Hp感染性胃病与TH1 / TH2细胞因子行为关系的研究

目的研究Hp感染性胃部疾病患者TH1/TH2型细胞因子的含量变化,藉以探讨其细胞免疫致病机制.方法分别对37例慢性浅表性胃炎,20例慢性萎缩性胃炎(CAG),32例消化性溃疡(PU)及20例胃癌患者与30例Hp阴性患者的外周血单个核细胞(PBMC)进行分离、培养,取上清液后,对其TH1型细胞因子IFN-γ,IL-2及TH2型细胞因子IL-4,IL-10的含量采取双抗体夹心ELISA法进行检测.结果①Hp感染者慢性浅表性胃炎,PU,CAG,胃癌患者IFN-γ(pg/mL54.92±21.15,63.65±15.36,98.95±32.14,194.66±39.33)均明显高于Hp阴性患者(41.63pg/mL±18.50pg/mL,P<0.05～0.001,具有显著性差异;IL-2在慢性浅表性胃炎、PU,CAG,胃癌患者(pg/mL37.38±15.02,40.41±16.57,47.11±7.46,68.69±19.97)均亦明显高于Hp阴性患者(28.58pg/mL±15.52pg/mL),P<0.05～0.001,均具有显著性差异;而Hp感染性胃部疾病与Hp阴性患者IL-4和IL-10的含量大部分无显著差异,P>0.05.仅慢性浅表性胃炎、PU患者IL-4(pg/mL19.75±15.99,19.87±18.59vs35.21±22.22)明显低于Hp阴性患者,P<0.05,有显著性差异.胃癌患者IL-10(pg/mL57.77±16.85vs28.80±20.69)明显高于阴性患者,P<0.001,有显著性差异.②PU与慢性浅表性胃炎患者相比,除IL-10前者(28.16pg/mL±18.52pg/mL)明显低于后者(39.92pg/mL±24.10pg/mL),P<0.05,有显著性差异外,余细胞因子含量均P>0.05,无显著性差异;CAG与慢性浅表性胃炎和PU细胞因子含量相比,除IFN-γ前者(98.95pg/mL+32.14pg/mL)明显高于后两者(54.92pg/mL±21.15pg/mL,63.65pg/mL±15.36pg/mL),P<0.001,具有显著性差异,余细胞因子含量均P>0.05,无显著性差异;胃癌患者与CAG相比,前者IFN-γ,IL-10含量(pg/mL194.66±39.33,57.77±16.85)明显高于后者(pg/mL98.95±32.14,26.89±5.25),均P<0.001具显著性差异,IL-2,IL-4的含量(pg/mL,IL-268.69±19.97vs47.11±7.46,IL-432.42±11.76vs21.34±6.26),前者明显高于后者,均P<0.05,具显著性差异.结论①Hp感染性胃部疾病患者尤其是PU,慢性萎缩性胃炎、胃癌患者均存在细胞免疫功能亢进,即均表现为TH1优势应答.②PU患者IL-10的含量明显低于慢性浅表性胃炎患者,IL-10的降低促使了IFN-γ的产生增加,加重了粘膜的炎症变化,增加了促胃液素的释放,促使溃疡形成.提示IL-10含量的变化可能与PU的发生有关.③慢性萎缩性胃炎患者较慢性浅表性胃炎和PU患者IFN-γ明显增高,提示长期、持续性Hp感染加重了TH1优势应答,使IFN-γ分泌明显增加,结果加速了上皮细胞的凋亡,此与加重胃粘膜炎症导致萎缩性胃炎的发生可能有关.④Hp特别是CagA+的Hp感染可致较高的IFN-γ分泌继而引起胃粘膜上皮细胞过度凋亡和增殖,过度增殖过程中,增加了基因突变而致癌的可能性.     

