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.     

Kinetics of adaptive immunity to cationic bovine serum albumin-induced membranous nephropathy

Membranous nephropathy is an autoimmune-mediated glomerulonephritis and a major cause of nephrotic syndrome. We studied the kinetics of adaptive immunity in the pathogenesis of membranous nephropathy in T1/T2 double transgenic mice (T1/T2 TG mice) that express human Thy1 protein under the control of interferon-gamma (INF-gamma) and mouse Thy1.1 protein under the control of interleukin (IL)-4. Nephropathy was induced by cationic bovine serum albumin. We found that splenocytes expressed a progressive Th2 response and a subsequent compensatory T-helper 1 (Th1) response, with a gradual augmentation of IL-4-producing Th2 cells and INF-gamma-producing Th1 cells. Increased Th2 marker expression was seen in peripheral blood and kidney cells, with the immunoglobulin G1 (IgG1) antibody isotype predominant in the serum and kidneys. We found that CD8+ T cells contribute more to the augmented INF-gamma production than CD4+ T cells. Moreover, CD19+ B cells demonstrated a greater production of IL-4 than the CD4+ T cells. Cytokine-related gene expression in kidneys and splenocytes showed an upregulation of proinflammatory Th1 and Th2 cytokines. Th2 cells but not Th1 cells were significantly correlated with serum cholesterol and proteinuria. Our study shows that both peripheral and renal immune reactions are strongly polarized toward Th2-type immune responses during the course of membranous nephropathy. The T1/T2 mouse model may help decipher the kinetic changes of adaptive immunity in glomerulonephritis.     

晚期肺癌患者化疗前后细胞免疫功能及T细胞分泌细胞因子的变化

目的:探讨晚期肺癌患者化疗前后外周血中T细胞免疫功能及其分泌细胞因子水平的变化.方法:通过流式细胞仪检测57例晚期肺癌患者和22例健康体检者CD4+T淋巴细胞和CD8+T淋巴细胞在外周血单核细胞(PBMC)中所占比例以及外周血CD4+T细胞分泌细胞因子水平的变化.结果:晚期肺癌患者化疗前CD4+T细胞在PBMC中所占比例、CD4+T细胞与CD8+T细胞比值均低于健康人(P＜0.001),化疗后均较化疗前明显升高(P＜0.01或P＜0.001):化疗前CD8+T细胞在PBMC中所占比例高于健康人(P＜0.001),而于化疗后较化疗前明显下降(P＜0.05).治疗前后患者血清中CD4+T细胞分泌Th1细胞因子(IFN-y、TNF-α、IL-2)和Th2细胞因子(IL-4、IL-6、IL-10)水平均无明显变化(P＞0.05).结论:化疗能够增加肺癌患者外周血中CD4+T细胞所占比例,但对Th1、Th2所分泌的相关细胞因子水平影响不大.     

Melanoma skews dendritic cells to facilitate a T helper 2 profile

BACKGROUND: Patients with progressing melanoma have a circulating cytokine profile reflecting a T helper cell type 2 (Th2) imbalance, while patients responding to therapy favor a Th1 profile. The aim of this study was to determine the role of circulating dendritic cells (DCs) in mediating this imbalance. METHODS: Isolated human peripheral blood mononuclear cells (PBMCs) were exposed to cell-free melanoma-conditioned medium (MCM) or control fibroblast-conditioned medium before stimulation. In separate experiments, isolated circulating DCs were exposed to MCM before addition of T cells. DC maturation and function were determined. Mixed leukocyte response T-cell proliferation was quantified and supernatants were assayed for Th1 (interleukin [IL]-2 and interferon gamma) and Th2 (IL-4, IL-5, and IL-10) cytokines. RESULTS: PBMCs exposed to MCM produced significantly more Th2-type cytokines (IL-4, IL-5, and IL-10) over time than those exposed to control medium. DCs exposed to MCM before addition of T cells, produced a similar pattern of a sustained longer term Th2 response after an initial burst of IL-2. Exposure to MCM did not significantly affect DC maturation or IL-12 production. T-cell proliferation did not change significantly in the mixed leukocyte response, however, the percentage of viable CD4+ T cells in the MCM-treated group was significantly less than control (37 vs 50%, P < .05). CONCLUSIONS: Exposure of PBMCs to melanoma produces a Th2-type cytokine profile, which may be, in part, facilitated by DCs.     

