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siRNA Knockdown of PD-L1 and PD-L2 in Monocyte-Derived Dendritic Cells only Modestly Improves Proliferative Responses to Gag by CD8+ T Cells from HIV-1-Infected Individuals

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Abstract

Introduction

Due to their capacity to elicit and regulate immunity, dendritic cells (DCs) are important targets to improve vaccination. Knowing that programmed death-1 (PD-1) high virus-specific T cells become functionally exhausted during chronic exposure to human immunodeficiency virus-1 (HIV-1), the development of a therapeutic DC-based HIV-1 vaccine might include strategies that downregulate PD-L1 and PD-L2 counter-receptors.

Methods

After showing that monocyte-derived DCs rapidly upregulated PD-L1 and PD-L2 expression upon maturation with a variety of stimuli, e.g., Toll-like receptor ligands and cytokines, we determined that PD-L1 and PD-L2 expression could be knocked down by electroporation of a single small interfering RNA (siRNA) sequence twice at the monocyte and immature stages of DC development. This knockdown approached completion and was specific and lasting for several days.

Results

We then added the PD-L1 and PD-L2 silenced monocyte-derived DCs to peripheral blood mononuclear cells from HIV-1-infected individuals along with pools of 15-mer HIV-1 Gag p24 peptides. However, in cultures from six patients, there was only a modest enhancing effect of PD-L1 and PD-L2 silencing on CD8+ T cell proliferative responses to the DCs.

Discussion

These findings suggest that, in monocyte-derived DCs, additional strategies than PD-L1 or PD-L2 blockade will be needed to improve the function of PD-1 high T cells.

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Abbreviations

APCs:

antigen-presenting cells

CFSE:

5-(and-6)-carboxyfluorescein diacetate succinimidyl ester

DCs:

dendritic cells

mDCs:

mature dendritic cells

iDCs:

immature dendritic cells

PBMCs:

peripheral blood mononuclear cells

siRNA:

small interfering RNA

ssRNA:

single-strand RNA

TLRs:

Toll-like receptors

PD-1:

programmed death-1

LTNPs:

long-term nonprogressors

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Acknowledgements

We thank Henry A. Zebroski (Proteomics Resource Center, The Rockefeller University) for synthesizing the Ova peptide library. This work was supported by a grant awarded to Argos Therapeutics from the National Institutes of Health (NIAID-DIADS-BAA-06-19). R.-P.S. is the Canada Research Chair in Human Immunology. J.-P.R. is a clinician–scientist supported by Fonds de Recherche en Santé du Québec.

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Authors and Affiliations

  1. Laboratory of Cellular Physiology and Immunology, Chris Browne Center for Immunology and Immune Diseases, The Rockefeller University, 1230 York Avenue, New York, NY, 10065, USA

    Gaëlle Breton, Lillian Cohn & Ralph M. Steinman

  2. Laboratoire d’Immunologie, Centre de Recherche du Centre Hospitalier de l’Université de Montréal (CR-CHUM) Saint-Luc, Montréal, QC, H2X 1P1, Canada

    Bader Yassine-Diab & Rafick-Pierre Sékaly

  3. Laboratoire d’Immunologie, Département de Microbiologie et d’Immunologie, Université de Montréal, Montréal, QC, H3T 1J4, Canada

    Bader Yassine-Diab & Rafick-Pierre Sékaly

  4. INSERM U743, CR-CHUM, Université de Montréal, Montréal, QC, H2X 1P1, Canada

    Bader Yassine-Diab & Rafick-Pierre Sékaly

  5. Department of Microbiology and Immunology, McGill University, Montréal, QC, H3A 2B4, Canada

    Jean-Pierre Routy & Rafick-Pierre Sékaly

  6. Immunodeficiency Service and Division of Hematology, Royal Victoria Hospital, McGill University Health Centre, McGill University, Montréal, QC, H3A 1A1, Canada

    Mohamed-Rachid Boulassel & Jean-Pierre Routy

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  1. Gaëlle Breton
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  7. Ralph M. Steinman
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Correspondence to Ralph M. Steinman.

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Breton, G., Yassine-Diab, B., Cohn, L. et al. siRNA Knockdown of PD-L1 and PD-L2 in Monocyte-Derived Dendritic Cells only Modestly Improves Proliferative Responses to Gag by CD8+ T Cells from HIV-1-Infected Individuals. J Clin Immunol 29, 637–645 (2009). https://doi.org/10.1007/s10875-009-9313-9

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  • DOI: https://doi.org/10.1007/s10875-009-9313-9

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