脾动脉缩窄对门静脉高压大鼠脾脏iNOS表达及Th1/Th2平衡的影响

目的 观察脾动脉缩窄对肝硬化门静脉高压大鼠脾脏iNOS、Th1/Th2型细胞因子表达的影响,并探讨机制.方法 肝硬化门静脉高压大鼠随机分3组(n=10):假手术组(SOG)、脾动脉缩窄术组(SAC)和脾动脉结扎术组(SAL);正常大鼠10只行假手术作为对照组(NCG).免疫组化法测脾脏iNOS表达,RT-PCR法测脾脏IFN-γ、IL-4mRNA表达,对iNOS与IFN-γ、IL-4表达量作相关分析.结果 SOG脾脏iNOS明显高于NCG(P＜0.01),SAC和SAL明显低于SOG(P＜0.01).SOG脾脏IFN-γmRNA和IFN-γ/IL-4明显低于NCG(P＜0.01),IL-4mRNA明显高于NCG(P＜0.01);SAC脾脏IFN-γmRNA高于SOG(P＜0.05),SAC和SAL脾脏IL-4mRNA低于SOG(P＜0.05),而IFN-γ/IL-4高于SOG(P＜0.05).iNOS与IFN-γ负相关(r=-0.672,P＜0.01),与IL-4正相关(r=0.634,P＜0.01).结论 脾动脉缩窄术后门静脉高压大鼠脾脏iNOS表达降低,IFN-γ/IL-4升高,脾脏Th1/Th2失衡改善可能与术后iNOS表达降低有关.     

Th1/Th2细胞因子mRNA的表达与心脏移植免疫耐受的关系

目的探讨Th1/Th2细胞因子信使核糖核酸(mRNA)表达改变与心脏移植免疫耐受的关系. 方法采用大鼠腹部心脏移植模型,将30只大鼠随机分成对照组、排斥反应组、免疫耐受组,每组10只.观察移植心脏存活时间,供心病理学改变,受者脾和心脏中Th1/Th2细胞因子白细胞介素2(IL-2)、γ干扰素(IFN-γ)、白细胞介素4(IL-4)、白细胞介素10(IL-10) mRNA表达水平. 结果免疫耐受组供心存活时间为85.28±7.48天,较排斥反应组的7.33±1.03天显著延长(P＜0.01);排斥反应组供心见大量炎性细胞浸润,免疫耐受组供心仅见少量炎性细胞浸润;排斥反应组Th1细胞因子IL-2、IFN-γ mRNA表达较对照组增强,免疫耐受组减弱;排斥反应组Th2细胞因子IL-4、IL-10 mRNA表达较对照组减弱,免疫耐受组增强. 结论 Th1/Th2细胞因子的动态平衡在移植耐受中起重要作用,Th1向Th2偏离是移植耐受的机制之一.     

PIAS调控PPARγ的SUMO化修饰在小鼠内毒素性急性肺损伤内源性保护机制中的作用

目的评价小泛素相关修饰物(SUMO) E3连接酶(PIAS)调控过氧化物酶体增殖物激活受体γ(PPARγ)的SUMO化修饰在小鼠内毒素性急性肺损伤(ALI)内源性保护机制中的作用。方法实验Ⅰ清洁级野生型雄性C57BL/6小鼠24只, 6～8周龄, 体质量18～22 g, 采用随机数字表法分为4组(n=6):对照组(C组)、ALI组、ALI+ PPARγ诱导剂TZD组(ALI+T组)、ALI+TZD+SUMO化抑制剂漆树酸组(ALI+T+A组)。尾静脉注射LPS 15 mg/kg制备内毒素性ALI模型。ALI+T+A组注射LPS前1 h时腹腔注射漆树酸5 mg/kg;ALI+T组和ALI+T+A组注射LPS前30 min时腹腔注射TZD 50 mg/kg。给予LPS 12 h后处死小鼠取肺组织, 测定湿重/干重(W/D)比值, 光镜下观察病理学结果, 并行肺损伤评分;分别采用Western blot法和PCR法测定PIAS1、PIAS2、PIAS3和PIASy及其mRNA的表达。实验Ⅱ 体外培养的小鼠肺泡巨噬细胞(MH-S细胞)采用随机数字表法分为4组(n=5):对照组(C组)、LPS...     

