雷公藤内酯醇对系统性红斑狼疮患者树突状细胞Toll样受体-9表达的影响

目的:检测雷公藤内酯醇对体外培养的系统性红斑狼疮(SLE)患者树突状细胞(DC)Toll样受体-9(TLR9)表达的影响。方法:体外培养SLE患者DC,用终浓度分别为20、40、60μg/L的雷公藤内酯醇刺激细胞3、6、12h,混合淋巴细胞反应观察DC刺激T细胞后T细胞增殖的变化,RT-PCR检测TLR9 mRNA在DC中的表达情况。结果:与健康对照者相比,SLE患者DC表达CD11c+、CD123+的百分率明显下降(P<0.05);雷公藤内酯醇以剂量依赖的形式抑制SLE患者DC刺激同种T细胞增殖;以剂量和时间依赖的形式下调SLE患者DCTLR9 mRNA表达。结论:雷公藤内酯醇可抑制SLE患者DC刺激同种T细胞增殖,雷公藤内酯醇可能通过下调SLE患者DCTLR9的表达而发挥免疫抑制作用。     

bcl-x和bcl-2基因在急性白血病患者中的表达及其临床意义

目的　观察凋亡调控基因bcl x和bcl 2在急性白血病 (AL)患者中的表达 ,探索AL的病理和化疗反应的分子机制。方法　应用半定量逆转录 聚合酶链反应 (RT PCR)技术检测bcl x(bcl xL、bcl xS)和bcl 2在 34例AL患者中mRNA水平的表达。结果　抑凋亡基因bcl xL 和bcl 2在AL细胞中的表达比在完全缓解和正常对照者骨髓细胞中的表达明显增高 (P <0 0 1) ,同时在复发患者AL细胞中的表达水平分别是初治患者的 1 8倍和 1 9倍 (P <0 0 1)。bcl xL 和bcl 2的表达以及bcl xL 和bcl xS 的表达均呈正相关 (P <0 0 1) ,但个体间有较大差异。未发现bcl xS 表达水平与AL的复发和疗效有关 ;治疗无效患者bcl xL 和bcl 2的表达比治疗有效者高 (P <0 0 1) ,bcl xL、bcl 2和此两基因同时高表达的患者临床治疗无效率分别为 80 0 %、91 7%和10 0 0 %。结论　AL的发病可能与抑凋亡基因bcl xL 和bcl 2的高表达有关 ;bcl xL 或bcl 2的高表达可以降低AL的化疗敏感性 ,而且是AL复发的高危因素     

Nonleukemic myeloid dendritic cells obtained from autologous stem cell products elicit antileukemia responses in patients with acute myeloid leukemia

BACKGROUND: Dendritic cell (DC)‐based immunotherapeutic protocols are being developed to treat acute myeloid leukemia (AML). So far, DCs for clinical use are obtained from leukemic blasts or from monocytes, after 6 to 10 days of ex vivo culture. However, DC precursors are easily driven to DCs in short‐term culture. We tested if DC precursors contained in peripheral blood stem cell (PBSC) products obtained from AML patients can be used to induce antileukemia responses. STUDY DESIGN AND METHODS: PBSCs obtained from 30 consecutive AML patients were tested. Myeloid DCs (MDCs) were purified by immunomagnetic selection and screened for cytogenetic and/or molecular abnormalities by fluorescence in situ hybridization (FISH) or polymerase chain reaction (PCR) assays. MDCs were matured and pulsed with autologous blast lysates and tested for stimulatory capability against AML cells. RESULTS: A median of 0.62 × 10⁶ MDCs (range, 0.04‐3.25)/mL were quantified in PBSC products. Isolated MDC expressed Class I and II HLA but CD86, CD54, and CCR5 partially. By FISH or PCR assay, these MDCs lacked cytogenetic or molecular abnormalities detected in leukemia cells at diagnosis. MDCs achieved a maturated stage (mature‐MDCs) after 24‐hour ex vivo culture with tumor necrosis factor‐α and autologous blast lysates. These mature‐MDCs were capable of stimulating autologous peripheral blood effectors to exert cytotoxicity against autologous leukemia cells and HL‐60 cell line. CONCLUSION: We conclude that PBSCs obtained for autologous stem cell transplantation can constitute a novel source of MDCs to design feasible vaccination trials.     

