Inadvertent Transfer of Murine VL30 Retrotransposons to CAR-T Cells

Figure 2

VL30 transduction of 293T cells is reverse transcription dependent. The effects of a reverse-transcriptase inhibitor on VL30 transduction. (a) Physical map of the -retroviral vector (-RVV) pTK2151. The 5 and 3 non-SIN LTRs are shown. The packaging signal (y), the SV40 promoter, and the GFP reporter gene are shown. (b) Bar graph showing the genome copy number (GCN) of VL30 in 293T cells following exposure to conditioned media collected from either PG13 or PG13 cells transduced with the -RVV pTK2151. Exposure to the abovementioned conditioned media was done either in the presence or in the absence of the reverse transcriptase inhibitor azidothymidine (AZT 10 μM). The experiment was performed in triplicate. (c) Bar graph showing the vector copy number (VCN) of the -RVV pTK2151 in 293T cells following exposure to conditioned media collected from either PG13 cells transduced with the -RVV pTK215 or 293T cells transiently transfected with the pTK2151 vector cassette, a VSV-G envelope, and -RVV packaging cassettes. Exposure to the abovementioned conditioned media was done either in the presence or in the absence of the reverse transcriptase inhibitor AZT (). The experiment was performed in triplicate. Significance of the AZT effect on VL30 GCN and pTK2151 VCN was determined by 2-way ANOVA, and .

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