Analysis of graft-infiltrating CD8+ T cell profile after CD3 antibody therapy

We performed a global gene expression analysis comparing intragraft tolerant CD8+ T cells from CD3 antibody-treated mice showing permanent islet graft survival to intra-graft effector CD8+ T cells isolated from untreated mice showing acute rejection of islet allografts. The objective was to emphasize the anergic profile of CD8+ T cells residing within the pancreatic islet allograft of mice rendered tolerant following CD3 antibody therapy. Five hundreds graft-infiltrating CD8+ T cells from individual tolerant CD3 Ab-treated recipients (n=8) or untreated recipients (n=8) were FACS sorted, lysed using SuperAmpTM lysis buffer (Miltenyi Biotec) and stored at -80°C until all samples were collected. Amplification of RNA, sample hydridization (Agilent whole mouse genome oligo microarrays), scanning and data acquisition were performed by Miltenyi Biotec (Cologne, Germany).