Th2 predominance at the single-cell level in patients with IgA nephropathy.

BACKGROUND: Abnormalities of lymphocyte function have been reported to be involved in the pathogenesis of IgA nephropathy (IgA-N). The aim of this study was to investigate helper T (Th) predominance at the single-cell level, one of the abnormalities of lymphocyte function in IgA-N. METHODS: Using flowcytometry, we assessed the levels of circulating Th cells in IgA-N patients (n=30), and in normal individuals (n=30) based on the expression of intracellular Th1 cytokines for interleukin-2 (IL-2) and interferon-gamma (IFN-gamma), and of intracellular Th2 cytokines for IL-4, IL-10, and IL-13. Because the production of each cytokine had a specific time course, we examined cytokine synthesis at 3, 6, 9, and 12 h after stimulation. RESULTS: The percentages of IL-2-positive Th cells from IgA-N patients were significantly lower than in normal individuals at 6, 9, and 12 h, with the difference becoming greater with time. The number of IFN-gamma-positive Th cells in IgA-N patients was significantly lower than in normal individuals at 9 h, and the number of IFN-gamma-positive Th cells increased more at 12 h than at 3 h in both groups. IL-4 and IL-13 expression was increased in patients with IgA-N at 6 h compared with normal individuals. In IgA-N patients, the percentage of IL-10-positive Th cells was significantly higher than that in normal individuals at each time-point. CONCLUSION: A polarization toward Th2 response at the stimulated lymphocyte level may lead to immune abnormalities in IgA-N.     

BCG dose reduction by decreasing the instillation frequency: effects on local Th1/Th2 cytokine responses in a mouse model

OBJECTIVES: Based on the requirement of a Th1 immune response for clinical efficacy, and incited by the arbitrary induction scheme, frequent side effects and the empirical approach in improving BCG immunotherapy for superficial bladder cancer, an alternative intravesical BCG treatment schedule for dose reduction was investigated without compromising Th1 cytokine induction in the bladder in a mouse model. METHODS: Mice were submitted to 6 weekly BCG instillations and treatment schedules omitting intermediate instillations during this standard scheme. Th1 (IFN-gamma, IL-2, IL-12p40), and Th2 (IL-10, IL-4) cytokine responses in individual mouse bladders were measured by a semiquantitative RT-PCR based method. RESULTS: A schedule of only two BCG instillations, administered in week 1 and week 6, resulted in induction of at least the same levels of IFN-gamma, IL-2 and IL-12p40 Th1 cytokine mRNA compared to 6 weekly instillations, whereas significantly lower levels of Th2 cytokines IL-10 and IL-4 mRNA were observed. CONCLUSIONS: During the 6-week period the intermediate weekly BCG instillations 2, 3, 4, and 5 do not contribute to Th1 cytokine upregulation in the bladder, provided that the BCG dose is sufficient. Whether such a reduced BCG frequency schedule has immune stimulating capacity and therapeutic efficacy associated with less side effects in patients remains to be investigated.     

Cytokine modifications after tension-free hernioplasty or open conventional inguinal hernia repair

BACKGROUND: The purpose of this study was to evaluate the involvement of proinflammatory cytokines (interferon-gamma [INF-gamma], interleukin [IL]-6) and anti-inflammatory cytokines (IL-4, IL-l0, IL-13) in patients undergoing Lichtenstein tension-free hernioplasty (LH) using polypropylene prosthetic materials or conventional Bassini hernia (BH) repair. METHODS: Thirty-five male patients (age range 25 to 60 years) with unilateral inguinal hernia without complications or recurrence were included in this study. Randomly, patients underwent conventional operation and had their inguinal hernia repair performed with polypropylene mesh. Peripheral venous blood samples were collected 24 hours prior to surgery and then 6, 24, 48, and 168 hours postoperatively. Fifteen healthy controls were included. RESULTS: We present evidence that LH patients showed both an increased serum level of Thelper 1 (Th1)-like cytokines (IFN-gamma) and an increase in Thelper 2 (Th2)-like cytokines (IL-6 and IL-l0), associated with a slight reduction of peripheral blood mononuclear cells (PBMC) producing IL-6 and a normal level of PBMC producing IFN-gamma, IL-l0, IL-13, and IL-4. Whereas BH patients showed in part an amplification of Th2-like cells, characterized by the sustained serum production of IL-6 and IL-l0, associated with an increase in IL-l0 secreted by in vitro stimulated PMBC. CONCLUSIONS: Our data show that LH is associated with a higher production of inflammatory cytokines (IFN-gamma and IL-6) compared with BH, likely induced by the presence of the polypropylene prostheses.     

