The Effect of PD-1 Inhibitor Combined with Irradiation on HMGB1-Associated Inflammatory Cytokines and Myocardial Injury

Figures & data

Figure 1 HE staining of heart tissue 1 month after irradiation and/or PD-1 inhibitors treatment in each group. “↑” shows cardiac structural impairments and/or lymphocyte aggregation. (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation.

Figure 2 Masson staining of heart tissue 1 month after irradiation and/or PD-1 inhibitors treatment in each group. “↑” shows collagen fibers. (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation.

Figure 3 Collagen volume fraction in heart tissue in the four groups of mice (n = 5). (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation. nsP>0.05, *P<0.05, ***P<0.0001.

Figure 4 TUNEL staining of heart tissue 1 month after irradiation and/or PD-1 inhibitors treatment in each group. (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation.

Figure 5 Apoptotic index in heart tissue in the four groups of mice (n = 5). (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation. nsP>0.05, *P<0.05, ***P<0.0001.

Table 1 Absolute Count of CD3+, CD4+ and CD8+ T-Lymphocytes in Each Group of Mice (Cells/μL, Mean±SD)

Groups	CD3+	CD4+	CD8+
Control	153±10.01	6±0.47	22±2.49
PD-1 inhibitors	429±7.41	12±1.63	117±5.73
Irradiation	465±16.11	26±2.16	171±8.96
PD-1 inhibitors+irradiation	534±11.43	32±3.40	234±12.28

Abbreviations: HMGB1, high mobility group box 1; qPCR, quantitative real-time PCR; CVF, collagen volume fraction; AI, apoptotic index; RIMI, radiation-induced myocardial injury; IR, ionizing radiation.

Figure 6 Distribution of T lymphocytes subsets in heart tissue in the four groups of mice. (A) Distribution of CD3+ T lymphocytes; (B) Distribution of CD4+, CD8+ T lymphocytes. (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation.

Figure 7 Relative percentage of T lymphocytes subsets in heart tissue in the four groups of mice (n = 3). (A) Relative percentage of CD3+ lymphocytes; (B) Relative percentage of CD4+ lymphocytes; (C) Relative percentage of CD8+ T lymphocytes. (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation. nsP>0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Figure 8 Concentrations of cytokine in heart tissue in te four groups of mice (n = 3). (A) IL-1β concentration; (B) IL-6 concentration; (C) TNF-α concentration. (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation. nsP>0.05, *P<0.05, ***P<0.001, ****P<0.0001.

Figure 9 Blot and expression of relative protein in heart tissue in the four groups of mice (n = 3). (A) HMGB1, TLR4, NF-κB p65 protein; (B) expression of HMGB1 protein; (C) expression of TLR4 protein; (d) expression of NF-κB p65 protein. (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation. nsP>0.05, *P<0.05, ***P<0.001, ****P<0.0001.

Figure 10 Expression of relative mRNA in heart tissue in the four groups of mice (n = 3). (A) Expression of HMGB1 mRNA; (B) Expression of TLR4 mRNA; (C) Expression of NF-κB p65 mRNA. (A) Control; (B) PD-1 inhibitors; (C) Irradiation; (D) PD-1 inhibitors + irradiation. nsP>0.05, *P<0.05, ***P<0.001, ****P<0.0001.