白细胞介素1受体Ⅱ型在子宫内膜异位症发生机制中的作用

目的:研究白细胞介素1受体Ⅱ型(IL-1RⅡ)在子宫内膜异位症(EMs)发生机制中的作用.方法:体外培养子宫内膜细胞经过可溶性IL-1RⅡ或IL-1RⅡ重组腺病毒(rAd-RⅡ)作用12 h后收集细胞培养液,酶联免疫吸附试验(ELISA)检测细胞培养液中IL-6和IL-8的水平.另外应用二维电泳和质谱分析鉴定rad-RⅡ作用后的差异表达蛋白.结果:2.μg/mL的可溶性IL-1RⅡ显著抑制了内膜细胞IL-6和IL-8的分泌.rAd-RⅡ感染内膜细胞后IL-1β刺激IL-8的分泌比磷酸盐缓冲液(PBS)和LacZ重组腺病毒(rAd-LacZ)对照组显著降低,而IL-6表达无显著差异.二维电泳分析有62个差异蛋白点,质谱分析鉴定出34个差异蛋白,大多数蛋白和细胞代谢与增殖相关.结论:IL-1RⅡ能阻断IL-1β对内膜基质细胞的作用,可能为EMs治疗提供新策略.     

子宫内膜异位症患者在位内膜细胞凋亡特性

目的:探讨子宫内膜异位症(EMs)在位内膜细胞凋亡特性的改变。方法:①应用免疫组化方法对32例EMs在位子宫内膜组织及24例对照组子宫内膜组织的Survivin蛋白及Ki-67蛋白的表达进行测定;②应用TUNEL技术监测两组子宫内膜细胞的凋亡特性。结果:EMs在位内膜组织Survivin蛋白表达高于对照组,差异有统计学意义(P<0.05);EMs在位内膜细胞分泌期凋亡指数低于同期对照组子宫内膜,差异有统计学意义(P<0.05);EMs在位内膜组织Ki-67蛋白表达与对照组差异无统计学意义(P>0.05)。结论:EMs在位子宫内膜细胞因凋亡能力下降并非高度增殖导致细胞活性升高,在位内膜细胞的生物学特性改变在EMs的病理生理中可能起着重要作用。     

血浆miR-451水平与子宫内膜异位症的关系

目的：探讨血浆微小RNA451（miR-451）与子宫内膜异位症（EMs）的关系,并观察miR-451对体外培养的子宫内膜基质细胞的增殖和迁移的作用。方法：选取2014年1月—2015年5月于深圳市罗湖区人民医院就诊的EMs患者（EMs组）54例,正常对照组59例,采血并提取血浆总RNA。采用实时荧光定量聚合酶链反应（real-time qPCR）检测血浆miR-451的表达强度。通过CCK-8实验检测miR-451对异位子宫内膜基质细胞增殖的调控作用;采用损伤愈合实验探讨过表达miR-451后异位子宫内膜基质细胞的迁移变化。结果：EMs患者血浆miR-451表达水平低于对照组[（0.53±0.14） vs. （1.00±0.20）,t=14.50,P＜0.01];过表达miR-451后EMs基质细胞的相对迁移活性下降[（1.00±0.12） vs. （0.74±0.02）,t=4.73,P=0.001]。结论：EMs患者血浆miR-451水平减低;miR-451可能通过抑制异位内膜基质细胞增殖和迁移发挥作用。     

ER、PR和AR在子宫内膜异位症组织中的表达及其意义

目的：研究ER、PR及AR在内异症异位内膜、在位内膜和正常对照子宫内膜组织中的表达,阐述性激素及其受体与子宫内膜异位症发生机制的关系,探究治疗EMs的新途径。方法：利用免疫组织化学的方法检测50例EMs患者异位内膜、在位内膜及20例正常对照子宫内膜组织中ER、PR及AR的表达情况。结果：ER在EMs患者在位子宫内膜组织中的表达均明显高于异位内膜及正常对照子宫内膜,比较差异均有统计学意义（P〈0.05）;PR在EMs患者异位内膜组织中的表达明显低于在位内膜及正常对照子宫内膜,比较差异均有统计学意义（P〈0.05）;AR在EMs患者异位内膜组织中的表达明显低于在位内膜及正常对照子宫内膜,比较差异均有统计学意义（P〈0.05）。结论：EMs患者异位内膜PR及AR表达的相对增高可能在内异症的发生与发展中起着一定的作用。     

