Validation Group An external validation set was analysed for CD8+

Validation Group An external validation set was analysed for CD8+ and TIA-1+ TILs. In this group of MMR-proficient colorectal cancer, only 7 patients had tumors with CD8+/TIA-1- TILs In univariable analysis, survival time differences for patients with CD8+/TIA-1+ TILs were significantly selleck chemical Seliciclib higher compared to patients with absence of CD8+ TILs (p<0.0001). Survival time differences between patients with different CD8/TIA-1 phenotypes were evaluated in multivariable analysis with pT stage, pN stage, pM and adjuvant therapy. Among patients with CD8-positive tumors, TIA-1 positivity led to a relative risk of 0.79 (95%CI: 0.7-0.9) compared to patients with TIA-1-negative cancers (p=0.006).

Discussion The novel findings of this study on 1406 MMR-proficient colorectal cancer patients suggest first, that higher numbers of TIA-1+ TILs represent a favourable and independent prognostic parameter and second that in addition to CD8, TIA-1 improves the prognostic stratification of patients by 35%. Although the prognostic significance of abundant CD8+ TILs has previously been established [3], [5], [15], [16], [17], this study goes one step further to identify the marker TIA-1 as a highly relevant prognostic parameter in colorectal cancer and particularly in tumors with marked cytotoxic CD8+ TILs. TIA-1 is a cytoplasmic granule-associated RNA binding protein reportedly expressed in cells with cytolytic potential, including 50-60% of CD8+ T-lymphocytes [18]. TIA-1 has been shown to be involved in Fas-mediated apoptosis in a variety of human malignancies, and to sensitize endothelial cells to pro-apoptotic stimuli while also enhancing NK cell cytotoxic activity [19], [20], [21].

Our results are in agreement with these findings. Here, we show that 64% of CD8+ TILs co-express TIA-1. Additionally, we identify a small population of CD8- but TIA-1+ tumor infiltrating cells representing populations of TCR�æ� cells and neutrophils and to a lesser extent CD4+ T cells, macrophages, NK cells, and NK/T cells. Investigations on the clinical and prognostic implications of TIA-1+ lymphocytes is mostly restricted to haematological malignancies with both favourable and unfavourable effects attributed to over-expression of this protein [22], [23]. Only a handful of studies have evaluated TIA-1 expression on TILs in colorectal cancer, relating up-regulation to apoptosis and an increased number of TIA-1+ TILs in MMR-deficient tumors [24], [25].

Interestingly, in our MMR-proficient cases, we could not find any correlation between higher numbers of TIA-1+ cells and tumor expression of the anti-apoptotic marker Bcl-2, pro-apoptotic markers Apoptosis Activating GSK-3 Factor-1 (APAF-1) and Mammalian Sterile20-like kinase 1 (MST-1), nor with p53 or the proliferation marker Ki67 (data not shown).

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