title: Woodchuck dendritic cells generated from peripheral blood mononuclear cells and transduced with recombinant human adenovirus serotype 5 induce antigen-specific cellular immune responses.
abstract: Woodchucks infected with the woodchuck hepatitis virus (WHV) is the best available animal model for testing the immunotherapeutic effects of dendritic cells (DCs) in the setting of a chronic infection, as woodchucks develop a persistent infection resembling that seen in humans infected with the hepatitis B virus. In the present study, DCs were generated from woodchuck peripheral blood mononuclear cells (wDCs) in the presence of human granulocyte macrophage colony-stimulating factor (hGM-CSF) and human interleukin 4 (hIL-4). After 7 days of culture, cells with morphology similar to DCs were stained positively with a cross-reactive anti-human CD86 antibody. Functional analysis showed that uptake of FITC-dextran by wDCs was very efficient and was partially inhibited after LPS-induced maturation. Furthermore, wDCs stimulated allogenic lymphocytes and induced proliferation. Moreover, wDCs were transduced efficiently with a human adenovirus serotype 5 for the expression of beta-galactosidase. Following transduction and in vivo administration of such DCs into woodchucks, an antigen-specific cellular immune response was induced. These results demonstrate that wDCs can be generated from the peripheral blood. Following transfection with a recombinant adenovirus wDCs can be used as a feasible and effective tool for eliciting WHV-specific T-cell responses indicating their potential to serve as prophylactic and therapeutic vaccines.
authors: Laboratory of Gene Therapy of Viral Hepatitis, Division of Gene Therapy and Hepatology, Center for Applied Medical Research (CIMA), Pamplona, Navarra, Spain.
authors: Laura
authors: L
authors: Ochoa-Callejero
authors: Pedro
authors: P
authors: Berraondo
authors: Julien
authors: J
authors: Crettaz
authors: Cristina
authors: C
authors: Olagüe
authors: Africa
authors: A
authors: Vales
authors: Juan
authors: J
authors: Ruiz
authors: Jesús
authors: J
authors: Prieto
authors: Bud C
authors: BC
authors: Tennant
authors: Stephan
authors: S
authors: Menne
authors: Gloria
authors: G
authors: González-Aseguinolaza
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D051940:B7-2 Antigen / Q000276:immunology / Q000378:metabolism; D002455:Cell Division; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D004195:Disease Models, Animal; D005822:Genetic Vectors; D019694:Hepatitis B, Chronic / Q000276:immunology / Q000628:therapy; D006801:Humans; D007111:Immunity, Cellular; D007114:Immunization; D007115:Immunization Schedule; D007167:Immunotherapy / Q000379:methods*; D007279:Injections, Subcutaneous; D008214:Lymphocytes / Q000276:immunology; D008392:Marmota / Q000276:immunology; D016865:Reassortant Viruses / Q000276:immunology; D013045:Species Specificity; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000276:immunology
chemical_list: D051940:B7-2 Antigen; D001616:beta-Galactosidase