免疫与子宫内膜异位症

研究表明子宫内膜异位生长与免疫异常有关。子宫内膜异位症（EMs）患者的自然杀伤（NK）细胞、巨噬细胞、淋巴细胞的数量及功能发生异常，对异位内膜的清除能力降低，从而促进EMs的发生与发展。自身抗体的产生、多种细胞因子和生长因子（IL-1，-6,-8，-12、MCP-1）含量和活性的改变都与EMs的发生发展有关。从免疫角度阐述EMs的发生机制，就其相关的免疫异常作综述。     

MCM5与子宫内膜癌发生发展的关系

目的：探讨MCM5在增殖期子宫内膜、分泌期子宫内膜和子宫内膜样腺癌组织中的表达和临床意义。方法：采用免疫组织化学链菌素亲生物素基因过氧化物酶连接法（sP法）检测15例增殖期子宫内膜、15例分泌期子宫内膜和70例子宫内膜样腺癌标本中MCM5蛋白的表达。结果：子宫内膜癌组织和增殖期宫内膜组织中MCM5蛋白高度表达，而正常分泌期宫内膜组织中低表达或不表达，子宫内膜癌组织和增殖期子宫内膜组织与分泌期宫内膜组织相比MCM5蛋白的表达具有显著差异（P〈0．05），有统计学意义。子宫内膜癌组织和增殖期子宫内膜组织中MCM5蛋白的表达没有显著差异（P〉0．05）。结论：MCM5蛋白在子宫内膜癌组织中高度表达，在正常增殖期子宫内膜组织中表达也明显增高。检测MCM5对子宫内膜癌进行高风险筛选及预后评估有一定意义。     

Bcl-2蛋白在子宫内膜异位症的表达及意义

目的 :检测正常子宫内膜及内膜异位症的在位、异位内膜 Bcl-2蛋白表达及细胞凋亡率 ,探讨其在子宫内膜异位症发生机制中的作用。方法 :采用免疫组化方法对 50例同期因宫颈癌 I期 ,输卵管节育术患者及盆腔子宫内膜异位症患者 47例的在位、异位内膜 Bcl-2蛋白表达及应用流式细胞技术检测细胞凋亡率。结果 :1对照组 :子宫内膜的 Bcl-2蛋白主要表达于子宫内膜腺上皮细胞浆和 /或细胞膜上。增生期 Bcl-2蛋白的表达显著高于分泌期 (P<0 .0 1)。 2内膜异位症组 :在位内膜的 Bcl-2蛋白亦主要表达于子宫内膜腺上皮细胞胞浆和 /或胞膜上 ,增生期 Bcl-2蛋白表达与分泌期有显著性差异 (P<0 .0 1)。异位症组异位内膜增生期、分泌期无显著性差异 (P>0 .0 5)。 3异位症组 :异位内膜与在位内膜分泌期比较 Bcl-2蛋白表达有显著性差异 (P<0 .0 5)。 4内膜异位症组与对照组比较 :其分泌期 Bcl-2蛋白表达显著高于对照组的分泌期 (P<0 .0 1)。内膜异位症在位内膜细胞凋亡率低于正常子宫内膜。异位内膜细胞 AP峰明显低于异位症在位内膜细胞和正常子宫内膜细胞。结论 :1子宫内膜周期性生理变化与 Bcl-2蛋白表达水平有关。子宫内膜的周期性变化可能与凋亡有关。 Bcl-2蛋白的表达水平越高 ,组织的抗凋亡性越强。 2异位内膜细胞凋?     

子宫内膜异位症患者腹腔液及血清中IL-8水平的测定及意义

目的检测子宫内膜异位症患者腹腔液及血清中IL-8水平的变化,探讨其在子宫内膜异位症发病中的作用。方法采用酶联免疫吸附试验（ELISA）,检测39例子宫内膜异位症患者（内异症组）和21例非子宫内膜并位症患者（对照组）腹腔液及血清中IL-8的水平,并比较其在月经周期中的变化。结果①内异症组腹腔液中IL-8水平明显高于对照组（P〈0．05）：而血清中IL-8水平与对照组比较无明显差异（P〉0．05）。②在月经周期中两组患者腹腔液IL-8水平在分泌期表达显著高于增生期（P〈0．05）;而内异症组腹腔液IL-8水平明显高于同期对照组腹腔液中水平（P〈0．05）。结论①腹腔液中IL-8的水平明显升高,导致腹腔局部微环境的改变,在子宫内膜异位症的发病中可能起重要作用。②子宫内膜异位症患者在分泌期IL-8水平显著升高,提示孕激素桔抗剂及IL-8拮抗剂治疗有可能用于子宫内膜异位症的治疗。     

