儿童支气管哮喘患儿外周血DC细胞数量和功能的分析研究

目的 观察支气管哮喘患儿外周血树突状细胞(DC)数量和亚型细胞的变化,研究DC细胞在支气管哮喘发病中的作用及可能的发病机理.方法 38例支气管哮喘患儿作为哮喘组,将哮喘组患儿随机分为控制组(21例)及新诊断组(17例),选取17例同期本院保健科体检健康儿童作为对照组,利用流式细胞技术先分选DCs细胞,后分选出亚型细胞:...     

肺炎支原体感染小儿支气管哮喘42例临床研究

目的:探讨小儿肺炎支原体(MP)感染的支气管哮喘临床特点.方法:对42例MP感染的小儿支气管哮喘临床特点、治疗及转归进行归纳总结.结果:42例MP感染的小儿支气管哮喘患儿Mp-Ab均为阳性,其发病向小年龄患儿发展,临床表现不典型,胸片以条絮状阴影为表现,肺部大片阴影或叶性改变不明显,病程迁延,病程>1个月20例(占47.6%),所有病例青霉素或头孢菌素加正规哮喘治疗方案无效,而改用阿奇霉素治疗后哮喘得以控制.结论:MP感染诱发的哮喘息儿发病临床特征与其他病毒或细菌感染所诱发哮喘临床特征不易区分,病程迁延,区分主要以血清学检查为主.确诊后早期用阿奇霉素治疗.     

阿奇霉素配伍氨茶碱治疗咳嗽变异哮喘疗效观察

目的对咳嗽变异哮喘患儿实施阿奇霉素配伍氨茶碱治疗,观察其临床治疗效果。方法 84例咳嗽变异哮喘患儿,随机分为研究组和对照组,每组42例。对照组患儿实施常规扩张支气管综合治疗,研究组患儿实施阿奇霉素配伍氨茶碱治疗,对两组患儿的治疗效果进行对比。结果研究组患儿治疗总有效率为90.5%高于对照组73.8%,差异具有统计学意义(P<0.05)。结论在对咳嗽变异哮喘患儿实施治疗的过程中,阿奇霉素配伍氨茶碱治疗效果显著,临床价值值得推广。     

阿奇霉素对喘息性疾病患儿免疫功能的影响

目的 探讨阿奇霉素对喘息性疾病患儿免疫功能的影响.方法 选取毛细支气管炎、支气管哮喘和哮喘性支气管炎患儿各80例,每组随机分为干预组与未干预组,干预组在常规治疗基础上接受阿奇霉素一日10 mg/kg,共4个疗程;未干预组仅接受常规治疗.检测并比较干预组与未干预组治疗前后血清T淋巴细胞亚群及白介素(IL)-8、IL-6、肿瘤坏死因子(TNF)-α水平.结果 与治疗前相比,治疗后干预组患儿血清IL-8、IL-6、TNF-a、CD4+T细胞、NK和总T淋巴细胞均降低,CD8+T细胞升高;未干预组患儿治疗前后血清T淋巴细胞亚群及细胞因子水平无显著变化.结论 阿奇霉素可改善喘息性疾病患儿的免疫功能.     

红霉素用药后阿奇霉素序贯疗法对支原体肺炎患儿伴哮喘的临床疗效评价

目的:评价红霉素用药后阿奇霉素序贯疗法对支原体肺炎患儿伴哮喘的临床疗效,以及支原体肺炎与哮喘的关系。方法:选取2014年3月—2016年3月间收治的支原体肺炎伴支气管哮喘患儿148例,将其分为治疗组和对照组,每组74例;对照组患儿给予常规治疗,治疗组患儿给予红霉素与阿奇霉素联用治疗,比较两组患儿治疗后总有效率,以及治疗后肺通气功能各指标测得值的变化情况。结果:治疗组患儿治疗后的总有效率明显高于对照组(P<0.05),肺通气功能各指标测得值明显优于对照组(P<0.05)。结论:采用红霉素与阿奇霉素联用治疗支原体肺炎患儿伴哮喘患者,对支原体肺炎所致哮喘的临床疗效较为显著,改善了其肺通气功能各指标。     

阿奇霉素治疗小儿支气管哮喘疗效观察

目的观察阿奇霉素治疗小儿支气管哮喘的临床疗效。方法将60例支气管哮喘病人随机分为阿奇霉素治疗组30例和对照组30例,比较两组的治疗效果。结果治疗组总有效率为96.97%显著优于对照组:60.00%,P0.05;在呼吸困难消失时间、肺部哮鸣音消失时间、咳嗽消失时间方面治疗组较对照组有显著性差异(P0.01)。两组均无明显的不良反应发生。结论阿奇霉素治疗小儿支气管哮喘疗效满意,值得临床推广。     

