Forced expression of HLA-DM at the surface of dendritic cells increases loading of synthetic peptides on MHC class II molecules and modulates T cell responses

Abdul Mohammad Pezeshki; Marie-Hélène Côté; Georges A Azar; Jean-Pierre Routy; Mohamed-Rachid Boulassel; Jacques Thibodeau

doi:10.4049/jimmunol.1002747

Forced expression of HLA-DM at the surface of dendritic cells increases loading of synthetic peptides on MHC class II molecules and modulates T cell responses

J Immunol. 2011 Jul 1;187(1):74-81. doi: 10.4049/jimmunol.1002747. Epub 2011 May 27.

Authors

Abdul Mohammad Pezeshki¹, Marie-Hélène Côté, Georges A Azar, Jean-Pierre Routy, Mohamed-Rachid Boulassel, Jacques Thibodeau

Affiliation

¹ Laboratoire d'Immunologie Moléculaire, Département de Microbiologie et Immunologie, Université de Montréal, Montréal, Québec H3C 3J7, Canada.

PMID: 21622867
DOI: 10.4049/jimmunol.1002747

Abstract

Adoptive transfer of autologous dendritic cells (DCs) loaded with tumor-associated CD4 and CD8 T cell epitopes represents a promising avenue for the immunotherapy of cancer. In an effort to increase the loading of therapeutic synthetic peptides on MHC II molecules, we used a mutant of HLA-DM (DMY) devoid of its lysosomal sorting motif and that accumulates at the cell surface. Transfection of DMY into HLA-DR(+) cells resulted in increased loading of the exogenously supplied HA(307-318) peptide, as well as increased stimulation of HA-specific T cells. Also, on transduction in mouse and human DCs, DMY increased loading of HEL(48-61) and of the tumor Ag-derived gp100(174-190) peptides, respectively. Interestingly, expression of DMY at the surface of APCs favored Th1 differentiation over Th2. Finally, we found that DMY(-) and DMY(+) mouse APCs differentially stimulated T cell hybridomas sensitive to the fine conformation of peptide-MHC II complexes. Taken together, our results suggest that the overexpression of HLA-DMY at the plasma membrane of DCs may improve quantitatively, but also qualitatively, the presentation of CD4 T cell epitopes in cellular vaccine therapies for cancer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Antigen Presentation / genetics*
Antigen Presentation / immunology
Cancer Vaccines / administration & dosage
Cancer Vaccines / genetics
Cancer Vaccines / immunology
Cell Line
Cell Membrane / genetics
Cell Membrane / immunology
Cell Membrane / metabolism
Coculture Techniques
Dendritic Cells / cytology
Dendritic Cells / immunology*
Dendritic Cells / metabolism
Epitopes, T-Lymphocyte / biosynthesis
Epitopes, T-Lymphocyte / genetics*
Epitopes, T-Lymphocyte / immunology
HEK293 Cells
HLA-D Antigens / biosynthesis*
HLA-D Antigens / genetics*
HLA-D Antigens / metabolism
HeLa Cells
Humans
Jurkat Cells
Lysosomes / genetics
Lysosomes / immunology
Lysosomes / metabolism
Mice
Mice, Inbred BALB C
Mice, Inbred C3H
Mice, Transgenic
Molecular Sequence Data
Peptides / chemical synthesis
Peptides / genetics*
Peptides / immunology
Protein Sorting Signals / genetics*
T-Lymphocyte Subsets / cytology
T-Lymphocyte Subsets / immunology*
T-Lymphocyte Subsets / metabolism
Transduction, Genetic

Substances

Cancer Vaccines
Epitopes, T-Lymphocyte
HLA-D Antigens
HLA-DM antigens
Peptides
Protein Sorting Signals