Bcl-2蛋白在子宫内膜异位症的表达及意义

目的检测正常子宫内膜及内膜异位症的在位、异位内膜Bcl-2蛋白表达及细胞凋亡率,探讨其在子宫内膜异位症发生机制中的作用.方法采用免疫组化方法对50例同期因宫颈癌Ⅰ期,输卵管节育术患者及盆腔子宫内膜异位症患者47例的在位、异位内膜Bcl-2蛋白表达及应用流式细胞技术检测细胞凋亡率.结果①对照组子宫内膜的Bcl-2蛋白主要表达于子宫内膜腺上皮细胞浆和/或细胞膜上.增生期Bcl-2蛋白的表达显著高于分泌期(P＜0.01).②内膜异位症组在位内膜的Bcl-2蛋白亦主要表达于子宫内膜腺上皮细胞胞浆和/或胞膜上,增生期Bcl-2蛋白表达与分泌期有显著性差异(P＜0.01).异位症组异位内膜增生期、分泌期无显著性差异(P＞0.05).③异位症组异位内膜与在位内膜分泌期比较Bcl-2蛋白表达有显著性差异(P＜0.05).④内膜异位症组与对照组比较其分泌期Bcl-2蛋白表达显著高于对照组的分泌期(P＜0.01).内膜异位症在位内膜细胞凋亡率低于正常子宫内膜.异位内膜细胞AP峰明显低于异位症在位内膜细胞和正常子宫内膜细胞.结论①子宫内膜周期性生理变化与Bcl-2蛋白表达水平有关.子宫内膜的周期性变化可能与凋亡有关.Bcl-2蛋白的表达水平越高,组织的抗凋亡性越强.②异位内膜细胞凋亡率低于在位内膜,更低于正常子宫内膜即子宫内膜异位症的异位内膜及在位内膜分泌期抗凋亡能力明显增强.     

激活素A对子宫内膜异位来源间质细胞IL-6、IL-8以及VEGF因子表达及分泌的影响

目的评估激活素A(ACT)以及其拮抗剂卵泡抑素(follistatin),对白介素-6(IL-6)、白介素-8(IL-8)和血管内皮生长因子(VEGF)等在上述2种细胞内的表达与释放的影响。方法来源于患有子宫内膜异位症的女性(n=10)和正常对照女性(n=10)的人表皮间质细胞,以不同剂量ACT处理或ACT合并卵泡抑素双处理。实时荧光定量PCR检测IL-6、IL-8、VEGF表达,ELISA法检测蛋白水平。结果患病组细胞中IL-6分泌基线水平是正常组的2.04倍,IL-8的分泌基线水平是正常组的6.35倍(P0.05),而ACT处理则显著上调正常组中IL-8的表达与分泌以及VEGF的分泌水平,并显著下调患病组中IL-6和VEGF的分泌水平,而且ACT的作用效果可被卵泡抑素抵消(P0.05)。结论 ACT能够调控体外培养的表皮间质细胞中IL-6、IL-8和VEGF的表达与分泌,而该机制似乎在患有子宫内膜异位女性的细胞中存在缺陷。     

Genetic polymorphisms related to interleukin-1 beta production and disease risk

PURPOSE: Interleukin (IL)-1A C-889T, IL-1B C-511T, IL-1B C-31T, IL-1B C3954T, and IL-1RN 86-bp VNTR (variable number of tandem repeats) are polymorphisms potentially influencing IL-1 beta production. This review summarizes 1) the biological roles of IL-1 beta, 2) allele frequencies of the polymorphisms, and 3) the reported associations between these polymorphisms and disease risk. METHODS: Papers were obtained from PubMed with keywords "IL-1, polymorphism", as well as from the references in each paper. The most relevant papers were then selected. In this review, a narrative approach was adopted. RESULTS: IL-1 beta is a multifunctional proinflammatory cytokine, whose signal is transduced through IL-1 receptor I (IL-1RI) on the cell surface. Binding levels are influenced by the IL-1 receptor antagonist (IL-1Ra), IL-1RII (decoy receptor with no signal transduction), soluble IL-1RI, and soluble IL-1RII. IL-1B encoding IL-1 beta is located on chromosome 2q14, along with IL-1A encoding IL-1 alpha and IL-1RN encoding IL-1Ra. The minor alleles, IL-1A-889T, IL-1B 3954T, and IL-1RN 2R, are rarer in Japanese than in Caucasians, while IL-1B-511T and IL-1B-31C are more frequent. These polymorphisms have been reported to have potential associations with the risk of diseases, such as stomach cancer, breast cancer, inflammatory bowel, Alzheimer's, and osteoporosis. DISCUSSION: Although there are many inconsistent findings on associations with the polymorphisms, IL-1B C-511T and the tightly linked T-31C may be useful for predicting the risk of diseases with an inflammation basis among Japanese.     

