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1.
目的:探讨谷氨酰胺(Gln)抑制人外周血单个核细胞(PBMC)细胞因子过度表达的分子机制。方法:用Ficoll密度梯度离心法提取健康志愿者新鲜外周血单个核细胞,在培养液中加入Gln、P38MAPK通路阻滞剂(SB203580)和HSP70阻断剂(Quercetin),用内毒素刺激4 h后,留取细胞和上清液,ELISA法测定内毒素刺激下单个核细胞肿瘤坏死因子(TNF-α)和IL-10的表达,Western blot方法检测细胞内磷酸化P38的表达情况。结果:Gln能明显增加HSP70的表达,同时TNF-α和IL-10表达受抑;使用Quercetin和SB203580后,磷酸化P38减少的同时TNF-α和IL-10也减少了。结论:Gln下调LPS刺激下PBMC细胞因子的过度表达依赖于P38MAPK信号蛋白的抑制。  相似文献   

2.
Theoretically, vitamin supplements may either enhance or reduce protein synthesis and proliferation in peripheral blood mononuclear cells (PBMC). In the present study, we determined whether administration of a pharmacologic dose of biotin affects proliferation rates of PBMC and cytokine release. Healthy adults (n = 5) ingested 3.1 micromol biotin/d for 14 d; blood and urine were collected pre- and postsupplementation. PBMC were isolated by density gradient and incubated with the mitogen concanavalin A for up to 3 d. At timed intervals during mitogen stimulation, we measured the following: 1) cellular uptake of [(3)H]thymidine to determine proliferation rates; 2) concentrations of various cytokines released into the medium; and 3) the percentages of PBMC subsets as judged by CD surface markers. Biotin supplementation caused a significant decrease of PBMC proliferation. At 2 d after mitogen stimulation, [(3)H]thymidine uptake by postsupplementation PBMC was 66 +/- 21% of the uptake by presupplementation PBMC (P < 0.05). Similarly, concentrations of interleukin-1beta (2 d after mitogen) and interleukin-2 (1 d after mitogen) in media from postsupplementation PBMC were 65 +/- 28% and 44 +/- 23%, respectively, of those for presupplementation PBMC (P < 0.01). Percentages of PBMC subsets were not affected by 14 d of biotin supplementation. Overall, this study provides evidence that administration of pharmacologic doses of biotin for 14 d decreases PBMC proliferation and synthesis of interleukin-1beta and interleukin-2.  相似文献   

3.
BACKGROUND: Dietary fish or fish oil rich in n-3 fatty acids (n-3 FAs), eg, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), ameliorate inflammatory reactions by various mechanisms. Whereas most studies have explored the effects of predominantly EPA-based n-3 FAs preparations, few have addressed the effects of n-3 FAs preparations with DHA as the main FA. OBJECTIVE: The objective was to determine the effects of 6 mo of dietary supplementation with an n-3 FAs preparation rich in DHA on release of cytokines and growth factors from peripheral blood mononuclear cells (PBMCs). DESIGN: In a randomized, double-blind, placebo-controlled trial, 174 Alzheimer disease (AD) patients received daily either 1.7 g DHA and 0.6 g EPA (n-3 FAs group) or placebo for 6 mo. In the present study blood samples were obtained from the 23 first randomized patients, and PBMCs were isolated before and after 6 mo of treatment. RESULTS: Plasma concentrations of DHA and EPA were significantly increased at 6 mo in the n-3 FAs group. This group also showed significant decreases of interleukin (IL)-6, IL-1beta, and granulocyte colony-stimulating factor secretion after stimulation of PBMCs with lipopolysaccharide. Changes in the DHA and EPA concentrations were negatively associated with changes in IL-1beta and IL-6 release for all subjects. Reductions of IL-1beta and IL-6 were also significantly correlated with each other. In contrast, this n-3 FA treatment for 6 mo did not decrease tumor necrosis factor-alpha, IotaL-8, IL-10, and granulocyte-macrophage colony-stimulating factor secretion. CONCLUSION: AD patients treated with DHA-rich n-3 FAs supplementation increased their plasma concentrations of DHA (and EPA), which were associated with reduced release of IL-1beta, IL-6, and granulocyte colony-stimulating factor from PBMCs. This trial was registered at clinicaltrials.gov as NCT00211159.  相似文献   

