DataSheet_1_IFNγ at the early stage induced after cryo-thermal therapy maintains CD4+ Th1-prone differentiation, leading to long-term antitumor immunity.docx

IntroductionRecently, more and more research illustrated the importance of inducing CD4+ T helper type (Th)-1 dominant immunity for the success of tumor immunotherapy. Our prior studies revealed the crucial role of CD4+ Th1 cells in orchestrating systemic and durable antitumor immunity, which contributes to the satisfactory outcomes of the novel cryo-thermal therapy in the B16F10 tumor model. However, the mechanism for maintaining the cryo-thermal therapy-mediated durable CD4+ Th1-dominant response remains uncovered. Additionally, cryo-thermal-induced early-stage CD4+ Th1-dominant T cell response showed a correlation with the favorable prognosis in patients with colorectal cancer liver metastasis (CRCLM). We hypothesized that CD4+ Th1-dominant differentiation induced during the early stage post cryo-thermal therapy would affect the balance of CD4+ subsets at the late phase.MethodsTo understand the role of interferon (IFN)-γ, the major effector of Th1 subsets, in maintaining long-term CD4+ Th1-prone polarization, B16F10 melanoma model was established in this study and a monoclonal antibody was used at the early stage post cryo-thermal therapy for interferon (IFN)-γ signaling blockade, and the influence on the phenotypic and functional change of immune cells was evaluated.ResultsIFNγ at the early stage after cryo-thermal therapy maintained long-lasting CD4+ Th1-prone immunity by directly controlling Th17, Tfh, and Tregs polarization, leading to the hyperactivation of Myeloid-derived suppressor cells (MDSCs) represented by abundant interleukin (IL)-1β generation, and thereby further amplifying Th1 response.DiscussionOur finding emphasized the key role of early-phase IFNγ abundance post cryo-thermal therapy, which could be a biomarker for better prognosis after cryo-thermal therapy.

引言近期，众多研究凸显了诱导CD4+辅助T细胞型（Th）-1优势免疫对于肿瘤免疫治疗成功的重要性。本课题组先前的研究揭示了CD4+ Th1细胞在调控全身性和持久性抗肿瘤免疫中的关键作用，这有助于新型冷冻热疗在B16F10肿瘤模型中取得令人满意的疗效。然而，维持冷冻热疗介导的持久CD4+ Th1优势反应的机制尚未揭晓。此外，冷冻热疗诱导的早期CD4+ Th1优势T细胞反应与结直肠癌肝转移（CRCLM）患者良好的预后相关。我们假设，冷冻热疗早期阶段诱导的CD4+ Th1优势分化将影响晚期CD4+亚群的平衡。方法为了理解Th1亚群的主要效应因子干扰素（IFN）-γ在维持长期CD4+ Th1倾向性极化中的作用，本研究建立了B16F10黑色素瘤模型，并在冷冻热疗早期阶段使用单克隆抗体进行干扰素（IFN）-γ信号通路阻断，评估其对免疫细胞表型和功能变化的影响。结果冷冻热疗早期阶段的IFNγ通过直接调控Th17、Tfh和Tregs极化，维持了持久的CD4+ Th1倾向性免疫，导致髓源抑制细胞（MDSCs）过度活化，表现为大量白细胞介素（IL）-1β的产生，从而进一步放大Th1反应。讨论我们的发现强调了冷冻热疗后期IFNγ丰度的重要性，这可能是冷冻热疗后良好预后的生物标志物。