Efficient generation of cytotoxic T lymphocytes against cervical cancer cells by adeno-associated virus/human papillomavirus type 16 E7 antigen gene transduction into dendritic cells.
Adeno-associated virus (AAV) is able to efficiently deliver a cytokine gene into dendritic cells (DC). Improvements in T cell priming by DC might be effected by the delivery of antigen genes into DC, resulting in continuous protein expression, as most proteins have short half-lives. In this study, a recombinant AAV vector containing the human papillomavirus (HPV)-16 E7 gene was used to pulse/infect DC and compared to the pulsing of DC by the lipofection of bacterially produced E7 protein. Pulsing of DC with AAV/antigen (Ag) gene was found to be superior to pulsing with protein in six different assay systems: (1) the level of antigen transfer into DC as determined by intracellular staining; (2) the level of MHC class I-restricted killing in cytotoxic T lymphocyte (CTL) assays; (3) the level of IFN-gamma expression; (4) the level of DC-T cell priming clusters generated; (5) the level of CD80 and CD83 expression on DC; and (6) in the resulting CD8:CD4 ratio. Finally, AAV/Ag gene pulsing resulted in strong CTL activity after only 7 days of priming. These data suggest that AAV vectors may offer advantages over the commonly used protein-pulsing technique and that AAV vectors may be useful for the stimulation of CTL activity and adoptive immunotherapy protocols.
Chiriva-Internati
Maurizio
M
Center for Immunology and Microbial Disease, Albany Medical College, Albany, USA.
Liu
Yong
Y
Salati
Emanuela
E
Zhou
Weiping
W
Wang
Zhiqing
Z
Grizzi
Fabio
F
Roman
Juan J
JJ
Lim
Seah H
SH
Hermonat
Paul L
PL
D000941:Antigens / Q000235:genetics / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology*; D000229:Dependovirus; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000276:immunology; D015847:Interleukin-4 / Q000276:immunology; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000276:immunology*; D050725:Papillomavirus E7 Proteins; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000276:immunology*
D000941:Antigens; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D015847:Interleukin-4; D007371:Interferon-gamma