在对抗 PD-1/PD-L1 治疗耐药的肿瘤中，无法释放 T 细胞控制，这似乎与抑制性巨噬细胞有关。 V 域 T 细胞激活免疫球蛋白抑制因子 (VISTA) 是一种非冗余免疫检查点，可诱导 T 细胞和骨髓细胞免疫抑制。我们确定高水平的 VISTA 免疫细胞与晚期相关，并预测接受或不接受卡介苗免疫治疗的膀胱癌患者的生存率较差。 VISTA 免疫细胞和 PD-L1 免疫细胞或 PD-1 T 细胞的高浸润组合预测最差的预后。流式细胞术和多重免疫荧光染色证实巨噬细胞中VISTA表达高于T细胞或中性粒细胞，并且只有VISTA CD163巨噬细胞密度才能预测膀胱癌患者的不良预后。 Toll 样受体 (TLR) 激动剂可以触发巨噬细胞的先天免疫反应。 VISTA mAb 13F3 增强了 TLR3 特异性佐剂在体外诱导巨噬细胞活化的能力。在 MB49 同基因小鼠模型中，与 TLR3 特异性佐剂联合使用 VISTA mAb 13F3 治疗可抑制肿瘤生长并延长小鼠存活时间。联合治疗减少了 CD206 抗炎巨噬细胞和免疫抑制分子 TGF-β1，但上调了肿瘤微环境中的免疫刺激分子（Ifna、Ifnb 和 Trail）并增加了 CD8 T 细胞/Treg 比率。这些发现表明，免疫细胞（尤其是巨噬细胞内）的 VISTA 表达升高与膀胱癌患者的不良预后密切相关。将 VISTA 与 TLR3 特异性佐剂协同作用，可触发膀胱癌小鼠的有效抗肿瘤免疫并延迟肿瘤生长。

T cell control could not be unleashed in tumors resistant to anti-PD-1/PD-L1 treatment, which appears to be associated with inhibitory macrophages. V-domain immunoglobulin suppressor of T-cell activation (VISTA) is a non-redundant immune checkpoint that can induce both T cell and myeloid cell immunosuppression. We determined high levels of VISTA+ immune cells were associated with advanced stage, and predicted poor survival in bladder cancer patients with or without bacillus Calmette-Guerin immunotherapy. Combination of high infiltration of VISTA+ immune cells and PD-L1+ immune cells or PD-1+ T cells predicted the worst prognosis. Flow cytometry and multiplex immunofluorescence staining confirmed that VISTA expression was higher in macrophages than that in T cells or neutrophils, and only VISTA+CD163+ macrophage density predicted poor prognosis in bladder cancer patients. Toll-like receptor (TLR) agonists can trigger the innate immune response in macrophages. VISTA mAb 13F3 augmented the ability of a TLR3-specific adjuvant to induce macrophage activation in vitro. In a MB49 syngeneic mouse model, treatment with VISTA mAb 13F3 curbed tumor growth and prolonged mice survival when combined with TLR3-specific adjuvant. The combination treatment reduced CD206+ anti-inflammatory macrophages and immunosuppressive molecule TGF-β1, but it up-regulated immunostimulatory molecules (Ifna, Ifnb and Trail) and increased CD8+ T cell/Treg ratio in the tumor microenvironment. These findings indicated that elevated VISTA expression in immune cells, particularly within macrophages, is closely associated with an unfavorable prognosis in patients with bladder cancer. Targeting VISTA with TLR3-specific adjuvant synergy triggered an effective anti-tumor immunity and delayed tumor growth in mice with bladder cancer.