Evaluation of airway inflammation by quantitative Th1/Th2 cytokine mRNA measurement in sputum of asthma patients

BACKGROUND: Asthma is a chronic inflammatory disorder of the airways driven by T cell activation. Th2 cells and their cytokines are thought to play a role in the pathophysiology of allergic as well as non-allergic asthma. METHODS: Airway cells were obtained by sputum induction from 15 healthy and 39 asthmatic individuals and the airway T cell cytokine profiles (interleukin (IL)-4, IL-5, IL-13, IL-10 and interferon (IFN)-gamma) at the mRNA level were studied by real time RT-PCR. RESULTS: Asthma patients had increased expression of IL-5 (p = 0.001) and IL-13 (p = 0.03) mRNA in sputum compared with non-asthmatic controls. IL-4 mRNA and IFN-gamma mRNA were detectable in the sputum of 44% and 21% of patients, respectively, but not in controls. Sputum IL-10 mRNA levels did not differ significantly between patients and controls. Sputum mRNA expression levels of IL-4, IL-5, and IL-13 were significantly correlated with the percentage of eosinophils and were higher in subjects with allergic asthma than in those with non-allergic asthma (p = 0.03, p = 0.02 and p = 0.0002, respectively); they did not differ between mild asthmatic subjects and those with moderate to severe asthma. In contrast, IFN-gamma mRNA expression was higher in non-allergic than in allergic patients (p = 0.04) and higher in patients with moderate to severe asthma than in those with mild asthma (p<0.01). Sputum IL-5 mRNA levels (but not the other cytokine mRNA levels) were also correlated with exhaled nitric oxide (eNO) and with bronchial hyperreactivity expressed as the histamine concentration resulting in a 20% decrease in forced expiratory volume in 1 second. CONCLUSION: Real time RT-PCR analysis of mRNA in induced sputum confirms a predominance of Th2 cytokines in both allergic and non-allergic asthma. IL-5 levels reflect eosinophil infiltration as well as eNO levels and hyperreactivity, and levels of the Th1 cytokine IFN-gamma indicate asthma severity. The technique is a promising tool for use in further studies of asthma severity and disease activity.     

Th2 subset dominance among peripheral blood T lymphocytes in patients with digestive cancers

BACKGROUND: Two types of helper T cells (Th), which are categorized as Th1 and Th2 on the basis of cytokine production, have been reported. Th1 cells produce interleukin (IL)-2 and interferon (IFN)-gamma, while Th2 cells secrete IL-4, IL-6, and IL-10. We assessed the intracellular cytokine profiles of CD3/CD4 positive lymphocytes (CD4+ T-cells) in peripheral blood in patients with digestive cancers. METHODS: Peripheral blood samples were collected from 50 patients with digestive cancers and 35 healthy volunteers. The proportions of CD4+ T-cells producing intracellular cytokines were determined using flow cytometry. RESULTS: The percentages (mean +/- SD) of CD4+ T-cells producing IL-4, IL-6, and IL-10 in the cancer group (73.9% +/- 13.0%, 73.0% +/- 16.6%, and 58.0% +/- 21.0%, respectively) were significantly higher than in the healthy group (37.4% +/- 12.4%, 37.8% +/- 13.5%, and 34.0% +/- 14.1%, respectively; P <0.01). Proportions of CD4+ T-cells producing IL-4, IL-6, and IL-10 in 10 patients undergoing curative resection had decreased significantly 1 month after surgery (P <0.01). No significant difference was noted between groups in the percentages of CD4+ T-cells producing IFN-gamma. CONCLUSIONS: Th2-dominant status develops in cancer patients. Such lymphocyte evaluations could find applications in diagnosis and therapeutic monitoring of cancer patients.     

