahl
  • Michael Rasmussen
  • Kasper Lamberth
  • William T. Golde
  • Ole Lund
  • Morten Nielsen
  • Soren Buus
Original Paper

Abstract

In all vertebrate animals, CD8+ cytotoxic T lymphocytes (CTLs) are controlled by major histocompatibility complex class I (MHC-I) molecules. These are highly polymorphic peptide receptors selecting and presenting endogenously derived epitopes to circulating CTLs. The polymorphism of the MHC effectively individualizes the immune response of each member of the species. We have recently developed efficient methods to generate recombinant human MHC-I (also known as human leukocyte antigen class I, HLA-I) molecules, accompanying peptide-binding assays and predictors, and HLA tetramers for specific CTL staining and manipulation. This has enabled a complete mapping of all HLA-I specificities (“the Human MHC Project”). Here, we demonstrate that these approaches can be applied to other species. We systematically transferred domains of the frequently expressed swine MHC-I molecule, SLA-1*0401, onto a HLA-I molecule (HLA-A*11:01), thereby generating recombinant human/swine chimeric MHC-I molecules as well as the intact SLA-1*0401 molecule. Biochemical peptide-binding assays and positional scanning combinatorial peptide libraries were used to analyze the peptide-binding motifs of these molecules. A pan-specific predictor of peptide–MHC-I binding, NetMHCpan, which was originally developed to cover the binding specificities of all known HLA-I molecules, was successfully used to predict the specificities of the SLA-1*0401 molecule as well as the porcine/human chimeric MHC-I molecules. These data indicate that it is possible to extend the biochemical and bioinformatics tools of the Human MHC Project to other vertebrate species.

Keywords

Recombinant MHC Peptide specificity Binding predictions 

Notes

Acknowledgments

We thank Lise Lotte Bruun Nielsen, Anne Caroline Schmiegelow, and Iben Sara Pedersen for their expert experimental support. This work was in part supported by the Danish Council for Independent Research, Technology and Production Sciences (274-09-0281) and by the National Institute of Health (NIH) (HHSN266200400025C).

Supplementary material

251_2011_555_MOESM1_ESM.pdf (63 kb)
Online Resource 1 Nucleotide sequence used to encode SLA-1*0401 fused to an enzyme restricted cleavage site (Fxa, purple), a biotinylation site (BSP, pink), and a histidine affinity tag (HAT, dark blue) (PDF 62 kb)
251_2011_555_MOESM2_ESM.pdf (73 kb)
Online Resource 2 Primers designed to induce the changes in the DNA coding for human β2m to get the DNA encoding porcine β2m (PDF 72 kb)
251_2011_555_MOESM3_ESM.pdf (107 kb)
Online Resource 3 Comparison of pseudo-sequences for the heavy chain molecules HLA-A*01:01, SLA-1*0401, HLA-A*11:01, HPP (hα123), and PHP (pα123). Orange and purple residue numbers have an impact on binding position 9 and position 3 of the peptide, respectively. Blue residue numbers have an impact on binding position 2 of the peptide, whereas green residue numbers have an impact on binding position 1 of the peptide. Black residues show similarities between the different molecules, whereas red residues illustrate differences when compared to HLA-A*11:01. White residue numbers on a black background indicate a difference in the pseudo-sequence residues between SLA-1*0401 and either HLA-A*01:01 or HLA-A*11:01 or both (PDF 106 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Lasse Eggers Pedersen
    • 1
  • Mikkel Harndahl
    • 1
  • Michael Rasmussen
    • 1
  • Kasper Lamberth
    • 1
  • William T. Golde
    • 2
  • Ole Lund
    • 3
  • Morten Nielsen
    • 3
  • Soren Buus
    • 1
  1. 1.Laboratory of Experimental Immunology, Faculty of Health SciencesUniversity of CopenhagenCopenhagenDenmark
  2. 2.Plum Island Animal Disease Center, Agricultural Research ServiceUSDAGreenportUSA
  3. 3.Center for Biological Sequence AnalysisTechnical University of DenmarkCopenhagenDenmark

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