Large-scale generation of autologous dendritic cells for immunotherapy in patients with acute myeloid leukemia

BACKGROUND: Mononuclear cells (MNCs) of severely impaired acute myeloid leukemia (AML) patients may be collected by leukapheresis for large-scale generation of dendritic cells (AML-DCs) under good manufacturing practice (GMP) conditions for adoptive immunotherapy. STUDY DESIGN AND METHODS: In five end-stage AML patients, a leukapheresis procedure was performed with a cell separator (either COBE Spectra [Gambro BCT] or Amicus [Baxter]). For large-scale AML-DC generation, the MNCs of a single leukapheresis concentrate were isolated by density gradient and plated into a cell factory under GMP conditions. The AML-DCs were harvested on Day 8 of culture, and their viability, the mature morphology, and the phenotype were evaluated. The AML-DCs were injected subcutaneously into five AML patients up to four times at a biweekly interval. RESULTS: All AML patients entered the leukapheresis procedure with a highly pathologic blood count. In a mean separation time of 198 +/- 33 minutes, a mean of 1.3 +/- 0.2-fold the total blood volume was processed with a white blood cell (WBC) yield of 9 x 10(9) to 70 x 10(9) per collection dependent on the precollection WBC count. After density gradient a mean of 2.2 x 10(9) +/- 0.3 x 10(9) MNCs were plated into a cell factory. This resulted in a mean viable and mature DC yield of 0.01 x 10(9) of MNCs. CONCLUSION: The leukapheresis procedure is a feasible and safe procedure even in patients with hematologic malignancies and highly pathologic blood counts. Sufficient amounts of MNCs can be collected in leukopenic patients and the large-scale generation of AML-DCs in cell factories under GMP conditions yields in an adequate quantity of viable and mature AML-DCs.     

bcl—x和bcl—2基因在急性白血病患者的表达及其临床意义

观察凋亡调控基因ｂｃｌ－ｘ和ｂｃｌ－２在急性白血病患中的表达,探索ＡＬ的病理和化疗反应的分子机制。方法应用半定量逆转录－聚合酶链反应技术检测ｂｃｌ－ｘ和ｂｃｌ－２在３４例ＡＬ患中ｍＲＮＡ水平的表达。结果抑凋亡基因ｂｃｌ－ｘＬ和ｂｃｌ－２在ＡＬ细胞中的表达比在完全缓解和正常对照组骨髓细胞中的表达明显增高（Ｐ〈０．０１）,同时在复发患ＡＬ细胞中的表达水平分别是初治患的１．８倍和１．９倍（Ｐ〈０     

De novo acute myeloid leukemia with monocytoid blasts and erythrophagocytosis

Acute myeloid leukemia (AML) with t(8:16) is an infrequent acute leukemia subtype. It can occur de novo or more frequently therapy‐related. The presence of blasts with monocytoid morphology and erythrophagocytosis suggest the presence of the t(8;16).     

急性髓系白血病患者外周血单个核细胞Toll样受体9以及血清肿瘤坏死因子和凋亡基因FAS的表达

背景：Toll样受体9（TLR9）、肿瘤坏死因子a、Fas可能共同参与白血病的发生发展过程。目的：测定TLR9在急性髓系白血病患者外周血单个核细胞中的表达及血清肿瘤坏死因子α、Fas水平。方法：从急性髓系白血病患者与正常对照组中分离出外周血单个核细胞,采用反转录-聚合酶链反应法检测外周血单个核细胞中TLR9mRNA的表达水平,采用酶联免疫吸附试验法检测血清肿瘤坏死因子α、Fas水平。结果与结论：在急性髓系白血病患者初治组和难治复发组中,外周血单个核细胞中TLR9mRNA表达高于正常对照组（P〈0.01）,化疗后完全缓解组与正常对照组差异无显著性意义（P〉0.05）;各病例组血清肿瘤坏死因子α、Fas水平显著高于正常对照组（P〈0.01）。TLR9mRNA的表达与血清肿瘤坏死因子α、Fas水平均呈正相关。     

