Th1型和Th2型细胞因子对人中性粒细胞吞噬结核杆菌活性的影响

Objective To establish a flowcytometry method for detecting phagocysis of Mycobacterium tuberculosis(Mth) by the human peripheral blood neutrophils( PMNs), and to explore the effects of Thl and Th2 cytokine on phagocytotic activity of PMNs. Methods By using acid-fast staining the phagocytosis of Mtb by human neutrophil was observed by microscopy, and the phagocytosis of FTTC labeled Mtb by human neutrophils was detected under confocal microscope. The whole fresh peripheral blood of healthy adults was incubated with FTTC labeled Mtb and the phagocytosis were measured by flow cytometry. The altered phagocytosis of FTTC-Mtb by neutrophils pretreated with IL-2, IFN-γ, GM-CSF or IL-4 were measured. Results Human peripheral blood neutrophils activity of Mtb phagocytosis was observed by acid-fast staining and confocal microscope. The percentage of phagocytosis of Mtb was dependent on the time of incubation with Mtb. The percentages were 47% at 5 min and reached plateau about 66% ～72% at 15 min to 20 min.Pretreatment with different concentrations of IL-2 or IFN-γincreased the phagocytosis of Mtb by 76.7% and 75.2%, respectively; but pretreatment with IL-4 decreased the phagocytosis by 31.7%. Conclusion IL-2and IFN-γcan increase phagocytosis of Mth by neutrophils; while IL-4 can reduced neutrophil activity of phagocytosis of Mtb by human neutrophils, and demonstrate that Th1/Th2 type cytokins involve in regulating the acitvities of neutrophils anti-Mtb infection.     

Th1型和Th2型细胞因子对人中性粒细胞吞噬结核杆菌活性的影响

Objective To establish a flowcytometry method for detecting phagocysis of Mycobacterium tuberculosis(Mth) by the human peripheral blood neutrophils( PMNs), and to explore the effects of Thl and Th2 cytokine on phagocytotic activity of PMNs. Methods By using acid-fast staining the phagocytosis of Mtb by human neutrophil was observed by microscopy, and the phagocytosis of FTTC labeled Mtb by human neutrophils was detected under confocal microscope. The whole fresh peripheral blood of healthy adults was incubated with FTTC labeled Mtb and the phagocytosis were measured by flow cytometry. The altered phagocytosis of FTTC-Mtb by neutrophils pretreated with IL-2, IFN-γ, GM-CSF or IL-4 were measured. Results Human peripheral blood neutrophils activity of Mtb phagocytosis was observed by acid-fast staining and confocal microscope. The percentage of phagocytosis of Mtb was dependent on the time of incubation with Mtb. The percentages were 47% at 5 min and reached plateau about 66% ～72% at 15 min to 20 min.Pretreatment with different concentrations of IL-2 or IFN-γincreased the phagocytosis of Mtb by 76.7% and 75.2%, respectively; but pretreatment with IL-4 decreased the phagocytosis by 31.7%. Conclusion IL-2and IFN-γcan increase phagocytosis of Mth by neutrophils; while IL-4 can reduced neutrophil activity of phagocytosis of Mtb by human neutrophils, and demonstrate that Th1/Th2 type cytokins involve in regulating the acitvities of neutrophils anti-Mtb infection.     

IL-12增强结核病患者中性粒细胞吞噬和杀伤结核杆菌的活性

目的:观察IL-12对结核病患者外周血中性粒细胞(PMN)对结核分枝杆菌(M.tb)吞噬和杀伤功能的影响.方法:结核病患者和正常人外周血与异硫氰酸荧光素( FTTC)标记的M.tb共同孵育不同时间后,用流式细胞术(FCM)检测PMN对M.tb的吞噬率;结核病患者和正常人外周血均加入2'7’二氯二氢荧光素二乙酸酯(DCFH-DA)作用,同时加或者不加M.tb共同孵育不同时间后,用FCM检测PMN的活化率并计算活性氧(ROS)产生量;不同浓度IL-12预先作用外周血后,再用上述方法检测PMN对M.tb吞噬率,活化率及产生ROS的变化.结果:结核病患者和正常人PMN对M.tb吞噬率,PMN活化率和ROS产生均随时间延长而增加,但结核病患者5 min吞噬率(51.82±6.93)％高于正常人(47.20±4.26)％(P＜0.05);IL-12预先作用后,结核病患者和正常人PMN吞噬率、PMN活化率和ROS的产生均随浓度增大而显著增加,两者之间无显著性差异.结论:结核病患者外周血PMN对M.tb的吞噬功能以及ROS产生与正常人无显著性差异,仅早期吞噬率较高.IL-12略增强PMN吞噬和产生ROS的反应能力.     

