A multi-trimeric fusion of CD40L and gp100 tumor antigen activates dendritic cells and enhances survival in a B16-F10 melanoma DNA vaccine model. Issue 38 (11th September 2015)

• Record Type: Journal Article
• Authors: Gupta, Sachin; Termini, James M.; Rivas, Yaelis; Otero, Miguel; Raffa, Francesca N.; Bhat, Vikas; Farooq, Amjad; Stone, Geoffrey W.
• Is Part Of: Vaccine. Volume 33:Issue 38(2015)
• Page Start: 4798
• Page End: 4806
• Publication Date: 2015-09-11
• Subjects: CD40L -- Dendritic cell -- Cancer vaccine -- DNA vaccine -- Surfactant protein D -- B16-F10 melanoma; ALS analytical light scattering -- APC antigen presenting cell -- BMDC bone marrow derived dendritic cells -- CD40L CD40 ligand -- CTL cytotoxic T lymphocytes -- DC dendritic cell -- DLS dynamic light scattering -- GM-CSF granulocyte macrophage colony-stimulating factor -- GVAX B16-F10 cells expressing GM-CSF -- IRES internal ribosome entry site -- MHC major histocompatibility complex -- Mn number-average molar mass -- Mw weight-average molar mass -- Rg radius of gyration -- Rh hydrodynamic radius -- Ro Raleigh ratio -- SEAP secreted alkaline phosphatase -- SLS static light scattering -- SPD surfactant protein D -- TAA tumor associated antigen -- TNFSFL TNF superfamily ligand; Vaccines -- Periodicals; 615.372
• DOI: 10.1016/j.vaccine.2015.07.081
• Languages: English
• ISSNs: 0264-410X

Abstract

Highlights: DNA vaccine technology fusing cancer antigens to CD40L soluble multi-trimers. Allows targeting of vaccine antigens to dendritic cells in situ. Activates and matures dendritic cells. Vaccine induces an anti-tumor immune response when combined with IL-12 and GM-CSF. Abstract: Vaccination with tumor-associated antigens can induce cancer-specific CD8+ T cells. A recent improvement has been the targeting of antigen to dendritic cells (DC) using antibodies that bind DC surface molecules. This study explored the use of multi-trimers of CD40L to target the gp100 melanoma tumor antigen to DC. The spontaneously-multimerizing gene Surfactant Protein D (SPD) was used to fuse gp100 tumor antigen and CD40L, creating the recombinant protein SPD-gp100-CD40L. This "third generation" DC-targeting vaccine was designed to both target antigen to DC and optimally activate dendritic cells by aggregating CD40 trimers on the DC membrane surface. SPD-gp100-CD40L expressed as a 110 kDa protein. Analytical light scattering analysis gave elution data corresponding to 4-trimer and multi-trimer SPD-gp100-CD40L oligomers. The protein was biologically active on dendritic cells and induced CD40-mediated NF-κB signaling. DNA vaccination with SPD-gp100-CD40L plasmid, together with plasmids encoding IL-12p70 and GM-CSF, significantly enhanced survival and inhibited tumor growth in a B16-F10 melanoma model. Expression of gp100 and SPD-CD40L as separate molecules did not enhance survival, highlighting the requirement to encode gp100 within SPD-CD40L for optimal vaccine activity. These data support a model where DNA vaccination with SPD-gp100-CD40L targets gp100 to DC in situ, induces activation of these DC, and generates a protective anti-tumor response when given in combination with IL-12p70 and GM-CSF plasmids.