Transcriptional profiling of LNGFR.FOXP3-expressing scurfy CD4+ T cells

We reported transcriptional characterization of LNGFR.FOXP3-expressing scurfy CD4+ T cells from lymph nodes, at day 35 after transfer in scurfy males, where they were able to rescue mice from scurfy autoimmune disease. Overall design: We describe development and implementation of a full preclinical strategy for transferring FOXP3 into scurfy (Foxp3 deficient) CD4+ T cells. Them, we have established a novel in vivo assay of Treg functionality, based on adoptive transfer of these cells into scurfy mice and a combination of cyclophosphamide conditioning and interleukin-2 treatment. This model highlighted the possibility of rescuing scurfy disease after the latter's onset. By using this in vivo model and an optimized lentiviral vector expressing human Foxp3 and as a reporter a truncated form of the 5 low-affinity nerve growth factor receptor (LNGFR), we demonstrated that the adoptive transfer of FOXP3-transduced scurfy CD4+ T cells enabled the long-term rescue of scurfy autoimmune disease. The efficiency was similar to that seen with wild-type Treg. After in vivo expansion, the converted CD4FOXP3 cells recapitulated the transcriptomic core signature for Treg.