Distribution of glomerular IgG subclass deposits in malignancy-associated membranous nephropathy.

BACKGROUND: Several studies have shown a predominant glomerular deposition of IgG4 in patients with idiopathic membranous nephropathy (MN), whereas significant depositions of other IgG subclasses have been shown in patients with lupus-associated MN and bucillamine-induced MN. METHODS: We examined the distribution patterns of glomerular IgG subclass deposits in 10 patients with malignancy-associated MN (M-MN) and in 15 patients with idiopathic MN by immunofluorescence (IF) microscopy. RESULTS: The glomerular IF intensities of IgG1 and IgG2 were significantly stronger in the malignancy group than in the idiopathic group (P<0.05). In contrast, there were no differences in glomerular IF intensities of IgG3 and IgG4 between the two groups. CONCLUSION: Our findings suggest that the distribution patterns of glomerular IgG subclass deposits are different in idiopathic MN and M-MN. The strong IF intensity of glomerular IgG1 and IgG2 in M-MN may provide a possible predictor for this condition.     

稳定存活肝移植受者血清细胞因子水平的研究

目的 研究稳定存活的肝移植受者血清细胞因子的表达状况及意义。方法ELISA检测22例原位肝移植患者、13例原发性肝癌患者及12例正常人血清细胞因子的表达情况。流式细胞术分析外周血T细胞表型。结果外周血CD3^ 、CD8^ T细胞百分比以及CD4^ ／CD8^ 比值,各组间比较无统计学差异。肝移植组CD3^ CD25^ T细胞百分比高于正常人组(P=0．022)。血清IL-2、IFN-γ、IL-10、IL-4及INF-α水平,各组间比较差异无显著性。IL-6、ICAM-1和P-seleetin水平肝移植组显著高于正常人组(P值分别为0．048、0．000和0．025)。结论稳定存活的肝移植患者体内效应T细胞仍处于低活化状态,炎症性细胞因子(TNF-α、IL-6)和黏附分子(ICAM-1、P-Seleetin)可能在移植物的慢性损伤过程中发挥了作用。     

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.     

86例膜性肾病回顾性分析及对CD4^＋CD^25＋调节性T细胞的初步研究

目的：回顾性总结、分析86例膜性肾病患者的临床表现、实验室检查及肾活检病理的特点及相互联系,认识膜性肾病的发病和流行病学特点。通过对特发性膜性肾病（idiopathic membranous nephropathy,IMN）患者和正常人外周血CD4＋CD2＋5调节性T细胞（Treg细胞）数量的检测,了解Treg在IMN患者外周血的变化规律,探讨其在IMN发病中的作用。方法：2004年3月～2008年12月间病理确诊为膜性肾病患者86例,分析患者一般资料、病理类型和临床特征。选择2007年～2008年IMN患者10例,随机选取与IMN患者年龄相匹配的健康志愿者10例,检测所有对象外周血Treg细胞数量。结果：（1）86例膜性肾病患者,其中IMN68例,占80%,4例患者随访后确诊为恶性肿瘤;继发性膜性肾病18例（其中乙肝病毒相关性肾炎5例,狼疮性肾炎4例,移植肾肾小球肾炎1例）,占20%。（2）IMN免疫荧光以IgG沉积为主,乙肝病毒相关性肾炎C1q沉积较IMN多（P〈0.05）,并均存在HBsAg沉积,与IMN相比狼疮性肾炎C1q沉积明显增多,C4也多于IMN（P〈0.05）。（3）病理分期分布特点：Ⅱ期膜性肾病多见。（4）IMN患者治疗前外周血Treg细胞占CD4＋淋巴细胞的百分比为（7.46±0.94）%,正常对照组为（6.54±1.0）%。结论：（1）根据病因分为IMN及继发性膜性肾病两种,男性发病率大于女性,中老年多发,继发性膜性肾病的年龄及性别分布根据病因的不同而有所不同,临床表现均以肾病综合征表现为主;IMN发病率明显大于继发性膜性肾病。（2）免疫荧光检查：IMN以IgG及C3沉积为主,乙肝病毒相关性肾炎均存在乙肝表面抗原,狼疮性肾炎与乙肝病毒相关性肾炎的C1q沉积较特发性膜性肾病明显增多（P〈0.05）。（3）IMN患者外周血Treg细胞数量较正常人增多。     