4.
Micronutrient deficiencies are frequently associated with tuberculosis (TB) worldwide. We tested the effect of exogenous vitamin E on proliferation and cytokine production of peripheral blood mononuclear cells (PBMC) from TB patients and healthy purified protein derivative (PPD)+ volunteers. Proliferation was stimulated with mycobacterial antigen (PPD) and evaluated by the incorporation of tritiated thymidine in PBMC cultured with or without 50 microm-vitamin E for 6 d. Cytokine production (IL-2 and interferon (IFN)-gamma) was determined by intracellular cytokine staining and by ELISA in the supernatant of PBMC stimulated for 24 h with phytohaemagglutinin or PPD. Our results show that culture with vitamin E increased (P < or = 0.05 ) the antigen-induced proliferation of PBMC in TB patients but not in healthy PPD+ volunteers. No significant changes in the number of cytokine-producing cells or in the production of IFN-gamma were observed with vitamin E treatment. These results indicate that vitamin E may enhance the antigen-specific in vitro response of PBMC from TB patients.  相似文献   

5.
Lipid peroxidation of polyunsaturated fatty acids (PUFA) generates reactive products that may cause DNA damage. To examine the possible relationship between DNA damage in peripheral blood mononuclear cells (PBMC) and the concentration of PUFA in red blood cells (RBC), endogenous DNA strand breaks, formamidopyrimidine DNA glycosylase (FPG) sites, and hydrogen peroxide (H2O2) sensitive sites were evaluated by the comet assay in blood samples from 98 Icelandic women. Fatty acid composition of RBC was analyzed by gas chromatography. Endogenous DNA strand breaks in PBMC correlated positively with the concentration of total PUFA, total n-3 PUFA, docosahexaenoic acid, linoleic acid, oleic acid, and palmitic acid in RBC. However, there was no association between FPG sites or H(2)O(2) sensitive sites in DNA in PBMC and the concentration of total PUFA or total saturated fatty acid in RBC. As there was no association between oxidative DNA damage or sensitivity of DNA to oxidative stress and the concentration of PUFA in RBC, the positive association between endogenous DNA strand breaks in PBMC and the concentration of total PUFA in RBC is probably not related to oxidative stress.  相似文献   

6.
BACKGROUND: Atherosclerosis is a chronic inflammatory disease. We previously reported that a diet high in alpha-linolenic acid (ALA) reduces lipid and inflammatory cardiovascular disease risk factors in hypercholesterolemic subjects. OBJECTIVE: The objective was to evaluate the effects of a diet high in ALA on serum proinflammatory cytokine concentrations and cytokine production by cultured peripheral blood mononuclear cells (PBMCs) from subjects fed the experimental diets. DESIGN: A randomized, controlled, 3-diet, 3-period crossover study design was used. Hypercholesterolemic subjects (n = 23) were assigned to 3 experimental diets: a diet high in ALA (ALA diet; 6.5% of energy), a diet high in linoleic acid (LA diet; 12.6% of energy), and an average American diet (AAD) for 6 wk. Serum interleukin (IL)-6, IL-1beta, and tumor necrosis factor-alpha (TNF-alpha) concentrations and the production of IL-6, IL-1beta, and TNF-alpha by PBMCs were measured. RESULTS: IL-6, IL-1beta, and TNF-alpha production by PBMCs and serum TNF-alpha concentrations were lower (P < 0.05 and P < 0.08, respectively) with the ALA diet than with the LA diet or AAD. PBMC production of TNF-alpha was inversely correlated with ALA (r = -0.402, P = 0.07) and with eicosapentaenoic acid (r = -0.476, P = 0.03) concentrations in PBMC lipids with the ALA diet. Changes in serum ALA were inversely correlated with changes in TNF-alpha produced by PBMCs (r = -0.423, P < 0.05). CONCLUSIONS: Increased intakes of dietary ALA elicit antiinflammatory effects by inhibiting IL-6, IL-1beta, and TNF-alpha production in cultured PBMCs. Changes in PBMC ALA and eicosapentaenoic acid (derived from dietary ALA) are associated with beneficial changes in TNF-alpha release. Thus, the cardioprotective effects of ALA are mediated in part by a reduction in the production of inflammatory cytokines.  相似文献   

7.
BACKGROUND: The incidence of cardiovascular diseases increases after menopause, and soy consumption is suggested to inhibit disease development. OBJECTIVE: The objective was to identify biomarkers of response to a dietary supplementation with an isoflavone extract in postmenopausal women by proteome analysis of peripheral blood mononuclear cells. DESIGN: The study with healthy postmenopausal woman was performed in a placebo-controlled sequential design. Peripheral mononuclear blood cells were collected from 10 volunteers after 8 wk of receiving daily 2 placebo cereal bars and after a subsequent 8 wk of intervention with 2 cereal bars each providing 25 mg of isoflavones. The proteome of the cells was visualized after 2-dimensional gel electrophoresis, and peptide mass fingerprinting served to identify proteins that by the intervention displayed altered protein concentrations. RESULTS: Twenty-nine proteins were identified that showed significantly altered expression in the mononuclear blood cells under the soy-isoflavone intervention, including a variety of proteins involved in an antiinflammatory response. Heat shock protein 70 or a lymphocyte-specific protein phosphatase and proteins that promote increased fibrinolysis, such as alpha-enolase, were found at increased intensities, whereas those that mediate adhesion, migration, and proliferation of vascular smooth muscle cells, such as galectin-1, were found at reduced intensities after soy extract consumption. CONCLUSION: Proteome analysis identified in vivo markers that respond to a dietary intervention with isoflavone-enriched soy extract in postmenopausal women. The nature of the proteins identified suggests that soy isoflavones may increase the antiinflammatory response in blood mononuclear cells that might contribute to the atherosclerosis-preventive activities of a soy-rich diet.  相似文献   

