Ⅲ型前列腺炎辅助性T细胞亚群的分化

目的 了解Ⅲ型前列腺炎前列腺液中CD4^＋T辅助性T细胞（Th细胞）亚群的分化情况。方法 按照美国国立卫生院（NIH）分类方法将病例分成ⅢA型／组（47例）、ⅢB型／组（29例），另设健康对照组（16例）。采用双抗体夹心酶联免疫法检测前列腺液（EPS）中Th；类细胞因子（IFN-γ）、Th2类细胞因子（IL-4）水平以及Th1／Th2比值（IFN-γ／IL-4）。结果 和对照组比较，ⅢA组、ⅢB组IFN-γ水平明显上调（P〈0．05），ⅢA组较ⅢB组升高更明显（P〈0．05）；和对照组比较，ⅢA组IL-4水平无明显变化（P〉0．05），ⅢB组IL-4水平上调（P〈0．05）：和对照组比较，ⅢA组IFN-γ／IL-4明显升高（P〈0．05），ⅢB组IFN-γ／IL-4无明显改变（P〉0．05）。结论 ⅢA型前列腺炎Th；细胞分化占优势，Th1／Th2平衡向Th1漂移，以细胞免疫反应为主；Th细胞分化也参与了ⅢB型前列腺炎的发病，但Th1／Th2处于相对平衡状态。     

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.     

自身免疫性甲状腺病Th1／Th2细胞因子平衡的偏离

目的探讨自身免疫性甲状腺病中Graves病和桥本甲状腺炎患者外周血Th1／Th2细胞因子平衡偏离的方向。方法应用酶联免疫吸附法分别检测50例Graves病患者，20例桥本甲状腺炎患者和20例健康献血者外周血γ干扰素（IFN-γ）和白介素-4（IL-4）。以IFN—γ/IL-4作为反映Th1/Th2细胞因子的指标。结果与正常对照组对比，Graves病和桥本甲状腺炎患者外周血IFN-γ无明显变化（P〉0．05），IL-4明显下降（P〈0．01），而IFN—γ/IL-4比值升高（P〈0．05）。结论自身免疫性甲状腺病中Graves病和桥本甲状腺炎外周血Th1/Th2细胞因子平衡显现Th2细胞因子低下型，而Th1／Th2相对升高。     

血液净化治疗多器官功能障碍综合征引起Th1/Th2漂移的研究

目的研究连续性血液净化治疗（CBP）对多器官功能障碍综合征（multiple organ dysfunction syndrome，MODS）患者循环Th1／Th2的影响及其引起Th1／Th2漂移的机制。方法18例多器官功能障碍综合征患者行CBP治疗。采用中心静脉留置单针双腔导管建立血管通路，采用连续性血液净化系统进行前稀释连续性静脉-静脉血液透析滤过（CVVHDF）。治疗前及治疗后24、48、72h采血检测Th1／Th2，同时检测血液中IL4和IFN-1浓度。结果MODS患者经CBP治疗后，Thl表达逐渐增加，而Th2表达逐渐下降，IFN-γ和IL-4的浓度均下降；Th1／Th2和IFN-γ／IL-4比值随治疗时间的增加升高；Th1／Th2和IFN-γ/IL-4比值呈正相关。结论CBP治疗可能可改善MODS患者的Th1／Th2平衡漂移，可能与改善机体内IFN-γ/IL-4比值有很大的相关性。     

血栓痔病人外周血Th1／Th2分析

为探讨血栓痔发病与T淋巴细胞活化及释放的Th1／Th2细胞因子失衡状态的关系,对32例血栓痔（实验组）和30例健康人（对照组）的外周血单个核细胞（PBMC）,经PMA诱导培养后,用流式细胞术测定CD3^＋标记T细胞内自细胞介素-4（IL-4）和干扰素-γ（IFN-γ）含量和CD8^-标记T细胞内IL-4和IFN-γ的含量,进行比较分析,从而估计Th1／Th2细胞的漂移状态。结果显示,实验组的IL-4和IFN-γ含量均显著高于对照组（P〈0．001）;且实验组IL-4^＋含量高于IFN-γ含量（P〈0．05）。结果表明,血栓痔病人发病时,体内的T淋巴细胞免疫系统被激活,且以Th2细胞为主,这种Th1／Th2细胞因子失衡可能在血栓痔发病过程中起到重要作用。     

