Eoxins are proinflammatory arachidonic acid metabolites produced via the 15-lipoxygenase-1 pathway in human eosinophils and mast cells

Human eosinophils contain abundant amounts of 15-lipoxygenase (LO)-1. The biological role of 15-LO-1 in humans, however, is unclear. Incubation of eosinophils with arachidonic acid led to formation of a product with a UV absorbance maximum at 282 nm and shorter retention time than leukotriene (LT)C4 in reverse-phase HPLC. Analysis with positive-ion electrospray tandem MS identified this eosinophil metabolite as 14,15-LTC4. This metabolite could be metabolized to 14,15-LTD4 and 14,15-LTE4 in eosinophils. Because eosinophils are such an abundant source of these metabolites and to avoid confusion with 5-LO-derived LTs, we suggest the names eoxin (EX)C4, -D4, and -E4 instead of 14,15-LTC4, -D4, and -E4, respectively. Cord blood-derived mast cells and surgically removed nasal polyps from allergic subjects also produced EXC4. Incubation of eosinophils with arachidonic acid favored the production of EXC4, whereas challenge with calcium ionophore led to exclusive formation of LTC4. Eosinophils produced EXC4 after challenge with the proinflammatory agents LTC4, prostaglandin D2, and IL-5, demonstrating that EXC4 can be synthesized from the endogenous pool of arachidonic acid. EXs induced increased permeability of endothelial cell monolayer in vitro, indicating that EXs can modulate and enhance vascular permeability, a hallmark of inflammation. In this model system, EXs were 100 times more potent than histamine and almost as potent as LTC4 and LTD4. Taken together, this article describes the formation of proinflammatory EXs, in particular in human eosinophils but also in human mast cells and nasal polyps.     

一个新的保护心脏TGF-β超家族成员——GDF15

GDF15基因是TGF-β超家族成员之一,具有抗肿瘤、抗器官损伤等多种功能。最新文献报道,GDF15基因抑制心肌肥厚,对缺血／缺血再灌注中的心脏损伤亦有保护作用,是一个新的心脏保护因子。本文就GDF15基因在多种心血管疾病中的功能及其相关的信号通路做一综述。     

IL-15对人γδT细胞杀伤结核杆菌感染THP-1细胞的影响

目的建立结核杆菌(Mtb)感染单核细胞THP-1细胞系模型;用流式细胞术检测人γδT细胞对Mtb感染THP-1细胞的细胞毒活性;并观察IL-15对人γδT细胞杀伤Mtb感染THP-1细胞的细胞毒活性的影响。方法 Mtb以10∶1的比例感染佛波醇酯(PMA)分化的THP-1细胞,建立Mtb感染细胞模型。人外周血单个核细胞用Mtb耐热性低分子多肽类抗原刺激优势扩增γδT细胞,作为效应细胞用于细胞毒活性实验。用羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)标记Mtb感染THP-1细胞,作为靶细胞。效应细胞和靶细胞以不同比例孵育4 h,用碘化丙啶(PI)染色后在流式细胞仪上检测,CSFE和PI双阳性细胞为被杀伤靶细胞。IL-15作用于γδT细胞24 h后,再检测γδT细胞对Mtb感染THP-1细胞的细胞毒活性。结果人γδT细胞与Mtb感染THP-1细胞以1∶1至50∶1的比例作用4 h后,人γδT细胞对结核杆菌感染的THP-1细胞的细胞毒活性从22.5%增加至80.7%;而γδT细胞与未感染Mtb的THP-1细胞的细胞毒活性为16.1%至47.2%。IL-15作用γδT细胞后的细胞毒活性(64.06%)与对照组(46.81%)相比明显增加(P0.05)。结论人γδT细胞对Mtb感染THP-1细胞的杀伤活性明显高于对未感染Mtb的THP-1细胞的作用,杀伤活性随效靶比的增加而增加。IL-15可以增强人γδT细胞对Mtb感染巨噬细胞的细胞毒活性。     

子宫内膜癌中p15及MDM2蛋白表达

目的 采用流式细胞术并结合免疫组化法检测p15蛋白和MDM2蛋白在正常子宫内膜、良、恶性子宫内膜肿物组织中表达,研究其与原发性妇科恶性肿瘤的关系及三者在原发性妇科恶性肿瘤表达的相关关系.方法 采用石蜡切片,提取各个实验组的单细胞悬液,利用流式细胞术并结合免疫组织化学链霉素抗生物素蛋白-过氧化酶结合法,分别检测子宫系列病变组织p15、MDM2表达情况.结果 从子宫肌瘤到子宫内膜癌,p15的表达量有明显进行性增高(P<0.01).而p15过度表达时(FI>1.30)5年生存率很低.两组生存率比较有显著性差异(P<0.01);MDM2的表达在子宫内膜样癌为48.28% 、非子宫内膜样癌为41.5%,都明显低于功能性子宫出血诊断性刮宫标本的60.2%(P<0.01).结论 p15、MDM2蛋白表达异常可成为子宫内膜癌危险性评估、子宫内膜癌早期诊断及预后和复发判定的新途径.     

