Hyperthermic isolated limb perfusion (ILP) with high-dose melphalan is a treatment option for melanoma patients with metastasis confined to limbs (in-transit metastasis). The therapy entails a complete response (CR) rate of 50-70%. Cellular immunity is proposed to impact on the clinical efficacy of ILP, but the detailed aspects of ILP-induced immune activation remain to be explored. For this study, we explored the potential role of interferon-stimulated gene (ISG) products, including CXCL10, CCL2, PD-L2 and IFN-γ along with expression of their cognate receptors CXCR3, CCR4, CCR5 and PD-1 on lymphocytes, for the clinical efficacy of ILP. Patients with high serum levels of CXCL10, CCL2, PD-L2 and IFN-γ were more likely to achieve CR after ILP. Additionally, the expression of CXCR3, CCR4 and CCR5 on T cells and/or natural killer (NK) cells was enhanced by ILP. Peripheral blood mononuclear cells (PBMCs) secreted high levels of CXCL10, CCL2 and IFN-γ in response to co-culture with melphalan-exposed melanoma cells in vitro. Activated T cells migrated toward supernatants from these co-cultures. Furthermore, melphalan-exposed melanoma cells triggered upregulation of CXCR3, CCR4, CCR5 and PD-1 on co-cultured T cells and/or NK cells. Our results suggest that constituents released from melphalan-exposed melanoma cells stimulate the ISG axis with ensuing formation of chemokines and upregulation of chemokine receptor expression on anti-neoplastic immune cells, which may contribute in ILP-induced tumor regression.

中文翻译：

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单独使用美法仑灌注肢体会激活干扰素刺激的基因，从而在黑色素瘤转移患者中诱导肿瘤消退。

高剂量美法仑的高温隔离肢体灌注（ILP）是转移局限于肢体的黑色素瘤患者的治疗选择（转运中）。该疗法的完全缓解率（CR）为50-70％。有人提出细胞免疫会影响ILP的临床疗效，但是ILP诱导的免疫激活的详细方面仍有待探索。在这项研究中，我们探讨了干扰素刺激基因（ISG）产品（包括CXCL10，CCL2，PD-L2和IFN-γ）及其在淋巴细胞上相关受体CXCR3，CCR4，CCR5和PD-1的表达的潜在作用，用于ILP的临床疗效。血清CXCL10，CCL2，PD-L2和IFN-γ较高的患者在ILP后更容易达到CR。此外，CXCR3的表达 ILP增强了T细胞和/或自然杀伤（NK）细胞上的CCR4和CCR5。外周血单核细胞（PBMC）分泌高水平的CXCL10，CCL2和IFN-γ，这是由于与暴露于美法仑的黑色素瘤细胞在体外共培养所致。活化的T细胞从这些共培养物中移向上清液。此外，暴露于美法仑的黑色素瘤细胞触发了共培养的T细胞和/或NK细胞上CXCR3，CCR4，CCR5和PD-1的上调。我们的结果表明，从暴露于马法兰的黑色素瘤细胞释放的成分刺激了ISG轴，继而形成了趋化因子并上调了抗肿瘤免疫细胞上趋化因子受体的表达，这可能有助于ILP诱导的肿瘤消退。CCL2和IFN-γ响应与美法仑暴露的黑色素瘤细胞在体外共培养。活化的T细胞从这些共培养物中移向上清液。此外，暴露于美法仑的黑色素瘤细胞触发了共培养的T细胞和/或NK细胞上CXCR3，CCR4，CCR5和PD-1的上调。我们的结果表明，从暴露于马法兰的黑色素瘤细胞释放的成分刺激了ISG轴，继而形成了趋化因子并上调了抗肿瘤免疫细胞上趋化因子受体的表达，这可能有助于ILP诱导的肿瘤消退。CCL2和IFN-γ响应与美法仑暴露的黑色素瘤细胞在体外共培养。活化的T细胞从这些共培养物中移向上清液。此外，暴露于美法仑的黑色素瘤细胞触发了共培养的T细胞和/或NK细胞上CXCR3，CCR4，CCR5和PD-1的上调。我们的结果表明，从暴露于马法兰的黑色素瘤细胞释放的成分刺激了ISG轴，继而形成了趋化因子并上调了抗肿瘤免疫细胞上趋化因子受体的表达，这可能有助于ILP诱导的肿瘤消退。