Combined TIM-3 and PD-1 blockade restrains hepatocellular carcinoma development by facilitating CD4+ and CD8+T cellmediated antitumor immune responses

BACKGROUND Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and T cell immunoglobulin and mucin domain-containing protein 3(TIM-3)are beneficial to the resumption of anti-tumor immunity response and hold extreme potential as efficient therapies for certain malignancies.However,ICIs with a single target exhibit poor overall response rate in hepatocellular carcinoma(HCC)patients due to the complex pathological mechanisms of HCC.AIM To investigate the effects of combined TIM-3 and PD-1 blockade on tumor development in an HCC mouse model,aiming to identify more effective immunotherapies and provide more treatment options for HCC patients.METHODS The levels of PD-1 and TIM-3 on CD4+and CD8+T cells from tumor tissues,ascites,and matched adjacent tissues from HCC patients were determined with flow cytometry.An HCC xenograft mouse model was established and treated with anti-TIM-3 monoclonal antibody(mAb)and/or anti-PD-1 mAb.Tumor growth in each group was measured.Hematoxylin and eosin staining and immunohistochemical staining were used to evaluate T cell infiltration in tumors.The percentage of CD4+and CD8+T cells in tissue samples from mice was tested with flow cytometry.The percentages of PD-1+CD8+,TIM-3+CD8+,and PD-1+TIM-3+CD8+T cells was accessed by flow cytometry.The levels of the cytokines including tumor necrosis factor alpha(TNF-α),interferon-γ(IFN-γ),interleukin(IL)-6,and IL-10 in tumor tissues were gauged with enzyme-linked immunosorbent assay kits.RESULTS We confirmed that PD-1 and TIM-3 expression was substantially upregulated in CD4+and CD8+T cells isolated from tumor tissues and ascites of HCC patients.TIM-3 mAb and PD-1 mAb treatment both reduced tumor volume and weight,while combined blockade had more substantial anti-tumor effects than individual treatment.Then we showed that combined therapy increased T cell infiltration into tumor tissues,and downregulated PD-1 and TIM-3 expression on CD8+T cells in tumor tissues.Moreover,combined treatment facilitated the production of T cell effector cytokines TNF-α and IFN-γ,and reduced the production of immunosuppressive cytokines IL-10 and IL-6 in tumor tissues.Thus,we implicated that combined blockade could ameliorate T cell exhaustion in HCC mouse model.CONCLUSION Combined TIM-3 and PD-1 blockade restrains HCC development by facilitating CD4+ and CD8+T cell-mediated antitumor immune responses.

Preoperative predictors of short-term survival after hepatectomy for multinodular hepatocellular carcinoma

AIM:To investigate preoperative factors associated with poor short-term outcome after resection for multinodular hepatocellular carcinoma(HCC) and to assess the contraindication of patients for surgery.METHODS:We retrospectively analyzed 162 multinodular HCC patients with Child-Pugh A liver function who underwent surgical resection.The prognostic significance of preoperative factors was investigated by univariate analysis using the log-rank test and by multivariate analysis using the Cox proportional hazards model.Each independent risk factor was then assigned points to construct a scoring model to evaluate the indication for surgical intervention.A receiver operating characteristics(ROC) curve was constructed to assess the predictive ability of this system.RESULTS:The median overall survival was 38.3 mo(range:3-80 mo),while the median disease-free survival was 18.6 mo(range:1-79 mo).The 1-year mortality was 14%.Independent prognostic risk factors of 1-year death included prealbumin < 170 mg/L [hazard ratio(HR):5.531,P < 0.001],alkaline phosphatase > 129 U/L(HR:3.252,P = 0.005),fetoprotein > 20 g/L(HR:7.477,P = 0.011),total tumor size > 8 cm(HR:10.543;P < 0.001),platelet count < 100 × 109/L(HR:9.937,P < 0.001),and-glutamyl transpeptidase > 64 U/L(HR:3.791,P < 0.001).The scoring model had a strong ability to predict 1-year survival(area under ROC:0.925,P < 0.001).Patients with a score ≥ 5 had significantly poorer short-term outcome than those with a score < 5(1-year mortality:62% vs 5%,P < 0.001;1-year recurrence rate:86% vs 33%,P < 0.001).Patients with score ≥ 5 had greater possibility of microvascular invasion(P < 0.001),poor tumor differentiation(P = 0.003),liver cirrhosis with small nodules(P < 0.001),and intraoperative blood transfusion(P = 0.010).CONCLUSION:A composite preoperative scoring model can be used as an indication of prognosis of HCC patients after surgical resection.Resection should be considered with caution in patients with a score ≥ 5,which indicates a contraindication for surgery.