白介素-15与支气管哮喘

白介素 15属TH1相关性细胞因子 ,与IL 2共用β和γ链 ,有许多相似生物活性。促进T细胞、B细胞、单核 /巨噬细胞等多种细胞的增殖、分化和分泌各种细胞因子 ,参与哮喘气道炎症的形成。研究IL 15在哮喘中的作用 ,有利于进一步阐述哮喘的发病机制。     

白细胞介素-15抗寄生虫感染的研究进展

白细胞介素-15(IL-15)是一种可溶性细胞因子,作为多种免疫细胞的趋化因子参与并调节机体炎症反应及免疫反应。目前,IL-15的分子结构与受体得到广泛研究,其信号转导方式也取得了较大进展,然而其在寄生虫学上的研究起步较晚。本文对近年来IL-15在抗蠕虫及原虫感染等方面的最新研究进展作一综述,并对其免疫增强作用机制和应用前景进行了展望,为进一步探索IL-15抗寄生虫免疫作用及其在寄生虫学的相关应用研究提供参考。     

Ethanol Self-Administration in Deprived Rats: Effects of Ro15-4513 Alone, and in Combination with Flumazenil (Ro15-1788)

Previous work in our laboratory demonstrated that Ro15-4513, a partial inverse benzodiazepine agonist, decreases self-administration of ethanol (ETOH) in rats maintained on a two-bottle regmine of a saccharin ethanol solution (ES) and water over a 35-day consumption period. The present study extended the consumption period to 60 days and examined the effects of Ro15-4513 (2.5 mg/kg), flumazenil (Ro15-1788) (8.0 mg/kg), and Ro15-4513 in combination with Ro15-1788 on the time course of ETOH self-administration. High initial intake of ES observed during the first 4 weeks declined significantly over subsequent weeks. Ro15-4513 pretreatment, however, resulted in significant reduction of ES, while significantly preventing the "normal" reduction of consumption as was observed under control conditions. The antagonistic actions of Ro15-4513 were blocked/attenuated by the benzodiazepine receptor antagonist, Ro15-1788, independent of whether consumption of the ES was low or high. Both Ro15-4513 and Ro15-1788 affected water intake differentially compared with vehicle-injected controls. The results suggest that GABA-benzodiazepine mechanisms may be important in altering chronic ETOH drinking patterns depending upon experience with ETOH, tolerance, or learning.     

克罗恩病相关的NOD2及β-防御素-2基因多态性对β-防御素-2表达的影响

目的探讨克罗恩病（Crohndisease,CD）相关的NOD2及人β-防御素-2（humanbeta—defensin2,hBD-2）基因多态性对hBD-2转录活性的影响及主要机理。方法将hBD-2报告基因质粒和NOD2真核表达载体共转染至HEK293T细胞,用脂多糖（LPS）和TNF—α分别孵育刺激后测定hBD-2转录活性变化。结果LPS对hBD-2转录活性有抑制作用（P=0．020）,TNF—α可相对升高hBD-2转录活性,呈剂量依赖性（P=0．004）;LPS作用时,NOD2（P268S）改变前后hBD-2的转录活性差异有统计学意义（P=0．008）;hBD-2（-233）改变前后hBD-2的转录活性差异无统计学意义（P=0．053）。在TNF-α（5ng／m1）刺激时,NOD2（P268S）改变前后hBD-2的转录活性差异无统计学意义（P=0．064）;hBD-2（-233）改变前后hBD-2的转录活性差异有统计学意义（P=0．006）;NF—κB抑制剂可显著下调hBD-2转录的激活（P〈0．001）。结论NOD2（P268S）改变能够降低hBD-2的内源性表达;hBD-2（-233）改变导致hBD-2诱生性表达的减少;NF—κB通路可能是hBD-2诱导表达的主要路径。     