Th1/Th2类细胞因子转换对小鼠心脏移植物存活的影响

目的 探讨Th1/Th2类细胞因子转换对小鼠心脏移植物存活时间的影响。方法 采用小鼠腹部心脏移植模型，分为同种异体移植组（A组）、同种异体移植+免疫抑制处理组（B组）和同系移植组（C组），每组20对。观察移植物存活时间、供心病理改变、受鼠脾和供心内IFN-γ，IL-2，IL-4及IL-10 mRNA的表达水平。结果 A组及B组移植物平均存活时间分别为（7.8±0.77）d和（14.80±1.01）d，C组移植物存活均超过28d；3组间差异均有显著性（P<0.05）。A组与B组、C组比较，移植物的心肌细胞变性坏死严重，并有大量炎性细胞浸润。A组受鼠脾脏及供心内IFN-γ和IL-10 mRNA表达比其余两组明显增强；3组移植心组织IL-2及IL-4 mRNA均无表达；A组脾脏IL-2 mRNA表达最强；B组脾脏IL-4 mRNA表达明显强于其余两组。结论 Th1/Th2转换在延长移植物存活过程中起重要作用；IL-10也参与移植物排斥反应过程。     

慢性乙型肝炎肝纤维化患者Th1／Th2细胞及其细胞因子的变化

目的探讨Th1／Th2细胞及其分泌的细胞因子在慢性乙型肝炎（CHB）肝纤维化患者发生发展中的作用。方法选取2011年3—10月在河北医科大学第三医院感染科接受肝组织病理活检的CHB肝纤维化患者46例,根据纤维化分期分为S0～1期15例,S2～3期20例,S4期11例三组,另取同期健康志愿者10名作为健康对照组。用流式细胞术和实时荧光定量PCR检测外周血Th1、Th2细胞频率和单个核细胞（PBMCs）中干扰素γ（IFN-γ）、白细胞介素4（IL-4）基因表达;用ELISA法检测血清IFN-γ和IL-4水平;用免疫组织化学染色检测肝组织IFN-γ和IL4表达。应用Spearman相关分析、Kruskal—WallisH检验、Logistic逐步回归方法分析Th1／Th2细胞及其细胞因子在CHB肝纤维化患者外周血和肝组织中的变化。结果不同肝纤维化分期的CHB患者外周血Th1／Th2细胞频率的比值、PBMCs中IFN-γ／IL-4mRNA、血清中IFN-γ／IL-4及肝内IFN-γ／IL-4差异均有统计学意义,并随肝纤维化程度加重逐渐下降（χ2=36．259、40．822、26．321和31．852,P〈0．05）。血清及肝组织中IFN-γ,／IL-4比值与肝纤维分期均呈负相关（r=-0．616和-0．531,P〈0．01）。Logistic回归分析显示：AST、凝血酶原时间（PT）和血清IFN-γ／IL4比值是发生显著肝纤维化（S2-4）的三个独立危险因素（OR=5．933,95％CI：1．324～26．586,P=0．02;OR=12．866,95％CI：1．746～94．788,P=0．01;OR=4．755,95％CI：1．034～21．862,P：0．04）。结论CHB患者体内存在Th1／Th2失衡,随着肝纤维化进展,外周血及肝脏内均呈现Th2应答的极化现象,这可能是CHB肝纤维化发生发展的一种机制。     

肝癌合并肝硬化患者脾脏Th1/Th2细胞因子免疫状态的研究

目的：探讨肝癌合并肝硬化患者脾脏Th1/Th2细胞因子分泌状态的变化。方法：以肝硬化患者为对照组，用酶联免疫吸附（ELISA）方法分析了31例肝癌合并肝硬化患者外周血有脾静脉血γ－干扰素（IFN－γ），白细胞介素（IL）－2，IL－10蛋白水平的表达。结果：6例Ⅰ期肝癌患者外周血IL－2，脾静脉血ITNF－γ高于肝硬化组（P＜0．01或P＜0．05），外周及脾静脉IL－10低于肝硬化组（P＜0．05）。12例Ⅱ期13例Ⅲ期肝癌患者外周血及脾静脉血IFN－γ，IL－2明显低于肝硬化组（P＜0．01或P＜0．05），而IL－10明显高于肝硬化组（P＜0．05），且脾静脉血IFN－γ，IL－2低于外周血，IL－10高于外周血水平（P＜0．05）。结论：Ⅰ期肝癌合并肝硬化患者机体免疫系统通过强势分泌Th1细胞因子发挥抗肿瘤免疫作用，而脾脏在维持Th1/Th2细胞因子平衡中起重要作用。随着肿瘤进展，机体细胞免疫功能受损，中晚期肝癌患者Th1细胞因子水平降低，Th2细胞因子水平增高，机体处于免疫抑制状态，而脾脏Th1/Th2细胞因子紊乱更为明显，呈现负性抗肿瘤免疫作用，加重了机体免疫抑制。     