消化性溃疡患者转化生长因子α、表皮生长因子和前列腺素E_2的关系

目的:研究转化生长因子α(TGFα)在慢性胃粘膜病变患者血液、胃液中的含量变化及其与表皮生长因子(EGF)和前列腺素E_2(PGE_2)的关系,初步探讨TGFα在胃粘膜病变发病过程中的作用。 方法:选择经胃镜和病理组织学检查证实的活动期消化性溃疡(PU)患者44例,其中十二指肠球溃疡(DU)22例,胃溃疡(GU)22例;慢性浅表性胃炎(CSG)患者18例;另选择20例胃镜观察胃粘膜基本正常者作为对照组。胃镜下经活检孔抽取胃液,并采集空腹静脉血,用放免分析法检测胃液、血液中TGFα,EGF,PGE_2的含量。 结果:GU和DU两组的血清TGFα(ng·L~(-1)),EGF(ug·L~(-1))含量均明显低于对照组和CSG组(3.5±1.1,3.4±1.3 vs (5.9±1.6.5.0±1.7,p<0.01;0.3±0.1,0.3±0.1 vs 0.6±0.2,0.5±0.2 p<0.01)。GU和DU血浆PGE_2(ng·L~(-1))含量与对照组和CSG组相比,差异无显著性(24.7±8.4,26.0±8.7 vs 25.2±8.0,20.9±7.7,p<0.05).GU,DU和CSG三组胃液中TGFα(ng·L~(-1)),EGF(ng·L~(-1)),PGE_2(ng·L~(-1))含量均明显低于对照组(1.7±0.7,1.6±0.7,2.1±0.7 vs 2.7±0.8,p<0.05;109±47.121±67.113±48 vs 373±78,p<0.01;15.8±6.6, 14.1±7.3,16.4±6.9 vs 21.9±7.5,p<0.05)。GU组胃液中TGFα与PGE_2间存在直线正相关(r=0.55,p<0.05),GU组     

白细胞介素12对结核病患者TH1/TH2平衡的影响

目的探讨白细胞介素12(IL-12)对结核病患者外周血单个核细胞(PBMC)分泌干扰素γ(IFN-γ)、白细胞介素4(IL-4)的影响.方法随机将25例结核患者和15名健康人PBMC分为RPMI1 640组、PPD组、PPD+rhIL-12组、PPD+anti-IL-12组,采用ELISA法检测各组培养上清液中IFN-γ、IL-4的水平.结果与PPD组相比,加入rhIL-12能有效增加结核患者及健康人PBMC分泌IFN-γ(分别为321.6±87.7至452.5±111.4, 387.0±70.8至515.4±44.1), 减少IL-4的分泌(54.6±11.0至41.3±13.5,55.1±9.5至38.2±12.7);而加入anti-IL-12可抑制IFN-γ的分泌,促进IL-4合成,且结核患者各组IFN-γ水平均低于健康人各对应组,而IL-4水平无差异.复治组IFN-γ、IL-4水平均低于初治组(P<0.05).结论 IL-12通过诱导IFN-γ分泌,抑制IL-4合成,从而调节TH1/TH2平衡,对结核分支杆菌感染患者产生保护性的免疫反应.IL-12可作为结核免疫调节剂,并可用于开发结核新疫苗.     

胃癌中幽门螺杆菌感染与癌基因蛋白表达及细胞凋亡研究

目的　研究幽门螺杆菌 (Hp )感染的胃癌 (GC)发展中c -met蛋白表达及细胞凋亡的关系和对胃癌预后意义。方法　 14 5例经病理证实 ,不同胃黏膜病变采用免疫组化检测c -met基因表达及Warthin -starry法检测Hp感染。采用原位末端标记法 (TUNEL)检测细胞凋亡。结果　在浅表性胃炎 (CSG)、萎缩肠化生胃炎 (CAG +IM)、异型增生 (DYS)、早期GC和进展期GC中 ,c-met基因表达率分别为 2 5 5 3% ,5 1 2 8% ,6 1 5 4 % ,6 6 6 7%和 6 8 4 2 % ,CAG +IM、DYS、GC均显著高于CSG(P <0 0 5 )。凋亡指数 (AI)分别为 (4 5 5± 2 33) %、(6 4 3± 5 6 0 ) %、(6 4 5± 5 12 ) %、(6 5 5± 4 80 ) %、(8 84±5 6 3) % ,进展期GC显著高于CSG(P <0 0 5 )。胃黏膜凋亡指数与c -met表达强度有密切相关 (P <0 0 5 )。c -met阳性表达与胃癌组织类型、浆膜浸润和淋巴结转移密切相关 ,而且BorrmannIV明显高于早期胃癌和BorrmannⅠ ,Ⅱ (P <0 0 5 )。Hp阳性者 5年存期显著短于Hp阴性者。 结论　Hp感染和c-met表达与胃黏膜增殖和恶化有关 ,且与凋亡有相关性。Hp感染与胃癌预后有关。     