T-helper and T-cytotoxic cell subsets monitoring during active cytomegalovirus infection in liver transplantation

OBJECTIVE: The objectives of this study was to analyze the peripheral blood T-lymphocyte subsets of the Th1-related versus Th2-related cytokines of CD4+ cells, and the Tc1 versus Tc2 cytokines of CD8+ cells liver transplant recipients with versus without active cytomegalovirus (CMV) infection. METHODS: Isolated peripheral blood mononuclear cells (PBMC) were stimulated using PMA/Ionomycin/Monensin. Interleukin (IL)-4 and interferon gamma (IFN-gamma) production by CD4+ and CD8+T cells were determined using fluorescence-activated cell sorter (FACS) analysis. RESULTS: The ratios of CD4/CD8 were significantly lower among active CMV-infected patients. The levels of Th2 and Tc2 cytokines (IL-4) were similar between CMV-infected and uninfected patients. However, the levels of Th1-type and Tc1-type cytokines (IFN-r) were significantly lower among active CMV-infected patients. CONCLUSIONS: Low levels of Th1-type cytokines seem to correlate with active CMV infection in liver transplant recipients.     

Interleukin-18 restores immune suppression in patients with nonseptic surgery, but not with sepsis

BACKGROUND: We investigated cellular immune responses, in particular interferon gamma (IFN-gamma) production, by peripheral blood mononuclear cells (PBMCs) in patients with septic and nonseptic surgical stress, focusing on interleukin (IL)-18 and its receptor (IL-18R). METHODS: Thirty-two patients with alimentary tract carcinoma who underwent elective surgery (OP) and 26 septic patients (SP) with peritonitis were enrolled in this study. Blood was collected on the first postoperative day (POD1), POD5, POD10, and POD15 in the OP group and on the emergency admission in the SP group. Ten healthy volunteers served as controls. PBMCs were cultured in the presence of anti-CD3 antibody or IL-2 and IL-12, with or without additional IL-18 stimulation, to measure IFN-gamma production. IL-18R expression on CD56+ NK (natural killer) cells was evaluated by flow cytometry. RESULTS: IL-2- and IL-12-induced IFN-gamma production by PBMCs was suppressed significantly in both the OP (POD5) and SP groups compared with that in healthy controls. Interestingly, additional IL-18 stimulation up-regulated IFN-gamma production by PBMCs in the OP group as well as the control group, but not in the SP group. IL-18R expression on CD56+ NK cells was maintained consistently in the OP group as well as the control group, but decreased in the SP group. CONCLUSIONS: IFN-gamma production induced by cytokines (IL-2 and IL-12) was suppressed in PBMCs from both patients with sepsis and those who had undergone elective surgery. However, IL-18R expression on CD56+ NK cells was different between patients with sepsis and nonseptic surgical stress. Our results suggest that exogenous IL-18 administration may be effective in preventing immune suppression in patients with nonseptic elective surgery.     

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.     

Up-regulated interleukin-4 production by peripheral T-helper cells in idiopathic membranous nephropathy.