A prospective, randomized clinical trial on perioperative feeding with an arginine-, omega-3 fatty acid-, and RNA-enriched enteral diet: effect on host response and nutritional status

BACKGROUND: The use of immune-enhancing enteral diets in the postoperative period has given contrasting results. The purpose of this prospective, randomized, double-blinded clinical study was to evaluate the effect of immunonutrition given perioperatively on cytokine release and nutritional parameters. METHODS: Patients with cancer of the stomach or colo-rectum were eligible. Subjects consumed 1 L/d of either a control enteral formula (n = 25; control group) or a formula supplemented with arginine, omega-3 fatty acids, and RNA (n = 25; verum group) for 1 week before surgery. Both formulas were given by mouth. Six hours after the operation, jejunal infusion with the same diets was started and maintained for 7 days. Blood was drawn at different time points to assess albumin, prealbumin (PA), transferrin, cholinesterase activity, retinol binding protein (RBP), interleukin-2 receptors alpha (IL-2Ralpha), IL-6, and IL-1 soluble receptors (IL-1RII). The composite score of delayed hypersensitivity response (DHR) to skin test also was determined (the higher the score, the lower the immune response). RESULTS: During the 7 days of presurgical feeding, none of the above parameters changed in either group. Eight days after operation, in the control group, the concentration of PA and RBP was lower than in the verum group (0.18 vs 0.26 g/L for PA and 30.5 vs 38.7 mg/L for RBP; p < .05). IL-2Ralpha concentration was 507 pg/mL in the verum group vs 238 pg/mL in the control group (p < .001), whereas IL-6 and IL-1RII were higher in the control group than in the verum group (104 vs 49 and 328 vs 183 pg/mL, respectively; p < .01). The DHR score was 0.68 in the control group vs 0.42 in the verum group (p < .05). CONCLUSIONS: Perioperative feeding with a supplemented enteral diet modulates cytokine production and enhances cell-mediated immunity and the synthesis of short half-life proteins.     

TGF-β受体在子宫内膜癌组织中的表达及其意义

目的:探讨转化生长因子-β(TGF-β)受体(TβRⅠ、TβRⅡ)在子宫内膜癌组织中的表达情况、与临床病理参数之间的关系及其与膜细胞骨架连接蛋白ezrin、血管内皮生长因子受体-1(flt-1)等的相关性。方法:采用免疫组织化学方法检测79例子宫内膜癌和12例转移灶组织中TβRⅠ和TβRⅡ蛋白的表达,选择不典型增生14例,单纯、复合性增生20例以及正常增生期子宫内膜组织15例作为对照,并在子宫内膜癌中检测了ezrin和flt-1。结果:TβRⅠ和TβRⅡ在子宫内膜癌组织中的表达低于正常增生期子宫内膜组织(P<0.05,P<0.01),并且深肌层浸润组的表达低于无肌层浸润及浅肌层浸润组(P=0.000,P=0.008),中、低分化癌中的表达低于高分化癌(P=0.003,P=0.000)。子宫内膜癌中,TβRⅠ与ezrin呈负相关(P=0.001),TβRⅡ与flt-1呈负相关(P=0.003)。结论:TβRⅠ、TβRⅡ表达下降可能参与了子宫内膜癌的发生和演进过程。     