Predominance of type-2 immune response in idiopathic membranous nephropathy. Cytoplasmic cytokine analysis

BACKGROUND: The Th1/Th2 paradigm is proving increasingly useful in the understanding of infectious diseases and many autoimmune diseases. Th1 cells predominantly produce interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) and are instrumental in initiating delayed-type hypersensitivity and activating macrophages. Th2 cells secrete other cytokines, such as IL-4, IL-5, IL-10 and IL-13 that trigger B-cell activation and immunoglobulin synthesis. It has been shown that in patients with membranous nephropathy, there may be a predominance of Th2, because of the presence of IgG, particularly IgG4, which belongs to a subclass of the type-2 immune response, and complement deposits in glomeruli. In this study, we investigated the immunoresponse of helper T cells, i.e. Th predominance in patients with idiopathic membranous nephropathy. METHODS: We used flow cytometry to assess the levels of circulating Th cells in patients with idiopathic membranous nephropathy (n = 8) and in normal individuals (n = 23) based on the expression of intracellular type-1 and type-2 cytokines. Because the production of each of these cytokines has a specific time course, we observed the cytokine synthesis at 3, 6, 9 and 12 h after stimulation. RESULTS: The percentages of IL-2+/CD4+ cells from patients with idiopathic membranous nephropathy were significantly lower than those from normal individuals at 6, 9 and 12 h, with the difference becoming more significant over time. IFN-gamma+/CD4+ cells and IL-4+/CD4+ cells were not significantly different between the two groups. In patients with idiopathic membranous nephropathy, the percentages of IL-10+/CD4+ cells were significantly higher than those in normal individuals at each point in time. CONCLUSION: Increased IL-10-producing Th cells may lead to suppression of delayed-type hypersensitivity and activate suppressor cells and IgG4 synthesis, resulting in idiopathic membranous nephropathy.     

T-cell activation follows Th1 rather than Th2 pattern in haemodialysis patients.

BACKGROUND: Patients on chronic intermittent haemodialysis (HD) show an impaired cellular and humoral immune response that clinically appears with frequent infectious complications and low vaccination responses. This immune defect strongly correlates with reduced in vitro proliferative responses of T cells. The defect is localized in antigen presenting cells, which show a decreased co-stimulatory activity. Furthermore, the impaired immune response correlates with an increased production of pro-inflammatory cytokines. In response to primary activation, CD4 positive T helper (Th) cells mainly differentiate into either Th1 or Th2 cells. Th1 cells support cell mediated immunity whereas Th2 cells enhance humoral immune responses. Since both types of responses mutually inhibit each other, the impaired humoral immune response seen in HD patients could either be due to a reduced number of Th2 cells or to a predominant Th1 response. METHODS: We analysed the Th cell profile in HD patients using flow cytometry. Monocytic cytokine expression was analysed using both flow cytometry and enzyme linked immunoadsorbant assays. RESULTS: Our data demonstrate that the cytokine differentiation profile in circulating T cells from HD patients is dysregulated and characterized by an increase in Th1 cells, but a normal amount of Th2 cells. Moreover, the skewed helper cell responses correlate with a higher percentage of monocytes capable of secreting the Th1 promoting cytokine interleukin 12 (IL-12). CONCLUSIONS: Our findings contribute to a better understanding of the pathogenesis of impaired cellular immune functions in dialysis patients and, in particular, the decreased antibody production after vaccination. They provide a link between overproduction of pro-inflammatory cytokines (IL-12) and imbalanced T-cell activation.     

抑制应激对烧伤血浆中LPS、前炎症细胞因子和TH1/TH2细胞因子的影响

目的 探讨抑制机体应激反应的措施,对严重烧伤后血浆中LPS、前炎症细胞因子和Th1/Th2细胞因子的影响。方法 将30％TBSA深Ⅱ度烧伤模型的SD大鼠分为烧伤立即复苏组（A）和烧伤立即复苏合剂组（B）,于伤后不同时相点取外周血分离血浆检测LPS、IL－1α、IL-6、TNFα、IL-8和IL－2、IFN－γ、IL－4、IL－10含量,另设假烧组作为对照（C）。结果 伤后LPS、IL－1α、IL－4、IL－10水平逐渐升高,IL－6、TNF－α、IL－8、IL－2和IFN－γ水平先升高后下降。A组炎症细胞因子和LPS升高幅度较大,其中IL－1α、IL－6和IL－4等升高时间（6h）较B组（12h）早。结论 及时液体复苏和采取抑制应激的措施,能延缓并减轻严重烧伤后炎症反应的发生,降低Th2细胞因子水平并使烧伤48h后Th1细胞因子得到部分恢复。     