15-Hydroxyprostaglandin dehydrogenase inactivation as a mechanism of resistance to celecoxib chemoprevention of colon tumors

Pharmacologic inhibitors of the prostaglandin-synthesizing COX-2 oncogene prevent the development of premalignant human colon adenomas. However, resistance to treatment is common. In this study, we show that the adenoma prevention activity of the COX-2 inhibitor celecoxib requires the concomitant presence of the 15-hydroxyprostaglandin dehydrogenase (15-PGDH) tumor suppressor gene, and that loss of 15-PGDH expression imparts resistance to celecoxib''s anti-tumor effects. We first demonstrate that the adenoma-preventive activity of celecoxib is abrogated in mice genetically lacking 15-PGDH. In FVB mice, celecoxib prevents 85% of azoxymethane-induced tumors >1 mm in size, but is essentially inactive in preventing tumor induction in 15-PGDH-null animals. Indeed, celecoxib treated 15-PGDH null animals develop more tumors than do celecoxib naive WT mice. In parallel with the loss of tumor prevention activity, celecoxib-mediated suppression of colonic PGE₂ levels is also markedly attenuated in 15-PGDH-null versus WT mice. Finally, as predicted by the murine models, humans with low colonic 15-PGDH levels also exhibit celecoxib resistance. Specifically, in a colon adenoma prevention trial, in all cases tested, individuals who developed new adenomas while receiving celecoxib treatment were also found as having low colonic 15-PGDH levels.     

p15蛋白表达及其基因甲基化对急性白血病预后的影响

观察53例急性白血病患者(观察组,其中ALL22例,AML31例;完全缓解者16例,初治者23例,复发者14例)。骨髓单个核细胞p15蛋白表达情况及p15基因甲基化情况,并与5例缺铁性贫血患者进行比较,随访治疗效果及其预后。p15蛋白阳性表达率为ALL 31.8%,AML 29.0%;p15基因甲基化阳性率为ALL59.1%,AML 58.1%,明显高于对照组(均呈阴性),P均<0.01。阳性率完全缓解者>初发者>复发者;p15甲基化完全缓解者<初发者<复发者。相关分析显示p15蛋白与p15基因甲基化呈明显负相关,二者与疗效均有相关性。认为p15基因甲基化与p15蛋白失活关系密切;p15蛋白表达及p15基因甲基化情况可预测急性白血病的疗效及预后。     

TGF-β1对乙肝疫苗免疫应答的影响及与HLA-DRB1*15基因的关系

目的探讨TGF-β1对乙肝疫苗免疫应答的影响以及与HLA-DRB1*15基因的关系。方法选取完成重组乙型肝炎疫苗标准全程接种的健康大学生中抗-HBsS/N值10mIU/ml者77名为无、弱应答组,选取抗-HBsS/N值10mIU/ml者101名为中、强应答组,应用PCR-SSP进行HLA-DRB1*15等位基因的检测,采用ELISA法检测血清中TGF-β1水平。结果①无、弱应答组的HLA-DRB1*15基因表达频率为14.29%(11/77),低于中、强应答组的28.71%(29/101)(P=0.022);②无、弱应答组中TGF-β1的平均表达水平为(24.04±14.74)ng/ml,高于中、强应答组(17.80±14.42)ng/ml;差异有统计学意义(P=0.005);③HLA-DRB1*15阳性组TGF-β1平均表达水平为(17.81±17.38)ng/ml,阴性组为(21.28±14.00)ng/ml,差异无统计学意义(P=0.194)。结论①HLA-DRB1*15基因可能是促进人群乙肝疫苗免疫应答的相关基因;②TGF-β1的表达水平可能影响机体乙肝疫苗的免疫效果;③HLA-DRB1*15基因可能不是通过影响TGF-β1的表达水平来影响乙肝疫苗的免疫应答。     

