Abstract
Genomic and plasmid DNA can cross the cell membrane by natural means, but the characteristics and functional relevance of this phenomenon have not been fully elucidated. We discovered that human and murine B lymphocytes internalize plasmid DNA in a totally spontaneous manner. To characterize this type of spontaneous transfection we used plasmid DNA coding for a functional immunoglobulin (Ig) heavy chain gene under the control of a B cell-specific promoter. Spontaneous transfection was effective in Burkitt's lymphoma cell lines, Epstein Barr Virus- (EBV) transformed cell lines, and peripheral blood B lymphocytes of the mature naive phenotype (IgM+/IgD+/CD27-). Transcription and expression of the transgene product were documented by reverse transcriptase-polymerase chain reaction (RT-PCR) and flow cytometry. With a reproducibility of 100% the efficiency of spontaneous transfection is on average 1.3% of the total unseparated PBMC population or ~13% of B lymphocytes in circulating blood.
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