中药对过敏性哮喘大鼠模型Th1/Th2型细胞因子的影响

目的:探讨中药治疗对哮喘大鼠IL- 4 mRNA和IFN-γ mRNA表达及IgE含量的影响.方法:24只Wistar大鼠, 随机分为中药治疗组、空白对照组和哮喘模型组, 利用卵白蛋白(OVA)及氢氧化铝制作大鼠哮喘模型, 在雾化激发期, 中药组用杏仁、甘草等中药制成的溶液雾化大鼠, 其余组用生理盐水代替.采用免疫放射法、逆转录聚合酶链反应(RT-PCR)半定量法分别检测各组大鼠血清和肺泡灌洗液(BALF)中免疫球蛋白E(IgE)含量以及肺组织IL- 4 mRNA和IFN-γ mRNA表达.结果:与空白对照组比较, 模型组大鼠IgE含量和IL- 4 mRNA的表达明显升高(P<0.05).与模型组比较, 中药治疗血清IgE的含量和IL- 4 mRNA的表达明显降低(P<0.05), 而IFN-γ mRNA表达则升高(P<0.05).结论:IL- 4可能参与哮喘的发生, 而中药治疗可通过抑制IL- 4 mRNA和上调IFN-γ mRNA的合成, 改善哮喘炎症状态.     

pHGF对人中性粒细胞吞噬功能的影响

我们曾报道，从乳猪肝脏提取的促肝细胞生长因子（ｐＨＧＦ）能增强小鼠中性粒细胞的吞噬功能，本文进而用吞噬杀菌试验、化学发光技术和促凝血活性试验证明，ｐＨＧＦ也能显著增强人中性粒细胞的吞噬和杀菌功能。     

过敏小鼠模型Th1/Th2漂移和纠正

目的：研究炒紫苏子醇提物对过敏小鼠模型Th1／Th2漂移和纠正作用。方法：设正常对照组、过敏模型组、炒紫苏子醇提取物0．32、0．64和1．28g／kg各剂量组小鼠共5组。利用流式细胞仪技术测定Th1型细胞因子IL-2、IFN-γTNF-α和Th2型细胞因子IL4、IL-5水平。结果：过敏模型小鼠IFN-γ／IL4为0．87，而正常小鼠IFN-γ／IL4为3．93，说明过敏小鼠Th1／Th2异常偏向Th2漂移。0．32、0．64和1．28g／kg各剂量组能明显提高IFN-γ水平（P〉0．05、P〈0．05和P〈0．01），降低IL-4水平（P〉0．05、P〉0．05和P〈0．05），其相应的IFN-γ／IL-4分别为1．92、2．85和3．14。结论：炒紫苏子醇提物能够纠正过敏小鼠Th1／Th2异常偏向Th2漂移，恢复正常，其作用呈剂量依赖关系。     

新生儿感染性疾病中性粒细胞CD64分子变化及临床意义

目的观察新生儿感染性疾病中性粒细胞CD64表达水平及其临床意义。方法将34例新生儿感染性疾病患儿根据其临床表现及实验室检查结果分为败血症12例,非败血症感染22例,以同期收治的非感染性新生儿28例为对照组。采用全血流式细胞术、放射酶联免疫吸附试验及特定蛋白分析仪比浊法分别检测三组患儿CD64指数、IL-6、CRP水平。结果败血症组CD64指数、IL-6及CRP水平均高于非败血症感染组和对照组,差异有显著性意义(P<0.05),非败血症感染组CD64指数、IL-6及CRP水平高于对照组(P<0.05);感染组CD64指数与IL-6呈显著的正相关(P<0.01);败血症组CD64指数随病情好转而逐渐降低。结论新生儿周血中性粒细胞CD64表达水平在感染性疾病时显著升高,并随病情而变化,早期检测意义较大。     

细胞因子与中性粒细胞的凋亡

中性粒细胞（ＰＭＮ）是机体防御系统的主要组成部分。多种细胞因子能调节ＰＭＮ的凋亡，如ＴＮＦ－α能加速ＰＭＮ的凋亡，ＧＭ－ＣＳＦ，ＩＬ－２，ＩＬ－６和ＩＦＮ－γ等能延缓其凋亡，这些因子对ＰＭＮ凋亡的影响是通过不同机理而实现的。ＰＷＮ以凋亡的方式被机体清除，对于控制炎症组织的损伤和正常情况下炎症的消退都有很重要的意义。     

HIV-1感染患者中性粒细胞吞噬及细胞内杀菌功能的变化

AIDS是世界性传染病,近几年其发病率呈上升趋势,死亡率很高,严重威胁着人类生命和健康.这些患者大多数不是死亡于AIDS本身,而是死亡于后期的机会致病微生物感染和其他微生物的重复感染或合并感染.     