急性髓系白血病患者外周血单个核细胞Toll样受体9以及血清肿瘤坏死因子和凋亡基因FAS的表达

背景:Toll样受体9(TLR9)、肿瘤坏死因子a、Fas可能共同参与白血病的发生发展过程.目的:测定TLR9在急性髓系白血病患者外周血单个核细胞中的表达及血清肿瘤坏死因子α、Fas水平.方法:从急性髓系白血病患者与正常对照组中分离出外周血单个核细胞,采用反转录-聚合酶链反应法检测外周血单个核细胞中TLR9 mRNA的表达水平,采用酶联免疫吸附试验法检测血清肿瘤坏死因子α、Fas水平.结果与结论:在急性髓系白血病患者初治组和难治复发组中,外周血单个核细胞中TLR9 mRNA表达高于正常对照组(P < 0.01),化疗后完全缓解组与正常对照组差异无显著性意义(P > 0.05);各病例组血清肿瘤坏死因子α、Fas水平显著高于正常对照组(P < 0.01).TLR9 mRNA的表达与血清肿瘤坏死因子α、Fas水平均呈正相关.     

Increased expression of toll-like receptor-2 and -4 on leukocytes from patients with sepsis

The reduced responsiveness of monocytes or granulocytes toward endotoxin (endotoxin tolerance) during sepsis may depend on Toll-like receptors (TLR). The expression of TLR-2 and TLR-4 was measured on neutrophils (PMN) and monocytes from patients with sepsis (n = 21) or healthy controls (n = 12). Leukocytes (1 x 10/mL) were incubated at 37 degrees C with or without a TLR-4 (LPS 1 microg/mL) or a TLR-2 ligand (MALP-2 2 nM). Surface expression of TLR-2 and TLR-4 at 0, 4, and 16 h was determined in FACS after staining with specific antibodies. The release of IL-8 and TNF-alpha was measured by ELISA. Freshly isolated PMN from patients with sepsis exhibited significantly (P < 0.05) higher mean fluorescence for TLR-2 (78.0 +/- 18.6) and TLR-4 (11.4 +/- 2.3) than controls (12.8 +/- 2.2 and 2.3 +/- 0.4). Similarly, monocytes from patients exhibited higher TLR-2 and TLR-4 expression (300.8 +/- 40.6 and 92.7 +/- 12.1) than cells from controls (149.5 +/- 27.1 and 52.2 +/- 7.6). In patients with sepsis, expression of TLR-2 and TLR-4 on PMN increased during 16 h of incubation (106.2 +/- 22.1 and 34.5 +/- 5.3), whereas it remained unchanged in controls (19.3 +/- 6.1 and 5.4 +/- 1.9). Incubation with LPS or MALP-2 had no effect on TLR-4 or TLR-2 expression in cells from either controls or patients. Despite increased TLR expression in cells from patients with sepsis, the endotoxin-induced release of TNF-alpha and IL-8 was indistinguishable from that in controls. Therefore, the endotoxin tolerance seen in patients with sepsis does not depend solely on TLR-2 or TLR-4 expression, and other mechanisms must be involved.     