反应停对重症肝炎外周血单个核细胞细胞因子的影响

目的观察反应停(Thal)对重症肝炎外周血单个核细胞(PBMCs)细胞因子的影响及其对重症肝炎的治疗作用.方法用淋巴细胞分离液从11例重症肝炎患者的外周血中分离PBMCs,体外培养,用酶联免疫吸附、免疫组化和半定量斑点杂交法,观察Thal对PBMCsTNFα和其他细胞因子的影响.结果Thl可抑制PBMCs TNFα和其他细胞因子的产生(从1923ng·L-1±697ng·L-1下降至713 ng·L-1±224ng·L-1,P＜0.05)以及TNFα mRNA的表达(相对灰度值从13.6±4.5下降至8.3±2.7,P＜0.05).结论Thal对重症肝炎的治疗有重大的意义.     

Effect of cholecystokinin on cytokines during endotoxic shock in rats

AIM To study the effect of cholecystokinin-octapeptide(CCK-8)on systemic hypotension and cytokine productionin lipopolysaccharide(LPS)-induced endotoxic shock(ES)rats.METHODS The changes of blood pressure were observedusing physiological record instrument in four groups ofrats:LPS(8mg·kg~(-1),iv)induced ES;CCK-8(40μg·kg~1,iv)pretreatment 10min before LPS(8mg.kg~(-1));CCK-8(40μg·kg~(-1),iv)or normal saline(control)groups.Differences in tissue and circulating specificity of theproinflammatory cytokines(TNF-α,IL-1β and IL-6)wereassayed with ELISA kits.RESULTS CCK-8 reversed LPS-induced decrease of meanartery blood pressure(MABP)in rats.Compared withcontrol,LPS elevated the serum level of IL-6 significantly(3567±687ng·L~(-1) vs 128±22ng·L~1,P<0.01),whilecontents of TNF-α and IL-1β elevated significantly(277±86ng.L~1 vs not detectable and 43±9ng·L~1 vs notdetectable,P<0.01)but less extent than IL-6.CCK-8significantly inhibited the LPS-induced increase in serumTNF-α,IL-1β and IL-6.LPS elevated spleen and lungcontent of IL-1β significantly(5184±85ng·L~1 vs 1047±21ng·L~1 and 4050±614ng·L~1 vs not detectable.P<0.01),while levels of TNF-α and IL-6 also rosesignificantly but in less extent than IL-1β.CCK-8 inhibitedthe LPS-induced increase of the cytokines in spleen andlung.In the heart,CCK-8 significantly inhibited LPS-induced increase of TNF-α(864±123ng·L~1 in CCK-8 LPS group vs 1599±227ng·L~1 in LPS group,P<0.01),and IL-1β(282±93ng·L~1 in CCK-8 LPS group vs 621±145ng·L~1 in LPS group,P<0.01).CONCLUSION CCK-8 reverses ES,which may be relatedto its inhibitory effect on the overproduction of cytokines.     

可溶性Fas,可溶性Fas配体与细胞因子在1型糖尿病发病中的意义

目的　研究可溶性Fas(sFas)和可溶性Fas配体(sFasL)与细胞因子在1型糖尿病发病中的作用。方法　32名1型糖尿病患者和20名正常人的血清,采用夹心BAS-ELISA法分别检测sFas,sFasL,γ干扰素(IFN-γ)及白细胞介素-1(IL-1)含量。结果　1型糖尿病血清中sFas,IFN-γ及IL-1含量分别为(1　　546±685,1　　074±451与1　　406±721)ng/L,显著高于正常组;sFasL为(211±73)mg/L,低于正常组但差异无显著性。在1型糖尿病患者中高浓度sFas伴高IFN-γ者共12例,低浓度sFas伴低IFN-γ者共9例。结论　1型糖尿病患者血清中的sFas,IFN-γ及IL-1高于正常人,sFas与IFN-γ浓度呈正相关(r=0.79,P＜0.05)。对1型糖尿病患者血清进行sFas,IFN-γ及IL-1等检测可作为反映胰岛细胞病变的辅助指标,有助于对疾病的诊断与治疗。     

