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Background The T cell cytokine profile is a key prognostic indicator of post-surgical outcome for colorectal cancer (CRC). Whilst T(H)1 (IFN-gamma(+)) cell-mediated responses generated in CRC are well documented and are associated with improved survival, antigen-specific T(H)17 (IL-17A(+)) responses have not been similarly measured. Methods We sought to determine the cytokine profile of circulating tumour antigen-(5T4/CEA) specific T cells of 34 CRC patients to address whether antigen-specific IL-17A responses were detectable and whether these were distinct to IFN-gamma responses. Results As with IFN-gamma-producing T cells, anti-5T4/CEA T(H)17 responses were detectable predominantly in early stage (TNM I/II) CRC patients. Moreover, whilst IL-17A was always produced in association with IFN-gamma, this release was mainly from two distinct T cell populations rather than by dual producing T cells. Patients mounting both tumour-specific T(H)1(+)/T(H)17(+) responses exhibited prolonged relapse-free survival. Conclusions Tumour antigen-specific T(H)17 responses play a beneficial role in preventing post-operative colorectal tumour recurrence., Funding Agencies|Cancer Research UK (CRUK)Cancer Research UK [C16731/A21200] Funding Source: Medline

BackgroundThe relationship between intestinal epithelial integrity and the development of intestinal disease is of increasing interest. A reduction in mucosal integrity has been associated with ulcerative colitis, Crohns disease and potentially could have links with colorectal cancer development. The Ussing chamber system can be utilised as a valuable tool for measuring gut integrity. Here we describe step-by-step methodology required to measure intestinal permeability of both mouse and human colonic tissue samples ex vivo, using the latest equipment and software. This system can be modified to accommodate other tissues.MethodsAn Ussing chamber was constructed and adapted to support both mouse and human tissue to measure intestinal permeability, using paracellular flux and electrical measurements. Two mouse models of intestinal inflammation (dextran sodium sulphate treatment and T regulatory cell depletion using C57BL/6-FoxP3(DTR) mice) were used to validate the system along with human colonic biopsy samples.ResultsDistinct regional differences in permeability were consistently identified within mouse and healthy human colon. In particular, mice showed increased permeability in the mid colonic region. In humans the left colon is more permeable than the right. Furthermore, inflammatory conditions induced chemically or due to autoimmunity reduced intestinal integrity, validating the use of the system.ConclusionsThe Ussing chamber has been used for many years to measure barrier function. However, a clear and informative methods paper describing the setup of modern equipment and step-by-step procedure to measure mouse and human intestinal permeability isnt available. The Ussing chamber system methodology we describe provides such detail to guide investigation of gut integrity., Funding Agencies|Cancer Research Wales [506719]; Wellcome Trust [504182]; Cancer Research UK [C16731/A21200]; Cardiff University