Figure 6.

CD30⁺ T cells were lost from cultures of 5F11-28Z T cells. (A) Plots show T cells 6 days after transduction with LSIN-5F11-28Z or untransduced T cells from the same donor; both untransduced and 5F11-28Z-transduced T cells were cultured in the same manner. Cells were stained with antibodies for CD3, CD4, CD8, and CD30. The plots are gated on live CD3⁺ lymphocytes and then gated on either CD4⁺ or CD8⁺ cells. Both CAR⁺ and CAR-negative cells are included on the plots. (B) The plot is gated on live CD3⁺ lymphocytes from the same LSIN-5F11-28Z-transduced culture as in A. The plot shows that residual CD30⁺ T cells were found predominantly among the CAR⁺ cells in the culture. CAR staining was by Protein L. This is 1 of 4 cultures with similar results. The percentages are shown of (C) CD4⁺CD3⁺ lymphocytes that were CD30⁺ and (D) CD3⁺CD8⁺ lymphocytes that were CD30⁺. Cells were cultured and stained as in A. Lines connect paired results of LSIN-5F11-28Z-tansduced or untransduced cultures from the same donor. Statistics were by two-tailed paired t-test; n = 4. p < 0.05 was considered statistically significant. (E) Accumulation of LSIN-5F11-28Z-transduced T cells was compared with accumulation of untransduced T cells in culture. This is one of three experiments that had similar results. For the LSIN-5F11-28Z-transduced culture, 94.4% of the T cells were CAR⁺ on day 7 of culture. Counts started on day 3, when lentiviral transductions were complete.