A23:卵巢癌调节性T细胞中Neuropilin-1的表达

A. Cillo, T. Bruno, F. Modugno, R. Edwards, D. Vignali
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Intracellular NRP1 was expressed on a median of 1.8% (interquartile range [IQR]: 0.69% to 4.8%) of Treg from healthy donors compared with a median of 66% (IQR: 28% to 90%; p Conclusions and Future Directions: Both intracellular and surface NRP1 are expressed more frequently on tumor-infiltrating Treg and Treg from ascites fluid compared to Treg from healthy donor peripheral blood. Although the number of patients currently included in this study is small, there was also evidence that intracellular NRP1 expression on Treg was higher on malignant compared with benign tissue. Additionally, Treg from ascites fluid were capable of suppressing CD8 cells ex vivo. Future studies will evaluate whether blockade of NRP1 on human Treg limits either the survival or suppressive capacity of Treg. Note: This abstract was not presented at the conference. Citation Format: Anthony R. Cillo, Tullia Bruno, Francesmary Modugno, Robert Edwards, Dario Vignali. 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引用次数: 0

摘要

调节性T细胞(Treg)是CD4+ T细胞的一个亚群,它抑制自身免疫反应,但也阻止肿瘤和慢性病毒感染的清除。在卵巢癌中,肿瘤浸润Treg的频率较高与预后不良有关,但在这种情况下,对Treg存活和抑制活性的机制的了解有限。我们的研究小组最近在小鼠癌症模型中描述了一个通过嗜神经蛋白-1 (NRP1)作用于Treg的信号轴,该信号轴促进Treg的存活和抑制功能。NRP1基因敲除小鼠Treg导致肿瘤生长减少和生存增加,强调了NRP1+ Treg在抑制抗肿瘤免疫中的重要性。鉴于NRP1+ Treg在小鼠肿瘤模型中的重要性,我们试图探索NRP1在卵巢癌患者肿瘤浸润性Treg中的表达模式。材料和方法:卵巢肿瘤通过匹兹堡大学健康科学组织库获得,卵巢腹水通过Magee妇女研究所获得。外周血通过红十字会从健康的献血者处获得。采用密度梯度离心分离外周血单个核细胞(PBMC),先机械破坏肿瘤,分离肿瘤浸润淋巴细胞,然后用50 ug/mL Liberase DL在37℃下处理15分钟。流式细胞术分析Treg时,首先对血液和肿瘤细胞进行表面标记物染色,然后进行活/死区分、固定和渗透,最后进行细胞内标记物染色。细胞在希尔曼癌症中心流动核心的LSR Fortessa上进行分析。为了进行功能测试,从腹水中选择Treg,并测定其体外抑制幼稚CD8+ T细胞的能力。采用Wilcoxon秩和检验评估组间差异,双侧alpha值小于5%。结果:共对12例健康供体和20例卵巢癌患者进行了研究。通过流式细胞术检测健康供者外周血CD4+CD25+FOXP3+ Treg和卵巢癌肿瘤或卵巢癌患者腹水的肿瘤浸润淋巴细胞,评估细胞内和表面NRP1的表达。细胞内NRP1在健康供者Treg中表达的中位数为1.8%(四分位数范围[IQR]: 0.69%至4.8%),而在健康供者Treg中表达的中位数为66% (IQR: 28%至90%;结论和未来方向:细胞内和表面NRP1在肿瘤浸润的Treg和来自腹水的Treg上的表达比来自健康供者外周血的Treg更频繁。虽然目前纳入本研究的患者数量较少,但也有证据表明,恶性组织中细胞内NRP1在Treg上的表达高于良性组织。此外,来自腹水的Treg能够在体外抑制CD8细胞。未来的研究将评估NRP1对人类Treg的阻断是否会限制Treg的存活或抑制能力。注:本摘要未在会议上发表。引文格式:Anthony R. Cillo, Tullia Bruno, Francesmary Modugno, Robert Edwards, Dario Vignali。Neuropilin-1在卵巢癌调节性T细胞中的表达。[摘要]。AACR会议论文集:解决卵巢癌研究和治疗中的关键问题;2017年10月1-4日;宾夕法尼亚州匹兹堡。费城(PA): AACR;临床肿瘤杂志,2018;24(15 -增刊):摘要nr - A23。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Abstract A23: Neuropilin-1 expression on regulatory T cells in ovarian cancer
Introduction: Regulatory T cells (Treg) are a subpopulation of CD4+ T cells that suppress autoimmune responses, but also prevent clearance of tumors and chronic viral infections. In ovarian cancer, a higher frequency of tumor-infiltrating Treg is associated with poor prognosis, but insights into the mechanisms governing the survival and suppressive activity of Treg in this context are limited. Our group has recently described a signaling axis through neurophilin-1 (NRP1) on Treg in murine models of cancer that promotes the survival and suppressive function of Treg. Genetic knockout of NRP1 on murine Treg leads to reduced tumor growth and increased survival, underscoring the importance of NRP1+ Treg in suppressing antitumor immunity. Given the importance of NRP1+ Treg in murine cancer models, we sought to explore the expression pattern of NRP1 on tumor-infiltrating Treg from patients with ovarian cancer. Materials and Methods: Ovarian tumors were obtained through the health sciences tissue bank at the University of Pittsburgh, and ovarian ascites fluid was obtained through Magee Women’s Research Institute. Peripheral blood was obtained from healthy donors through the Red Cross. Peripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation, and tumor-infiltrating lymphocytes were isolated by first mechanically disrupting the tumor, followed by treatment for 15 minutes at 37°C with 50 ug/mL Liberase DL. For flow cytometric analysis of Treg, cells from blood and tumors were first stained for surface markers, followed by live/dead discrimination, fixation and permeabilization, and staining for intracellular markers. Cells were analyzed on an LSR Fortessa at the Hillman Cancer Center Flow Core. For functional assays, Treg were selected from ascites fluid and assayed for their ability to suppressive naive CD8+ T cells ex vivo. Wilcoxon rank sum tests were used to assess differences between groups, and a two-sided alpha less than 5% was considered significant. Results: A total of 12 healthy donors and 20 ovarian cancer patients were studied. Intracellular and surface NRP1 expression was assessed by flow cytometry on CD4+CD25+FOXP3+ Treg from healthy donor peripheral blood and tumor-infiltrating lymphocytes from either ovarian cancer tumors or ascites fluid from patients with ovarian cancer. Intracellular NRP1 was expressed on a median of 1.8% (interquartile range [IQR]: 0.69% to 4.8%) of Treg from healthy donors compared with a median of 66% (IQR: 28% to 90%; p Conclusions and Future Directions: Both intracellular and surface NRP1 are expressed more frequently on tumor-infiltrating Treg and Treg from ascites fluid compared to Treg from healthy donor peripheral blood. Although the number of patients currently included in this study is small, there was also evidence that intracellular NRP1 expression on Treg was higher on malignant compared with benign tissue. Additionally, Treg from ascites fluid were capable of suppressing CD8 cells ex vivo. Future studies will evaluate whether blockade of NRP1 on human Treg limits either the survival or suppressive capacity of Treg. Note: This abstract was not presented at the conference. Citation Format: Anthony R. Cillo, Tullia Bruno, Francesmary Modugno, Robert Edwards, Dario Vignali. Neuropilin-1 expression on regulatory T cells in ovarian cancer. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr A23.
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