PanCancer Immune panel on infiltrating memory CD8 T cells purified from the allografts subjected to prolonged cold ischemic storage in mice cardiac heart transpalnatation model.

We testd differences in hetelogous memory CD8 T cells and donor specific memory CD8 T cells infiltrating A/J heart allografts from of CTLA-4Ig non- or conditioned B6 recipients.Transcroptome analysis using memory CD8 T cell lysates isolated from the allografts showed differential expression of transcripts in this model. To test differences in heterologous memory CD8 T cells and donor specific memory CD8 T cells infiltrating A/J heart allografts subjected to 8 hours cold ischemic storage (CIS), we purified CD45.2+CD44+CD8 cells from the grafts on day2 post-transplant or at the time of rejection by flow sorting. To generate donor reactive memory CD8+ T cells, wild type (CD45.2+) B6 recipients received A/J skin allografts. After 8 weeks, recipient spleen suspensions were prepared from the mice and total CD8+ T cells were enriched by a negative selection mouse T cell isolation kit. Then, purified CD8 T cell populations were transferred intravenously (2x106/mouse) to congenic (CD45.1+) B6 recipients. After 72 hours, recipient mice received cardiac allografts subjected to prolonged CIS, were injected CTLA-4 Ig at 0.25 mg i.p. daily on days 0 and +1 and the allografts were harvested on days 2 post-transplant. Then, we performed transcriptomic analysis of heterologous or donor specific memory CD8 T cells using a PanCancer immune-profiling gene panel for the NanoString platform.