细胞黏附分子在高容量血液滤过防治多器官功能障碍中的表达变化及意义

目的研究细胞黏附分子在高容量血液滤过（HVHF）干预多器官功能障碍（MODS）猪模型过程中的表达变化情况。方法18头实验动物被随机分为2组：模型组（M组）实施失血性休克＋内毒素注射“二次打击”，而治疗组（HF组）在二次打击后接受HVHF干预。检测两组中性粒细胞（PMN）表面CD11b、L-选择素（selectin）的表达和血浆中可溶性E选择素（sE-seleetin）、可溶性细胞间黏附分子（sICAM-1）的浓度。结果HVHF治疗开始后，HF组CD11b表达出现下降，在各个时间点与M组比较，差异有统计学意义P〈0．01；治疗组中L-selectin的表达在治疗后24、48h处表达高于M组的表达（P〈0．05）。治疗开始后sE-selectin浓度就出现明显下降，治疗后24h与M组比较差异有统计学意义（P〈0．01），到治疗后48h（P〈0．05）和72h（P〈0．01）已经高于M组水平；治疗后24h，HF组中sICAM-1水平低于M组（P〈0．05），此后维持于较高水平，至治疗后72h高于M组（P〈0．01）。结论HVHF可以间接、双向调节MODS发生过程中细胞黏附分子的表达水平以及PMN、内皮细胞的活化状态。     

慢性非细菌性前列腺炎辅助性T细胞亚群分化及其对炎症的影响

目的：了解慢性非细菌性前列腺炎（CP）前列腺液（EPS）中Th类细胞亚群分化及其和局部炎症的关系。方法：对47例CP患者（CP组）,根据炎症程度分成Ⅰ组（轻度炎症）26例和Ⅱ组（重度炎症）21例;另以16例健康者为对照组。采用双抗体夹心酶联免疫法法检测各组EPS中Th1类细胞因子（IFN-γ）、Th2类细胞因子（IL-4）水平,以及Th1／Th2比值（IFN-γ／IL-4）。结果：与对照组比较,CP组IFN-γ、IFN-γ／IL-4明显升高（P〈0．05）,IL-4水平无明显变化;其中,Ⅰ组IL-4水平无明显变化,而Ⅱ组IL-4水平显著性下调（P〈0．05）;Ⅱ组和Ⅰ组IFN-γ水平均明显上调（P〈0．05）,但此两组间水平无显著性差别;Ⅱ组IFN-γ／IL-4明显高于Ⅰ组（P〈0．05）。结论：CP患者Th1细胞分化占优势,以细胞免疫反应为主;Th1的极化可能是导致前列腺局部炎症发展原因之一。     

