Evaluating tumor-infiltrating lymphocytes in hepatocellular carcinoma using hematoxylin and eosin-stained tumor sections

Figure 1 The distribution and recurrence association of tumor infiltrating lymphocytes in the tumor center, invasive front, and peritumor. A: The spectrum of tumor infiltrating lymphocytes in the tumor center (TILs^CT), TILs in the invasive front (TILs^IF), and TILs in the peritumor (PILs) of 204 cases. TILs^CT were lower than TILs^IF and PILs. TILs^IF was the most flaming part than the other two areas; B: Dot map of TILs^CT, TILs^IF, and PILs, indicating a heterogeneous distribution of inflammation; C: Comparison of the mean of TILs^CT, TILs^IF and PILs from patients with tumor recurrence (black bars) or without tumor recurrence (white bars); D: Median survival time for all patients, with high densities (red bars) or low densities (black bars) of TILs^CT, TILs^IF and PILs; E, F and G: Patients with high TILs^CT, TILs^IF, and PILs had a lower recurrence rate. TILs^CT: Tumor infiltrating lymphocytes in the tumor center; TILs^IF: Tumor infiltrating lymphocytes in the invasive front 1 mm spacing from the malignant nests; PILs: infiltrating lymphocytes in the peritumor.

Figure 2 Representative hematoxylin and eosin images of the three immune subtypes (200 ×). A and B: Tumors with high infiltrating lymphocytes in the tumor center, invasive front and peritumor (Immune^high) subtype; C and D: Tumors other than immune^high and immune^low (Immune^mod) subtype; E and F: Tumors with low infiltrating lymphocytes in the tumor center, invasive front and peritumor (Immune^low) subtype; A, C, and E: Immune cells in tumor center; B, D, and F: Immune cells in the peritumor region.

Figure 3 The comparison of immune^high, immune^mod and immune^low subtypes. A: Immune subtypes can predict patients’ progression-free survival. Immune^high patients had a low recurrence rate, and immune^low patients experienced a high recurrence rate; B: The median survival time for all patients divided into three categories: Immune^high (red bars), immune^mod (green bars), and immune low (purple bars); C: The incidence rate of microvascular invasion (MVI) in three immune subtypes, immune^high subtype had a lower rate of MVI compared to immune^mod and immune^low subtypes; D: The presence of neutrophils in the three immune subtypes, a high incidence of neutrophils was detected in immune^high and immune^mod subtypes. Immune^high: Tumors with high infiltrating lymphocytes in the tumor center, invasive front and peritumor; Immune^mod: Tumors other than immune^high and immune^low; Immune^low: Tumors with low infiltrating lymphocytes in the tumor center, invasive front and peritumor; PFS: Progression-free survival; MVI: Microvascular invasion.

Figure 4 The pathological picture and recurrence association of programmed cell death-ligand 1 SP142 expression in tumor cells and immune cells (200 ×). A and B: Hematoxylin and eosin (H&E) and immunohistochemistry (IHC) picture of programmed cell death-ligand 1 (PD-L1) SP142 in tumor cells of one case; C and D: H&E and IHC picture of PD-L1 SP142 in immune cells of another case; E: Patients with high expression of PD-L1 (SP142) on tumor infiltrating lymphocytes tend to have less recurrence; F: Expression of PD-L1 (SP142) on tumor cells was not statistically significant. PD-L1: Programmed cell death-ligand 1; IHC: TILs: Tumor infiltrating lymphocytes; TCs: Tumor cells; PFS: Progression-free survival.