化学修饰的糖脂4′″-dh-iGb3对NKT细胞分泌Th1/Th2型细胞因子的影响

为研究两种i Gb3类似物(化学修饰的糖脂4′″-dh-i Gb3和4-HO-i Gb3)对NKT细胞Th1/Th2型细胞因子分泌的影响。采用流式细胞术检测经腹腔注射两种i Gb3类似物后C57BL/6小鼠脾脏NKT细胞数量的变化以及NKT细胞胞内IFN-γ和IL-4的表达水平;用Real-ti me PCR方法检测体外培养的脾脏淋巴细胞与i Gb3类似物共孵育后IFN-γ和IL-4的mRNA表达水平,并用ELISA方法检测孵育上清中IFN-γ和IL-4的含量。结果显示:与i Gb3组相比,经两种i Gb3类似物体内刺激后脾脏NKT细胞的数量都没有显著性变化。糖脂4-dh-i Gb3能够较i Gb3更强地诱导脾脏NKT细胞胞内IFN-γ的表达,也能够上调体外培养的脾脏淋巴细胞IFN-γ的mRNA水平及IFN-γ的分泌,而IL-4在所检测的各个水平上都没有显著性变化。提示化学修饰的糖脂4′″-dh-i Gb3能够诱导C57BL/6鼠脾脏NKT细胞Th1型细胞因子的分泌,而并不显著影响Th2型细胞因子的分泌,从而诱导Th1/Th2型细胞因子平衡向Th1方向偏移。     

华支睾吸虫感染患者血清Th1/Th2细胞因子水平检测及意义

目的研究华支睾吸虫病患者血清中Th1/Th2细胞因子IL-2和IL-4的水平变化,并探讨其在肝吸虫致病机制中的作用。方法分别采集华支睾吸虫病人和健康人的血清,用ELISA方法检测血清中Th1/Th2细胞因子IL-2和IL-4的水平。结果华支睾吸虫病人血清IL-2的水平与正常对照组相比(P<0.01)显著降低,IL-4的水平与正常对照组相比显著升高(P<0.05)。结论华支睾吸虫感染病人血清细胞因子水平异常,表现为Th1型细胞因子水平下降,Th2型细胞因子水平升高,病人细胞免疫功能下降,体液免疫功能增强升高,本研究结果表明肝吸虫感染后引起的细胞因子紊乱参与肝吸虫致病过程。     

Chronic experimental colitis induced by dextran sulphate sodium (DSS) is characterized by Th1 and Th2 cytokines

Oral administration of DSS has been reported to induce an acute and chronic colitis in mice. The aim of our study was to evaluate if the chronic phase of DSS-induced colitis was characterized by a Th1/Th2 response and how this would relate to mucosal regeneration. Swiss Webster mice were fed 5% DSS in their drinking water for 7 days, followed by 2–5 weeks consumption of water. Control mice received only water. The animals were killed at 3 and 6 weeks after induction. Their colons were isolated for histology and immunohistochemistry, using specific MoAbs for T and B cells, macrophages, interferon-gamma (IFN-γ), IL-4 and IL-5. Colons were scored for inflammation, damage and regeneration. Two weeks after stopping DSS the colonic epithelium had only partially healed. Total colitis scores were still increased, especially in the distal colon, which was due to more inflammation, damage and less regeneration. In areas of incomplete colonic healing the basal parts of the lamina propria contained macrophages and CD4⁺ T cells. These CD4⁺ T cells showed a focal increase of IFN-γ and IL-4 staining compared with control animals. These findings were still observed 5 weeks after stopping DSS in some mice, albeit less extensive. Chronic DSS-induced colitis is characterized by focal epithelial regeneration and a Th1 as well as Th2 cytokine profile. We postulate that chronic immune activation mediated by both populations of Th cells can interfere with colonic healing and can play a role in the pathogenesis of chronic colitis.     

