Efficient generation of antigen-specific cytotoxic T cells using retrovirally transduced CD40-activated B cells.
The development of rapid, efficient, and safe methods for generating Ag-specific T cells is necessary for the clinical application of adoptive immunotherapy. We show that B cells stimulated with CD40 ligand and IL-4 (CD40-B cells) can be efficiently transduced with retroviral vectors encoding a model Ag, CMV tegument protein pp65 gene, and maintain high levels of costimulatory molecules after gene transfer. CTL lines specific for pp65 were readily generated in all four healthy CMV-seropositive donors by stimulating autologous CD8(+) T cells with these transduced CD40-B cells, both of which were derived from 10 ml peripheral blood. ELISPOT assays revealed that the CTL lines used multiple HLA alleles as restricting elements. Thus, CD40-B cells transduced retrovirally with Ag-encoding cDNA can be potent APC and facilitate to generate Ag-specific CTL in vitro.
Kondo
Eisei
E
Division of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Topp
Max S
MS
Kiem
Hans-Peter
HP
Obata
Yuichi
Y
Morishima
Yasuo
Y
Kuzushima
Kiyotaka
K
Tanimoto
Mitsune
M
Harada
Mine
M
Takahashi
Toshitada
T
Akatsuka
Yoshiki
Y
D000483:Alleles; D017951:Antigen Presentation; D000956:Antigens, Viral / Q000235:genetics; D001402:B-Lymphocytes / Q000276:immunology*; D019013:CD40 Antigens / Q000378:metabolism*; D023201:CD40 Ligand / Q000494:pharmacology; D003586:Cytomegalovirus Infections / Q000276:immunology; D000939:Epitopes / Q000235:genetics; D006680:HLA Antigens / Q000235:genetics; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014161:Transduction, Genetic; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology
D000956:Antigens, Viral; D019013:CD40 Antigens; D000939:Epitopes; D006680:HLA Antigens; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D023201:CD40 Ligand; D015847:Interleukin-4