阿奇霉素辅助治疗小儿咳嗽变异性哮喘疗效观察

目的探讨阿奇霉素在治疗小儿咳嗽变异性哮喘中的临床疗效。方法将196例咳嗽变异性哮喘患儿随机分为对照组和治疗组,每组各98例。对照组常规给予扩张支气管药物综合治疗,治疗组加用阿奇霉素治疗。结果治疗组和对照组总有效率分别是93.9%和71.4%。经统计学处理两组有极显著性差异(P<0.05)。未发现明显不良反应。结论阿奇霉素治疗小儿咳嗽变异性哮喘疗效显著。     

支原体感染与咳嗽变异性哮喘的关系

目的 探讨肺炎支原体感染与咳嗽变异件哮喘的关系及阿奇霉素治疗的效果.方法 随机选择咳嗽变异性哮喘患儿42例(观察组),间期就诊年龄相仿的急性上呼吸道感染患儿50例(对照绀);用支原体培养法测定两组患儿肺炎支原体感染率,同时应用阿奇霉素进行治疗.结果 观察组肺炎支原体阳性20例(47.6%),对照组阳性7例(14.0%)(P<0.01).42例咳嗽变异性哮喘的患儿在常规治疗的基础上,加用阿奇霉素口服,有效率90.5%.结论 咳嗽变异性哮喘与肺炎支原体感染有密切关系,阿奇霉素疗效明显.     

孟鲁司特钠联合阿奇霉素治疗小儿支原体肺炎并继发性哮喘的临床研究

目的:探讨孟鲁司特钠和阿奇霉素联合治疗小儿肺炎支原体继发性哮喘发作的临床效果.方法:本研究纳入200例诊断为继发性哮喘发作的支原体肺炎患儿,依据随机抽签法分为观察组与对照组,各100例.对照组接受阿奇霉素和支气管扩张药或糖皮质激素的联合治疗.观察组在对照组基础上加用孟鲁司特钠联合治疗.比较两组治疗前后的肺功能指标,T淋...     

阿奇霉素联合布地奈德福莫特罗在支气管哮喘治疗中的作用

目的:观察临床使用阿奇霉素联合布地奈德福莫特罗治疗支气管哮喘治疗中的疗效。方法:将某院100例支气管哮喘患者随机分为实验组和对照组,每组各50例。对照组吸入布地奈德福莫特罗治疗,实验组阿奇霉素联合布地奈德福莫特罗治疗。记录两组治疗后症状改善、肺功能变化情况、不良反应等。结果:实验组症状改善和肺功能变化明显优于对照组,且二者差异具有统计学意义(P0.05)。结论:临床治疗支气管哮喘使用阿奇霉素联合布地奈德福莫特罗疗效较好,同时不良反应轻微,值得临床推广应用。     

黄芪多糖对哮喘小鼠气道炎症启动因子TSLP和DCs表达的影响

摘要：目的 研究黄芪多糖(APS)对支气管哮喘模型小鼠气道上皮中胸腺间质淋巴细胞生成素(thymic stromal lymphopoietin,TSLP)和树突状细胞(dendritic cell,DCs)表达的影响。方法 饲养雌性BALB/c小鼠30只,随机将其分为对照 组、哮喘模型组和黄芪多糖组,各组采取相应的干预措施。通过支气管哮喘激发试验、肺组织病理学和酶联免疫吸附试验 (ELISA)评价哮喘模型造模是否成功;肺组织中TSLP mRNA的相对表达水平采用实时荧光定量PCR(qRT-PCR)检测;蛋白免疫印 迹实验(Western blot)检测肺组织中TSLP蛋白的表达;流式细胞术检测支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中 DCs表面CD40、CD80和CD86的表达水平。结果 哮喘模型组小鼠气道反应性增高和肺组织HE染色结果均为哮喘的典型表现且 哮喘模型组BALF中IL-13、IL-5和IL-4的表达水平显著高于黄芪多糖组和对照组(P<0.001);黄芪多糖组和对照组小鼠肺组织中 TSLP mRNA的表达水平与哮喘模型组相比较低(P<0.001),TSLP mRNA的表达水平在黄芪多糖组与对照组之间差异无统计学意 义(P>0.05);黄芪多糖组与对照组小鼠肺组织中TSLP蛋白的表达与哮喘模型组相比较弱;黄芪多糖组与对照组BALF中DCs表面 CD54、CD80、CD86的表达明显低于哮喘模型组(P<0.001),黄芪多糖组与对照组之间差异无统计学意义(P>0.05)。结论 黄芪 多糖可明显抑制哮喘模型小鼠气道上皮细胞TSLP水平和DCs表面CD54、CD80、CD86的表达,并减轻气道炎症反应。     