腹腔镜和开腹胆囊切除术对患者血清细胞因子IL-6、IL-10和IL-18的影响

目的:探讨腹腔镜胆囊切除术(LC)和开腹胆囊切除术(OC)对慢性结石性胆囊炎患者血清IL-6、IL-10和IL-18影响,比较两种方法对机体肝功能的损伤和机制。方法:选择LC患者和OC患者各30例,分别于术前、术后1d、5d抽取静脉血测定血清ALT、AST、IL-6、IL-10和IL-18含量,并进行对比研究。结果:LC组和OC组术后血清ALT、AST、IL-6和IL-18含量均较术前增加(P<0.01),血清IL-10含量较术前降低(P<0.01),上述指标术后1d变化最为明显,LC组上述血清指标变化小于同期OC组(P<0.05)。结论:LC对患者肝功能损伤较OC小,可能与LC刺激机体产生IL-6和IL-18较少,维持较高血清IL-10水平,维护适度机体细胞因子平衡有关。     

No effect of transfusion transmitted virus viremia on the distribution and activation of peripheral lymphocytes in hemodialyzed patients

AIM: We aimed to examine the distribution and activation of peripheral T cells in TTV positive (n = 32) and negative (n = 17) hemodialyzed patients. The control group (n = 20) consisted of healthy blood donors. METHOD: TTV-DNA was detected by seminested PCR. CD3, CD4, CD8, CD19, CD56, CD3/HLA-DR, CD3/CD69 and the Th1/Th2 ratio of T cells were analyzed by flow cytometry. Circulating IFN-gamma, IL-2, IL-4, IL-6, IL-10, IL-13, TNF-alpha, TGF-beta levels were measured by ELISA in the sera. RESULTS: There was no difference between the CD3, CD4, CD8 and CD19 values of HD subjects. In addition, the expression of both activation markers, HLA-DR and CD69, was significantly elevated in the TTV-positive and -negative HD groups compared to the controls, but not showing any difference from each other. The measurements of intracellular cytokines showed the enhanced occurrence of INF-gamma + CD4 T cells, and decreased appearance of IL-4 + CD4 lymphocytes in the HD groups without any significant difference between the TTV virus positive and negative patients. In addition, HD also elevated the expression of IL-10 in CD4 and CD8 (Th2) cells. There were only two significant changes in the levels of circulating cytokines: (a) IL-2 increased; (b) IL-13 decreased in both groups of HD patients compared to the controls, independently of TTV positivity or negativity. CONCLUSIONS: We assume that transfusion-transmitted virus does not cause any specific change in the distribution and activation of lymphocytes in the peripheral blood of hemodialyzed patients. Hemodialysis itself, however, results in a significant activation of peripheral T cells with the domination of increased production of Th1 type cytokines, IFN-gamma, IL-2, in contrast to the decreased synthesis of Th2 type cytokines, IL-4 and IL-13. Furthermore, the increased expression of IL-10 in the CD4 and CD8 cells of HD patients can be the sign of a contraregulatory Th2 activation as an answer on the Th1 effect.     