急性髓细胞性白血病患者外周血单个核细胞CD28 mRNA的表达

背景：大量研究显示,肿瘤患者外周血T细胞表面共刺激分子CD28蛋白表达存在差异,提示共刺激通路异常可能与恶性肿瘤的发生进展有关。目的：观察急性髓细胞性白血病外周血单个核细胞共刺激信号分子CD28 mRNA在中的表达。方法：急性髓细胞性白血病患者80例,其中M0型7例,M1型6例,M2型18例,M3型15例,M4型17例,M5型9例,M6型8例。并根据急性白血病疗效标准将80例患者分为完全治愈组、缓解组、未缓解组。采用Taqman探针实时荧光定量PCR检测80例患者及76名健康人群外周血单个核细胞CD28 mRNA的表达。结果与结论：急性髓细胞性白血病外周血单个核细胞M1,M3和M4亚型中的CD28 mRNA表达量低于健康人群（P〈0.05）;急性髓细胞性白血病未缓解组中CD28 mRNA低于健康人群（P〈0.05）,完全治愈组和缓解组中CD28 mRNA表达与健康人群差异无显著性意义。说明急性髓细胞白血病患者外周血单个核细胞存在CD28 mRNA表达缺陷,并与临床分期、病情进展及预后有关。     

急性髓细胞性白血病患者外周血单个核细胞CD28mRNA的表达

背景:大量研究显示,肿瘤患者外周血T细胞表面共刺激分子CD28蛋白表达存在差异,提示共刺激通路异常可能与恶性肿瘤的发生进展有关.目的:观察急性髓细胞性白血病外周血单个核细胞共刺激信号分子CD28 mRNA在中的表达.方法:急性髓细胞性白血病患者80例,其中M0型7例,M1型6例,M2型18例,M3型15例,M4型17例,M5型9例,M6型8例.并根据急性白血病疗效标准将80例患者分为完全治愈组、缓解组、未缓解组.采用Taqman探针实时荧光定量PCR检测80例患者及76名健康人群外周血单个核细胞CD28 mRNA的表达.结果与结论:急性髓细胞性白血病外周血单个核细胞M1,M3和M4亚型中的CD28 mRNA表达量低于健康人群 (P < 0.05);急性髓细胞性白血病未缓解组中CD28 mRNA低于健康人群 (P < 0.05),完全治愈组和缓解组中CD28 mRNA表达与健康人群差异无显著性意义.说明急性髓细胞白血病患者外周血单个核细胞存在CD28 mRNA表达缺陷,并与临床分期、病情进展及预后有关.     

组合性细胞因子与钙离子载体诱导急性髓系白血病细胞向树突状细胞分化中的作用

目的：探讨钙离子载体能否诱导急性髓系白血病细胞分化为树突状细胞及其抗肿瘤免疫。方法：选择2004-01／09解放军广州军区广州总医院血液科住院的初诊或复发的急性髓系白血病患者7例。分离急性髓系白血病患者的急性髓系白血病细胞，在体外给予钙离子载体100μg/L培养3-5d（钙离子载体组），或1000U／mL重组人粒细胞-巨噬细胞集落刺激因子、1000U／mL白细胞介素4和500U／mL肿瘤坏死因子α培养7-10d（组合细胞因子组）。通过细胞形态的观察及细胞免疫表型的检测鉴定树突状细胞，用同种异体混合淋巴细胞实验及细胞毒实验检测树突状细胞的功能。结果：①形态学观察和免疫表型变化：钙离子载体组及组合细胞因子组诱导急性髓系白血病细胞均可获得具有典型树突状细胞形态及免疫表型的成熟树突状细胞，但钙离子载体组诱导的树突状细胞表面CD80，CD86，CD83，CD40，CD54等分子的表达较组合细胞因子组明显增高。②混合淋巴细胞反应和胞毒性T淋巴细胞活性：钙离子载体组所诱导的急性髓系白血病细胞刺激同种异体混合淋巴细胞反应的能力及激发细胞毒性T淋巴细胞的活力比组合细胞因子组强。结论：钙离子载体能高效诱导急性髓系白血病细胞分化成树突状细胞，比组合细胞因子诱导分化的树突状细胞功能更强。     