Ciglitazone inhibits oxidized-low density lipoprotein induced immune maturation of dendritic cells

BACKGROUND: The peroxisome proliferator-activated receptor (PPAR) activation has generally been shown to have anti-inflammatory effects and dendritic cells (DCs) are the most efficient antigen presenting cells that play an active role in the development of atherosclerosis. The effects of PPARgamma on DCs maturation and immune function remain unknown now and we, therefore, studied the influence of PPARgamma agonist ciglitazone on the maturation and immune function of DCs. METHODS: Human monocytes were purified and immature DCs derived; ciglitazone (25 micromol/L) was added to the medium for 24 hours; ox-LDL (50 microg/ml) was then added to the medium for another 24 hours. The immunophenotypic expressions (CD1a, CD40, CD86, and HLA-DR) were analyzed by FACS and endocytosis function by FITC-dextran and the cytokines secretions of culture supernatants (IL-12,IL-10,TNFalpha, and IL-2) were measured with ELISA. RESULTS: Ciglitazone reduced ox-LDL induced immunophenotypic expressions of DCs (CD40, CD1a, and HLA-DR). Ox-LDL inhibited the endocytosis of DCs, which was prevented by ciglitazone; ciglitazone attenuated ox-LDL induced cytokine secretions of DCs (IL-12, 116 +/- 29 versus 34 +/- 3 pg/ml*; IL-10, 49 +/- 1 versus 28 +/- 9 pg/ml*; TNFalpha, 46 +/- 16 versus 24 +/- 8 pg/ml*, *P < 0.05 compared with ox-LDL, respectively). CONCLUSION: Our study suggested that one of the anti-inflammatory mechanisms of PPAR-gamma agonist ciglitazone was mediated by inhibiting the ox-LDL induced maturation and immune function of DCs.     

牙龈卟啉单胞菌脂多糖对树突状细胞成熟及功能影响的体外研究

目的 研究牙龈卟啉单胞菌脂多糖(P.gingivalis-LPS)的刺激对大鼠树突状细胞(DCs)成熟及功能的影响,为探索DCs在牙周炎的发生发展中的作用机制提供实验依据.方法 采用流式细胞学方法检测P.gingivalis-LPS和大肠杆菌脂多糖(E.coli-LPS)刺激下,CD11c+MHCⅡ+、CD11c+CD80+、CD11c+CD86+和CD11c+CD40+DCs的比率;采用ELISA法检测DCs分泌白介素-12(IL-12)、干扰素-γ(IFN-γ)、白介素-10(IL-10)和白介素-13(IL-13)的量.采用CCK8法检测与上述DCs共培养的CD4+T细胞的增殖;采用ELISA法检测T细胞分泌IL-2、IFN-γ、IL-10和IL-13的量.在上述的培养系统中加入Toll样受体4(TLR4)抑制剂(polymyxin B,PmB)或TLR2/TLR4抑制剂(oxidation of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphoryl-choline,OxPAPC),观察TLR抑制剂对上述DCs成熟及功能的影响.结果 P.gingivalis-LPS与E.coli-LPS均能刺激DCs成熟.TLR4抑制剂明显抑制E.coli-LPS组DCs成熟和抗原提呈功能,对P.gingivalis-LPS组DCs成熟和抗原提呈功能没有显著抑制.TLR2/TLR4抑制剂对P.gingivalis-LPS组DCs成熟和抗原提呈功能显著抑制.P.gingivalis-LPS组DCs分泌IL-12和IFN-γ 的量低于E.coli-LPS组(P<0.05);P.gingivalis-LPS组DCs分泌IL-10和IL-13的量高于E.coli-LPS组(P<0.05).与P.gingivalis-LPS和E.coli-LPS共培养的DCs均能促进CD4+T细胞增殖.与P.gingivalis-LPS组DCs共培养的T细胞分泌IL-2和IFN-γ 的量低于E.coli-LPS组(P<0.05);其分泌IL-10的量高于E.coli-LPS组(P<0.05).结论 P.gingivalis-LPS能促进DCs的成熟和抗原提呈功能.P.gingivalis-LPS刺激下的DCs促进Th2型免疫应答;E.coli-LPS刺激下的DCs促进Th1型免疫应答.P.gingivalis-LPS通过TLR2通路刺激DCs成熟;E.coli-LPS通过TLR4通路刺激DCs成熟.     