8.
Neonates are thought to be generally deficient in production of Th-1-associated cytokines at birth, and thereby more susceptible to bacterial infections. Using neonatal foals as a model, this study examined the age-dependent maturation of both basal and stimulus-induced immune responses, as reflected by the expression of a panel of Th-1-associated and pro-inflammatory cytokines. Results showed that although the basal production of IFN-γ and IL-6 was impaired (P < 0.05) in PBMCs of neonatal foals at birth, the basal production of IL-8, IL-12(p35/p40) and IL-23(p19/p40) were either in excess of or comparable to that of older foals. In response to Rhodococcus equi and CpG-ODN stimulation in vitro, PBMCs of neonatal foals showed increased (P < 0.05) expression of IFN-γ and IL-6, and preferentially increased expression of either IL-23(p19/p40) with R. equi stimulation or IL-12(p35/p40) with CpG-ODN stimulation. The magnitude of these stimulus-induced responses (except for IL-23p19), were significantly (P < 0.05) less for newborn foals than for older foals. The selective impairment of age-dependent basal and stimulus-induced cytokine expression by newborn foals may reflect the different functional state of various TLR pathways in newborns, and be directly associated with their age-dependent susceptibility to infection. Our results indicate that CpG-ODNs can selectively stimulate deficient cytokines (P < 0.05) from PBMCs in newborn foals in vitro, suggesting immunoprophylactic or therapeutic potential of CpG-ODNs.  相似文献   

9.
OBJECTIVE: The present study examined the effect of zinc at concentrations of the apoptotic signaling pathway and immune function of peripheral blood mononuclear cells (PBMCs). METHODS: PBMCs from healthy subjects were treated in vitro with various zinc concentrations to imitate different serum statuses of physiologic (2 to 15 microM) and pharmacologic (15 to 100 microM) concentrations to higher than 100 microM and analyzed their expressions of cytokines and apoptotically related factors. RESULTS: Although a normal physiologic concentration of zinc had no effect on immunologic function or apoptosis of PBMCs, a pharmacologic concentration (100 microM) or higher affected both functions. Zinc decreased cell proliferation at concentrations higher than 100 microM and stimulated cytokine expression at concentrations of at least 100 microM. Further, at concentrations of at least 100 microM, apoptosis was induced, and expressions of caspase-3 and proapoptotic genes, including Fas (FasL) and c-fos, which trigger apoptosis through receptor-mediated extrinsic and mitochondrion-mediated apoptotic pathways, respectively, were increased. At concentrations at least 300 microM, expressions of antiapoptotic factors nuclear factor-kappaB, Bcl-2, and Bcl-X(L) were markedly decreased. CONCLUSIONS: Zinc stimulates cytokine expression and induces apoptosis of PBMCs from healthy subjects only at concentrations equal to or greater than the serum pharmacologic range. Receptor-mediated extrinsic and mitochondrial-mediated intrinsic pathways are involved in this zinc-induced apoptosis.  相似文献   

10.
To better characterize the cytokine response to measles virus vaccine, we examined the levels of IL-2, IL-4, IL-5, IL-10, IL-12 and γ-interferon (γ-IFN) in measles virus-stimulated peripheral blood mononuclear cells from 18 donors before and 2 weeks after vaccination. Donors were grouped as seropositive or seronegative on the basis of measles-specific IgM antibody present at 2 weeks postvaccination. After vaccination, similar levels of upregulation of IL-2 and γ-IFN mRNA were observed in the two groups. The majority of donors in both groups did not exhibit an increase in measles specific IL-4 or IL-10 mRNA after vaccination. IL-12 mRNA was not induced by measles virus in any of the donors. A statistically significant upregulation of IL-5 mRNA was observed among seropositive (9/13) compared with seronegative (1/5) donors after vaccination (P=0.09, one tailed Fisher's test). The observed measles specific induction of IL-5 mRNA is suggestive of a possible association between IL-5 production and an antibody response to measles virus.  相似文献   

