Abstract In colorectal cancer (CRC) large scale tissue microarray (TMA) based quantitative immune cell counts using immune cell surface molecules (CD3, CD8, Granzyme B, and CD45RO) have identified the number of infiltrating immune cells to be potentially better predictors for patient survival than the classical TNM system. The spatial heterogeneity of immune cells may not be well reflected in the highly selected, and typically small (0,6-1 mm2) tissue cores of the TMA. This represents an obstacle in the individual prognosis prediction or classification of a single patient. To investigate this aspect, the localization and distribution of immune cell subpopulations based on the analysis of complete tissue sections by a dedicated novel staining and imaging system were performed. Using a specialized staining platform and whole slide imaging & analysis by virtual microscopy (VM), immunological “tumor maps” were generated. These tumor maps are based on cell densities in fields of 1mm2 size, visualizing intratumoral heterogeneity for the surface markers CD3, CD8, Granzyme B, and CD45RO. In total, an area of 867 mm2 was automatically evaluated with an average of 48 mm2 of evaluated tumor tissue per patient slide. Cell counts varied within a patient significantly, ranging from 0 to up to 2550 cells / mm2. Further analyses revealed, that sampling of single field counts within the tumor can only yield clear diagnostic decisions for a fraction of the analyzed patients, with ambiguous decisions for 11 out of 20 patients. Interestingly, the overall degree of heterogeneity also varied between patients, with lower heterogeneity found only in samples with lower cell counts. No samples with a homogeneous high cell density distribution were observed. The observed variability has implications for the individual prognosis prediction and represents the first spatial quantitative study of immune cells in a set of CRC primary tumors. The presented tumor maps therefore are a suitable tool to visualize heterogeneity. Furthermore, whole slide imaging & analysis by VM is essential in the identification of prognostic markers as well as in their subsequent application. In the future, spatial marker signatures could contribute to individual patient classification. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1917.
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