IL-12 suppression,enhanced endocytosis and up-regulation of MHC-II and CD80 in dendritic cells during experimental endotoxin tolerance

Aim:

The aim of this study was to investigate endocytosis, MHC-II expression and co-stimulatory molecule expression, as well as interleukin-12 (IL-12) production, in bone marrow dendritic cells (DCs) derived from endotoxin tolerant mice.

Methods:

Endotoxin tolerance was induced in C57BL/10J mice through four consecutive daily intraperitoneal injections of 1.0 mg/kg of 055:B5 Escherichia coli lipopolysaccharide (LPS). Bone marrow DCs were isolated in the presence of GM-CSF and IL-4 and purified by anti-CD11c Micro beads. FITC–dextran uptake by DCs was tested by flow cytometric analysis and the percentage of dextran-containing cells was calculated using a fluorescence microscope. The expression of surface MHC-II, CD40, CD80, and CD86 was also detected by flow cytometric analysis. An ELISA was used for the measurement of IL-12 production by DCs with or without LPS stimulation.

Results:

Endotoxin tolerance was successfully induced in C57BL/10J mice, evidenced by an attenuated elevation of systemic TNF-α. DCs from endotoxin tolerant mice possessed enhanced dextran endocytosis ability. The expression of surface MHC-II and CD80 was higher in DCs from endotoxin tolerant mice than in DCs from control mice, whereas the expression of CD40 and CD86 was not altered. Compared with DCs from normal control mice, DCs from endotoxin tolerant mice produced less IL-12 after subsequent in vitro stimulation with LPS.

Conclusion:

These data suggest enhanced endocytosis, selective up-regulation of MHC-II and CD80 and IL-12 suppression in DCs during in vivo induction of endotoxin tolerance.     

Activation of human monocyte-derived dendritic cells in vitro by the biological response modifier arabinoxylan rice bran (MGN-3/Biobran)

Arabinoxylan rice bran (MGN-3/Biobran) is a potent biological response modifier (BRM) that activates natural killer (NK) cells, T cells and monocytes. Currently, little is known regarding the effects of MGN-3 on dendritic cells (DCs), the cell type that bridges innate and adaptive immunity. Therefore, we examined the stimulatory effects of MGN-3 on DCs. Human monocyte-derived DCs were treated with MGN-3 at different concentrations (5-20 microg/ml) for 24 hours in vitro. Activation of DCs was determined by assessing the expression of co-stimulatory and maturation markers (CD40, CD80, CD83, CD86 and HLA-DR) by flow cytometry, and production of cytokines by ELISA. DC function was determined by assessing their ability to activate na?ve T cells. Activation of T cells was assessed by measuring cell proliferation and cytokine production. MGN-3 treatment, in a dose-dependent manner, resulted in: 1) up-regulation of the surface expression of CD83 and CD86, on DCs; 2) an increase in the production of pro-inflammatory and immuno-regulatory cytokines (IL-1beta, IL-6, IL-10, TNF-alpha, IL-12p40 and low levels of IL-12p70 and IL-2) by DCs; and 3) MGN-3 stimulated DC induced CD4+T cell proliferation and their production of cytokines, IFN-gamma, IL-10, IL-17. Results suggest that MGN-3 functions as a natural adjuvant for DC activation and thus may be used in DC-based vaccine strategies against infections and cancer.     

干扰素抗病毒应答与慢性乙型肝炎患者外周血树突状细胞功能的关系

目的:探讨干扰素抗病毒应答与慢性乙型肝炎患者外周血树突状细胞(DCs)功能的关系。方法:分别采集23例慢性乙型肝炎患者干扰素治疗前和治疗满4mo时的抗凝外周静脉血,分离外周血单个核细胞(PBMCs),在重组人白细胞介素4和重组人粒细胞-巨噬细胞集落刺激因子的作用下培养7 d使DCs增殖、成熟。以间接免疫荧光流式细胞技术检测DCs的表型;以ELISA法检测DCs单独培养上清液中IL-12的水平;用DCs与HBsAg共同孵育,丝裂霉素C处理后,再与自体PBMCs共同培养,加入H-TDR,收集细胞测定cpm值。实验中以8例正常健康人作为对照。结果:干扰素完全应答组DCs表面CD,HLA-DR和ICAM-1的表达较治疗前均显著增加(均为P<0.05),且CD,CD,HLA-DR和ICAM-1的增高与无应答组相比具有显著性意义(分别为P<0.05、P<0.01、P<0.01和P<0.05);完全应答组DCs分泌IL-12的水平在治疗后显著增加(P<0.01);DCs的抗原提呈作用在治疗前、后,完全应答组比无应答组均显著增强(P<0.01和P<0.001),而部分应答与无应答组之间无显著性差异(P>0.05)。结论:慢性乙肝患者干扰素的抗病毒应答与外周血DCs的功能状态有关,干扰素能显著促进患者DCs功能的改善,进而可能增加患者对治疗的应答。     