微小隐孢子虫重组BCG疫苗免疫保护作用观察

目的研究微小隐孢子虫重组BCG疫苗的免疫保护作用。方法 100只BALB/c小鼠随机分为4组,BCG组免疫BCG,载体组免疫含PMV262质粒的BCG(BCG-PMV262),实验组免疫rBCG-CP15/60-23(各组均为每鼠尾静脉注射0.1mL,接种物浓度均为106细菌/mL),对照组每鼠尾静脉注射0.05mol/LpH7.2PBS0.1mL。免疫后,每组分别随机选6只小鼠检测其血清抗体和脾细胞培养液中的CD4+、CD8+以及细胞因子水平,各组剩余小鼠进行攻击实验。结果 rBCG-CP15/60-23能诱发小鼠产生细胞免疫和体液免疫反应,小鼠血清抗体水平于免疫后2周逐渐升高,与BCG组和载体组差异显著(P均<0.01);实验组CD4+T细胞增殖明显,CD4+/CD8+比值显著升高(P<0.05);脾细胞培养液中白细胞介素2(IL-2)、IL-12、γ干扰素(IFN-γ)均有不同程度的升高,实验组IFN-γ和IL-2含量与对照组和BCG组差异显著(P均<0.05),IL-12水平组间差异无统计学意义(P>0.05)。各组小鼠经隐孢子虫卵囊攻击感染后,实验组小鼠卵囊排泄数量减少40%～60%,初始排卵囊时间延长2d,卵囊持续排出时间缩短5d;组间差异有统计学意义(P<0.05)。结论微小隐孢子虫重组BCG-CP15/60-23疫苗免疫小鼠后,能产生良好的免疫保护作用。     

15-脂氧合酶在肿瘤中的作用

15-脂氧合酶(15-LOX)是催化不饱和脂肪酸代谢的关键酶之一.人类中存有两型15-LOX:15-LOX-1和15-LOX-2,两者在催化性质、组织分布及肿瘤生物学功能上有所差异.近年来大量研究发现15-LOX在结肠癌、前列腺癌、食道癌等恶性肿瘤表达较正常组织降低,能够抑制肿瘤生长,并使其发生凋亡,在肿瘤发生发展中起着重要作用.以15-LOX为靶向的肿瘤防治及其临床应用具有广阔的前景.     

原发性肝癌中p15、p16基因缺失和STK15基因过表达的研究

目的 探讨肿瘤组织p15、p16基因缺失和STK15基因过表达与原发性肝癌的关系。 方法 术中取原发性肝癌组织和相应癌旁正常组织标本30例,术前均未经化学疗法及放射疗法治疗。提取DNA并应用聚合酶链反应(PCR)技术检测p15第二外显子(p15E2)和p16第二外显子(p16E2)纯台缺失。提取RNA,逆转录合成cDNA,应用PCR技术检测STK15基因的表达。以β-肌动蛋白基因作为内对照。测定癌组织和癌旁正常组织中STK15基因和β-肌动蛋白基因的平均密度值(ADV)。 结果 在癌组织中p15E2缺失率为13.3％(4／30),p16E2缺失率为16.7％(5／30),p15E2和p16E2共缺失率为6.7％(2／30)。在30例癌组织中有19例(63.3％)STK15基因表达高于邻近正常组织。STK15基因ADV／β-肌动蛋白基因ADV比值,癌组织为1.53±0.31,癌旁正常组织为0.91±0.25,t=2.86,P<0.01。 结论 p15E2、p16E2纯合缺失和STK15过表达可能在原发性肝癌的发生发展中发挥一定作用。     

15-脂氧合酶-1在胃癌中的表达及其临床意义

目的检测15-脂氧合酶-1（15-LOX-1）mRNA及蛋白在胃癌及癌旁正常组织中的表达，并探讨15-LOX-1表达与胃癌临床病理因素的关系。方法采用RT-PCR、westernblot和免疫组织化学方法检测胃癌组织及相匹配的癌旁正常组织中15-LOX-1 mRNA和蛋白的表达。结果15-LOX-1 mRNA及蛋白在胃癌组织中的表达均显著低于癌旁正常组织（P＜0．05）。15-LOX-1蛋白表达水平与胃癌中肿瘤大小、淋巴结转移和TNM分期呈明显的负相关（P＜0．05）。结论15-LOX-1蛋白可能对胃癌的发生发展有一定抑制作用。检测胃癌中15-LOX-1的表达情况对评估患者的预后可能具有意义。     

