Th1 and CTL-based immune signature and elevated checkpoint expression in MSI colorectal cancer. Th1 and CTL-based immune signature and elevated checkpoint.

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Date of download: 6/3/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Novel Integrative Methods for Gene Discovery Associated.

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Th1 and CTL-based immune signature and elevated checkpoint expression in MSI colorectal cancer. Th1 and CTL-based immune signature and elevated checkpoint expression in MSI colorectal cancer. RNA was extracted from tissue samples laser-microdissected representing TIL in tumor nests (A), stroma surrounding tumor (B), and invasive front (C) areas of MSS (blue squares) and MSI (red circles) colorectal cancer specimens. Immune-related gene expression profiles were assessed using TaqMan-based qRT-PCR for selected genes. Sets of genes were defined by functional relevance (Th1/Tc1, CTL, Th17, Treg, proinflammation, immune checkpoints, and metabolism). The Y-axis represents an arbitrary unit of expression 2−ΔCt, ΔCt representing cycle threshold (Ct) of the gene of interest normalized by Ct of ubiquitous genes (GUSB and GAPDH). The graphs display the geometric means. Their differential representation between MSS and MSI specimens was analyzed using adjusted Wilcoxon–Mann–Whitney test as described in Methods. *, Wilcoxon P < 0.05. D, gene group comparison in TIL, tumor stroma, and invasive front areas between MSS and MSI specimens. Permutation test results based on the maximum Wilcoxon–Mann–Whitney test statistic within the gene groups Th1/Tc1, CTL, Th17, and immune checkpoints. *, statistically significant differences between MSS and MSI (P < 0.05). Nicolas J. Llosa et al. Cancer Discovery 2015;5:43-51 ©2015 by American Association for Cancer Research