胃癌中幽门螺杆菌感染与癌基因蛋白表达及凋亡

目的　研究幽门螺杆菌 (Hp)感染的胃癌 (GC)发展过程中c met蛋白表达与细胞凋亡的关系及对胃癌预后的意义。方法　对 14 5例经病理证实的不同胃粘膜病变采用免疫组化法检测c met基因表达及Warthin starry法检测Hp感染。采用原位末端标记法 (TUNEL)检测细胞凋亡。结果　在浅表性胃炎 (CSG)、萎缩肠化生胃炎 (CAG +IM )、异型增生 (DYS)、早期GC和进展期GC中 ,c met基因表达率分别为 2 5 .5 3 %、5 1.2 8%、6 1.5 4%、6 6 .6 7%和 6 8.42 % ,CAG +IM、DYS、GC的c met基因表达率均显著高于CSG (P <0 .0 5 )。凋亡指数 (AI)分别为( 4 .5 5± 2 .33) %、( 6 .43± 5 .6 0 ) %、( 6 .45± 5 .12 ) %、( 6 .5 5± 4.80 ) %和 ( 8.84± 5 .6 3) % ,进展期GC显著高于CSG(P <0 .0 5 )。胃粘膜凋亡指数与c met表达强度密切相关 (P <0 .0 5 )。c met阳性表达与胃癌组织类型、浆膜浸润和淋巴结转移密切相关 ,而且BorrmannⅣ明显高于早期胃癌和BorrmannⅠ ,Ⅱ (P <0 .0 5 )。Hp阳性者 5年生存期显著短于Hp阴性者。 结论　Hp感染和c met表达与胃粘膜增殖和恶化有关 ,且与凋亡有相关性。Hp感染与胃癌预后有关。     

姜黄素对三硝基苯磺酸诱导的结肠炎模型细胞因子的影响

目的探讨姜黄素对三硝基苯磺酸(TNBS)诱导的实验性结肠炎的疗效,并研究肠黏膜、脾细胞和血清中细胞因子的变化。方法45只Sprague-Dawley大鼠经直肠注入TNBS(100 mg/kg)诱导肠炎模型。实验动物分为阴性对照组、模型组(TNBS)和姜黄素治疗组(30 mg·kg~(-1)·d~(-1),腹腔注射)。RT-PCR测定肠黏膜细胞因子表达水平,流式细胞术测定脾细胞内干扰素(IFN)-γ和白细胞介素(IL)-4,ELISA法测定血清中IFN-γ和IL-4。结果姜黄素治疗前后肠黏膜大体评分(3．9±1．0比2．2±0．7),髓过氧化物酶(MPO)活性[每毫克蛋白(15．0±2．6)U比(7．3±1．4)U],肠黏膜中Th1类细胞因子IFN-0γ(1．02±0．07比0．06±0．02)、IL-12 mRNA相对表达水平(0．29±0．05比0．11±0．01)和IFN-γ/IL-4比值(11．44±0．97比0．38±0．10),脾细胞中IFN-γCD4~+细胞百分比(31．7±7．5比21．1±3．7)和IFN-γ/IL-4 CD4~+比值(19．9±5．1比6．1±1．8),血清中IFN-γ质量浓度[(1528±159)pg/ml比(513±14)pg/ml]和IFN-γ/IL-4比值(19．5±4．1比4．2±0．6)均降低(P＜0．05或P＜0．01)。且姜黄素治疗前后肠黏膜中Th2类细胞因子IL-4 mRNA(0．09±0．01比0．15±0．04)和IL-10 mRNA相对表达水平(0．28±0．08比0．63±0．12),脾细胞中IL-4 CD4~+细胞百分比(1．6±0．5比3．4±1．1)和血清IL-4质量浓度[(81±1 5)pg/ml比(124±20)pg/ml]升高(P＜0．05或P＜0．01)。结论姜黄素能治疗TNBS诱导的肠炎,其机制可能与调节肠道局部和机体Th1/Th2平衡有关。     

