共查询到20条相似文献,搜索用时 125 毫秒
1.
目的:检测抗小鼠CD4+T淋巴细胞活化后表面白细胞介素10受体I(IL-10R1)表达变化,为进一步研究白细胞介素10(IL-10)相关免疫疾病的发病机制及进展提供有力支持.方法:取C57BL/6小鼠脾细胞,溶血后采用尼龙毛去除B淋巴细胞后,流式细胞仪无菌分选CD4+T淋巴细胞,利用anti-CD3和anti-CD28多克隆抗体刺激分选后细胞,分别在第0、12、24、48和72小时流式细胞术检测IL-10R1表达情况.结果:0小时时小鼠CD4+T淋巴细胞不表达IL-10R1,随着刺激时间的延长IL-10R1表达增加,24小时时达到最高,随后表达水平开始下降,72小时时表达情况与0小时基本相同.结论:CD4+T淋巴细胞活化后IL-10R1表达短暂,以24小时时表达最高. 相似文献
2.
目的 探讨过敏性鼻炎(AR)患者外周血2型辅助T(Th2)细胞白细胞介素18(IL-18)、 IL-18结合蛋白亚型a(IL-18BPa)和IL-18受体α(IL-18Rα)的表达水平及过敏原对其表达的影响。方法 采用大籽蒿花粉过敏原粗提液(ASWE)和梧桐花粉过敏原粗提液(PPAE)、室尘螨过敏原粗提液(HDME)激发外周血单个核细胞和免疫磁珠分选后的CD4+ T细胞,流式细胞术检测上述标本CD4+ Th2细胞中IL-18、 IL-18BPa和IL-18Rα的表达,BioPlex检测血浆IL-4的水平,并分析其与IL-18+ Th2细胞比例的相关性。结果 与正常对照相比,AR患者外周血CD4+ Th2细胞中IL-18+细胞的比例增加,IL-18的平均荧光强度(MFI)增强;IL-18Rα MFI降低。过敏原可直接诱导分选后的HC CD4+ Th2细胞表达IL-18和IL-18Rα, AR患者CD4+ Th2细胞表达IL-18Rα。A... 相似文献
3.
目的:检测抗小鼠CD4+T淋巴细胞活化后表面白细胞介素10受体I(IL-10R1)表达变化,为进一步研究白细胞介素10(IL-10)相关免疫疾病的发病机制及进展提供有力支持。方法:取C57BL/6小鼠脾细胞,溶血后采用尼龙毛去除B淋巴细胞后,流式细胞仪无菌分选CD4+T淋巴细胞,利用anti-CD3和anti-CD28多克隆抗体刺激分选后细胞,分别在第0、12、24、48和72小时流式细胞术检测IL-10R1表达情况。结果:0小时时小鼠CD4+T淋巴细胞不表达IL-10R1,随着刺激时间的延长IL-10R1表达增加,24小时时达到最高,随后表达水平开始下降,72小时时表达情况与0小时基本相同。结论:CD4+T淋巴细胞活化后IL-10R1表达短暂,以24小时时表达最高。 相似文献
4.
目的建立荧光定量-逆转录-聚合酶链反应(FQ-RT-PCR)测定TH细胞中IL-2 mRNA和IL-4 mRNA方法,并作临床初步应用。方法制备IL-2 cDNA和IL-4 cDNA,分别构建含有人IL-2基因和叫基因片段的pMD18-T质粒,克隆后作为定量阳性模板;设计和制备用于FQ-RT-PCR的引物和FAM、TAMRA标记的探针,优化实验条件,建立FQ-RT-PCR方法;用固相单抗方法从健康人、妇科良性疾患和恶性肿瘤患者以及慢性肾功能衰竭(cRF)患者PBMC中富集CD4(TH)细胞,经PMA和calcium ionophore诱导,提取总RNA,继用所建FQ-RT-PCR方法,作IL-2 mRNA和IL-4 mRNA定量测定。实验设阻actin为内控基因以监测RNA的质量。结果根据系列模板浓度的对数与CT值作直线回归建立标准曲线,线性范围为10^2~10^7copies/μl;不精密度试验显示,高值样品的批内、批间CV分别为7.8%和12.5%,低值样品的批内、批间CV分别为10.8%和19.5%;妇科恶性肿瘤患者与健康对照组和妇科良性疾患组比较,CRF患者与健康对照组比较,IL-2 mRNA表达均显著降低,IL-4 mRNA表达均显著升高(P〈0.001)。结论用所建立的FQ-RT-PCR法可对TH细胞内IL-2 mRNA和IL-4 mRNA作定量测定,方法简便、敏感、准确;妇科恶性肿瘤患者和CRF患者TH细胞内IL-2mRNA和IL-4 mRNA表达有明显的极化现象,呈TH2反应,提示恶性肿瘤患者和CRF患者TH细胞功能处于失衡状态。可通过改善和调整TH细胞的失衡提高恶性肿瘤患者的免疫监视功能和纠正CRF患者免疫紊乱。 相似文献
5.