BACKGROUND: Helper T (Th) cells are classified into Th1 and Th2 subsets based on cytokine production and the Th1/Th2 paradigm explains differences in inflammatory effector pathways in various human diseases. Membranous nephropathy (MN) is an immune complex disease associated with Th2 nephritogenic immune response. However, overproduction of interleukin (IL)-4, a principal Th2 cytokine, has not been demonstrated. We investigated Th1/Th2 cytokine production by peripheral Th cells and its association with the degree of proteinuria in MN. METHODS: We analysed production of Th1/Th2 cytokines, interferon (IFN)-gamma and IL-4 by peripheral Th cells, using an intracellular cytokine detection method with flow cytometry in patients with MN (n = 24). The data were compared with data from healthy subjects (n = 51), subjects with minimal change nephrotic syndrome (MCNS; n = 13) and subjects with focal segmental glomerulosclerosis (FSGS; n = 12). We compared the percentages of IFN-gamma+ and IL-4+ Th cells and the peripheral Th1/Th2 ratio (IFN-gamma/IL-4 ratio) among the four groups. We also examined the association of IFN-gamma and IL-4 production with clinical parameters of MN. RESULTS: The mean percentage of IL-4+ cells in MN (3.9+/-1.2%) was significantly higher than in the control (2.4+/-1.0%), MCNS (2.3+/-1.4%) and FSGS (2.3+/-1.2%) groups (P<0.001, respectively). The Th1/Th2 ratio was significantly lower in MN (5.3+/-2.0) than in the control (8.2+/-4.2, P<0.05), MCNS (10.0+/-5.3, P<0.01) and FSGS (10.2+/-5.3, P<0.01) groups. Moreover, the percentage of IL-4+ cells correlated significantly with the amount of proteinuria in MN (r = 0.57, P<0.01). CONCLUSIONS: IL-4 production by peripheral Th cells is up-regulated in patients with MN and correlated with the severity of proteinuria. Intracellular cytokine analysis could be a useful index in idiopathic MN.     

Inhibition of CD1d activation suppresses septic mortality: a role for NK-T cells in septic immune dysfunction

BACKGROUND: Studies indicate that following septic insult there is development of generalized immune dysfunction in T cells, B cells and phagocytes, which is thought to contribute to morbidity and mortality. Specifically, there is a shift in the lymphocytes of septic animals toward an increased release of Th2 cytokines. NK-T cells have been shown to contribute to propagation of the Th2 response. The influence of NK-T cells on the immune response to septic challenge is poorly understood. In this study, we examine whether NK-T cells contribute to the immune dysfunction seen following the onset of polymicrobial sepsis, as produced by cecal ligation and puncture (CLP). MATERIALS AND METHODS: Male 129S1/SvImJ mice were pretreated with either rat IgG (isotypic control) or monoclonal antibody to CD1d (clone 1B1) (0.5 mg), which blocks signaling/antigen presentation via the CD1d cell surface receptor, thereby, ablating the activation and differentiation of the NK-T cells. Septic survival with and without anti-CD1d (CLP/CD1d) pretreatment was assessed. Mice sacrificed 24 h after CLP were assessed for change in splenic %NK-T cell (via flourescense activated cell sector) and for splenic, hepatic, and lymphoid/macrophage production of pro-inflammatory or anti-inflammatory cytokines (via enzyme-linked immunosorbent assay). RESULTS: Administration of anti-CD1d reduced septic mortality 35% at 6-10 d (n = 23 mice/group) (P <.05). There was a consistent increase in the %CD3(+) NK1.1(+) cell population (NK-T cells) in septic mice (1.706%), which was markedly suppressed by pretreatment with anti-CD1d (0.592%). IL-6 and IL-10 levels were suppressed by anti-CD1d in the spleen and blood. CONCLUSIONS: Together these findings imply not only that NK-T cells may play a role in mediating the immune suppression seen in bacterial sepsis, but that inhibition of their activation promotes survival to septic challenge.     

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.     

