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Highly Efficient and Consistent Gene Transfer into Dendritic Cells Utilizing a Combination of Ultraviolet-irradiated Adenovirus and Poly(L-Lysine) Conjugates¹

Urologic Oncology Research Laboratories, Jonsson Comprehensive Cancer Center, Department of Urology [P. M., S. P., J. D., R. K., A. H., K. M., C-L. T., A. B.], and Department of Pulmonary Medicine [M. R.], University of California-Los Angeles School of Medicine, Los Angeles, California 90095

Dendritic cells (DCs) are capable of presenting tumor-associated antigens and subsequently play an essential role in T-cell activation. The aim of this study was to develop an efficient method for gene transfer into DCs. These genetically transduced DCs can then be used as potent inducers of specific cell-mediated immune response. When compared with physical methods for gene transfer (lipofection and calcium phosphate precipitation), adenovirus (AdV) vectors proved to be highly efficient for gene transfer into DCs. To overcome concomitant AdV gene expression and potential immunogenicity, AdVs were irradiated with UV. The UV dose was optimized to block AdV transcription without altering AdV receptor binding and endocytosis capacity. We subsequently used a polycationic amino acid compound, poly(L-lysine), to conjugate the irradiated AdVs to transgenes. The resulting complexes were found to mediate a highly efficient transfer of transgenes into DCs, without concomitant expression of AdV gene products. Low titers of irradiated AdVs were sufficient for a consistent gene transfer into DCs. This is the first study to demonstrate efficient, consistent, and practical gene transfer using an UV approach to irradiated AdV-poly(L-lysine) conjugates and should be useful for the development of DC-based tumor vaccine therapies.

¹ This work was supported by grants from the Jonsson Comprehensive Cancer Center, UCLA, the Dutch Cancer Foundation, and NIH (CA66022).

² To whom requests for reprints should be addressed, at Department of Urology, UCLA School of Medicine, 10833 Le Conte Avenue, Los Angeles, CA 90095. Phone: (310) 206-5406; Fax: (310) 206-5343; E-mail: abelldeg@ucla.edu.

Received 9/ 3/97. Accepted 1/ 5/98.

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