目的:研究白藜芦醇(RSV)对活化的小鼠T淋巴细胞分泌IL-2和CD25的影响,并探讨其免疫抑制机制。方法:无菌分离小鼠淋巴结细胞,与不同浓度的白藜芦醇共同孵育1小时后,用多克隆刺激剂佛波醇酯和离子霉素刺激T细胞活化,继续培养6小时后收获细胞,进行胞内细胞因子染色,流式细胞仪检测IL-2的分泌情况,RT-PCR检测IL-2 mRNA的表达:24小时后检测CD25表达。结果:RSV对IL-2的分泌具有抑制作用,并呈剂量依赖性,同时RSV也能抑制T细胞表面活化分子CD25。结论:RSV对活化T细胞分泌的细胞因子IL-2及IL-2α链CD25的表达可能是RSV对T细胞具有免疫抑制作用的机制之一,并可能与T细胞活化通路的PKC-NF-κB信号传导途径相关。 相似文献
6.
目的:探讨牛膝多糖免疫调节作用的机制。 方法: (1)以牛膝多糖 (800 mg/L)分别对哮喘和肺癌病人的(PBMC)进行体外诱导培养,在18 h以RT-PCR检测IFN-γ和IL-4的基因表达率。(2)用牛膝多糖在不同时间内和不同剂量分别对Th细胞进行体外诱导其分化,并分别收集细胞悬液和抽提细胞中RNA,分别采用RT-PCR和ELISA检测Th细胞所分泌的IFN-γ和IL-4的含量。 结果: (1)哮喘和肺癌患者PBMC中IFN-γ mRNA表达率分别由6/25升到14/25(P<0.01)、3/22升到10/22(P<0.05),IL-4 mRNA的表达率由17/25降到9/25(P<0.05)、14/22降到5/22(P<0.05)。 (2)Th细胞培养至6 h,即可检到IFN-γ mRNA分泌,18 h达到高峰,之后迅速下降,48 h已难以检出。而IL-4被抑制。(3) IFN-γ蛋白表达量存在时间依赖性,从18-24 h时段开始,蛋白表达量开始显著增加(P<0.05),但在24 h、36 h和48 h的3个时段内,蛋白表达量增加不明显(P>0.05),进入平台期。IL-4蛋白表达量被抑制。(4)IFN-γ基因水平和蛋白表达量分别与牛膝多糖的诱导浓度相关(r=0.979)。400 mg/L和800 mg/L是较好的的诱导浓度。而IL-4的蛋白表达被抑制。 结论: (1)应用牛膝多糖能初步纠正肺癌和哮喘患者Th1和Th2细胞因子的失平衡。(2)牛膝多糖能在转录水平和翻译水平促进Th1类细胞因子的分泌,而抑制Th2类细胞因子的分泌。 相似文献
7.
IL-2和IL-15协同调节T细胞的增殖和LAK细胞的杀伤活性 总被引:2,自引:6,他引:2
目的 探讨IL-2和IL-15在免疫调控和应答中的协同作用。方法 IL-2、IL-15和IL-2 IL-l5组刺激CTLL细胞的增殖,^3H—TdR掺入法测cpm值;IL-2、IL-15和IL-2 IL-l5组诱导PBMC中NK和LAK细胞的发育,4h^51Cr释放实验检测对K562和LiBr细胞的杀伤活性。结果 IL-2和IL-15都能诱导CTLL认细胞的增殖和NK、LAK细胞的杀伤活性,IL-2和IL-15可明显协同上调CTLL认的增殖和LAK细胞的杀伤活性。结论 IL-2和IL-15在免疫调控和应答中有一定的协同作用,为细胞因子联合应用于临床治疗肿瘤等疾病提供了实验依据。 相似文献
8.