Dendritic cells of IgA nephropathy patients have an impaired capacity to induce IgA production in naïve B cells

BACKGROUND: IgA nephropathy (IgAN) is the most common primary glomerulonephritis worldwide, characterized by mesangial IgA1 deposits. We have previously demonstrated that IgAN patients have a hampered IgA immune response after mucosal challenge with a neoantigen. Dendritic cells are critically involved in the initiation of humoral immune responses, not only via activation of T-helper cells, but also via direct effect on na?ve B cells. The aim of this study was to investigate the capacity of dendritic cells from IgAN patients to regulate IgA production. METHODS: Dendritic cells were generated by culturing monocytes for 7 days in the presence of interleukin (IL)-4 and granulocyte macrophage-colony-stimulating factor (GM-CSF). Dendritic cells from either IgAN patients (N= 12) or controls (N= 12) were cultured for 14 days with na?ve B cells in the presence of CD40L-transfected mouse fibroblasts (L-CD40L cells) and medium with or without IL-2 or IL-10. Supernatants were tested for the presence of immunoglobulins by specific enzyme-linked immunosorbent assay (ELISA). RESULTS: In the presence of CD40L and IL-10, dendritic cells were able to increase immunoglobulin production by na?ve B cells. Dendritic cells of IgAN patients induced significantly (P= 0.026) less IgA production than dendritic cells of control persons (2.30 microg/mL vs. 5.24 microg/mL), whereas no differences were found in the IgG and IgM production. When dendritic cells were replaced by supernatant of CD40L-stimulated dendritic cells of patients and controls, IgA production was increased, but no difference was seen between the two groups. CONCLUSION: In the present study we show that dendritic cells of IgAN patients have an impaired capacity to induce IgA production in na?ve B cells, which might explain the observed IgA hyporesponse upon mucosal challenge with a neoantigen.     

Systemic administration of interleukin-4 expressing plasmid DNA delays the development of glomerulonephritis and prolongs survival in lupus-prone female NZB x NZW F1 mice.

BACKGROUND: T helper (Th)1/Th2 balance determines the direction of some kinds of autoimmune diseases. Th1 cytokines, especially interferon (IFN)-gamma has been proven important in the pathogenesis in lupus. The present study examined the effects of administration of interleukin (IL)-4 (Th2 cytokine) expressing plasmid DNA (IL-4pDNA) on the development of glomerulonephritis and survival in lupus-prone female NZB x NZW (B/W)F1 mice. METHODS: B/WF1 mice were administrated intraperitoneally either with IL-4pDNA (100 microg/mouse), plasmid (100 microg/mouse) or saline at 4 and 6 weeks of age and at 4 week intervals from 8 to 32 weeks of age. RESULTS: Compared to the saline and plasmid groups (controls), the IL-4pDNA-treatment drastically delayed the development of glomerulonephritis with deposits of IgG2a and C3 leading to excretion of urine protein, and prolonged survival. Clinical improvement was associated with the reduction in productions of IgG anti-dsDNA autoantibody. Also, compared to the other two controls the IL-4pDNA-treatment reduced production of IFN-gamma and increased IL-4 production from splenic cells. CONCLUSIONS: The present study suggests that systemic IL-4pDNA administration may delay lupus onset by suppressed IFN-gamma production due to shifting from Th1 to Th2 responses.     

干预协同刺激信号诱导不明原因早期复发性流产患者外周血Th1/Th2转换的体外研究

目的观察体外干预细胞毒性T细胞相关抗原4(CTLA-4)对不明原因早期复发性流产(URSA)患者外周血辅助性T细胞(Th)1/Th2转换的影响。方法用人绒毛膜癌细胞株JEG-3制备的滋养细胞抗原刺激30例URSA患者外周血单核细胞,分为实验组A:加重组蛋白CTLA-4Ig(10和1 mg/L)组,实验组B:加CTLA-4抗体(10和1 mg/L)组;对照组:加IgG(10和1 mg/L)组。采用酶联免疫吸附试验(ELISIA)测上清液中白细胞介素(IL)-2、干扰素γ(IFN-γ)和IL-4水平。结果与相应浓度的对照组相比,CTLA-4Ig组IL-4水平显著升高(P<0.05),IL-2显著降低(P<0.05),IFN-γ两组间无显著性差异(P>0.05);而CTLA-4抗体组IL-4水平均显著低于对照组(P<0.01),而IFN-γ、IL-2水平均明显高于对照组(P<0.05)。CTLA-4Ig 10 mg/L组与CTLA-4Ig 1mg/L组比较,IL-4显著升高(P<0.05),IL-2显著降低(P<0.05),IFN-γ两组间无显著性差异(P>0.05)。与同浓度的对照组及CTLA-4抗体1 mg/L组相比,CTLA-4抗体10 mg/L组IL-4较低,而IFN-γ和IL-2稍高,但两组间亦无显著性差异(P>0.05)。结论CTLA-4Ig使URSA患者Th1/Th2型细胞因子向Th2型细胞因子偏移,有利于URSA的治疗;而CTLA-4抗体可以上调Th1型细胞因子,下调Th2型细胞因子,不利于妊娠。     

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.     