骨髓移植小鼠急性移植物抗宿主病早期弥漫性肺损伤中TH17细胞的作用

目的 探讨骨髓移植小鼠急性移植物抗宿主病(aGVHD)早期肺损伤中TH 17细胞的作用和机理.方法 Balb/c小鼠为受鼠,经致死量全身照射(TBI)后,输注C57BL/6小鼠来源的骨髓细胞和脾细胞,建立aGVHD模型.实验分为3组:TBI组小鼠仅接受TBI,异基因骨髓移植(BMT)组小鼠TBI后输注供者骨髓细胞和脾细胞,常山酮(HF)组小鼠TBI后输注骨髓细胞和脾细胞,并注射HF.动态观察小鼠GVHD的表现,并进行肺组织病理学、T淋巴细胞亚群及相关细胞因子的检测.结果 移植后小鼠出现典型GVHD的表现.移植后6d时HF组肺组织病理学评分为(2.00±0.35)分,BMT组为(0.67±0.07)分.BMT组TH 1细胞占CD4+T淋巴细胞的比例为(5.53±0.11)％,TH 17细胞占(1.04±0.34)％;HF组TH1细胞占(8.61±0.21)％,TH 17细胞占(0.49±0.07)％;组间比较,差异有统计学意义(P＜0.05).两组均未检测到TH2细胞.移植后6d,BMT组白细胞介素(IL)-17A为(2.81±0.19)pg/ml,γ干扰素(IFN-γ)为(42.97±0.23) pg/ml; HF组IL-17A＜0.8 pg/ml,IFN-γ为(9.89±0.51)pg/ml;组间比较,差异有统计学意义(P＜0.05).两组均未检测到IL-10.结论 在异基因造血干细胞移植早期阻断TH17细胞及其细胞因子的分泌,导致炎症因子分泌失衡,加重肺损伤.     

白细胞介素-7促进CD8+T细胞干扰素-γ的分泌增强小鼠抗乳腺肿瘤的免疫效应

目的 观察白细胞介素(IL)-7是否通过促进CD8+T细胞干扰素-γ(IFN-γ)分泌来增强小鼠抗乳腺肿瘤的免疫效应.方法 荷瘤小鼠瘤体内直接注射IL-7真核表达质粒(pcDNA3-IL-7);免疫磁珠分选CD4+T、CD8+T细胞并体外培养;流式细胞仪检测细胞内干扰素(IFN)-γ的分泌量;酶联免疫吸附试验(ELISA)检测培养上清IFN-γ的量;噻唑蓝(MTT)法检测CD8+T细胞体外杀伤活性;小鼠体内预先注射anti-CD8抗体阻断活化.结果 pcDNA3-IL-7注射组CD8+T细胞内IFN-γ(38.6±4.4)％、CD8+T细胞培养上清IFN-γ( 136.0±11.2) ng/L,与对照组比较,pcDNA3-IL-7明显促进CD8+T细胞IFN-γ表达量(P＜0.05);IL-7处理过的荷瘤小鼠CD8+T细胞杀伤活性显著增强,尤其效靶比为100∶1;小鼠体内预先注射anti-CD8抗体阻断CD8+T细胞活化,显著抑制IL-7抗瘤效应(P＜0.05).结论 IL-7通过促进小鼠体内CD8+T细胞分泌IFN-γ介导抗瘤效应,从而增强小鼠机体抗乳腺肿瘤的免疫反应.     

己酮可可碱对大鼠内毒素性急性肺损伤炎症反应的影响

目的探讨己酮可可碱对大鼠内毒素(LPS)诱导急性肺损伤(ALI)炎症反应的影响。方法腹腔注射0.01%LPS1mg/kg,16h后在机械通气下气管内滴注1.5mg/kg(0.5ml)LPS建立大鼠内毒素性ALI模型。24只大鼠随机分为生理盐水对照组(C组)、急性肺损伤组(LPS组)和己酮可可碱组(PTX组),每组8只。7h后处死大鼠。酶联免疫吸附法(ELISA)测定支气管肺泡灌洗液(BAL)中肿瘤坏死因子α(TNF-α)、白细胞介素-10(IL-10)的含量,测肺湿/干重比,并观察BAL白蛋白浓度。结果与LPS组相比,PTX组大鼠BAL中TNF-α浓度、肺湿/干重比以及BAL白蛋白浓度显著降低(P<0.05),而IL-10显著升高。结论己酮可可碱能显著抑制内毒素性急性肺损伤大鼠的炎症反应,具有一定的肺保护作用。     