Th1 and Th17 Cells Induce Proliferative Glomerulonephritis

Th1 effector CD4+ cells contribute to the pathogenesis of proliferative and crescentic glomerulonephritis, but whether effector Th17 cells also contribute is unknown. We compared the involvement of Th1 and Th17 cells in a mouse model of antigen-specific glomerulonephritis in which effector CD4+ cells are the only components of adaptive immunity that induce injury. We planted the antigen ovalbumin on the glomerular basement membrane of Rag1^−/− mice using an ovalbumin-conjugated non-nephritogenic IgG1 monoclonal antibody against α3(IV) collagen. Subsequent injection of either Th1- or Th17-polarized ovalbumin-specific CD4+ effector cells induced proliferative glomerulonephritis. Mice injected with Th1 cells developed progressive albuminuria over 21 d, histologic injury including 5.5 ± 0.9% crescent formation/segmental necrosis, elevated urinary nitrate, and increased renal NOS2, CCL2, and CCL5 mRNA. Mice injected with Th17 cells developed albuminuria by 3 d; compared with Th1-injected mice, their glomeruli contained more neutrophils and greater expression of renal CXCL1 mRNA. In conclusion, Th1 and Th17 effector cells can induce glomerular injury. Understanding how these two subsets mediate proliferative forms of glomerulonephritis may lead to targeted therapies.Although proliferative and crescentic glomerulonephritides occur in different primary renal diseases and are an important component of several systemic diseases, features of human renal biopsies suggest some common effector pathways. In most cases of rapidly progressive GN there is evidence for an important role for cellular immune effectors: T cells, macrophages, and neutrophils,^1–3 a role confirmed in animal models.^4–7 CD4+ T cells are key components of renal injury.^4,8 When activated, CD4+ cells tend to differentiate into subsets (T helper cells—Th1, Th2 and Th17) that engage immune effectors in different ways. In proliferative forms of GN, T cells direct adaptive immune responses that drive glomerular disease, but also, in rapidly progressive GN, CD4+ cells themselves accumulate in glomeruli as effectors. These effector T helper cells activate innate effector cells, predominantly neutrophils and macrophages, and activate and damage intrinsic renal cells.In GN, the variable Th1-Th2 predominance of responses influences the histologic patterns and severity of GN.⁹ Th1 cells, which secrete IFNγ and activate macrophages, are important in some forms of experimental proliferative GN. Th2 cells, characterized by IL-4 production, promote humoral immunity and are important in several forms of GN, but there is little evidence that Th2 cells play primary roles as effector cells within glomeruli in rapidly progressive GN. A binary Th1/Th2 model explains many of the differences in the patterns of immune responses in GN. However, there are discrepancies¹⁰ that might be explained by defining a role for a third major subset, Th17 cells, characterized by the production of IL-17A. Its biology has recently been comprehensively reviewed.¹¹ Th17 subset effects potentially relevant to rapidly progressive GN include direct effects on neutrophils and stimulating the production of neutrophil chemoattractants by tissue cells. Thus, in most rapidly progressive types of GN, cell-mediated injury, a key component of injury, may be directed by the Th17 subset, the Th1 subset, or both subsets.Although antigen-specific T cells are critical to adaptive immune responses, the cells themselves are relatively infrequent. T cell receptor (TcR) transgenic mice help define the contributions of different antigen-specific T helper cell subsets in organ-specific disease.^12–14 In the studies presented here we have established a new antigen-specific model of GN. Ovalbumin (OVA)-specific OT-II TcR transgenic CD4+ cells¹⁵ are polarized ex vivo under Th1- or Th17-inducing conditions. Effector cells are transferred into recombination activating gene-1 deficient (Rag1^−/−) mice with OVA planted in their glomeruli by injecting an OVA conjugate. OVA is conjugated to a mouse IgG1 mAb binding to α3(IV) collagen in murine glomerular basement membrane (GBM). The mAb-OVA conjugate dose is capable of planting significant OVA in glomeruli as an antigen to induce effector responses, but is insufficient to induce significant histologic or functional injury as an antibody. This model allows us to understand effector CD4+ T cell and Th subset-induced injury, with no effects from CD8+ cells or B cells.CD4+ cells, isolated from OVA-specific TcR transgenic (OT-II) mice and cultured under Th1 or Th17 priming conditions (see Concise Methods), were confirmed to be Th1 or Th17 by cytokine production before transfer. Th1 polarized cells expressed IFNγ, whereas no IL-17A or IL-4 production was detected (Figure 1A). Th17 polarized cells were strong IL-17A producers, showing only weak IFNγ production, with >20% of cells producing IL-17A, but few IFNγ or double-positive cells (Figure 1B). To plant OVA in glomeruli, the mAb 8D1, a non-nephritogenic, murine IgG1 binding to mouse α3(IV)NC1,¹⁶ was conjugated to OVA and purified by size-exclusion chromatography so that no free OVA or unconjugated 8D1 mAb remained, confirmed by Western blotting (Figure 1C). Antigen-specific CD4+ cells recognized OVA bound to the 8D1 anti-GBM mAb. Culture of CFSE-labeled naive OT-II cells incubated with the 8D1-OVA conjugate enhanced their survival (30% to 40% survival after 72 h versus 5% to 6% with unconjugated antibody) and induced OT-II cell proliferation (72 h: 27% of cells, up to 4 cycles, Figure 1D). OT-II cells incubated with unconjugated 8D1 did not proliferate (Figure 1E). After intravenous injection, 8D1-OVA conjugates bound to the GBM in a