幽门螺杆菌相关性胃癌患者局部Th1/Th2细胞免疫应答变化

背景:幽门螺杆菌(Hp)感染与胃癌发生相关,免疫应答可能是其主要致病机制之一。目的:探讨Hp相关性胃癌患者黏膜局部Th1/Th2细胞免疫应答变化。方法:选取2011年1～6月27例胃癌患者和28例对照者,采用14C-尿素呼气试验、Hp抗体检测Hp感染,免疫组化法检测胃黏膜局部干扰素(IFN)-γ和白细胞介素(IL)-4表达情况。结果:Hp总体感染率为61.8%。Hp阳性患者中,胃癌组IFN-γ表达明显低于对照组(P<0.05),IL-4表达明显高于对照组(P<0.05);Hp阴性患者中,胃癌组IFN-γ、IL-4表达与对照组均无明显差异(P>0.05)。Hp阳性胃癌患者IL-4表达明显高于Hp阴性胃癌患者(P<0.05),Hp阳性对照组IFN-γ表达明显高于Hp阴性对照组(P<0.05)。结论:Hp阳性对照者以Th1细胞介导的免疫应答为主,Hp阳性胃癌患者以Th2细胞为主。黏膜局部Th1/Th2细胞免疫应答变化可能与Hp感染相关疾病的进展,特别是胃癌的发生密切相关。     

Helper (TH1, TH2, TH17) and regulatory cells (Treg, TH3, NKT) in rheumatoid arthritis

The immune response foreign antigens require a perfect coordination of cells that participate in its different phases. The objective of the response is the rapid destruction of the microorganisms with a minimum repercussion on self-cells and tissues. The regulation of this process is carried out fundamentally by T lymphocytes. There are two main types of coordinator cells: helper cells, what organize the initial immune response, and regulatory cells, what avoid immune attack against self and once the infection is controlled, disassemble the response. There are three types of helper cells which coordinate answers to intracellular parasites (TH1), helmints (TH2) and extracellular bacteria and fungi (TH17). The hyperfunction of TH17 cells is associated with diseases as reumatoid arthritis, due to the hypersecretion of the proinflammatory citoquine IL17. The condition of helper or regulatory cell is the current object of review. TH1, TH2 and TH17 cells have helper and also regulatory functions. In addition, regulatory T cells play an important role in the coordination of the first moments of the response to viral infection in a direct and indirect way, inducing differentiation of TH17 cells.     

TH亚群细胞因子在儿童ITP发病中的变化及作用

目的：探讨TH亚群细胞因子在儿童特发性血小板减少性紫癜（ITP）发病中的变化及作用。方法：采用定量夹心酶联免疫测定技术检测40例ITP患儿血清中的TH1型细胞因子IL－2、IFN－γ和TH2型细胞因子IL－4、IL－10的变化，采用改良的血小板抗原单抗特异性固相化法（MAIPA）测定这些患儿血清中抗GPⅡb/Ⅲa和抗GPⅠ/Ⅰx自身抗体。结果：（1）与正常健康儿童相比，ITP患儿TH2型细胞因子IL－4和IL－10显著升高（P＜0．05和0．01），而TH1型细胞因子IL－2和IFN－γ显著降低（P＜0．05和0．01），慢性ITP患儿的TH1型和TH2型细胞因子变化较急性患儿更为显著。（2）ITP患儿血清抗GPⅡb/Ⅲa和抗GPIb/Ix抗体的阳性率分别为67．5％和57．5％，抗体滴度与患儿外周血小板计数呈负相关（P＜0．01），抗体阳性患儿的TH1型和TH2型细胞因子变化较抗体阴性患儿更为显著（P＜0．05）。结论：儿童ITP发病中存在TH亚群平衡的偏离，表现为TH2型细胞因子功能亢进和TH1型细胞因子功能不足，从而出现体液免疫功能增强，B细胞过度活化，产生多种抗血小板自身抗体，导致血小板破坏而减少。     

