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.     

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

目的 建立用流式细胞术检测人外周血中性粒细胞(PMNs)吞噬结核分枝杆菌(Mtb)的方法 ,并探讨Th1和Th2型细胞因子对PMNs吞噬Mtb活性的影响.方法 运用抗酸染色、激光共聚焦显微镜观察人PMNs吞噬Mtb,并用流式细胞术检测人PMNs对FTTC标记Mtb的吞噬活性.外周血预先分别与IL-2、IFN-γ、GM-CSF和IL-4等细胞因子孵育,再加FTTC标记Mtb后,用流式细胞术检测PMNs对Mtb吞噬率,并与对照组比较吞噬率的变化.结果 抗酸染色和激光共聚焦显微镜均能观测到人PMNs吞噬Mtb.用流式细胞术检测健康人外周血PMNs对Mtb的吞噬率在5 min时为47%,15～20 min达到平台期,为66%～72%.外周血预先加IL-2或IFN-γ作用后,PMNs对Mtb的吞噬率可分别增加76.7%和75.2%;而预先加IL-4作用后,吞噬率降低31.7%.结论 IL-2和IFN-γ对PMNs吞噬Mtb功能有增强作用,而IL-4有降低作用,表明Th1型和Th2型细胞因子参与调节PMNs抗结核杆菌感染的免疫作用.     

加味宣肺透解剂对流感病毒感染小鼠细胞因子的影响

目的：观察加味宣肺透解剂（JXT）对流感病毒感染小鼠细胞因子的影响。方法：复制流感病毒鼠肺适应株（FM1）小鼠肺炎模型，以加味宣肺透解剂灌胃治疗。ELISA法检测感染后第5天血清中IL-2、TNF-α、IL-6、IFN-γ的含量，肺组织切片HE染色。结果：加味宣肺透解剂明显升高血清中IL-2、IFN-γ水平，降低TNF-α、IL-6水平，减轻肺组织病变。结论：调节细胞因子的分泌，可能是加味宣肺透解剂减轻病毒感染小鼠肺组织损伤的机制之一。     

NOX1基因荧光素酶报告基因系统的建立

目的：对炎症细胞因子TNF-α及IFN-γ刺激下,人NOX1基因的表达调控进行初步分析。方法：将NOX1基因5′-端上游序列连接到无启动子的PGL3-BASIC质粒,构建了PGL3-BASIC／NOX1报告质粒。PGL3-BASIC／NOX1质粒转染A549细胞,用TNF-α、IFN-γ刺激12h,双荧光素酶报告基因系统检测基因表达情况。结果：克隆的NOX1片段具有较强的启动子活性,在TNF-α和IFN-γ共同刺激下,转染报告基因的A549细胞萤光素酶活性与对照相比有明显的增高（约4．3倍）。分析显示NOX1基因5′端上游序列片段含有NF-KB结合位点,提示细胞因子刺激的荧光素酶表达增强可能与NF-KB位点激活相关。结论：NOX1基因表达水平明显受到炎症细胞因子的调控,提示该基因可能参与机体免疫防御（特别是上皮细胞免疫防御）,值得进行深入研究。     

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.     

伤寒沙门氏菌诱导不同分化和活化状态的THP-1细胞产生TNF-α和IL-12的研究

为探讨伤寒沙门氏菌诱导宿主细胞产生细胞因子,以及细胞因子在宿主防御伤寒沙门氏菌感染中的调节作用,我们应用人类单核白血病细胞系—THP1细胞,对伤寒沙门氏菌诱导不同分化和活化状态的THP1细胞产生TNFα和IL12进行了研究。结果表明,伤寒沙门氏菌不能诱导THP1细胞产生TNFα,但可诱导PMA分化的THP1细胞产生TNFα和IL12;TNFγ既可增强伤寒沙门氏菌诱导THP1细胞产生IL12,又可增强伤寒沙门氏菌诱导的PMA分化的THP1细胞产生TNFα和IL12。以上结果提示,TNFα的产生可能与单核细胞的分化状态有关;TNFα和IL12的产生可能存在着不同的诱导机制;IFNγ对TNFα和IL12的产生具有调节作用。     

Th1/Th2类细胞因子在被动型Heymann肾炎发病中的作用

本文动态观测了被动型Ｈｅｙｍａｎｎ肾炎（ＰＨＮ）发病各期的Ｔｈ１类细胞因子（ＩＬ ２,ＩＦＮ γ）和Ｔｈ２类细胞因子（ＩＬ ４）的变化。结果表明在注射兔抗肾近曲小管刷状缘抗体（Ｔｕｂ Ａｂ）７ｄ后即开始出现蛋白尿,同时ＩＬ ２,ＩＦＮ γ较正常对照诱生水平降低（Ｐ＜００５）,而ＩＬ ４诱生水平开始升高;在注射Ｔｕｂ Ａｂ第１４天时,出现大量蛋白尿,此时ＩＬ ４进一步升高,与正常对照及注射第７天时ＩＬ ４水平均存在统计学差异,ＩＬ ４与蛋白尿定量之间并有一定相关性;第２８天时蛋白尿开始降低,ＩＬ ２,ＩＦＮ γ比第７天和第１４天时略有升高,此时ＩＬ ４水平与正常对照无差异。表明ＩＬ ２,ＩＦＮ γ的降低和ＩＬ ４升高（即Ｔｈ１／Ｔｈ２的失衡）与本病存在一定关系。     

