It remains elusive whether or not this difference is a result of the malignant selleck screening library disease or the higher median age of the patient cohort compared to the healthy control group. Corresponding significant TCR V��-family elevations in blood and tumor tissue were not found in the present study. From previous analyses in colorectal cancer, we know, that TAA-specific T-cells circulated with frequencies lower than 1% of CD8+ cells without proven clonality . As anticipated, the proportion of peripheral TAA-specific T-cells was too low to be detected by TCR V��-quantification due to physiological variation of the relative V��-family expressions. Assuming several TAAs, expansions of T-cell clones would affect various V��-families additionally reducing sensitivity of the relative V��-quantification regarding single family percentages.
It is purely speculative to assume that such a specific T cell clone might be trapped at the tumor site and is therefore, not detectable in the peripheral blood. In carcinoma tissues, we did not observe an elevated repertoire restriction compared to corresponding tissue of unaffected colon. Infiltration of CTLs in colon carcinoma tissue had been identified as prognostic factor suggesting TAA associated CTL activation . Interestingly, we found no difference in the expression of C�� comparing healthy colon and carcinoma tissue. These inconsistent observations could be explained by recent results of Salama et al., who showed, that CD8+ CTL density is reduced but CD4+/CD25+/FoxP3+ T regulator cells are elevated in colon cancer compared to normal colon tissue [46,47].
The fact that clonal CTL expansions are not principally associated with restrictions in V��-family usage of the T regulatory cell population of the same compartment [42,47] might explain the absence of TCR restriction differences between healthy colon and CRC tissue. Independently of these considerations, we have to conclude that the present molecular quantitative TCR analysis is not suitable for the identification of local expansions in colorectal cancer tissue (if they exist) compared to normal colon tissue. This may differ in other compartments and/or for other tumor entities. Regarding p-values comparing the different sample groups (blood of healthy controls, blood of carcinoma patients, unaffected colonic mucosa, carcinoma tissue) one has to keep in mind the differences in sample sizes and applied tests.
The highly significant difference in CD between blood and tissue samples but the absence of a significant difference between unaffected colon and tumor tissue does not necessarily mean hat there would be no difference to detect if we Brefeldin_A would compare the same sample size of tissue samples as we used for comparing tissue with blood samples (48 vs 42 instead of 24 vs 18).