In this retrospective study on 67 patients with hepatocellular carcinoma (HCC), after tumor resection, we evaluated the significance of CD3+ and CD8+ T-lymphocytes and CD20+ B-lymphocytes in tumor and non-tumor liver for time to recurrence (TTR), disease-free survival (DFS) and overall survival. After immunohistochemical staining, the density of nucleated lymphocyte profiles (QA) was estimated stereologically in the tumor center (TC), inner margin (inn M), outer margin (out M), peritumor and non-tumor liver. In TC, intermediate and high QA of CD8+ cells predicted longer TTR, whereas CD3+ and CD20+ were predictive only at high QA. DFS was predicted by high QA of CD3+, CD8+ and CD20+ cells in TC. The inn M harbored smaller QA of CD3+, CD8+ and CD20+ lymphocytes than out M. In contrast to out M, high T-cells' QA and intermediate and high B-cell QA in inn M predicted longer TTR and DFS. High inn M/out M QA ratios of CD3+ and CD20+ cells were associated with longer TTR and DFS, whereas high inn M/out M QA ratio of CD8+ was predictive only for DFS. Patients with intermediate-high QA of combined CD8+ and CD20+ cells in inn M showed longer TTR and DFS, compared to CD8+-high or CD20+-high alone. Our findings highlight overall heterogeneity of the tumor invasive margin, the importance of inn M, and the predictive role of B-cells.

Bioptická Laboratoř s r o Mikulášské Nám 4 32600 Pilsen Czech Republic

Department of Cancer Epidemiology German Cancer Research Center Im Neuenheimer Feld 280 69120 Heidelberg Germany

Department of Histology and Embryology Faculty of Medicine in Pilsen Charles University Karlovarská 48 30166 Pilsen Czech Republic

Department of Medical Chemistry and Biochemistry Faculty of Medicine in Pilsen Charles University Karlovarská 48 30166 Pilsen Czech Republic

Department of Pathology 3rd Faculty of Medicine Charles University Ruská 87 10000 Prague Czech Republic

Department of Surgery and Biomedical Center Faculty of Medicine in Pilsen Charles University Alej Svobody 80 32300 Pilsen Czech Republic

Laboratory of Cancer Treatment and Tissue Regeneration Biomedical Center Faculty of Medicine in Pilsen Charles University Alej Svobody 1665 76 32300 Pilsen Czech Republic

Laboratory of Quantitative Histology Biomedical Center Faculty of Medicine in Pilsen Charles University Alej Svobody 1665 76 32300 Pilsen Czech Republic

Laboratory of Translational Cancer Genomics Biomedical Center Faculty of Medicine in Pilsen Charles University Alej Svobody 1665 76 32300 Pilsen Czech Republic

Sikl's Institute of Pathology Faculty of Medicine and Teaching Hospital in Plzen Charles University ul Dr E Beneše 13 30599 Pilsen Czech Republic

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CTNNB1 mutations, TERT polymorphism and CD8+ cell densities in resected hepatocellular carcinoma are associated with longer time to recurrence