A template to quantify the location and density of CD3 + and CD8 + tumor-infiltrating lymphocytes in colon cancer by digital pathology on whole slides for an objective, standardized immune score assessment

Abstract

Background In colon cancer, the location and density of tumor-infiltrating lymphocytes (TILs) can classify patients into low and high-risk groups for prognostication. While a commercially available ‘Immunoscore®’ exists, the incurred expenses and copyrights may prevent universal use. The aim of this study was to develop a robust and objective quantification method of TILs in colon cancer. Methods A consecutive, unselected series of specimens from patients with colon cancer were available for immunohistochemistry and assessment of TILs by automated digital pathology. CD3 + and CD8 + cells at the invasive margin and in tumor center were assessed on consecutive sections using automated digital pathology and image analysis software (Visiopharm®). An algorithm template for whole slide assessment, generated cell counts per square millimeters (cells/mm2), from which the immune score was calculated using distribution volumes. Furthermore, immune score was compared with clinical and histopathological characteristics to confirm its relevance. Results Based on the quantified TILs numbers by digital image analyses, patients were classified into low (n = 83, 69.7%), intermediate (n = 14, 11.8%) and high (n = 22, 18.5%) immune score groups. High immune score was associated with stage I–II tumors (p = 0.017) and a higher prevalence of microsatellite instable (MSI) tumors (p = 0.030). MSI tumors had a significantly higher numbers of CD3 + TILs in the invasive margin and CD8 + TILs in both tumor center and invasive margin, compared to microsatellite stable (MSS) tumors. Conclusion A digital template to quantify an easy-to-use immune score corresponds with clinicopathological features and MSI in colon cancer.

Correlation between Lymphocyte-to-Monocyte Ratio (LMR), Neutrophil-to-Lymphocyte Ratio (NLR), Platelet-to-Lymphocyte Ratio (PLR) and Tumor-Infiltrating Lymphocytes (TILs) in Left-Sided Colorectal Cancer Patients

“…Mean values of CD3+ and CD8+ lymphocytes in our study were high compared to some analyses; however, even higher mean densities of lymphocytes, especially in early stages of the disease, and higher intrapatient variability of the density of lymphocytes have been reported [10][11][12][13]. In most studies, only semi-quantitative evaluations of TIL densities were conducted, making it difficult to perform conclusive comparisons in this regard.…”

Association of Epithelial Mesenchymal Transition, Stemness and Inflammation in Colorectal Carcinoma. Doctoral Thesis

“…T cells play one of main roles in a tumour invasion. Lea et al reported very similar results: Perineural invasion was associated with a low Immunoscore (p = 0.008), but lymphatic invasion had no significant association with its changes (p = 0.10)(Lea et al, 2021). These results could indicate the potential role of CD8+ T cells in tumour metastasis potential and local structure involvement, as well as local tumour growth.…”

“…lymphocytes are known to limit neoplastic cell growth, to suppress tumour infiltration, and to induce elimination of tumours(Steele et al, 2018). In CRC, the presence of CD8+ lymphocytes is associated with high MSI, although data about overall survival is controversial(Lea et al, 2021;Millen et al, 2020;Sudoyo et al, 2019). A high CD8+ T cell count has been associated with stage I-II CRC (p = 0.017), and the CD8+ T cell count is significantly higher in the tumour centre and invasive margin in MSI tumours compared with MSS tumours(Lea et al, 2021).Researchers have also suggested that a combined score of CD8+ T cells and tumour buds in CRC could be a better prediction marker for nodal metastases and survival(Dawson et al., transmembrane glycoprotein that normally participates in cell-cell interactions, adhesion of the cytoskeleton to the extracellular matrix, and cell migration.…”

“…In CRC, the presence of CD8+ lymphocytes is associated with high MSI, although data about overall survival is controversial(Lea et al, 2021;Millen et al, 2020;Sudoyo et al, 2019). A high CD8+ T cell count has been associated with stage I-II CRC (p = 0.017), and the CD8+ T cell count is significantly higher in the tumour centre and invasive margin in MSI tumours compared with MSS tumours(Lea et al, 2021).Researchers have also suggested that a combined score of CD8+ T cells and tumour buds in CRC could be a better prediction marker for nodal metastases and survival(Dawson et al., transmembrane glycoprotein that normally participates in cell-cell interactions, adhesion of the cytoskeleton to the extracellular matrix, and cell migration. In cancers, CD44 is associated with clonality, metastatic spreading ability, resistance to chemotherapy and radiotherapy, as well as lower overall survival (Mare et al, 2021; Simtniece et al, 2015; C. Wang et al, 2012; Z. Wang et al, 2019).…”

“…Figure 2.2 Immunohistochemical marker expression evaluation Atotal loss of E-cadherin expression in colorectal cancer (red arrow), original magnification (OM) 400×; Bweak E-cadherin expression in colorectal cancer cells, OM 400×; Cmoderate and strong E-cadherin expression in colorectal cancer, OM 100×. Images taken by Inese Briede.For CD8+ tumour-infiltrating lymphocytes, the cell number per mm 2 was counted via Diapath in the tumour centre and the invasive margin, as described in previous studies(Lea et al, 2021;Ueno et al, 2014).…”