组胺对T细胞IL-2产生及增殖活性影响的实验研究 总被引:3,自引:0,他引:3
目的:了解组胺对 CD4 和 CD8 T细胞IL-2产生和细胞增殖活性的影响。方法:密度梯度离心及吸附法分离PBMC和PBLC,采用抗CD4 和CD8 抗体分别制备CD8 和CD4 T细胞进行培养,然后采用ELISA法和MTT比色法测上清液IL-2含量及增殖活性。结果:①组胺 CD4 (CD8 )培养上清液中IL-2水平及MTT增殖指数与T细胞自然培养孔比较明显降低(P<0.05)。②组胺 CD4 (CD8 ) 西咪替丁培养孔上清液中IL-2水平及 MTT增殖指数明显高于未加西咪替丁孔(P<0.05)。③CD4 T细胞自然培养孔上清液中IL-2水平显著高于CD8 T细胞自然培养孔。结论:组胺可抑制T细胞IL-2产生及增殖。西咪替丁可阻断组胺对T细胞的抑制作用。CD8 T细胞也可产生IL-2,但其功能较 CD4 T细胞为低。 相似文献
9.
10.
本文用免疫酶标法测定IL-2分泌细胞和细胞膜IL-2受体。其结果表明急性非淋白血病细胞条件培养液(即含白血病细胞相关抑制活性)对PHA诱导的正常人外周血IL-2分泌细胞和细胞膜IL-2受体表达均有明显抑制作用,但对PHA诱导的正常人外周血单个核细胞培养上清的IL-2活性无明显影响。 相似文献
11.
Barthlott T Moncrieffe H Veldhoen M Atkins CJ Christensen J O'Garra A Stockinger B 《International immunology》2005,17(3):279-288
Maintenance of homeostasis in the immune system involves competition for resources between T lymphocytes, which avoids the development of immune pathology seen in lymphopenic mice. CD25+ CD4+ T cells are important for homeostasis, but there is as yet no consensus on their mechanisms of action. Although CD25+ CD4+ T cells cause substantial down-regulation of IL-2 mRNA in responder T cells in an in vitro co-culture system, the presence of IL-protein can be demonstrated by intracellular staining. As a consequence of competition for IL-2, CD25+ CD4+ T cells further up-regulate the IL-2R alpha chain (CD25), a process that is strictly dependent on IL-2, whereas responder T cells fail to up-regulate CD25. Similarly, adoptive transfer into lymphopenic mice showed that CD25+ CD4+ T cells interfere with CD25 up-regulation on co-transferred naive T cells, while increasing their own CD25 levels. IL-2 sequestration by CD25+ CD4+ T cells is not a passive phenomenon but instead initiates--in conjunction with signals through the TCR--their differentiation to IL-10 production. Although IL-10 is not required for in vitro suppression, it is vital for the in vivo function of regulatory T cells. Our data provide a link explaining the apparent difference in regulatory mechanisms in vitro and in vivo. 相似文献
12.
13.
14.
目的:研究脑不对称性对下丘脑中IL -1β、IL -6及细胞因子信号转导抑制因子 3(SOCS -3)表达的影响,从分子水平探讨脑不对称性在神经内分泌免疫调节网络中的作用。方法:运用伸爪取食法将BALB c小鼠区分为左利、右利和双利鼠。小鼠分组一周后断头杀死,迅速分离出下丘脑,一部分标本制备匀浆后采用ELISA法测定下丘脑中IL- 1β、IL- 6的蛋白水平;另一部分标本以RT- PCR法间接测定下丘脑SOCS -3mRNA水平。结果:(1)下丘脑IL 6蛋白水平:左利鼠水平明显高于右利鼠,差异有统计学意义(P <0 . 0 5 )。(2 )下丘脑IL -1β蛋白水平:左利、右利小鼠水平相近,差异无显著性(P >0 . 0 5 )。(3)下丘脑SOCS 3表达:右利鼠下丘脑中SOCS 3的表达显著高于左利鼠(P <0 . 0 5 )。结论:脑不对称BALB c小鼠下丘脑IL -1β、IL- 6及细胞因子信号转导抑制因子SOCS -3的表达存在差异,提示这可能是左、右利人群免疫紊乱性疾病发生率不同原因之一。 相似文献
15.
