Expansion of human T regulatory type 1 cells in the microenvironment of cyclooxygenase 2 overexpressing head and neck squamous cell carcinoma
Expansion of human T regulatory type 1 cells in the microenvironment of cyclooxygenase 2 overexpressing head and neck squamous cell carcinoma
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DOI：
10.1158/0008-5472.can-07-0767
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发表时间：
2007-09-15
期刊：
影响因子：
11.2
通讯作者：
Whiteside, Theresa L.
中科院分区：
文献类型：
--
作者：
Bergmann, Christoph;Strauss, Laura;Whiteside, Theresa L.
头颈部鳞状细胞癌（HNSCC）中环氧合酶2（考克斯-2）的过表达和前列腺素E-2（PGE（2））的产生诱导I型调节性T（TrI）细胞，并通过产生耐受性环境而促进癌的发生。为了验证这一假设，从10名正常供体的外周血中获得的CD 4（+）CD 25（-）T细胞与自体树突状细胞、辐照的HNSCC细胞和细胞因子白细胞介素2（IL-2）、IL-10和IL-15共培养。HNSCC细胞是考克斯-2阴性的、组成型表达考克斯-2的、用考克斯-2转染的、或具有被小干扰RNA敲低的考克斯-2表达。其他修改包括共培养加或减COX抑制剂，双氯芬酸，或合成的PGE 2在没有HNSCC。通过流式细胞术对在10天共培养物中增殖的淋巴细胞进行表型分析，通过ELISA研究细胞因子产生，并通过USE抑制试验加或减抗IL-10或抗转化生长因子-β（1）（TGF-β（1））单克隆抗体（mAb）研究抑制功能。考克斯-2（+）HNSCC或外源性PGE 2诱导具有CD 3（+）CD 4（+）CD 25（-）IL 2 R β（+）IL 2 R γ（+）FoxP 3（+）CTLA-4（+）IL 10（+）TGF-β +1 IL-4（-）表型和高抑制功能（范围，46-68%）的TrI细胞生长。HNSCC中考克斯-2基因的小干扰RNA敲除导致淋巴细胞生长，其具有降低的IL 2 R γ（P = 0.0001）、FoxP 3（P = 0.05）和IL-10（P = 0.035）表达和低抑制活性（范围，26-34%）。而考克斯-2（+）共培养物含有IL-10和TGF-β（I）（中位数，615和824 pg/mL），考克斯-2（-）共培养物中细胞因子水平降低（P < 0.0001）。抑制HNSCC中的考克斯-2酶活性可消除TrI细胞的生长。中和IL-10和/或TGF-β 1的mAb可消除Tr 1介导的抑制。HNSCC中考克斯-2过表达在肿瘤微环境中TrI细胞的诱导中起主要作用。
Cyclooxygenase 2 (COX-2) overexpression and production of prostaglandin E-2 (PGE(2)) by head and neck squamous cell carcinomas (HNSCC) induce type I regulatory T (TrI) cells and contribute to carcinogenesis by creating a tolerogenic milieu. To test this hypothesis, CD4(+)CD25(-) T cells obtained from the peripheral blood of 10 normal donors were cocultured with autologous dendritic cells, irradiated HNSCC cells and cytokines, interleukin 2 (IL-2), IL-10, and IL-15. HNSCC cells were either COX-2 negative, constitutively expressed COX-2, were transfected with COX-2, or had COX-2 expression knocked down by small interfering RNA. Other modifications included coculture plus or minus the COX-inhibitor, Diclofenac, or synthetic PGE2 in the absence of HNSCC. Lymphocytes proliferating in 10-day cocultures were phenotyped by flow cytometry, studied for cytokine production by ELISA and for suppressor function in USE inhibition assays plus or minus anti-IL-10 or anti-transforming growth factor-beta(1) (TGF-beta(1)) monoclonal antibodies (mAb). COX-2(+) HNSCC or exogenous PGE2 induced outgrowth of TrI cells with the CD3(+)CD4(+)CD25(-)IL2R beta (+)IL2R gamma(+)FoxP3(+)CTLA-4(+)IL10(+)TGF-beta+1IL-4(-) phenotype and high suppressor functions (range, 46-68%). Small interfering RNA knockout of COX-2 gene in HNSCC led to outgrowth of lymphocytes with decreased IL2R gamma (P = 0.0001), FoxP3 (P = 0.05), and IL-10 (P = 0.035) expression and low suppressor activity (range, 26-34%). Whereas COX-2(+) cocultures contained IL-10 and TGF-beta(I), (medians, 615 and 824 pg/mL), cytokine levels were decreased (P < 0.0001) in COX-2(-) cocultures. Inhibition of COX-2 enzymatic activity in HNSCC abrogated outgrowth of TrI cells. Neutralizing mAbs to IL-10 and/or TGF-beta(1) abolished Tr1-mediated suppression. COX-2 overexpression in HNSCC plays a major role in the induction of TrI cells in the tumor microenvironment.