Maturation of dendritic cells induced by Candida beta-D-glucan

We investigated whether Candida beta-D-glucan (CSBG) alters the maturation of dendritic cells (DCs). DC phenotypes were analyzed using FACScan. The expression of surface molecules, including major histocompatibility complex (MHC) classes I and II, as well as CD80 and CD86, increased on DCs that were stimulated with lipopolysaccaride (DCs/LPS), in comparison with unstimulated bone marrow-derived DCs (BM-DCs). Furthermore, the level of surface molecule expression on DCs stimulated with CSBG (DCs/CSBG) was between that of DCs and DCs/LPS. Phagocytosis was assessed by the uptake of FITC-dextran. There were no differences in the uptake of dextran among DCs/LPS and DCs/CSBG. The ability of BM-DCs to uptake dextran was higher than that of DCs/LPS and DCs/CSBG. We analyzed the concentration of IL-12 secreted by DCs using ELISA. BM-DCs secreted a low concentration of IL-12, while DCs/LPS and DCs/CSBG secreted higher levels of IL-12 than BM-DCs. There were no remarkable differences in the concentrations of IL-12 produced by DCs/LPS and DCs/CSBG. This data suggests that CSBG may augment DC maturation.     

哮喘患者外周血单个核细胞CD4~+CD25~+调节性T细胞及IL-4分泌水平的检测

目的:探讨CD4+CD25+调节性T细胞在哮喘发病机制中的作用,为哮喘的临床治疗提供新的思路和方法。方法:分离哮喘患者和正常人外周血单个核细胞(PBMC),用流式细胞仪分析CD4+CD25+调节性T细胞在外周血单个核细胞中的比例;酶联免疫吸附法(ELASA)检测外周血PBMC培养上清IL-4的分泌状况。结果:哮喘组外周血CD4+CD25+调节性T细胞的百分率为(3.2±1.5)%,健康对照组为(6.5±2.5)%,哮喘组明显低于对照组(P<0.05);哮喘组外周血PBMC培养上清IL-4分泌水平为(57.3±13.5)ng/L,健康对照组为(28.6±7.2)ng/L,哮喘组明显高于对照组(P<0.05)。结论:CD4+CD25+调节性T细胞可能参与了哮喘的发生与发展。     

IL-18对胶质瘤大鼠肿瘤组织中CD80、CD86及MHCⅠ类分子、Ⅱ类分子的影响

目的 观察转染IL-18基因的大鼠胶质瘤细胞疫苗9L/IL-18对大鼠胶质瘤肿瘤组织中CD80、CD86及MHCⅠ类分子、MHCⅡ类分子的影响,探讨IL-18在胶质瘤中的抗肿瘤作用.方法 F344大鼠随机分为3组,每组7只,分别接种9L/IL-18、9L/LXSN和9L细胞,流式细胞术检测CD80、CD86、MHCⅠ类分子、MHCⅡ类分子在肿瘤组织中的表达.结果 CD80、CD86、MHCⅠ类分子在接种9L/IL-18细胞组大鼠的肿瘤组织中的表达均明显高于接种9L/LXSN细胞组和接种9L细胞组大鼠的肿瘤组织中的表达,差异均有统计学意义(P<0殚.05);CD80、CD86、MHCⅠ类分子在接种9L/LXSN细胞组和接种9L细胞组大鼠的肿瘤组织中的表达,差异无统计学意义(P>0.05);MHCⅡ类分子在3组中的表达,差异无统计学意义(P>0.05).结论 IL-18通过促进肿瘤细胞表面CD80、CD86、MHCⅠ类分子的表达,促进树突状细胞分化成熟,进而激活T细胞,增强肿瘤细胞的抗原活性,提高免疫系统的监视功能,发挥抗肿瘤作用.