linear manner; no fluorescence signal was observed after transfer of Th1 cells without 8D1-OVA (Figure 1G). Western blotting showed OVA in the kidney after injection of 8D1-OVA conjugate (Figure 1H). Lungs from 8D1-OVA-injected mice were weakly positive for mouse IgG, whereas no IgG was detected in the spleen or liver (detecting antibody titer 1:100) 3 or 21 d after injection. Mouse IgG was not detected in sera (ELISA, dilution 1:100) at day 3 or day 21. As expected (given the transfer of only CD4+ cells to Rag1^−/− mice), no anti-OVA antibodies in sera were detected in recipient mice (data not shown).Open in a separate windowFigure 1.Differentiation of OVA-specific OT-II Th1 and Th17 cells, antibody-OVA conjugation, glomerular IgG and intrarenal OVA detection, and recipient immune responses after cell transfer. (A) After stimulating naive OT-II cells with OVA in a Th1 environment, IFNγ was produced and intracellular cytokine staining of CD4+ cells demonstrated strong IFNγ staining with minimal IL-17A or IL-4. (B) Culturing cells in a Th17-stimulating environment led to strong IL-17A production, whereas cells stained positive for IL-17A but not IL-4, and only 2% of cells produced IFNγ. (C) Chromatographic profile of 8D1-OVA conjugation. The numbers 1 to 7 represent fractions collected for analyses by Western blotting, which confirmed that all OVA-conjugated fractions contained OVA and IgG (lanes 1–6), whereas unconjugated fractions (represented as “Un”) contained IgG alone. The lane labeled “M” contained molecular weight markers. (D and E): 8D1-OVA was recognized by OT-II cells because multiple cycles of proliferation of cultured naive OT-II cells (D) were seen with 8D1-OVA conjugate and (E) not seen with unconjugated antibody. Strong linear IgG staining of glomeruli was seen after (F) the administration of 8D1-OVA to Rag1^−/− mice, but not after (G) the injection of Th1 cells without antibody. Western blotting of homogenized kidney (H) 24 h after the administration of 8D1-OVA demonstrated OVA in the kidneys (labeled as OVA-Ab); this was not seen after the administration of unconjugated antibody (labeled as Un Ab). (I) Systemic immune responses of recipient Rag1^−/− mice at 21 d assessed by splenic cytokine production demonstrated enhanced IFNγ production in mice given 8D1-OVA and Th1 cells, with enhanced IL-17A production by mice receiving 8D1-OVA and Th17 cells. (J) DTH to OVA (at 21 d) was induced only in mice given 8D1-OVA and Th1 cells. *P < 0.05, ***P < 0.001.To determine whether transfer of either Th1 or Th17 antigen-specific effector cells induces glomerular injury, 8D1-OVA conjugate was administered intravenously to Rag1^−/− mice (lacking adaptive immunity). Three hours later, 5 × 10⁶ Th1 or Th17 cells were injected intravenously. Groups of mice injected with 8D1-OVA alone (without cells) or Th1 cells alone (without 8D1-OVA) served as controls. At 21 d, the injected T cells largely maintained their initial phenotype, because host splenocytes from mice given Th1 cells showed enhanced OVA-stimulated IFNγ production whereas IL-17A production was enhanced in mice given Th17 cells (Figure 1I). Dermal-delayed-type hypersensitivity (DTH) was induced by footpad injection of OVA and measured after 24 h. Only mice that received the 8D1-OVA conjugate and Th1 polarized cells developed dermal DTH (Figure 1J), a classical Th1 response.¹⁷After planting OVA in glomeruli, administration of Th1 or Th17 cells induced glomerular disease. Urinary albumin excretion was not increased in mice given 8D1-OVA conjugate alone or Th1 cells alone, but Th1 or Th17 cells with 8D1-OVA induced significant albuminuria (Figure 2A). Albuminuria was consistent throughout the time course of the study in the Th17 group, whereas in the Th1 group there was a progressive increase in albuminuria until day 21 (Figure 2B). Control mice given Th1 cells alone or the 8D1-OVA conjugate alone exhibited only mild histologic changes (no crescent formation, fibrinoid necrosis, or hyalinosis). Analysis of histologic injury demonstrated substantially more abnormal glomeruli in the mice given 8D1-OVA conjugate with Th1 or Th17 cells compared with control groups (Figure 2C). Th1 and Th17 (+8D1-OVA) recipients developed proliferative GN, [glomerular hypercellularity: 8D1-OVA and Th1 cells: 32.1 ± 1.0 cells/glomerular cross section (c/gcs), 8D1-OVA and Th17 cells: 29.8 ± 1.1 c/gcs, 8D1-OVA alone: 21.3 ± 0.2 c/gcs, Th1 cells alone: 18.9 ± 2.0 c/gcs; P < 0.001]. Representative kidney sections from each group are shown (Figure 2, D through G). Crescent formation and fibrinoid necrosis, although seen in only a few glomeruli, was observed exclusively in mice given 8D1-OVA conjugate and Th1 cells (5.5 ± 0.9% at day 21; Figure 2, H and I). No crescent formation was observed in mice receiving 8D1-OVA conjugate and Th17 cells. Mice did not develop significant renal impairment (measured by BUN; data not shown).Open in a separate windowFigure 2.Renal injury in mice injected with 8D1-OVA conjugate, then either Th1 or Th17 cells. (A) Mice given 8D1-OVA conjugate or Th1 cells alone did not develop albuminuria above values for noninjected Rag1^−/− mice (dotted line). At 21 d, albuminuria was increased in mice given 8D1-OVA and Th1 cells or 8D1-OVA and Th17 cells. (B) In mice given 8D1-OVA and Th17 cells, albuminuria had plateaued by day 7 and did not progress. In mice given 8D1-OVA and Th1 cells there was a progressive rise in albuminuria. (C) Histologic injury was significant in mice given 8D1-OVA and either Th1 or Th17 cells. Representative glomeruli from mice given (D) 8D1-OVA alone, (E) Th1 cells alone, (F) 8D1-OVA and Th1 cells, and (G) 8D1-OVA and Th17 cells are shown. (H and I) Crescentic injury and fibrinoid necrosis were only seen in mice given 8D1-OVA and Th1 cells. ***P < 0.001Recruitment and activation of leukocyte subpopulations differed in mice administered Th1 or Th17 cells (Figure 3A). Although glomerular CD4+ cell and macrophage numbers were similarly increased in mice given 8D1-OVA conjugate and either Th1 or Th17 cells at day 21, more neutrophils were found in mice given 8D1-OVA and Th17 cells compared with mice given 8D1-OVA and Th1 cells. Interstitial leukocyte infiltrates followed a similar pattern (Figure 3B). Consistent with the finding of increased neutrophils in kidneys of mice receiving Th17 cells, renal mRNA expression of the primary neutrophil attracting chemokine CXCL1 was elevated (Figure 3C). Th17 cells attract neutrophils¹⁸ and in vitro studies have shown that neutrophil recruitment is achieved via production of CXCL8, the human homologue of CXCL1, by Th17 cells.