组合性细胞因子与钙离子载体诱导急性髓系白血病细胞向树突状细胞分化中的作用

目的:探讨钙离子载体能否诱导急性髓系白血病细胞分化为树突状细胞及其抗肿瘤免疫。方法:选择2004-01/09解放军广州军区广州总医院血液科住院的初诊或复发的急性髓系白血病患者7例。分离急性髓系白血病患者的急性髓系白血病细胞,在体外给予钙离子载体100μg/L培养3～5d(钙离子载体组),或1000U/mL重组人粒细胞-巨噬细胞集落刺激因子、1000U/mL白细胞介素4和500U/mL肿瘤坏死因子α培养7～10d(组合细胞因子组)。通过细胞形态的观察及细胞免疫表型的检测鉴定树突状细胞,用同种异体混合淋巴细胞实验及细胞毒实验检测树突状细胞的功能。结果:①形态学观察和免疫表型变化:钙离子载体组及组合细胞因子组诱导急性髓系白血病细胞均可获得具有典型树突状细胞形态及免疫表型的成熟树突状细胞,但钙离子载体组诱导的树突状细胞表面CD80,CD86,CD83,CD40,CD54等分子的表达较组合细胞因子组明显增高。②混合淋巴细胞反应和胞毒性T淋巴细胞活性:钙离子载体组所诱导的急性髓系白血病细胞刺激同种异体混合淋巴细胞反应的能力及激发细胞毒性T淋巴细胞的活力比组合细胞因子组强。结论:钙离子载体能高效诱导急性髓系白血病细胞分化成树突状细胞,比组合细胞因子诱导分化的树突状细胞功能更强。     

急性和慢性髓系白血病患者中螺旋酶抗原基因表达的研究(英文)

本研究旨在探讨螺旋酶抗原基因(HAGE)在急性髓系白血病(AML)和慢性髓系白血病(CML)中的表达状况和临床意义。应用实时定量PCR(RQ-PCR)方法检测AML和CML患者骨髓单个核细胞中HAGE cDNA的表达。结果表明:74例AML患者中11例(14.8%)存在HAGE过表达(117.12%-9842.70%,中位434.96%);HAGE过表达患者年龄显著高于HAGE阴性者(中位年龄分别为67和45岁,p=0.001)。HAGE表达在单核细胞亚型AML(M4和M5,7/20,35.0%)中明显高于其它亚型AML(4/54,7.4%)(p=0.007)。28例核型正常的AML患者中8例(28.6%)呈HAGE过表达,而40例核型异常的AML中仅3例(7.5%)存在HAGE表达(p=0.041)。9/26例(34.6%)CML患者存在HAGE过表达,加速期和急变期患者中HAGE表达率(4/4,100%)明显高于慢性期患者(5/22,22.7%)(p=0.008)。结论:HAGE cDNA表达与AML单核细胞亚型类别相关,且与CML疾病进展相关。     

Azacitidine for the treatment of patients with acute myeloid leukemia with 20%-30% blasts and multilineage dysplasia

Azacitidine is approved in the EU for the treatment of adult patients who are not candidates for allogeneic stem cell transplantation, and who have intermediate-2 risk or high-risk myelodysplastic syndromes, according to the International Prognostic Scoring System. The approval includes the treatment of patients with acute myeloid leukemia (AML) with 20%-30% blasts and multilineage dysplasia, according to the World Health Organization (WHO) classification. This review focuses on the outcomes with azacitidine in this latter group of patients, previously classified as refractory anemia with excess of blasts in transformation, as defined by the French-American-British classification criteria. The main clinical evidence is based on the results of two large phase III clinical trials (Cancer and Leukemia Group B 9221, and AZA-001). The AZA-001 trial shows azacitidine significantly prolongs median overall survival in older patients with low marrow blasts (20%-30%) according to WHO-defined AML, and significantly improved several patient morbidity measures, compared with conventional care regimens. In addition, the review examines the results of azacitidine in combination with other treatments currently used in AML.     

