A novel peptide targeting Clec9a on dendritic cell for cancer immunotherapy

Dendritic cells (DCs) are professional antigen-presenting cells with antigen recognition molecules on the surface. Clec9a is selectively expressed on mouse CD8a+ DCs and CD103+ DCs subsets, which are functionally similar to human BDCA3+ DCs. It is reported that Clec9a is responsible for the antigen cross-presentation of these DC subsets. In the present study, by using phage display technique, we discovered a novel peptide WH, which can selectively bind to mouse Flt3L induced Clec9a+ DCs or Clec9a over-expressed HEK-293T cells. Furthermore, by using computer-aided docking model and mutation assay, we observed that Asp248 and Trp250 are two key residues for Clec9a to bind with peptide WH. When coupled with OVA257-264 epitope, peptide WH can significantly enhance the ability of Clec9a+ DCs to activate OVA-specific CD8+ T cells, which elicit strong ability to secret IFN-γ, express perforin and granzyme B mRNA. In B16-OVA lung metastasis mouse model, WH-OVA257-264 fusion peptide can also enhance the activation of CD8+ T cells and decrease the lung metastasis loci. All these results suggested that peptide WH could be considered as an antigen delivery carrier targeting Clec9a+ DCs for cancer immunotherapy.

[1]  C. Figdor,et al.  Dendritic cell immunotherapy: mapping the way , 2004, Nature Medicine.

[2]  D. Underhill,et al.  Dectin-1 Stimulation by Candida albicans Yeast or Zymosan Triggers NFAT Activation in Macrophages and Dendritic Cells1 , 2007, The Journal of Immunology.

[3]  J. Taylor‐Papadimitriou,et al.  Targeting DNGR-1 (CLEC9A) with antibody/MUC1 peptide conjugates as a vaccine for carcinomas , 2014, European journal of immunology.

[4]  C. Reis e Sousa,et al.  The DC receptor DNGR-1 mediates cross-priming of CTLs during vaccinia virus infection in mice. , 2012, The Journal of clinical investigation.

[5]  L. Durant,et al.  DNGR‐1 is dispensable for CD8+ T‐cell priming during respiratory syncytial virus infection , 2014, European journal of immunology.

[6]  Yanfeng Gao,et al.  Identification of novel T cell epitopes from efflux pumps of Mycobacterium tuberculosis. , 2011, Immunology letters.

[7]  K. Shortman,et al.  Dendritic cell subtypes in mouse lymphoid organs: cross-correlation of surface markers, changes with incubation, and differences among thymus, spleen, and lymph nodes. , 1997, Journal of immunology.

[8]  Gordon D. Brown,et al.  CLEC9A Is a Novel Activation C-type Lectin-like Receptor Expressed on BDCA3+ Dendritic Cells and a Subset of Monocytes , 2008, Journal of Biological Chemistry.

[9]  Anna M. Keller,et al.  Characterization of human DNGR-1+ BDCA3+ leukocytes as putative equivalents of mouse CD8α+ dendritic cells , 2010, The Journal of experimental medicine.

[10]  Y. Hiasa,et al.  Phase I/II study of immunotherapy using tumor antigen-pulsed dendritic cells in patients with hepatocellular carcinoma , 2012, International journal of oncology.

[11]  N. Gow,et al.  Pattern recognition: recent insights from Dectin-1 , 2009, Current opinion in immunology.

[12]  M. Sioud,et al.  A novel peptide carrier for efficient targeting of antigens and nucleic acids to dendritic cells , 2013, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[13]  B. Thompson,et al.  F-actin is an evolutionarily conserved damage-associated molecular pattern recognized by DNGR-1, a receptor for dead cells. , 2012, Immunity.

[14]  R. Steinman,et al.  In Vivo Targeting of Antigens to Maturing Dendritic Cells via the DEC-205 Receptor Improves T Cell Vaccination , 2004, The Journal of experimental medicine.

[15]  Michel C. Nussenzweig,et al.  Dendritic Cells Induce Peripheral T Cell Unresponsiveness under Steady State Conditions in Vivo , 2001, The Journal of experimental medicine.

[16]  Karolina Palucka,et al.  Cancer immunotherapy via dendritic cells , 2012, Nature Reviews Cancer.

[17]  R. Steinman,et al.  DEC-205, a 205-kDa protein abundant on mouse dendritic cells and thymic epithelium that is detected by the monoclonal antibody NLDC-145: purification, characterization, and N-terminal amino acid sequence. , 1995, Cellular immunology.

[18]  K. Namba,et al.  Structure of the Complex of F-Actin and DNGR-1, a C-Type Lectin Receptor Involved in Dendritic Cell Cross-Presentation of Dead Cell-Associated Antigens. , 2015, Immunity.

[19]  Siamon Gordon,et al.  Dectin-1 Is A Major β-Glucan Receptor On Macrophages , 2002, The Journal of experimental medicine.

[20]  M. Shurin.  Dendritic cells presenting tumor antigen , 1996, Cancer Immunology, Immunotherapy.

[21]  M. Wright,et al.  The dendritic cell subtype-restricted C-type lectin Clec9A is a target for vaccine enhancement. , 2008, Blood.

[22]  Anna M. Keller,et al.  Identification of a dendritic cell receptor that couples sensing of necrosis to immunity , 2009, Nature.

[23]  M. Wright,et al.  The dendritic cell receptor Clec9A binds damaged cells via exposed actin filaments. , 2012, Immunity.

[24]  Lei Liu,et al.  Blocking of the PD-1/PD-L1 Interaction by a D-Peptide Antagonist for Cancer Immunotherapy. , 2015, Angewandte Chemie.

[25]  J. Ruland,et al.  Syk- and CARD9-dependent coupling of innate immunity to the induction of T helper cells that produce interleukin 17 , 2007, Nature Immunology.

[26]  M. Bros,et al.  Selective uptake of cylindrical poly(2-oxazoline) brush-antiDEC205 antibody-OVA antigen conjugates into DEC-positive dendritic cells and subsequent T-cell activation. , 2014, Chemistry.

[27]  Scott N. Mueller,et al.  Targeting Antigen to Mouse Dendritic Cells via Clec9A Induces Potent CD4 T Cell Responses Biased toward a Follicular Helper Phenotype , 2011, The Journal of Immunology.

[28]  Gordon D. Brown,et al.  Syk‐coupled C‐type lectin receptors that mediate cellular activation via single tyrosine based activation motifs , 2010, Immunological reviews.

[29]  D. Pennington,et al.  Tumor therapy in mice via antigen targeting to a novel, DC-restricted C-type lectin. , 2008, The Journal of clinical investigation.

[30]  R. Steinman.  Dendritic cells: versatile controllers of the immune system , 2007, Nature Medicine.

[31]  Michel C. Nussenzweig,et al.  Efficient Targeting of Protein Antigen to the Dendritic Cell Receptor DEC-205 in the Steady State Leads to Antigen Presentation on Major Histocompatibility Complex Class I Products and Peripheral CD8+ T Cell Tolerance , 2002, The Journal of experimental medicine.