Temperature-based MHC class-I multimer peptide exchange for human HLA-A, B and C.

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

T cell recognition of specific antigens presented by major histocompatibility complexes class-I (MHC-I) can play an important role during immune responses against pathogens and cancer cells. Detection of T cell immunity is based on assessing the presence of antigen-specific cytotoxic CD8+ T cells using MHC class-I (MHC-I) multimer technology. Previously we have designed conditional peptides for HLA-A*02:01, H-2Kb and HLA-E that form stable peptide-MHC-I-complexes at low temperatures and dissociate when exposed to a defined elevated temperature. The resulting conditional MHC-I complex can easily and without additional handling be exchanged with a peptide of interest, allowing to exchange peptides in a ready-to-use multimer and a high-throughput manner. Here we present data that this peptide-exchange technology is a general applicable, ready-to-use and fast approach to load many different peptides in MHC-I multimers for alleles of the HLA-A, HLA-B and HLA-C loci. We describe the development of conditional peptides for HLA-A*03:01, HLA-A*11:01, HLA-B*07:02 and HLA-C*07:02 that only form stable peptide-MHC-I complexes at low temperatures, allowing peptide exchange at higher defined temperature. We document the ease and flexibility of this technology by monitoring CD8+ T cell responses to virus-specific peptide-MHC complexes in patients.

Authors 13

1. Cilia R. Pothast Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

2. Ian Derksen Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

3. Anneloes van der Plas - van Duijn Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

4. Angela el Hebieshy Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

5. Wesley Huisman Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

6. Kees L. M. C. Franken Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

7. Jacques Neefjes Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

8. Jolien J. Luimstra Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

9. Marieke Griffioen Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center;

   Leiden University Medical Center

10. Michel G.D. Kester Aachen

    Leiden University Medical Center

    Affiliation as printed

    Leiden University Medical Center;

    Leiden University Medical Center

11. Maarten H. Vermeer Aachen

    Leiden University Medical Center

    Affiliation as printed

    Leiden University Medical Center;

    Leiden University Medical Center

12. Mirjam H.M. Heemskerk Aachen

    Leiden University Medical Center

    Affiliation as printed

    Leiden University Medical Center;

13. Ferenc A. Scheeren corresponding Aachen

    Leiden University Medical Center

    Affiliation as printed

    Leiden University Medical Center;