Generating MHC Class II+/Ii- phenotype after adenoviral delivery of both an expressible gene for MHC Class II inducer and an antisense Ii-RNA construct in tumor cells.
Tumor cells engineered by gene transduction to be MHC Class II+/Ii- are novel APCs capable of presenting endogenous tumor antigen epitopes to activate T helper cells. The MHC Class II+/Ii- tumor cell phenotype is created by transfecting genes for either CIITA or IFN-gamma, and inhibiting induced Ii mRNA by an Ii reverse gene construct (Ii-RGC). Adenoviral vectors are preferred for the delivery of such genes because of high infection efficiency and ubiquity of the adenoviral receptor on many cell types and tumors. Here we show that at 5 MOI (multiplicity of infection), recombinant adenoviruses with CIITA or IFN-gamma genes converted virtually all MC-38 colon adenocarcinoma cells and Renca renal carcinoma cells in culture to MHC Class II+/Ii+ cells. A single recombinant adenovirus with both genes for IFN-gamma and Ii-RGC (rAV/IFN-gamma/Ii-RGC) efficiently induced the MHC Class II+/Ii- phenotype. Injection of tumor nodules with rAV/Ii-RGC and rAV/CIITA/IFN-gamma combined with a suboptimal dose of rAV/IL-2 induced a potent antitumor immune response. The methods are adaptable for producing enhanced genetic vaccines, attenuated virus vaccines (eg, vaccinia), and ex vivo cell-based vaccines (dendritic and tumor cells).
Hillman
G G
GG
Department of Radiation Oncology, Barbara Ann Karmanos Cancer Institute at Wayne State University School of Medicine, Detroit, MI, USA.
Kallinteris
N L
NL
Li
J
J
Wang
Y
Y
Lu
X
X
Li
Y
Y
Wu
S
S
Wright
J L
JL
Slos
P
P
Gulfo
J V
JV
Humphreys
R E
RE
Xu
M
M
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics; D000951:Antigens, Neoplasm / Q000235:genetics; D018274:Electroporation; D005802:Genes, MHC Class II; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D000949:Histocompatibility Antigens Class II / Q000235:genetics; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007279:Injections, Subcutaneous; D007371:Interferon-gamma / Q000235:genetics; D051379:Mice; D009369:Neoplasms / Q000628:therapy*; D010641:Phenotype; D016372:RNA, Antisense / Q000008:administration & dosage*; D014161:Transduction, Genetic / Q000379:methods; D014407:Tumor Cells, Cultured
D000944:Antigens, Differentiation, B-Lymphocyte; D000951:Antigens, Neoplasm; D000949:Histocompatibility Antigens Class II; D016372:RNA, Antisense; C045084:invariant chain; D007371:Interferon-gamma