¹⁹ It is therefore likely that at least some of the Th17-induced renal injury is mediated by neutrophils. In mice receiving 8D1-OVA and Th1 cells, macrophages were likely to be more activated; only these mice developed dermal DTH and increased expression of mRNA for the macrophage chemoattractants CCL2 and CCL5 (Figure 3, D and E), which have been associated with experimental crescentic GN.²⁰ Furthermore, type 2 nitric oxide synthase (NOS2/iNOS) mRNA, a marker of macrophage activation²¹ and urinary nitrate, a marker of intrarenal macrophage NOS2 production, were increased in this group (Figure 3, F and G).Open in a separate windowFigure 3.Leukocytes in kidneys of mice with either Th1- or Th17-induced injury 21 d after cell transfer. (A) Glomerular CD4+T cells, neutrophils, and macrophages were increased in mice given 8D1-OVA and Th1/Th17 cells. Neutrophil recruitment was incrementally increased in mice given 8D1-OVA and Th17 cells compared with 8D1-OVA and Th1 cells. (B) A similar pattern of recruitment was seen in the cortical interstitium. Renal chemokine mRNA expression demonstrated (C) enhanced CXCL1 mRNA in mice given 8D1-OVA and Th17 cells, whereas (D) CCL2 and (E) CCL5 were increased in mice given 8D1-OVA and Th1 cells. (F and G) NOS2 and urinary nitrate, markers of macrophage activation, were increased in mice receiving 8D1-OVA and Th1 cells. For mRNA, values for the 8D1-OVA alone group are presented as 1. *P < 0.05, **P < 0.01, ***P < 0.001.Further studies were performed 3 d after cell transfer. At this time point, albuminuria was present in mice receiving 8D1-OVA conjugate and Th17 cells, but not 8D1-OVA conjugate and Th1 cells (Figure 4A), and a higher proportion of glomeruli were abnormal in mice that had received Th17 cells (Figure 4B). Therefore, Th17-induced glomerular injury occurred earlier than Th1-induced injury. Leukocytes were present in glomeruli (Figure 4C) with increased numbers of neutrophils in glomeruli of mice receiving Th17 cells (compared with Th1 cell recipients), whereas Th1 cell recipients exhibited more macrophages. At day 3, these findings were glomerulo-specific; differences between Th1 and Th17 cell recipients were not seen in the interstitium (Figure 4D).Open in a separate windowFigure 4.Renal disease in mice 3 d after injection with 8D1-OVA and either Th1 or Th17 cells. (A) Pathologic albuminuria (dotted line represents values for noninjected Rag1^−/− mice) and (B) increased numbers of abnormal glomeruli were evident in mice that received 8D1-OVA and Th17 cells. (C) Leukocyte recruitment to glomeruli demonstrated CD4+ cells (more in mice receiving Th1 cells), with comparatively more neutrophils in glomeruli of Th17 cell recipients and more macrophages in glomeruli of Th1 cell recipients. (D) Interstitial leukocytes were similar in Th1 and Th17 cell recipients 3 d after cell transfer. *P < 0.05, **P < 0.01, ***P < 0.001.These studies used Rag1^−/− mice as recipients of effector antigen-specific Th1 or Th17 cells. Because these mice do not possess T or B cells, OVA planted in glomeruli cannot induce CD8+ or B cell responses, and regulatory T cells are unable to influence the pattern of injury. A major advantage of this strategy is that Th1- and Th17-mediated injury can be assessed in a pure experimental system. However, T cells transferred into Rag1^−/− mice can undergo homeostatic expansion, and it is possible that the transferred Th1 cells might have expanded more rapidly than Th17 cells. Recently, studies in experimental type 1 diabetes induced by transfer of cells from a TcR transgenic mouse specific for an islet autoantigen showed conversion of Th17 cells to a Th1 phenotype after transfer.^22,23 Although our Th17 polarized OT-II cells, specific for a foreign antigen, showed some IFNγ production after 21 d, they were still capable of producing IL-17A. Furthermore, dermal DTH and renal disease were different in Th1 recipients compared with the Th17 recipients at 21 d, supporting the maintenance of separate phenotypes after transfer. Although the studies presented here are the first to demonstrate a role for Th17 and Th1 cells in the same experimental system, other studies^24–26 have used genetically deficient mice to implicate Th17 cells in experimental renal disease.These studies describe a novel model of cell-mediated proliferative GN for which adaptive components are only effector antigen-specific CD4+ T cells. They demonstrate that both Th1 and Th17 cells can induce proliferative GN. Th17 cells induce albuminuria early, with persistent accumulation of leukocytes. Administration of Th1 cells lead to a slower rise in albuminuria, but more macrophage activation and DTH-like injury, including, in some glomeruli, crescent formation and fibrinoid necrosis. It is likely that Th1 and Th17 responses play a role in proliferative forms of GN and both represent potential therapeutic targets.     

