Transduction of human dendritic cells with a recombinant modified vaccinia Ankara virus encoding MUC1 and IL-2.
The epithelial mucin MUC1 is considered an opportune target antigen for cancer immunotherapy, as it is over-expressed and exhibits aberrant glycosylation in malignant cells. Because dendritic cells (DC) are powerful initiators of immune responses, efforts have focused on tumor antigen-bearing DC as potent cancer vaccines. In this study we have characterized the transduction of monocyte-derived DC with a highly attenuated vaccinia virus vector [modified vaccinia Ankara (MVA)] encoding human MUC1 and the immunostimulatory cytokine IL-2. Analysis of transduced DC cultures generated from a number of donors revealed MUC1 expression in the range of 27-54% of the cells and a co-regulated secretion of bioactive IL-2. As shown by FACS analysis with MUCI-specific antibodies, the MVA-MUC1/IL-2-transduced DC predominantly expressed the fully processed glycoform of MUC1, typical of that displayed by normal epithelia. Over a 3-day period after transduction, transgene expression declined concurrent with an increase in MVA-induced cytopathic effects. The transduced DC stimulated allogeneic lymphocyte proliferation, indicating that DC immunostimulatory function is not impaired by vector transduction. In the presence of IL-2, MVA-transduced DC were able to enhance autologous lymphocyte proliferation. Also, vector expression was analyzed in DC cultures treated with TNF-alpha, a known DC maturation factor. As indicated by the up-regulation of several DC maturation markers, neither virus infection nor transgene expression influenced the maturation capacity of the cells. The MVA-MUC1/IL-2 vector effectively transduced both immature and TNF-alpha-matured DC. Overall, our results are encouraging for the clinical application of MVA-MUC1/IL-2-transduced DC.
Trevor
K T
KT
Arizona Cancer Center, Tucson 85724, USA. ktrevor@azcc.arizona.edu
Hersh
E M
EM
Brailey
J
J
Balloul
J M
JM
Acres
B
B
D017951:Antigen Presentation; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D017209:Apoptosis; D001782:Blood Donors; D002478:Cells, Cultured / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D004306:Dose-Response Relationship, Immunologic; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D006031:Glycosylation; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology*; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D018396:Mucin-1 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D014162:Transfection; D019076:Transgenes; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014616:Vaccinia virus / Q000235:genetics
D000951:Antigens, Neoplasm; D007376:Interleukin-2; D018396:Mucin-1; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha