Ion inThPOK in Colorectal CarcinogenesisFigure 6. RUNX3, CD8 and ThPOK triple fluorescence. Triple colocalization of RUNX3, CD8 and ThPOK in NM (panel A-D, Scale bar = 50 mm), MA (panel E-H, Scale bar = 30 mm) and CRC (panel I-L, Scale bar = 30 mm). RUNX3: green (panel A, E, I); CD8: red (panel B, F, J); ThPOK: blue (panel C, G, K). Merge (panel D, H, L): CD8+ cells expressing RUNX3: yellow (arrow in panel H); CD8+ cells coexpressing RUNX3 and ThPOK: white (arrows in panel L). doi:10.1371/journal.pone.0054488.gnormal mucosa, and the number of CD56+ cells became almost undetectable during neoplastic progression. However, the marked decrease of CD56+ cells, together with the action exerted by ThPOK in CD8+ T lymphocytes, may be the key mechanisms of tumour microenvironment modification, referred as immunoediting, which makes the immune system inefficient against neoplastic growth. The number of blood white cells which have been typed 
is currently growing. Recent studies performed by flow cytometry showed a great plasticity of the immune system in terms of patterns or networks assumed by various leucocytic lineages. The results of the present study suggest that a pattern of proteins might exist which could define an overall status of the immune system, not a subpopulation of leukocytes in particular. In other words, colorectal cancer development could somehow influence not only the type of infiltrating cells themselves, but also drive its plasticity. ThPOK may be considered one of the main regulators of suchplasticity, influencing the immune escape mechanisms since the early onset of neoplastic clones.HIV-RT inhibitor 1 site AcknowledgmentsWe thank the Fondazione Umberto Veronesi. For this study the confocal microscope Leica TCS SP2 of the C.I.G.S. (Centro Interdipartimentale Grandi Strumenti) of the University of Modena and Reggio Emilia has been used. A particular thank to Dr. Andrea Tombesi for the valuable technical support.Author ContributionsConceived and designed the experiments: LR FM PS. Performed the experiments: FM PB MP PM AM. Analyzed the data: LR MPDL CP. Contributed reagents/materials/analysis tools: CDG CP MPDL. Wrote the paper: FM PS LR.
Crohn’s Disease (CD) is a chronic relapsing inflammatory disorder of the gastrointestinal tract. The etiopathogenesis of CD is not fully understood, but genetic and environmental factors interact to promote an excessive and poorly controlled mucosal inflammatory response directed against components of the gut microflora. [1?] Functional abnormalities of many components of the immune system can be seen in the damaged gut of CD patients, but hyperactivity of T cells with excessive production of inflammatory cytokines is believed to be one of the major immunological hallmarks of this disorder. CD-associated destructive immune response is polarized along the T helper (Th)1 cell pathway, as indicated by the demonstration that mucosal CD4+T cells produce large quantities of interferon (IFN)-c [3] and overexpress T-bet, a transcription factor necessary for driving and sustaining Th1 cell get SPDP Crosslinker responses. [4] CD tissue also contains high interleukin (IL)-12, [5] the major Th1-inducing factor in humans, [6] and IL-18, a cytokine that expands Th1 cell responses. [7] Despite these observations and the demonstration that Th1-typecytokines are pro-inflammatory in murine models of CD, [8] blockade of IFN-c with a neutralizing antibody (i.e. Fontolizumab) was not beneficial in CD patients. [9?0] These disappointing results could rely.Ion inThPOK in Colorectal CarcinogenesisFigure 6. RUNX3, CD8 and ThPOK triple fluorescence. Triple colocalization of RUNX3, CD8 and ThPOK in NM (panel A-D, Scale bar = 50 mm), MA (panel E-H, Scale bar = 30 mm) and CRC (panel I-L, Scale bar = 30 mm). RUNX3: green (panel A, E, I); CD8: red (panel B, F, J); ThPOK: blue (panel C, G, K). Merge 
(panel D, H, L): CD8+ cells expressing RUNX3: yellow (arrow in panel H); CD8+ cells coexpressing RUNX3 and ThPOK: white (arrows in panel L). doi:10.1371/journal.pone.0054488.gnormal mucosa, and the number of CD56+ cells became almost undetectable during neoplastic progression. However, the marked decrease of CD56+ cells, together with the action exerted by ThPOK in CD8+ T lymphocytes, may be the key mechanisms of tumour microenvironment modification, referred as immunoediting, which makes the immune system inefficient against neoplastic growth. The number of blood white cells which have been typed is currently growing. Recent studies performed by flow cytometry showed a great plasticity of the immune system in terms of patterns or networks assumed by various leucocytic lineages. The results of the present study suggest that a pattern of proteins might exist which could define an overall status of the immune system, not a subpopulation of leukocytes in particular. In other words, colorectal cancer development could somehow influence not only the type of infiltrating cells themselves, but also drive its plasticity. ThPOK may be considered one of the main regulators of suchplasticity, influencing the immune escape mechanisms since the early onset of neoplastic clones.AcknowledgmentsWe thank the Fondazione Umberto Veronesi. For this study the confocal microscope Leica TCS SP2 of the C.I.G.S. (Centro Interdipartimentale Grandi Strumenti) of the University of Modena and Reggio Emilia has been used. A particular thank to Dr. Andrea Tombesi for the valuable technical support.Author ContributionsConceived and designed the experiments: LR FM PS. Performed the experiments: FM PB MP PM AM. Analyzed the data: LR MPDL CP. Contributed reagents/materials/analysis tools: CDG CP MPDL. Wrote the paper: FM PS LR.
Crohn’s Disease (CD) is a chronic relapsing inflammatory disorder of the gastrointestinal tract. The etiopathogenesis of CD is not fully understood, but genetic and environmental factors interact to promote an excessive and poorly controlled mucosal inflammatory response directed against components of the gut microflora. [1?] Functional abnormalities of many components of the immune system can be seen in the damaged gut of CD patients, but hyperactivity of T cells with excessive production of inflammatory cytokines is believed to be one of the major immunological hallmarks of this disorder. CD-associated destructive immune response is polarized along the T helper (Th)1 cell pathway, as indicated by the demonstration that mucosal CD4+T cells produce large quantities of interferon (IFN)-c [3] and overexpress T-bet, a transcription factor necessary for driving and sustaining Th1 cell responses. [4] CD tissue also contains high interleukin (IL)-12, [5] the major Th1-inducing factor in humans, [6] and IL-18, a cytokine that expands Th1 cell responses. [7] Despite these observations and the demonstration that Th1-typecytokines are pro-inflammatory in murine models of CD, [8] blockade of IFN-c with a neutralizing antibody (i.e. Fontolizumab) was not beneficial in CD patients. [9?0] These disappointing results could rely.