High-efficiency gene transfer to primary T lymphocytes by recombinant adenovirus vectors

Recombinant, replication-deficient adenoviruses are efficient vectors for gene transfer to a wide range of cell types, with the exception of T lymphocytes. Here, we show that primary T lymphocytes from peripheral blood, cord blood, and the Jurkat T cell line are efficiently transduced by recombinant adenovirus. Nearly 100% infection efficiency of primary T cells is obtained with high multiplicity of infection (MOI) (5000) of recombinant adenovirus coding for lacZ. Similar infection efficiency by adenovirus-mediated gene transfer was obtained at lower MOI (3000) by activating primary T cells with PHA and PMA. Addition of cationic liposomes together with RAdlacZ markedly enhanced the infection efficiency at lower MOI (1000) resulting in over 90% infection efficiency. Primary T cells express low levels of coxsackievirus and adenovirus receptor (CAR), a cell surface receptor for adenovirus fiber attachment, as well as _vβ₃ and _vβ₅ integrins, cellular receptors for adenovirus internalization. This suggests that adenovirus entry to T cells at high MOI is mediated by other mechanisms. In conclusion, these results demonstrate that genes can be efficiently transferred to primary lymphocytes by adenovirus vectors at high MOI or in combination with cationic liposomes.