纳米氧化锌诱导人肺泡Ⅱ型上皮细胞A549发生炎性凋亡的研究

目的用纳米氧化锌(ZnO-NPs)刺激体外培养的人肺泡Ⅱ型上皮细胞(A549),探讨其是否发生炎性凋亡。方法用10μg/ml和20μg/ml的ZnO-NPs分别刺激A549细胞8和24h,以磷酸盐缓冲液(PBS)为对照,用乳酸脱氢酶(LDH)活力测定试剂盒和白介素-1β(IL-1β)ELISA试剂盒分别检测细胞培养上清液中的LDH酶活力和IL-1β含量,用半胱氨酸天冬氨酸特异性蛋白酶-1(caspase-1)活性测定试剂盒检测细胞caspase-1的活性。结果纳米氧化锌作用于A549细胞8h,染毒组和对照组LDH酶活力差异无显著性,与对照组比较,20μg/ml ZnO-NPs染毒组caspase-1活性和IL-1β含量均升高(P<0.05)。ZnO-NPs作用于A549细胞24h时,染毒组和对照组caspase-1活性差异无显著性。染毒组IL-1β含量和LDH酶活力均显著高于对照组(P<0.05)。结论纳米氧化锌(20μg/ml)刺激A549细胞早期caspase-1活性升高,继之IL-1β含量和LDH酶活力升高,可能发生了炎性凋亡。     

Influence of glutamine-supplemented Caco-2 cells on cytokine production of mononuclear cells

BACKGROUND: Caco-2 cells, cultured with mononuclear cells, were used as an in vitro model of human intestinal cell function. This study shows the effect of glutamine (Gln) supplementation on the production of tumor necrosis factor alpha, interleukin-10 (IL-10), and interleukin-6 (IL-6). METHODS: Confluent Caco-2 cells were cultured in media with Gln at 0 mmol/L, 4 mmol/L, or 10 mmol/L +/- 1 microg/mL lipopolysaccharide (LPS), treated with fluorescein isothiocyanate- (FITC-) conjugated intercellular adhesion molecule-1 (ICAM-1) mononuclear antibody, and assessed for ICAM-1 expression levels via flow cytometry. Confluent Caco-2 cells alone in apical inserts, or mononuclear cells (MNCs) alone in basal chambers of transwells, were cultured in media with 0 mmol/L, 4 mmol/L, or 10 mmol/L Gln. Supernatants were taken to assess cytokine and endotoxin levels. Confluent Caco-2 cells in apical inserts of transwells were cultured in media containing Gln at 0 mmol/L, 4 mmol/L, or 10 mmol/L, whereas MNCs were cultured in the basal chamber in media containing Gln at 4 mmol/L +/- LPS. Supernatants were collected to determine cytokine levels in each chamber. RESULTS: With Gln supplementation of the media at 10 mmol/L, enterocytes displayed a decrease in ICAM-1 expression. MNCs showed a decrease in tumor necrosis factor alpha and IL-6 production and an increase in IL-10 production when incubated with Caco-2 cells in media supplemented with Gln at 10 mmol/L. CONCLUSIONS: Although cytokine production by Caco-2 or mononuclear cells incubated alone was not influenced by the Gln concentration of the media, cultured together, Gln levels affected cytokine production by mononuclear cells, which suggests that Caco-2 cells produce mediators in Gln-rich conditions that can influence mononuclear cell cytokine production.     

黄绿青霉素对肿瘤坏死因子α诱导的血管内皮细胞表达单核细胞趋化蛋白1和白细胞介素的影响

目的观察黄绿青霉素(CIT)对血管内皮细胞表达单核细胞趋化蛋白1(MCP-1)、白细胞介素1β(IL-lβ)、IL-6和IL-8等细胞因子的影响,以及CIT上调肿瘤坏死因子α(TNF-α)诱导的内皮细胞表达MCP-1、IL-lβ、IL-6和IL-8的作用。方法体外原代培养人脐静脉内皮细胞(HUVECs),选择第5～6代进行试验,待细胞融合80%后随机分组,加入TNF-α(10μg/L)、CIT(2μmol/L)建立TNF-α组、CIT组、TNF-α+CIT联合处理组和空白对照组,处理时间24h。用酶联免疫吸附法(ELISA)检测细胞上清液IL-lβ、IL-6、IL-8及MCP-1的含量;免疫荧光染色法测定细胞核转录因子κB(NF-κB)的激活表达;RT-PCR检测各组MCP-1 mRNA表达。结果在TNF-α组和TNF-α+CIT组,细胞上清液IL-lβ、IL-6、IL-8、MCP-1浓度,细胞MCP-1 mRNA表达量,NF-κB P65蛋白表达量均高于空白对照组(P<0.05);且TNF-α+CIT组均高于TNF-α组(P<0.05)。结论 CIT可明显上调TNF-α诱导的内皮细胞MCP-1、IL-lβ、IL-6、IL-8表达和NF-κB的激活。     