消退素E1对急性肺损伤小鼠炎症反应的影响

目的 观察消退素E1( RvE1)对急性肺损伤(ALI)小鼠炎症反应的影响.方法 建立小鼠ALI模型,12h后处死,观察RyE1治疗对肺组织形态、湿/干比、肺泡灌洗液(BALF)肿瘤坏死因子-α( TNF-α)含量、组织匀浆核因子-κB( NF-κB)蛋白水平的影响.结果 给予RvE1治疗后,肺组织损伤明显减轻;RYE1治疗组肺湿/干比值(4.637 ±0.125) g/g、TNF-α含量(217.2 ±30.1)ng/L较ALI模型对照组肺湿/干比值(4.906±0.176) g/g和TNF-α含量(372.2±20.6)ng/L均明显降低,差异有统计学意义(P ＜0.05);RvE1治疗组肺组织匀浆中NF-κB蛋白表达水平较ALI模型对照组明显下降.结论 RvE1能减轻肺部炎症反应,从而减轻肺组织损伤,对ALI具有保护作用.     

Toll样受体4对失血性休克小鼠肺组织血红素加氧酶和诱导型一氧化氮合酶的影响

目的 观察Toll样受体4(TLR4)对失血性休克小鼠所致急性肺损伤(ALI)中肺组织血红素加氧酶-1(HO-1)和诱导型一氧化氮合酶(iNOS)的影响.方法 TLR4基因突变型小鼠C3H/HeJ和野生型小鼠C3H/HeN(48只),随机分为假手术组和失血性休克(6、24、48 h)组.采用小鼠失血性休克致急性肺损伤模型,观察血气分析,HO-1和iNOS蛋白表达,白细胞介素-6(IL-6)水平,肺组织肺湿/干重比和病理形态学改变.结果 与假手术组比较,C3H/HeN和C3H/HeJ小鼠失血休克后24 h肺组织HO-1蛋白强阳性表达,IL-6含量明显增加为365.38±48.26和300.89±39.34;6 h肺组织iNOS蛋白强阳性表达(P＜0.01).与C3H/HeN小鼠比较,C3H/HeJ小鼠失血休克后24 h肺组织HO-1、IL-6含量和W/D明显降低(P＜0.05);6 h肺组织iNOS蛋白显著减少为0.049±0.013.病理学检查显示失血休克后各时点肺组织损伤程度较假手术组明显加重.结论 TLR4在失血性休克后ALI过程中被激活,通过影响HO-1和iNOS的表达参与了失血性休克致急性肺损伤的过程.     

白细胞介素-2、白细胞介素-15、α-干扰素、粒细胞-巨噬细胞集落刺激因子体外转录mRNA组合抑制小鼠乳腺癌的生长

目的本研究旨在探讨白细胞介素(IL)-2、IL-15、α-干扰素(IFN-α)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)细胞因子组合体外转录mRNA注射小鼠乳腺癌后的治疗效果。方法体外转录合成细胞因子IL-2、IFN-α、GM-CSF和IL-15 mRNA组合, 并分别在体外转染4T1乳腺癌细胞, 检测对应蛋白的表达。使用4T1乳腺癌细胞系构建BALB/c小鼠乳腺癌模型, 并对其注射4种mRNA组合, 通过酶联免疫吸附试验(ELISA)检测注射后第1、4、7、10天小鼠血液中4种细胞因子的水平;测量注射后小鼠肿瘤的大小;采用免疫组织化学方法检测肿瘤中细胞因子的表达。两组间比较采用独立样本t检验, 采用单因素方差分析进行多组间比较。结果 Western blot实验结果显示, 转染组IL-2、IFN-α、GM-CSF和IL-15的表达分别为均显著高于对照组(tIL-2=4.170, P<0.05;tIFN=9.648, P<0.01;tGM-CSF=16.250, P<0.01;tIL-15=13.550, P<0.01)。与对照组小鼠比较, 体外转录mRNA...     