Edgar Schmitt Petra Hoehn Christoph Huels Sigrid Goedert Norbert Palm Erwin Rüde Tieno Germann 《European journal of immunology》1994,24(4):793-798
It was observed in vitro and in vivo that both interferon (IFN)-γ and interleukin (IL)-12 can promote the development of T helper type 1 (TH1) cells. Since IL-12 was shown to be a costimulator for the production of IFN-γ by T or natural killer (NK) cells, IL-12 might play only an indirect role in TH1 differentiation by providing IFN-γ which represents the essential differentiation factor. Using anti-CD3 monoclonal antibody (mAb) for activation of naive CD4+ T cells in the absence of accessory cells we could demonstrate that costimulation by IFN-γ alone results only in marginal TH1 development. Similarly, IL-12 in the absence of IFN-γ is only a poor costimulator for inducing differentiation towards the TH1 phenotype. Our data indicate that both cytokines are required to allow optimal TH1 development and that IL-12 has a dual role, it promotes differentiation by direct costimulation of the T cells and also enhances the production of IFN-γ which serves as a second costimulator by an autocrine mechanism. Another cytokine that was reported to favor TH1 differentiation in certain experimental systems is transforming growth factor (TGF)-β. With naive CD4+ T cells employed in this study TGF-β strongly inhibited the production of IFN-γ triggered by IL-12 as well as the IL-12-induced TH1 development. When TGF-β was combined with anti-IFN-γ mAb for neutralization of endogenous IFN-γ the TH1-inducing capacity of IL-12 was completetly suppressed. 相似文献
16.
17.
Tumour growth promotes the expansion of CD4(+) CD25(+) FoxP3(+) regulatory T cells (Tregs) which suppress various arms of immune responses and might therefore contribute to tumour immunosurveillance. In this study, we found an inverse correlation between circulating Treg frequencies and phosphoantigen-induced gammadelta T-cell proliferation in cancer patients, which prompted us to address the role of Tregs in controlling the gammadelta T-cell arm of innate immune responses. In vitro, human Treg-peripheral blood mononuclear cell (PBMC) co-cultures strongly inhibited phosphoantigen-induced proliferation of gammadelta T cells and depletion of Tregs restored the impaired phosphoantigen-induced gammadelta T-cell proliferation of cancer patients. Tregs did not suppress other effector functions of gammadelta T cells such as cytokine production or cytotoxicity. Our experiments indicate that Tregs do not mediate their suppressive activity via a cell-cell contact-dependent mechanism, but rather secrete a soluble non-proteinaceous factor, which is independent of known soluble factors interacting with amino acid depletion (e.g. arginase-diminished arginine and indolamine 2,3-dioxygenase-diminished tryptophan) or nitric oxide (NO) production. However, the proliferative activity of alphabeta T cells was not affected by this cell-cell contact-independent suppressive activity induced by Tregs. In conclusion, these findings indicate a potential new mechanism by which Tregs can specifically suppress gammadelta T cells and highlight the strategy of combining Treg inhibition with subsequent gammadelta T-cell activation to enhance gammadelta T cell-mediated immunotherapy. 相似文献
18.
卵巢癌细胞培养上清诱导CD4+CD25-T细胞分化为CD4+CD25+调节性T细胞 总被引:2,自引:0,他引:2
目的研究卵巢癌细胞培养上清液是否能诱导外周血CD4^+CD25^- T细胞转变为CD4^+CD25^+调节性T细胞。方法将外周血CD4^+CD25^- T细胞分离后,对照组用CD3和CD28单抗活化,实验组在对照基础上加用卵巢癌细胞株SKOV3培养上清,72h后分离各组的CD25^+和CD25^-T细胞,溴化脱氧尿嘧啶掺入标记法测定增殖能力及对静息的自体同源CD4^+CD25^- T细胞的增殖抑制能力,流式细胞仪测定细胞糖皮质激素诱发型TNF受体(glucocorticoid-induced TNFR,GITR)与CTLA-4分子的表达,RT-PCR检测细胞卿mRNA的表达。结果与对照组相反,实验组的CD4^+CD25^+T细胞具有免疫抑制功能,自身增殖能力下降,GITR和CTLA-4分子的表达和CD4^+CD25^+调节性T细胞相似,并被诱导表达转录因子Foxp3 mRNA。结论卵巢癌细胞分泌的可溶性物质能诱导外周血CD4^+CD25^-T细胞转化为CD4^+CD25^+调节性T细胞。 相似文献
19.
