Translational profiling of in vivo virus-specific CD8 T cells

Translation is a critical cellular process to synthesize proteins from their transcripts. However, translational regulation in antigen-specific T cells in vivo has not been well defined. To address this question, we performed microarray analyses of both total and polysome-associated mRNAs in antigen specific CD8 T cells after acute viral infection. P14 transgenic CD8 T cells specific for GP33 epitope of lymphocytic choriomeningitis virus (LCMV) were adoptively transferred into B6 mice, followed by LCMV Armstrong infection. Day 5 effector and day 8 effector P14 CD8 T cells were purified from the LCMV-infected mice. Naive P14 CD8 T cells were directly purified from P14 transgenic mice. Total and polysome-associated (3 or more ribosomes) mRNAs were isolated from these purified P14 cells, and microarray analyses were performed.