Characterization of an adaptive immune response in microsatellite-instable colorectal cancer

Florence Boissière-Michot, Gwendal Lazennec, Hélène Frugier, Marta Jarlier, Lise Roca, Jacqueline Duffour, Emilie Du Paty, Daniel Laune, France Blanchard, Florence Le Pessot, Jean-Christophe Sabourin, Frédéric Bibeau, Florence Boissière-Michot, Gwendal Lazennec, Hélène Frugier, Marta Jarlier, Lise Roca, Jacqueline Duffour, Emilie Du Paty, Daniel Laune, France Blanchard, Florence Le Pessot, Jean-Christophe Sabourin, Frédéric Bibeau

Abstract

Sporadic or hereditary colorectal cancer (CRC) with microsatellite instability (MSI) is frequently characterized by inflammatory lymphocytic infiltration and tends to be associated with a better outcome than microsatellite stable (MSS) CRC, probably reflecting a more effective immune response. We investigated inflammatory mechanisms in 48 MSI CRCs and 62 MSS CRCs by analyzing: (1) the expression of 48 cytokines using Bio-Plex multiplex cytokine assays, and (2) the in situ immune response by immunohistochemical analysis with antibodies against CD3 (T lymphocytes), CD8 (cytotoxic T lymphocytes), CD45RO (memory T lymphocytes), T-bet (Th1 CD4 cells), and FoxP3 (regulatory T cells). MSI CRC exhibited significantly higher expression of CCL5 (RANTES), CXCL8 (IL-8), CXCL9 (MIG), IL-1β, CXCL10 (IP-10), IL-16, CXCL1 (GROα), and IL-1ra, and lower expression of MIF, compared with MSS CRC. Immunohistochemistry combined with image analysis indicated that the density of CD3+, CD8+, CD45RO+, and T-bet+ T lymphocytes was higher in MSI CRC than in MSS CRC, whereas the number of regulatory T cells (FoxP3+) was not statistically different between the groups. These results indicate that MSI CRC is associated with a specific cytokine expression profile that includes CCL5, CXCL10, and CXCL9, which are involved in the T helper type 1 (Th1) response and in the recruitment of memory CD45RO+ T cells. Our findings highlight the major role of adaptive immunity in MSI CRC and provide a possible explanation for the more favorable prognosis of this CRC subtype.

Keywords: chemokine; colorectal cancer; cytokine; inflammation; microsatellite instability.

Figures

https://www.ncbi.nlm.nih.gov/pmc/articles/instance/4121339/bin/onci-3-e29256-g1.jpg
Figure 1. Distribution of immune cells in MSS and MSI colorectal cancers. (A–D) Comparison of the density of immune cells in human colorectal cancers with microsatellite stability (MSS; black bars) and microsatellite instability (MSI; gray bars). (A) The intratumoral (ct) and peritumoral (im) densities of CD3+, CD8+, T-Bet+, CD45RO+, FoxP3+, CD68+, and CD20+ cells were assessed by image analysis of tissue microarray spots. (B–D) Representative examples of staining are shown for CD8 (B), T-Bet (C), and FoxP3 (D); images show immunoperoxidase staining (×100) with the corresponding digital images (stained cells are represented in red). Statistical analyses were performed by non-parametric Mann-Whitney test; *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant

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