Williams MA Porter M Horton M Guo J Roman J Williams D Breysse P Georas SN 《The Journal of allergy and clinical immunology》2007,119(2):488-497
BACKGROUND: Dendritic cells (DCs) translate environmental cues into T-cell activating signals, and are centrally involved in allergic airway inflammation. Ambient particulate matter (APM) is ubiquitous and associated with allergic diseases, but it is unknown whether APM directly activates DCs. OBJECTIVE: To study comprehensively the effects of APM on myeloid DC phenotype and function. METHODS: Development of DC was modeled using human CD34(+) progenitor cells. APM was collected from ambient outdoor air in Baltimore city. We studied the effects of APM on DC activation in vitro, compared with LPS. RESULTS: Ambient particulate matter enhanced DC expression of costimulatory receptors but suppressed the expression of both the endocytosis receptor CD206 and uptake of fluorescein isothiocyanate-conjugated dextran. The expression of the Toll-like pattern-recognition receptors Toll-like receptor 2 and Toll-like receptor 4 was also blunted. APM-exposed DCs secreted less IL-12 and IL-6 but exhibited increased secretion of IL-18 and IL-10 compared with LPS stimulation. A T(H)2-like pattern of cytokine production was seen in cocultures of APM-stimulated DCs and alloreactive naive CD4(+) T cells where the IL-13 to IFN-gamma ratio was reversed. This contrasted with the T(H)1 polarizing effects of LPS on DCs. CONCLUSION: We report for the first time that APM-exposed DCs direct a complex T(H)1/T(H)2-like pattern of T-cell activation by mechanisms that involve nonclassic activation of DCs. CLINICAL IMPLICATIONS: Inhaled APM can act directly on DCs as a danger signal to direct a proallergic pattern of innate immune activation. 相似文献
20.
Tanijiri T Shimizu T Uehira K Yokoi T Amuro H Sugimoto H Torii Y Tajima K Ito T Amakawa R Fukuhara S 《Journal of leukocyte biology》2007,82(3):576-584
A recent report revealed that a large population of Hodgkin's lymphoma-infiltrating lymphocytes (HLILs) consisted of regulatory T cells. In this study, we cocultured CD4+ naive T cells with KM-H2, which was established as a Hodgkin's Reed-Sternberg cell line, to clarify their ability to induce CD25+ Forkhead box P3+ (Foxp3+) T cells. The characteristic analyses of T cells cocultured with KM-H2 revealed the presence of CD4+CD25+ T cells. They expressed CTLA-4, glucocorticoid-induced TNFR family-related gene, and Foxp3 and could produce large amounts of IL-10. Conversely, KM-H2 also generated CD4+ CTLs, which expressed Granzyme B and T cell intracellular antigen-1 in addition to Foxp3+ T cells. They exhibit a strong cytotoxic effect against the parental KM-H2. In conclusion, KM-H2 promotes a bidirectional differentiation of CD4+ naive T cells toward Foxp3+ T cells and CD4+ CTLs. In addition to KM-H2, several cell lines that exhibit the APC function were able to generate Foxp3+ T cells and CD4+ CTLs. Conversely, the APC nonfunctioning cell lines examined did not induce both types of cells. Our findings suggest that the APC function of tumor cells is essential for the differentiation of CD4+ naive T cells into CD25+Foxp3+ T cells and CD4+ CTLs and at least partly explains the predominance of CD25+Foxp3+ T cells in HLILs and their contribution to a better prognosis. Therefore, in APC-functioning tumors, including classical Hodgkin lymphomas, which generate Foxp3+ T cells and CD4+ CTLs, these T cell repertories play a beneficial role synergistically in disease stability. 相似文献