Th1/Th2细胞与肿瘤复发

目的探讨T辅助淋巴细胞Ⅰ型（Th1）／T辅助淋巴细胞Ⅱ型（Th2）在肿瘤复发中的研究进展。方法复习国内、外相关文献并进行综述。结果肿瘤治疗后体内出现Th1向Th2漂移，使肿瘤细胞逃避机体的免疫监视，导致肿瘤的复发。结论Th1向Th2型漂移与肿瘤治疗后的复发有关，促使肿瘤治疗后的机体细胞因子由Th2向Th1逆转，重新达到平衡，成为肿瘤免疫治疗的新思路。     

The Th1 and Th2 paradigm in ANCA-associated vasculitis

In the pathogenesis of anti-neutrophil cytoplasm antibodies (ANCA)-associated vasculitis, T cell contribution is indicated by T cell-dependent ANCA production combined with the presence of T cells in inflammatory infiltrates. However, the exact pathogenic role of T cells in ANCA-associated vasculitis remains to be determined. The Th1/Th2 concept is useful for understanding T cell involvement in pathological processes. This review focuses on T cells and particularly the Th1/Th2 paradigm in ANCA-associated vasculitis. Most research has been done in Wegener's granulomatosis, where a shift in T cell response, from a Th1 pattern in localized disease towards a Th0/Th2 pattern in generalized disease, appears to occur. Although less thoroughly studied, data in Churg-Strauss syndrome and microscopic polyangiitis indicate that these diseases are predominantly associated with Th2 patterns. Further studies elucidating the true nature of the polarization towards Th1 or Th2 in ANCA-associated vasculitis are clearly needed.     

