Increased co-expression of PD1 and TIM3 is associated with poor prognosis and immune microenvironment heterogeneity in gallbladder cancer.

BACKGROUND: The effectiveness of immune checkpoint inhibitors in treating gallbladder cancer (GBC) remains unsatisfactory. Recently, several new immune checkpoints have been identified. However, investigations exploring these immune checkpoints in GBC are limited. In this study, we aim to investigate the expression patterns and clinical implications of various immune checkpoints, and further characterize the spatial and quantitative heterogeneity of immune components in GBC. METHODS: We employed single and multiplex immunohistochemistry to evaluate the expression of five immune checkpoint markers and four immune cell markers in the primary tumor core, hepatic invasion margin, and liver metastasis. Subsequently, we analyzed their interrelationships and their prognostic significance. RESULTS: We observed a robust positive correlation between PD1/TIM3 expression in GBC (R = 0.614, P < 0.001). The co-expression of PD1/TIM3 exhibited a synergistic effect in predicting poor prognosis among postoperative GBC patients. Further analysis revealed that the prognostic significance of PD1/TIM3 was prominent in the subgroup with high infiltration of CD8 + T cells (P < 0.001). Multiplex immunohistochemistry reveals that PD1 + TIM3 + FOXP3 + cells constitute a significant proportion of FOXP3 + TILs in GBC tissue. Moreover, the co-high expression of PD1 and TIM3 is positively correlated with the accumulation of CD8 + TILs at the hepatic invasion margin. Lastly, our findings indicated reduced expression levels of immune checkpoints and diminished immune cell infiltration in liver metastases compared to primary tumors. CONCLUSIONS: Increased co-expression of PD1/TIM3 is associated with poor prognosis in GBC patients and is related to the heterogeneity of immune microenvironment between GBC primary tumor and its hepatic invasion margin or liver metastases, which may be a potential target for future immunotherapy of GBC.

[Acute oral toxicity and 90 days feeding test of recombinant human lactoferrin].

OBJECTIVE: To study the edible safety of recombinant human lactoferrin( rh LF) expressed from transgenic cow mammary gland bioreactor. METHODS: According to the food additive safety toxicology evaluation procedures and method, acute oral toxicity in rats and 90 day subchronic toxicity test in mice were done to evaluate the edible safety of rh LF. RESULTS: Acute oral toxicity test indicated that rh LF was no toxic effect during the observation period, mouse acute oral LD50 of recombinant human lactoferrin was greater than 20 000 mg/kg. 90 days feeding test indicated that there was no-observedadverse-effect-level after givening 300 times rh LF recommended dose of animals body, toxicological parameters NOAEL was 10. 00 g/( kg·d). CONCLUSION: According to the acute toxic dose graduation standard, rh LF was nonpoisonous. Rh LF was no-observedadverse-effect-level and no subchronic toxicity after givening 300 times rh LF recommended dose of animals body. According to the result, rh LF was no potential food safety risk.