Dendritic cells infected with recombinant fowlpox virus vectors are potent and long-acting stimulators of transgene-specific class I restricted T lymphocyte activity

Abstract

The identification of dendritic cells (DC) as the major antigen-presenting cell type of the immune system, combined with the development of procedures for their ex vivo culture, has opened possibilities for tumour immunotherapy based on the transfer of recombinant tumour antigens to DC. It is anticipated that the most effective type of response would be the stimulation of specific, MHC class I restricted cytotoxic T lymphocytes capable of recognising and destroying tumour cells. In order to make this approach possible, methods must be developed for the transfer of recombinant antigen to the DC in such a way that they will initiate an MHC class I restricted response. Here, we demonstrate that murine DC infected with a recombinant fowlpox virus (rFWPV) vector stimulate a powerful, MHC class I restricted response against a recombinant antigen. A rFWPV containing the OVA gene was constructed and used to infect the DC line

Enhanced CD8+ T Cell Immune Responses and Protection Elicited against Plasmodium berghei Malaria by Prime Boost Immunization Regimens Using a Novel Attenuated Fowlpox Virus

“…rFPV vaccine strains have been used to elicit immune responses against HIV (28,30) and tumor Ags (24 -27). rFP9 expressing a model Ag has been used to infect dendritic cells leading to up-regulation of MHC and costimulatory molecules and recognition of the dendritic cells by a CD8 ϩ T cell clone in vitro (35). In addition, FP9 encoding HIV epitopes has recently been shown to elicit CD8 ϩ T cell responses against HIV Ags in mice (29).…”

Deletion of the Virion Host Shutoff Protein (vhs) from Herpes Simplex Virus (HSV) Relieves the Viral Block to Dendritic Cell Activation: Potential of vhs−HSV Vectors for Dendritic Cell-Mediated Immunotherapy

“…These include the mixing of DC with tumor extracts or peptides derived from tumor antigen sequences (6,18,20,25,26,43) and the direct fusion of DC with tumor cells (16). Adenovirus, alphavirus, and poxvirus vectors have also been used (3,8,24,35,44), although adenovirus requires high virus doses (MOI ϭ 30 to 1,000), and toxicity results following infection of DC with alphavirus or poxvirus vectors (8,12).…”

Processing of recombinant AAV genomes occurs in specific nuclear structures that overlap with foci of DNA-damage-response proteins

“…Cellular GFP fluorescence was analyzed by flow cytometry using a FACSCalibur (Becton Dickinson, San Jose, CA, USA) 36 hours after AAV-GFP addition. In the case of AAV-LacZ transduction, ␤-galactosidase activity was determined 36 hours post-infection by measuring o-nitrophenyl-D-galactopyranoside (ONPG) cleavage using a photometric assay, as described elsewhere (Brown et al, 2000). Relative light units (RLU) were expressed as the ratio between ONPG absorbance at 405 nm and the protein concentration (in ng/ml) of the sample, multiplied by an arbitrary factor of 1000.…”