Modulating effect of glutamine on IL-1beta-induced cytokine production by human gut

BACKGROUND & AIMS: Balance between pro-and anti-inflammatory mediators plays a key role in the pathogenesis and treatment of inflammatory bowel disease. Glutamine can modulate cytokine production by intestinal mucosa in healthy subjects, but studies in inflammatory states are still limited. The aim of this work was to evaluate the effects of glutamine on IL-1beta-induced cytokine production by human gut. METHODS: Duodenal biopsies from healthy volunteers were stimulated in vitro by IL-1beta in the presence of increasing glutamine concentrations. Cytokine production was assessed in culture media by ELISA and cytokine mRNA expression in biopsies by RT-PCR. Results, in pg/mg of tissue, (median [range]), were compared by non-parametric paired tests. RESULTS: IL-1beta stimulation increased IL-6 and IL-8, but did not affect IL-4 and IL-10 production. IL-8 and IL-6 production from stimulated biopsies significantly decreased with increasing glutamine concentration from 0.5 to 10mM, (2543 [828-3634] to 1499 [282-2617] for IL-8, 62 [22-117] to 24 [12-99] for IL-6, both P<0.05), whereas IL-10 production was increased (0.7 [0.2-1.6] to 1.2 [2.6-0.5],P<0.05). Glutamine also increased IL-10 mRNA level in biopsies (P<0.05). IL-4 production was not affected by glutamine. CONCLUSIONS: Glutamine was shown in human intestinal mucosa to reduce the production of the pro-inflammatory cytokines IL-6 and IL-8, and enhance the production of the anti-inflammatory cytokine, IL-10.     

大气PM_(10)对两种不同细胞分泌炎性细胞因子的影响

目的探讨大气可吸入颗粒物(PM10)对小鼠肺泡巨噬细胞(RAW264·7)和人肺成纤维细胞(HLF)分泌肿瘤坏死因子(TNF-α)、白细胞介素6(IL-6)和白细胞介素8(IL-8)的影响。方法颗粒物样品采自北京市城区采暖期。两种染毒途径:(1)不同剂量的PM10直接对HLF染毒;(2)PM10首先作用于RAW264·7细胞,染毒一定时间后的细胞上清液再作用于HLF细胞。采用四甲基偶氮唑盐微量酶反应比色(MTT)法测定细胞毒性;放射免疫法测定炎性因子TNF-α、IL-6和IL-8。结果PM10作用24h后,表现为低剂量时刺激细胞增殖,而高剂量时抑制细胞增殖,细胞存活率随浓度增加而降低(P<0·01);PM10能刺激HLF分泌炎性因子TNF-α、IL-6和IL-8,且有剂量-反应关系(P<0·05);巨噬细胞上清液亦能刺激HLF产生TNF-α、IL-6和IL-8,且其产生量高于PM10作用于HLF产生的量(P<0·05)。结论大气可吸入颗粒物PM10对HLF有一定毒性作用;PM10染毒24小时后能诱导人肺成纤维细胞分泌炎性因子TNF-α、IL-6和IL-8;巨噬细胞上清液亦能刺激人肺成纤维细胞释放上述三种炎性因子,其产生量高于PM10。     

Effect of Theobroma cacao flavonoids on immune activation of a lymphoid cell line

We analysed the effect of (-)-epicatechin and cocoa extract on the activation of a lymphoid cell line. Particularly the expression of IL-2 receptor alpha (IL-2Ralpha or CD25) and, the secretion of IL-2 and IL-4 were established after flavonoid treatment. Two media culture conditions (1 and 10 % of fetal calf serum supplementation) and the different moments of flavonoid addition (simultaneously or 2 h before cell-activation) were compared. IL-2Ralpha (CD25) expression on activated cells was significantly reduced by epicatechin and cocoa extract in a dose-dependent manner, achieving the highest inhibition of about 50 % when flavonoids were added 2 h before stimulation. IL-2 secretion was also inhibited by the presence of both epicatechin and cocoa extract, displaying 60 and 75 % of inhibition, respectively. Cocoa flavonoids were also able to enhance 3-4.5-fold IL-4 release. In summary, cocoa extract down-modulated T lymphocyte activation and therefore the acquired immune response. This fact could be important in some states of the immune system hyperactivity such as autoimmune or chronic inflammatory diseases.