IFN-γ和TNF-α协同对MIN6细胞凋亡的影响

目的研究细胞因子IFN-γ和TNF-α协同对小鼠胰岛瘤细胞株MIN6凋亡的影响。方法采用四甲基偶氮唑蓝(MTT)比色法观察比较了IFN-γ、TNF-α单独及其组合对MIN6细胞活性的影响,并进一步通过光学显微镜和激光共聚焦显微镜从形态学上对其毒性作用进行了验证,最后通过流式细胞术区分了其对MIN6细胞的凋亡和坏死的作用。结果 IFN-γ和TNF-α单独作用对MIN6细胞活性有轻微影响,但两者组合对MIN6细胞的活性具有显著的抑制作用,并且早期凋亡的细胞百分率明显高于IFN-γ单独处理组(69.91%&13.03%),而坏死或晚期凋亡细胞的百分率两组之间没有差异(2.14%&2.41%)。结论IFN-γ和TNF-α协同可促进MIN6细胞的凋亡。     

Modulation of the Th1/Th2 bias by lipopeptide and saponin adjuvants in orally immunized mice

We compared the adjuvanticity of the synthetic lipopeptide P3CSK4 of bacterial origin and the plant-derived adjuvant saponin using the wheat storage protein gliadin as antigen. Gluten sensitive BALB/c mice were orally immunized with gliadin in a mixture with either lipopeptide or saponin. The gliadin-specific serum IgG response was markedly enhanced by the saponin adjuvant. The lipopeptide adjuvant enhanced the IgG2a response, but reduced IgG1 production. In contrast, the saponin adjuvant enhanced both IgG2a and IgG1, and the sera showed elevated specific IgE concentrations. Enhanced specific IgA levels were detected in sera and in faeces especially after immunizations with gliadin in combination with P3CSK4 Enhanced specific IgG and IgA levels could also be detected in supernatants of cell cultures prepared from mesenteric lymph nodes and Peyer's patches of immunized mice. Our data suggest that both adjuvants enhance the mucosal as well as the systemic immune response; P3CSK4 predominantly elicits the activation of the Th1 subset, whereas saponin activates both the Th1 and Th2 subser. Our findings are of importance for the improvement of mucosal immunizations, and might be a tool for the immunotheraphy of food allergies.     

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。     

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。     

Th1/Th2细胞与纤维化疾病

纤维化可以发生在不同组织器官和系统,是细胞外基质成分过度积聚、组织瘢痕形成的过程,是致病和致死的主要原因。纤维化疾病中Th1细胞/Th2细胞对纤维增生的反应失衡有重要影响。随着IFNγ对Th1细胞的免疫反应作用研究的深入,IL-4拮抗剂、IL-5和IL-13已经被用于纤维化疾病的治疗。这些细胞因子拮抗剂在临床试验中发挥着重要作用。     

The effect of MS14 on Th2 cytokines pattern in Balb/C mice

The tendency of immune response toward either Th1 or Th2 cytokine pattern can cause a number of pathologic conditions. Multiple sclerosis is postulated to be a Th1-type cell-mediated autoimmune disease. MS14—an Iranian natural product—seems to possess anti-inflammatory properties. Thus, we studied the effect of orally administered MS14 on Th2 cytokines (IL-5 and IL-10) in normal Balb/C mice (100?mg/kg; 5 days). The result indicated that activated splenocytes of MS14 group produced significantly more IL-5 and IL-10 (3–4 times in comparison with control group mice). MS14 could upregulate Th2 cytokine and thereby it may possess immunoregulatory properties probably useful in treatment of some diseases.     

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。     

Dietary nucleic acids promote a shift in Th1/Th2 balance toward Th1-dominant immunity

BACKGROUND: Dietary sources of nucleic acids and their relative components are known to affect host immune function; however, it has not yet been clarified whether such dietary nucleic acids influence the pathogenesis of allergic reaction. OBJECTIVE: The purpose of this study is to elucidate the effect of dietary nucleic acids on Th1/Th2 balance. METHODS: Both human flora-associated and specific pathogen-free BALB/c mice were maintained on either nucleic acid-free, or -supplemented diets. The effects of nucleic acids on both in vivo antibody levels and in vitro splenocyte cytokine production were compared using these mice. RESULTS: Supplementation of nucleic acids caused a reduction in the serum antibody levels of total IgM, IgG, IgG1, and IgE in the human flora-associated mice without affecting the composition of intestinal flora. In contrast, there was no significant difference of the serum IgG2a levels between nucleic acid-free and -supplemented mice. Such a phenomenon as that, the supplementation of dietary nucleic acids reduces the serum IgE or IgG1 levels, but not the IgG2a level, was also seen in the specific pathogen free mice. Moreover, when the mice were systematically challenged with ovalbumin, the supplementation of nucleic acids also suppressed the serum ovalbumin-specific IgE and IgG1 antibody levels as well as in vitro IL-4 and IL-10 secretion, while enhancing both the serum ovalbumin-specific IgG2a antibody levels and in vitro IFN gamma secretion. CONCLUSION: These results suggested that dietary nucleic acids may play an important role in promoting a shift in Th1/Th2 balance toward Th1-dominant immunity.