The relevance of HPA-15 antigen expression for anti-HPA-15 antibody detection

Introduction: The HPA‐15 antigen system is characterized by a low antigen expression on platelets. The antibodies against this antigen are implied in fetal/neonatal alloimmune thrombocytopenia (F/NAIT), post‐transfusion purpura, and refractoriness to platelet transfusions. Detection of these antibodies appears to be related to the level of HPA‐15 expression on the platelets used in the monoclonal antibody–specific immobilization of platelet antigen (MAIPA) assay. Methods: We performed genotyping of 300 healthy blood donors for HPA‐15 by TaqMan real‐time PCR technology, and the HPA‐15 antigen expression was investigated in 13 HPA‐15aa and 19 HPA‐15bb individuals. We also investigated the relevance of HPA‐15 antigen expression on donor platelets used in MAIPA for antibody detection in 223 multitransfused hematological patients and 271 women with suspected F/NAIT. Results: In Polish donors, the HPA‐15a allele frequencies were lower than the HPA‐15b (0.480 vs. 0.515). We identified three HPA‐15 expression groups: high (36.7 ± 8.36 MFI – eight cases), medium (19.5 ± 6.2 MFI – 21 cases), and low (6.5 ± 5.9 MFI – three cases). The HPA‐15 expression was stable over time. The HPA‐15aa and HPA‐15bb platelets with high antigen expression were used for anti‐HPA‐15 antibody detection; anti‐HPA‐15 antibodies were detected in 4/223 (1.8%) patients receiving multiple transfusions but in none of the 271 women with suspected F/NAIT. Further examination of the four sera by MAIPA with various platelets revealed the optical density in the assay to be closely related to the level of HPA‐15 antigen expression. Conclusion: Anti‐HPA‐15 antibody detection should be based on carefully selected platelets with high HPA‐15 expression level.     

Development of a highly efficacious vaccinia-based dual vaccine against smallpox and anthrax, two important bioterror entities

Bioterrorism poses a daunting challenge to global security and public health in the 21st century. Variola major virus, the etiological agent of smallpox, and Bacillus anthracis, the bacterial pathogen responsible for anthrax, remain at the apex of potential pathogens that could be used in a bioterror attack to inflict mass casualties. Although licensed vaccines are available for both smallpox and anthrax, because of inadequacies associated with each of these vaccines, serious concerns remain as to the deployability of these vaccines, especially in the aftermath of a bioterror attack involving these pathogens. We have developed a single vaccine (Wyeth/IL-15/PA) using the licensed Wyeth smallpox vaccine strain that is efficacious against both smallpox and anthrax due to the integration of immune-enhancing cytokine IL-15 and the protective antigen (PA) of B. anthracis into the Wyeth vaccinia virus. Integration of IL-15 renders Wyeth vaccinia avirulent in immunodeficient mice and enhances anti-vaccinia immune responses. Wyeth/IL-15/PA conferred sterile protection against a lethal challenge of B. anthracis Ames strain spores in rabbits. A single dose of Wyeth/IL-15/PA protected 33% of the vaccinated A/J mice against a lethal spore challenge 72 h later whereas a single dose of licensed anthrax vaccine protected only 10%. Our dual vaccine Wyeth/IL-15/PA remedies the inadequacies associated with the licensed vaccines, and the inherent ability of Wyeth vaccinia virus to be lyophilized without loss of potency makes it cold-chain independent, thus simplifying the logistics of storage, stockpiling, and field delivery in the event of a bioterror attack involving smallpox or anthrax.     

抑癌基因P15在胃癌中的表达及其临床意义

采用ＳＰ免疫组化法测定了８０例胃癌及２６例胃良性病变患者胃组织石蜡标本的Ｐ＾１５基因蛋白表达情况,结合临床病理指标（肿瘤大小、生长方式、组织分化、浸润深度、转移及ＰＴＮＭ分期）进行分析。结果在胃癌组织中Ｐ＾１５阳性表达率为４３．８％,胃良性病变为６９．２％;Ｐ＾１５表达与肿瘤大小、生长方式、组织分化、浸润深度,转移及ＰＴＮＭ分期无关。认为Ｐ＾１５在胃癌发生中起重要调控作用,与临床病理因素无关。     

急性白血病P~(15)基因异常及甲基化研究

用聚合酶链反应 (PCR)方法扩增 P1 5基因外显子 1,再用限制性内切酶 - PCR方法检测 P1 5基因甲基化。结果 48例患者中 P1 5基因失活者共 2 7例 (5 8.92 % ) ,AL L2 1例中有 12例 (5 7.41% ) ,ANL L2 7例中 15例(5 5 .5 6 % )。 P1 5 基因以甲基化失活为主。 P1 5 Cp G岛甲基化在各型急性白血病中均有很高的发生率 ,可以作为各型急性白血病通用的微量残留病 (MRD)指标 ;白血病缓解期 P1 5 Cp G岛甲基化仍为阳性可能预示着复发。认为 P1 5基因失活的检测对于探讨急性白血病的发病机制、判断预后有重要意义