Characterization of TH1/TH2 profile in uremic patients

End-stage renal failure (ESRD) induces a clinical state of immunodeficiency with a higher incidence of infections and a higher mortality due to infectious complications compared with the normal population. Using a newly developed immunofluorescent staining of intracellular cytokines for flow cytometric analysis, we studied Th subsets in 22 healthy control subjects, 28 patients with compensated chronic renal failure (CRF), 25 patients on hemodialysis (HD), and 24 patients on continuous ambulatory peritoneal dialysis (CAPD). Our results demonstrate that the percentage of both interferon-gamma-positive cells and interleukin-4-positive cells increased in compensated CRF patients compared with those in healthy subjects. Moreover, a significantly higher percentage of CD4-positive cells is characterized by a Th1-type cytokine production pattern in HD patients and by a Th2-type cytokine secretion pattern in CAPD patients. These results suggest that the altered Th1/Th2 balance may be associated with the pathogenesis of ESRD.     

TH1/TH2 cytokine analysis in Iranian renal transplant recipients

The pretransplant cytokine profile of donor and recipient blood and tissues may be associated with transplant outcome. A Th1 response is generally associated with transplant rejection, while a Th2 response may lead to tolerance and stable graft survival. A total of 56 (37 male and 19 female) patients of mean 36 +/- 5 years were candidates for living unrelated kidney transplantation. Serum samples were collected 24 hours pretransplantation as well as at 1 and 2 weeks posttransplantation. Immunosuppression consisted of cyclosporine, prednisolone, and mycophenolate mofetil. Among the transplanted patients, 19 (33.9%) individuals experienced an acute rejection episode, as proven by biopsy, as well as an increased serum creatinine and blood urea nitrogen, within 14 days after transplantation. We determined serum concentrations of interleukin (IL) 2 and interferon (IFN)-gamma for Th1 and IL4 and IL10 for Th2 by an enzyme-linked immunosorbent assay method (Bender med system kits, Germany). Among Th1 cytokines, the mean concentration levels for groups with versus without acute rejection were: IL-2 pretransplant 15 pg/mL vs 6.8 pg/mL, respectively (P = .005); IL-2 at 1 week, 19 pg/mL vs 4.85 pg/mL, respectively (P = .001); IL-2 at 2 weeks, 21.1 pg/mL vs 4.65 pg/mL, respectively (P = .0001); IFN-gamma pretransplant 161.1 pg/mL vs 65.2 pg/mL, respectively (P = .001); IFN-gamma at 1 week, 175.6 pg/mL vs 66.5 pg/mL, respectively (P = .001); and IFN-gamma at 2 weeks, 173.7 pg/mL vs 77.1 pg/mL (P = .001). IL-2 and IFN-gamma levels were significantly higher in the group with acute rejection versus those without acute rejection. In conclusion, these data suggest that cytokine analysis, especially of Th1 cytokines, might be a valuable prognotic index of kidney transplant outcome.     

Role of TH1/TH2 Cytokines in Kidney Allograft Rejection

One of the major issues in contemporary kidney transplantation is prevention of acute allograft rejection episodes (AREs). Cytokines are crucial mediators of immune reactions leading to AREs. We correlated serum Th1/Th2 cytokine concentrations with AREs. The project included 44 patients undergoing kidney transplantation. During the 3-month period following the transplantation, ARE was diagnosed in 11 patients. Serum samples collected 1 day before and 2, 7, 14, and 30 days after transplantation were tested for interleukin (IL)-2, IL-4, IL-5, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α concentrations using flow cytometry. Nonrejection (NONAR) and rejection (ARE) groups of patients did not show significant differences in baseline demographic characteristics. We observed that higher pretransplantation serum levels of IFN-γ (P = .000003) and IL-10 (P = .000001) were associated with AREs. Our analysis also showed slightly higher IL-4 serum levels among NONAR patients up to 7 days posttransplantation, followed by a drop in concentrations in NONAR patients. In contrast, there was a continuous increase among ARE patients. No significant differences were observed in plasma levels of IL-2, IL-5, IL-10, or TNF-α between the two groups. Higher pretransplantation levels of IFN-γ and IL-10 observed in ARE patients indicated ongoing nondetected, probably nonspecific, inflammatory processes able to intensify an immune response directed against the transplanted organ leading to its acute rejection. Higher levels of IL-4 prior to and shortly after transplantation may have protective effects on graft survival. However, a prolonged, increased production of IL-4 after transplantation can also contribute to AREs.     