TH1 and TH2 Cytokine Production Among Asthmatic Children After Immunotherapy

Allergen immunotherapy results in a number of changes in clinical and immunological parameters. To study the kinetic influence of immunotherapy on cytokine production, we evaluated the ratios of Th1/Th2 for patients receiving mite-extract immunotherapy with 3-month intervals. Changes in Th1/Th2 ratio were calculated based on the intracellular cytokine and surface CD4 staining of peripheral blood mononuclear cells (PBMC). No significantly different Th1/Th2 ratios were detected after immunotherapy, although ratios were lower for asthmatic children when compared with normal controls. Cytokine production was also determined on supernatants from mite- or mitogen-activated PBMC cells. A significantly increased production of all cytokines was detected from activated PBMC from patients after immunotherapy. Thus hyposensitization with mite extract for 1 year might improve the cytokine production capacity of asthma patients' PBMC.     

TH1 and TH2 Cytokine Production Among Asthmatic Children After Immunotherapy

Allergen immunotherapy results in a number of changes in clinical and immunological parameters. To study the kinetic influence of immunotherapy on cytokine production, we evaluated the ratios of Th1/Th2 for patients receiving mite-extract immunotherapy with 3-month intervals. Changes in Th1/Th2 ratio were calculated based on the intracellular cytokine and surface CD4 staining of peripheral blood mononuclear cells (PBMC). No significantly different Th1/Th2 ratios were detected after immunotherapy, although ratios were lower for asthmatic children when compared with normal controls. Cytokine production was also determined on supernatants from mite- or mitogen-activated PBMC cells. A significantly increased production of all cytokines was detected from activated PBMC from patients after immunotherapy. Thus hyposensitization with mite extract for 1 year might improve the cytokine production capacity of asthma patients' PBMC.     

Role and development of TH1/TH2 immune responses in the lungs

The development of T helper 1 (Th1) and Th2 responses is dependent on the cells and early signals of the innate immune system. Following inhalation of pulmonary pathogens, lung antigen-presenting cells (APCs) ingest the microbe, begin to process antigen, and migrate to peripheral lymphoid tissues (i.e., LALNs). It is in the lymph node that the APC-T cell interaction takes place; therefore, the microenvironment of the lymph node significantly influences the developing T cell response (Th1 vs Th2). Several factors can determine the nature of the T cell response, including cytokines, chemokines, microbial virulence factors, and dendritic cell phenotype. A shift in the Th1/Th2 balance in the lungs can result in chronic infection, allergic disease, and immunopathology. This review discusses the mechanisms of developing Th1/Th2 pulmonary responses, the counterregulation of Th1/Th2 immunity, and the consequences of immune deviation in the lungs.     

Methotrexate selectively modulates TH1/TH2 balance in active rheumatoid arthritis patients

OBJECTIVE: The mechanism by which low dose methotrexate (MTX, the gold standard treatment for rheumatoid arthritis) exerts its anti-inflammatory effect in rheumatoid arthritis (RA) patients is still debated. Lately, the MTX immunosuppressive effect has been related to apoptosis, especially in active RA patients, with ROS involvement. METHODS: In the present research we investigated MTX oxidative effect and its ability to modulate immune balance in active versus non-active RA patients. RESULTS: Our results show that MTX induces IL-10 secretion (a TH2 cytokine) and significantly reduces TH1 profile in Peripheral Mononuclear Cells (PMNC) derived from active RA patients (n=28). Additionally, we found that MTX modulates the immune status towards TH2 dominance by decreasing the IL-12R and the CXCR3 receptors typical for the TH1 population. Moreover, MTX was found to inhibit the production of nitric oxide (NO) in these patients, a phenomenon that might contribute to MTX action toward cytokine homeostasis. A significant correlation was found between MTX IL-10 induction and NO inhibition in active RA patients. CONCLUSION: Our data suggest that, in active RA patients, apoptosis induction by MTX may be primarily due to IL-10 production via modulation of oxidative stress, which may restore the critically important immune balance. These findings may contribute to determining which group of RA patients may better respond to MTX therapy.