Th17细胞水平的变化与急性髓系白血病发病关系的探讨

目的 研究Th17细胞在急性髓系白血病(AML)患者外周血中的表达水平,并探讨Th17细胞的改变与疾病的相关性.方法 ①用酶联免疫吸附实验(ELISA)分别检测AML初诊组、化疗完全缓解或部分缓解组及正常对照组外周血中IL-17、TGF-β1的表达水平.②用流式细胞术分别检测AML初诊组、化疗完全缓解或部分缓解组及正常对照组外周血中Th17细胞所占比例.③用半定量RT-PCR分别检测各组外周血中Th17细胞的相关转录因子IL-17 mRNA的表达水平,并逐层分析比较.结果 AML初诊组和治疗后缓解组IL-17水平、Th17细胞比例及IL-17 mRNA的表达水平分别为(10.502±1.071)ng/L、(0.935±0.140)%、0.262±0.510和(11.345±0.987)ng/L、(1.091±0.159)%和0.307±0.031,均明显低于正常对照组的(16.852±1.198)ng/L、(2.586±0.235)%、0.501±0.060(P＞0.01),AML缓解组Th17细胞比例、IL-17及IL-17 mRNA的表达水平均高于AML初诊组,差异有统计学意义(P＞0.05),AML初诊组和缓解组中TGF-β1的表达水平分别为(29.963±1.588)ng/L、(25.163±1.848)ng/L,明显高于正常对照组(13.366±1.565)ng/L(P＞0.01),且AML初诊组TGF-β1的表达水平高于AML缓解组(P＞0.05).结论 Th17细胞可能与AML的发生发展呈负相关,提示AML患者体内TGF-β1的过度表达可能抑制Th17细胞的分化发育.     

Engraftment with chronic granulocytic leukemia cells in acute myeloid leukemia

A deliberate engraftment with nonirradiated chronic granulocytic leukemia (CGL) cells was performed in a patient with acute myeloid leukemia (AML) at a time when he was resistant to cytotoxic drug chemotherapy, pancytopenic and developed an infection. The CGL engraftment was confirmed by the presence of a Ph1-positive donor clone in the recipient's bone marrow and by the pattern of colony growth of the recipient's bone-marrow cells cultured in vitro. Bone marrow engraftment in the host helped in the resolution of infection and permitted the administration of further cytotoxic drugs, as a result of which a remission of AML occurred.     

急性髓细胞白血病患者DC-CIK治疗后T细胞亚群动态变化的临床观察

目的观察急性髓细胞白血病患者经自体外周血移植联合自身白血病细胞冻融抗原负载的DC-CIK输注治疗后体内的T细胞亚群动态变化。方法选择50名初发急性髓细胞白血病患者,抽取其未缓解的骨髓制备自身白血病细胞冻融抗原,在自体外周血干细胞采集时分离部分单个核细胞培养DC-CIK,以自身白血病冻融抗原共培养激活扩增DC-CIK。患者自体外周血造血干细胞移植30～60 d输注自身白血病细胞冻融抗原负载的DC-CIK,每例每疗程回输细胞总数>7×109,同时给予IL-2(20 WU皮下注射,1次/d,连续10 d)皮下注射。动态观察患者接受DC-CIK细胞输注治疗后不同时间点体内T细胞亚群动态变化,并与既往接受自体外周血移植但未输注DC-CIK细胞治疗的52名急性髓细胞白血病患者进行比较分析。结果 50名患者DC-CIK输注后患者体内CD8+、CD16+、CD56+淋巴细胞均显著高于输注治疗前水平(P<0.01),治疗组同一随访时间段CD3+、CD4+、CD8+、CD16+、CD56+淋巴细胞均显著高于未采用DC-CIK治疗组(P<0.01)。结论对急性髓细胞白血病患者采用自体外周血移植联合自身白血病细胞冻融抗原负载的DC-CIK输注治疗,能显著提高患者肿瘤杀伤性T细胞水平,有助于清除移植后微小残留病,改善患者无病生存率(PFS)。     