极高频电磁波对恶性肿瘤化疗患者外周血辅助淋巴细胞亚群Th1/Th2免疫应答平衡的影响

目的研究极高频电磁复合波对晚期恶性肿瘤患者外周血辅助淋巴细胞亚群Th1/Th2免疫应答平衡的影响。方法对32例恶性肿瘤患者化疗后进行极高频电磁复合波幅照,并采用ELISA法检测幅照治疗前、后外周血中的IFN-γ和IL-4水平变化;另对30例恶性肿瘤化疗后患者,同比进行细胞因子检测;再分别对上述患者外周血进行IFN-γ和IL-4水平变化的自身对照研究,以评价极高频电磁复合波对恶性肿瘤患者化疗后免疫功能的影响。结果①恶性肿瘤化疗后第8天,患者IFN-γ水平(24.66±12.85)pg·mL-1低于化疗后第3天水平(27.88±17.07)pg·mL-1,但未见显著性差异(P>0.05);而IL-4水平(54.80±28.56)pg·mL-1则明显地高于化疗后第3天水平(44.97±27.53)pg·mL-1,P<0.05。②极高频电磁复合波幅照的化疗患者,化疗后第8天,IFN-γ水平(34.79±27.23)pg·mL-1远高于化疗后第3天水平(20.39±12.67)pg·mL-1,P<0.05;IL-4水平变化研究结果显示,化疗后第8天,患者的IL-4水平(43.49±34.04)pg·mL-1高于化疗后第3天水平(35.77±22.23)pg·mL-1,但其间差异无显著性(P>0.05)。③恶性肿瘤患者化疗后第3天至第8天,细胞因子IFN-γ/IL-4水平比值降低,其间有显著性差异(P<0.05);经极高频电磁复合波幅照后,其比值明显升高[从(0.57±0.44)pg·mL-1升至(0.80±0.67)pg·mL-1],P<0.05。结论恶性肿瘤患者化疗后第3天至第8天,细胞因子IFN-γ水平降低,而IL-4水平则明显升高,反映恶性肿瘤化疗后患者Th细胞的存在异常(Th2)漂移;但极高频电磁复合波幅照治疗,可干预或阻抑恶性肿瘤患者化疗后Th细胞的异常漂移。     

The Th1 and Th2 cytokines IFN-γ and IL-4 antagonize the inhibition of monocyte IL-1 receptor antagonist by glucocorticoids: involvement of IL-1

Monocytes express IL-1 and IL-1 receptor antagonist (IL-1Ra) in response to lipopolysaccharide (LPS). IL-1 self-induction contributes to the increase in IL-1 following LPS stimulation. LPS-stimulated IL-1 and IL-1Ra production are inhibited by glucocorticoids. In the present work we examined the regulation of IL-1Ra by Th1 cytokine IFN-γ, Th2 cytokine IL-4, glucocorticoids and IL-1 in human monocytes. We demonstrate that IL-1 contributes to LPS-induced IL-1Ra expression as shown by IL-1 blockade in LPS-stimulated monocytes using a specific anti-IL-1β antibody or recombinant IL-1Ra. Glucocorticoids inhibited IL-1β-stimulated IL-1Ra mRNA expression and protein production. Glucocorticoids inhibited both IL-1-mediated and non-mediated LPS stimulation of IL-1Ra expression. Both IFN-γ and IL-4 reversed the inhibitory effect of glucocorticoids on IL-1Ra expression and secretion. The effect of IFN-γ was blocked by pretreatment of monocytes with an anti-IL-1β blocking antibody, whereas the effect of IL-4 could not be blocked, demonstrating that IFN-γ acts through a mechanism dependent on endogenous IL-1 production, whereas IL-4 acts through an IL-1-independent one. Consistent with this finding, IFN-γ (but not IL-4) failed to reverse the inhibitory effect of glucocorticoids when stimulated by IL-1, and only IL-4 combined with IL-1 showed synergism resulting in an increase in IL-1Ra production. The differential regulation and involvement of IL-1 in the expression of IL-1Ra by IFN-γ, IL-4 and glucocorticoids sets the level of monocyte responsiveness during the Th1 or Th2 responses.     

膜表达的HLA-G1对同种反应性PBMC分泌Th1/Th2型细胞因子的影响

目的研究人脐静脉内皮细胞系(ECV304)表达的HLA-G1对同种异体外周血单个核细胞(PBMC)Th1/Th2型细胞因子分泌的影响。方法采用脂质体介导的基因转染技术将pcDNA3-HLA-G1转入ECV304,以间接免疫荧光技术在蛋白质水平上检测HLA-G1分子在ECV304上的表达;以表达HLA-G1的ECV304作为刺激细胞,灭活后,与健康人PBMC共同培养,用ELISA检测上清液中Th1/Th2型细胞因子的浓度,观察HLA-G1对同种异体抗原激活的PBMC分泌Th1/Th2型细胞因子的影响。结果与转染pcDNA3空质粒的对照组相比,HLA-G1能使PBMC的IL-10分泌增加(P<0.05),而IL-2,IFN-γI、L-4分泌无明显影响。结论HLA-G1能引起由同种异体抗原激活的PBMC分泌IL-10增加,提示HLA-G1有可能使Th1/Th2型细胞因子向Th2型偏移。