Anti -PLA2R autoantibodies in serum and IgG subclass deposits on glomeruli in undetermined atypical membranous nephropathy

ObjectiveTo investigate the characteristic of autoantibodies of M – type phospholipase A2 receptor (PLA2R) in serum and the glomerular IgG subclass deposits in undetermined atypical membranous nephropathy (MN) patients. MethodsFrom Feb 2004 to Nov 2011, 53 cases diagnosed as MN by kidney puncture biopsy in our hospital were included into the study. There were 20 undetermined atypical membranous nephropathy (UAMN), 20 idiopathic membranous nephropathy (IMN) and 13 secondary membranous nephropathy (SMN) which were composed of lupus membranous nephropathy (LMN) and HBV related membranous nephropathy (HBV-MN). Clinlical and pathological characteristics were analyzed. The autoantibodies of PLA2R in serum were detected and the glomerular IgG subclass deposits were observed. Results(1) The average age underwent renal biopsy was (37.9±3.8) years of UAMN, (50.1±3.0) years of IMN and (49.5±4.5) years of SMN. The difference in onset average age at disease was significant between UAMN and IMN (P＝0.0178). The female/male ratio (F/M) in UAMN, IMN and SMN was 0.8∶1, 0.7∶1 and 0.6∶1(P＞0.05). (2) Compared with SMN, the level of 24-hours urinary protein excretion (3.47 g vs 7.89 g, P＝0.023), the ratio of amount urinary protein patients (50.0% vs 84.6%, P＝0.043), the level of serum IgG [(8.40±3.58) g/L vs (10.09±4.69) g/L, P＝0.025] and the positive rate of ANA in serum (10.0% vs 53.8%, P＝0.006) in UAMN were all much lower. There were no significant statistical differences in serum albumin, serum creatinine, eGFR, positive rate of HBsAg, HBeAg or HCV, as well as the ratio of hypo - albuminemia and nephritic syndrome among the three groups. (3) IF positive rate of IgA, IgM and C1q in UAMN were all significantly higher than that in IMN (P＜0.01). There were no significant differences in IF positive rate of IgA, IgM, C1q, IgG and C3 between UAMN and SMN. The IF strength of IgA, IgG, IgM, C3 and C1q in UAMN showed no significant differences between UAMN and SMN. (4) The serum autoantibodies of PLA2R were only detected in 10 cases of IMN group (50%) with all the other cases negative. This detection rate of serum autoantibodies of PLA2R showed significant statistical differences among the three groups (P＜0.01), but no differences between UAMN and SMN (the detection rate in both groups were 0%). (5) IgG1 deposits was the dominant IgG on the glomeruli in UAMN group (40%), as well as in SMN group (76.9%). IgG4 deposits was the dominant IgG on the glomeruli in IMN group (60%). The positive rate of IgG1 and IgG3 in UAMN showed no significant statistical differences when compared with IMN or SMN. The positive rate of IgG2 in UAMN was significantly lower than in SMN（30.0% vs 69.2%, P＜0.05). The positive rate of IgG4 in UAMN was significantly lower than in IMN (20% vs 60%, P＜0.05). The positive rate of IgG1, IgG2 and IgG3 in SMN were all significantly higher than in IMN. ConclusionsNone of the UAMN group had autoantibodies of PLA2R in serum, and IgG1 deposits was the dominant IgG subclass on the glomeruli which indicated the similarity with the SMN group. At the same time, UAMN was significantly different from SMN in clinical manifestations.