急性髓系白血病患者IDH1和IDH2基因突变分析

目的 初步探讨急性髓系白血病(AML)患者IDH基因(IDHI和IDH2)突变发生率、突变类型及其与FLT3基因内部串联重复(ITD)突变、NPMl基因突变和部分l临床参数间的相关性.方法 采用基因组DNA-PCR扩增产物直接测序法检测163例初诊患者IDHI和IDH2基因4号外显子突变、NPMl基因12号外显子突变和FLT3基因14、15号外显子中ITD的突变发生情况.结果 ①163例AML患者中共检测出IDH突变25例,且均为杂合突变.其中IDHl突变7例,突变类型分别为c.395G→A(p.R132H)4例;c.394C→A(p.R132S)1例;c.394C→G(p.R132G)1例;c.315C→T 1例.除c.315C→T为同义突变外,其余均为p.R132错义突变.共检测出IDH2突变18例,均为c.419G→A(p.R140Q)错义突变.IDH2基因突变发生率高于IDHl(11.0%和4.3%,P=0.022),其中1例患者同时检测到IDHl和1DH2基因突变,但IDHl系同义突变.②IDH突变在FLT3→ITD突变阳性组发生率为34.6%,高于阴性组的11.9%(P=0.003),在NPMl突变阳性组和阴性组的发生率分别为28.1%和12.7%,差异有统计学意义(P=O.033);其中IDH在FLT3→ITD和NPMl突变双阳性组中的突变率明显高于双阴性组(45.5%和11.7%,P=0.002).正常核型患者中IDH突变发生率高于异常核型患者(20.5%和5.8%,P=0.020).IDH突变型患者的中位年龄高于野生型患者,差异有统计学意义(P<0.001);但具有IDH突变的患者在性别、初诊外周血细胞水平方面与野生型患者相比差异无统计学意义.结论 IDH基因突变在初诊AMI.患者中有较高的发生率,其中IDH2突变更为频繁;发生IDH突变者年龄偏高,且与正常核型相关;该突变可与FLT3-ITD、NPMl突变共同存在并有一定相关性,提示IDH突变在促进白血病发生中可能和后两种基因起协同作用.     

卵巢癌患者TLR1、TLR2、TLR6信号通路活化诱导PBMC前炎症细胞因子分泌

目的观察卵巢癌(OC)患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)中Toll样受体1(Toll-like receptors 1,TLR1)、TLR2及TLR6信号通路活化后前炎症细胞因子分泌水平,研究其在OC发病中的可能机制。方法纳入OC患者、女性妇科良性疾病(BC)患者及同期体检健康女性(NC)各13例,用密度梯度离心法分离PBMC并用TLRs的相应配体刺激PBMC,用CBA免疫荧光法检测细胞中前炎症细胞因子白细胞介素-1β(IL-1β)、IL-6、IL-8、肿瘤坏死因子-α(TNF-α)的分泌水平;用western blot检测MyD88、TRAF6、TANK、NF-κB和P-NF-κB的表达水平。结果 OC组、BC组、NC组未加配体刺激前,PBMC中IL-1β、IL-6、IL-8、TNF-α分泌水平差异均无统计学意义(P>0.05)。与未经配体刺激PBMC相比,OC组PBMC经TLR1配体Pam3CSK4刺激24 h后,IL-1β分泌水平增高,差异具有统计学意义(t=-2.667,P<0.05);TLR2配体HKLM刺激24 h后,OC组IL-1β、TNF-α分泌水平亦增高,差异有统计学意义(t分别为-5.391和-3.564,P均<0.05);相较于未经配体刺激时,BC组、NC组的PBMC在相应TLRs配体刺激后,IL-1β、IL-6、IL-8、TNF-α分泌水平变化无统计学意义。OC患者PBMC经Pam3CSK4、HKLM、FSL-1刺激后,TLRs信号通路蛋白MyD88、TRAF6、TANK及P-NF-κB表达上调。结论 OC患者PBMC中TLR1、TLR2、TLR6经相应配体刺激后可活化TLR信号通路,并诱导前炎症细胞因子IL-1β、TNF-α分泌水平增加,在OC的发生发展中发挥作用。