Alloimmune responses of humanized mice to human pluripotent stem cell therapeutics

Temporal tracking of transcriptional changes of human T cells in humanized mice. In this experiment, we studied the ability of immunodeficient NOD-SCID IL2Rgamma-null mice with reconstituted human immune systems to recapitulate graft vs host disease against engrafted tissues. Interestingly, we found that the mice failed to fully reject engrafted tissues, and evaluation of T-cells isolated from grafts demonstrated decreased expression of inflammatory cytokines. We hypothesized this phenotype was due to the devlopment of an "exhausted phenotype." To assess this, we tracked the development of human T-cells in these mice beginning 8 weeks after engraftment of human immune components, and monitored their transcriptional profiles to assess for markers of anergy, exhaustion, and immune activation. Overall design: RNA-seq (3 samples + 1 control) - Samples are T cells pooled from 5 mice with reconstituted human immune systems at three time points. Control sample represents T cells from healthy human adult. Circulating T cells isolated from whole blood via flow cytometry.