共刺激后淋巴细胞产生Th1/Th2细胞因子的变化及免疫抑制剂的影响

目的 探讨CD28、CD40通路共刺激后淋巴细胞产生Th1(IL-2、IFN-γ、IL-12)及Th2细胞因子(IL-4、IL-10)的变化及免疫抑制剂环孢素(CsA)、雷帕霉素(RPM)及霉酚酸(MPA)对共刺激通路激活后淋巴细胞产生Th1及Th2细胞因子的影响.方法采用单克隆抗体(mAb)与淋巴细胞表面CD3、CD28及CD40L分子结合产生相应刺激信号,单刺激及共刺激组分为:a组,CD3 mAb单刺激;b组,CD3 mAb加CD28 mAb共刺激;c组,CD3 mAb加CD28 mAb加CD40 L mAb共刺激;d组,CD3 mAb加CD28 mAb加CTLA4 mAb共刺激.各mAb的终浓度均为100 ng/ml.干预组分别将终浓度为300 ng/ml的CsA、RPM、MPA加入上述4组.ELISA法测定上述细胞培养上清中的细胞因子值.结果 a、b、c 3组IFN-γ分别为(248.91±11.20)、(555.08±24.42)、(548.19±33.06)ng/ml,IL-2分别为(29.48±8.61)、(1100.82±99.29)、(842.23±29.31)ng/ml,IL-4分别为(32.29±6.76)、(116.02±15.03)、(147.28±18.07)ng/ml,IL-10分别为(147.01±10.47)、(291.79±12.47)、(302.52±35.18)ng/ml.b、c组与a组比较,差异均有统计学意义(P＜0.01);b、c组IL-2、IL-12、IL-4比较,差异均有统计学意义(P＜0.05).d组IFN-γ、IL-2及IL-10分别为(497.42±29.03)、(739.77±18.58)及(120.33±13.21)ng/ml,与b组相比,差异均有统计学意义(P＜0.05).CsA、RPM及MPA对共刺激后Th1/Th2细胞因子的产生均有抑制作用,CsA对4种细胞因子产生的抑制作用强于RPM和MPA,其中对IL-2及IL-4的抑制作用更为明显.CsA与CTLA4 mAb有协同作用.共刺激后IL-12产生升高,MPA可抑制单刺激和共刺激后IL-12的产生,CsA和RPM对IL-12的产生无明显抑制作用.结论 CD28、CD40共刺激通路在淋巴细胞活化中起关键作用.CsA、RPM、MPA及CTLA4 mAb对共刺激后Th1/Th2细胞因子的产生均有抑制作用,CsA的抑制作用更为明显.CD40 L mAb使Th1细胞因子及IL-12水平下降,又促进Th2细胞因子(以IL-4为主)产生,该作用可能是抗CD40 L抗体诱导移植物存活期延长的机制之一.     

Mesenchymal stem cells therapy protects fetuses from resorption and induces Th2 type cytokines profile in abortion prone mouse model

The imbalance of Th1/Th2 cytokines is well known in recurrent spontaneous abortion (RSA) mouse model. Mesenchymal stem cells (MSCs) possess potent immunoregulatory properties that could modulate the Th1 cytokine responses in benefit of Th2 types. In this study, we aimed to analyze the local and systemic balance of Th1/Th2 cytokines following MSCs therapy.Syngeneic adipose derived MSCs were administered to abortion prone mice during the implantation window. The abortion rate was determined and IL-4, IL-6, IL-12, IL-2, IFN-γ and GM-CSF gene expression was evaluated by Real-Time-PCR in decidual and placental tissues of pregnant mice at day 13.5 of pregnancy. Splenocytes of pregnant mice were co-cultured with mitomycin C treated paternal splenocytes and IL-2, IL-4, IL-10 and IFN-γ cytokines were measured in co-cultures supernatants by ELISA method. Proliferation response of female splenocytes to paternal antigens was also evaluated using the CFSE method.Our results showed a significant reduction in abortion rate following MSCs administration in abortion prone mice. We also observed a significant down-regulation of IL-2 and IFN-γ as well as up-regulation of IL-4 and IL-10 production from pregnant mouse splenocytes following MSCs therapy along with a significant reduction of splenocytes proliferation against paternal antigens. Our findings revealed that MSCs therapy increased the IL-4, IL-6, IL-10 and GM-CSF and at the same time decreased the IL-12, IL-2 and IFN-γ gene expression at feto-maternal interface.Here, we showed that MSCs therapy could modulate the systemic as well as local Th1/Th2 cytokines production along with protection of fetus from resorption in abortion prone mice. The fine balance of Th1/Th2 cytokine response could be considered as one of the possible mechanisms for fetal protection following MSCs therapy.