Th1/Th2 balance in childhood idiopathic nephrotic syndrome

AIMS: In view of the conflicting evidence of helper T cell type 1 (Th1) or type 2 (Th2) pattern of cytokine synthesis in childhood idiopathic nephrotic syndrome (INS) this study examined the balance of Th1 and Th2 which are characterized by intracellular cytokine production of interferon-gamma (IFNgamma) and interleukin-4 (IL-4), respectively. SUBJECTS AND METHODS: Sixteen children with steroid-sensitive INS (mean age 9.0 years) were included in this study, together with 15 healthy normal children (mean age 7.9 years) for the control group. Intracellular production of both IFNgamma and IL-4 in helper T cell (CD4+ cell) was investigated by a 3-color flow cytometry. RESULTS: The cross-sectional data showed no significant differences of percentages of Th0 (IFNgamma+ IL-4+ CD4+ cell), Th1 (IFNgamma+ lL-4- CD4+ cell) and Th2 (IFNgamma- IL-4+ CD4+ cell) in CD4+ cells (p > 0.05). The Th1/Th2 ratio during nephrotic relapse did not differ from those during nephrotic remission and in normal healthy children (p > 0.05). CONCLUSION: We conclude that there is no significant skew of Th1/Th2 balance in childhood INS and that the cardinal immunological abnormality does not lie in helper T cells but in other cells, such as suppressor/cytotoxic T cells, natural killer cells or monocytes/macrophage. To clarify the pathogenesis of INS, comprehensive studies for these cells would be worthwhile.     

Th1/Th2细胞因子对器官移植排斥反应的影响

目的:研究Th1/Th2细胞因子对同种异系小鼠心脏移植存活时间的影响.方法:使用野生型BALB/c小鼠作为供体,野生型B6小鼠、IL-4基因去除B6小鼠及IFN-γ基因去除B6小鼠作为受体,行腹部异位小鼠心脏移植.部分IL-4基因去除小鼠、IFN-γ基因去除小鼠联合应用α-半乳糖神经酰胺(α-galactosylceramide,α-GalCer),以获得更强的Th1/Th2偏移.比较移植物存活时间.向野生型、IL-4基因去除及IFN-γ基因去除B6小鼠腹腔内注射供体小鼠脾细胞,提取受体小鼠脾脏CD8<'8>T细胞行淋巴细胞毒试验.结果:IFN-γ基因去除组小鼠的移植物存活时间为(6.40±0.55)d,联合应用α-GalCer组移植物存活时间为(8.00±1.15)d.IL-4基因去除组小鼠的移植物存活时间为(8.00±1.00)d,联合应用α-GalCer组移植物存活时间为(8.60±1.34)d.淋巴细胞毒试验显示IFN-γ基因去除小鼠的CD8+T细胞毒性明显增强.结论:Th1/Th2细胞因子与排斥反应之间并不存在简单的对应关系.     

Influence of Th1, Th2, and Th3 cytokines during the early phase after liver transplantation

To investigate the change of Th1, Th2, Th3 cytokines during early liver transplantation. IFN-r, IL-4, TGF-beta production by CD4+ T cells were determined by fluorescence activated cell sorter analysis. Comparing the acute rejection with the non-acute rejection groups on the 7th, 14th, and 28th day showed that high interferon-gamma production associated with acute rejection in the early posttransplant period. There was no evidence of significant changes in interleukin-4 and transforming growth factor beta (TGF-beta) levels between non-acute rejection groups with acute rejection groups. Th1 cytokine high production is significantly associated with acute rejection in liver transplant recipients.     

儿童原发性肾病综合征与Th1/Th2细胞失衡

目的 探讨Th1/Th2细胞失衡在原发性肾病综合征（PNS）患儿发病中的作用。方法运用三色荧光标记法流式细胞术检测21例初发和16例缓解期NS患儿外周血Th1和Th2细胞百分率（％）,并以10例正常儿童作对照。结果 正常儿童外周血Th1、Th2和Th0细胞百分率分别为（13.42±4.36）％,（2.53±1.97）％和（1.25±0.92）％。初发的PNS患儿均明显减低,分别为（2.34±2.09）％,（1.02±0.96）％和（0.40±0.38）％（P<0.05）。缓解期PNS患儿,分别为（11.96±4.75）％,（2.87±2.46）％和（1.31±0.87）％,与正常儿童比,差异均无显著性意义。由于初发的PNS患儿Th1细胞减低相对于Th2细胞而言更显著,因而导致Th1/Th2比值也较缓解期PNS患儿和正常儿童明显下降（2.43±2.65比4.17±2.32和5.41±2.77,P均<0.05）。结论 儿童PNS是一种以Th2细胞占优势的免疫介导的肾小球疾病,源于Th1细胞减低所致的Th1/Th2细胞失衡在原发性肾病综合征发病过程中可能起着重要作用。