黄芪地黄汤对小儿过敏性紫癜肾炎TH1/TH2的影响

目的：观察黄芪地黄汤治疗小儿过敏性紫癜肾炎（HSPN）的临床疗效及其对免疫功能的影响,探讨临床疗效和免疫功能变化之间的关系。方法：收集HSPN患者80例。随机分成两组,西医组40例,中西医结合组40例（西医组用药基础上加用黄芪地黄汤）,并建立正常儿童对照组30例。观察疗程3个月。测定治疗前后相关细胞因子（IL-2、IL-4、IL-6、IL-12、TNF-α）水平及血尿常规、肝肾功能、免疫球蛋白、24h尿蛋白定量等变化。结果：与正常儿童对照组比较,西医组和中西医结合组的IL-12水平均下降,IL-4、IL-6水平上升（P均〈0.05）,TNF-α水平明显上升（P〈0.01）,而IL-2水平无统计学意义（P〉0.05）。3个月后,中西医结合组总缓解率87.5%,西医组疗效总缓解率60%,二者有显著差别（P〈0.05）;中西医结合组IL-12、IL-4、IL-6水平基本降至正常,TNF-α水平仅略高于正常对照组;西医组IL-12水平接近正常水平,IL-4、IL-6及TNF-α水平略有下降。结论：黄芪地黄汤结合西药常规治疗可更有效逆转TH2亢进状态,使病情得到改善和防止进展,其疗效机制与免疫功能调控相关。     

Evaluation of TH17 and TH1 Immune Response Profile in Patients After Renal Transplant

Background

Renal transplantation (RT) is the best treatment option for patients with end-stage renal disease (ESRD) because it improves both quality of life and survival. However, allograft rejection remains the most important barrier to successful transplantation. Underlying immunologic mechanisms should be understood to develop appropriate treatment strategies.

TH1/TH2 cytokine release pattern during in vivo cytomegalovirus disease in solid organ transplantation

BACKGROUND: Cytomegalovirus (CMV) disease is associated with an increased net immunosuppressive state in solid organ transplant recipients, leading to more bacterial and fungal infections. The release of pro- and anti-inflammatory cytokines could be one of the responsible factors. METHODS: We prospectively included all patients undergoing solid organ transplantation between April and November 2004. During follow-up, plasma samples were collected in the immediate postsurgical period, at the first and second months, at the time of maximum antigenemia during CMV disease, and at 6 months posttransplantation. We determine the levels of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-10. Log-transformed data were compared by a nonparametric Wilcoxon test for related variables. RESULTS: During the study period, we monitored 146 recipients of solid organ transplantation: 77 kidneys, 8 kidney-pancreas, 46 liver, 11 heart, 2 liver-kidney, and 2 heart-kidney. No differences were observed between the TNF-alpha and IL-10 levels in the immediate postsurgical period or during CMV disease. TNF-alpha and IL-10 levels during CMV disease were higher than levels during the first month (mean TNF-alpha first month = 12.71 pg/mL vs CMV disease = 22.71 pg/mL, P = .028; mean IL-10 first month = 3.47 pg/mL vs CMV disease = 19.2 pg/mL, P = .018). Th1/Th2 ratio (measured as TNF-alpha/IL-10) was 1.75 in the immediate postsurgical period, 7.5 during the first month, 1.86 at the time of CMV disease, and 4.61 at the sixth month. The difference in Th1/Th2 ratio during CMV disease and in the first month was statistically significant (P = .043). CONCLUSION: During CMV disease, we observed an increase in TNF-alpha and IL-10 release, which was similar to that during the postsurgical period. An imbalance toward an anti-inflammatory pattern was noted in these two periods. This could reflect a cooperative factor increasing the net state of immunosuppression during CMV disease.