11.
目的探讨杂色曲霉素(ST)对体外培养的人外周血单个核细胞表面(HPBMc)HLAⅠ分子表达的影响。方法采用流式细胞定量术(FCM)和免疫印迹(Westernblot)分析方法,研究不同浓度ST(0.125、0.25、0.5、1和2mgL)处理后人外周血单个核细胞表面HLAⅠ分子表达的变化。结果FCM定量分析结果表明,经不同浓度ST处理24小时后,与对照组相比,各组细胞HLAⅠ的平均荧光强度均降低,以较高浓度(0.5、1和2mgL)ST处理组降低更明显(P<0.05)。在0.125mgL到2mgL的浓度范围内,随ST处理浓度的升高,HLAⅠ荧光指数逐渐降低,两者呈明显的负相关(r=-0.841,P<0.01)。免疫印迹结果也表明,随ST浓度的增高,HLAⅠ分子降低越明显。结论提示在0.125mgL到2mgL的浓度范围内,ST抑制人外周血单个核细胞表面HLAⅠ分子的表达呈现出负的剂量-反应关系。  相似文献   

12.
杂色曲霉素是我国肿瘤高发区粮食中的优势污染霉菌毒素。为探讨杂色曲霉素 (ST)对人体免疫机能的影响 ,采用双抗体夹心ELISA方法对ST作用后人外周血单核细胞 (HPBMc)培养上清液中白血细胞介素Ⅱ (IL 2 )的分泌水平进行了检测。结果表明 ,不同浓度 ( 0 0 3 12 5~ 2mg L)ST处理 2 4h后 ,体外培养的HPB Mc的IL 2分泌均受到一定程度的抑制 ,其中以较低浓度ST( 0 0 3 12 5~ 0 12 5mg L)和较高浓度ST( 1~ 2mg L)抑制作用最明显 (P <0 0 5 )。在ST 1mg L作用 1~ 64h的时间范围内 ,ST对HPBMcIL 2的分泌总体表现抑制作用。ST处理后 8~ 64h ,随ST处理时间的延长 ,对IL 2分泌的抑制作用逐渐增强 (r =0 82 2 ,P <0 0 5 )。本研究结果提示 ,ST对HPBMcIL 2的分泌有一定的抑制作用  相似文献   

13.
Farming environment and environmental exposure to microbial agents have been suggested to promote favorable development of immune system in children and protect against allergic diseases. However, effects of farm exposure on adult immune responses are less clear. Aim of the present study was to examine associations of farm related factors and measured microbial exposure with stimulated production of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) in peripheral blood samples among farming women. Whole peripheral blood samples were obtained from 112 women living on farms and stimulated with phorbol myristate acetate/ionomycin, lipopolysaccharide and staphylococcal enterotoxin B. Following 24h stimulation, protein levels of IFN-γ and IL-4 in the supernatants were measured by ELISA. From house dust, concentrations of 3-hydroxy fatty acids (C10:0-C14:0, marker for Gram-negative bacteria), muramic acid (Gram-positive bacteria) and ergosterol (fungal biomass) were analyzed with GC-MS/MS and viable microbes by culturing. Information on farm related factors and allergic diseases were collected from self-administered questionnaires. We found that household pets or other current or childhood farm-related factors had only few associations with stimulated cytokine production among studied farming women. Similarly, no strong associations were observed between markers of microbial exposure measured in house dust and cytokine levels. Atopic sensitization, allergic rhinitis and recent respiratory infections were, however, associated with reduced IFN-γ production. Our results suggest that the capacity of the studied environmental factors to modulate immune system is relatively weak in adulthood.  相似文献   

14.
目的探讨嗜肺军团菌活菌刺激对人外周血单个核细胞(PBMC)表达TLR2 mRNA的影响。方法采用不同浓度的嗜肺军团菌活菌悬液刺激体外培养的健康人PBMC,收集细胞运用RT-PCR方法测定TLR2 mRNA的表达水平。结果析因分析结果显示,时间的主效应有统计学意义(F=26.06,P<0.05);时间与浓度的交互作用有统计学意义(F=11.39,P<0.05);24 h时,MOI 1、MOI 10刺激组TLR2 mRNA的表达量均高于对照组(P<0.05),呈现随菌液浓度升高表达量也逐渐升高,48 h时,各组之间TLR2 mRNA的表达量差异虽无统计学意义(P>0.05),但表达量随浓度升高呈现下降趋势,72 h时,各组之间TLR2 mRNA的表达量差异有统计学意义(P<0.05),但随菌液浓度升高表达量反而降低。结论用军团菌体外刺激人PBMC,其TLR2 mRNA的表达在一定范围内呈一定的时效与量效关系,可为今后的研究提供有益的时间和剂量参考。  相似文献   

15.
16.
BackgroundSyphilis is a sexually transmitted disease that threatens human health worldwide. However, the immune regulation cascade caused by treponemia pallidum (TP) infection remains still largely unclear.MethodsTo investigate the expression of ISGs in secondary syphilis (SS), we recruited 64 patients with SS and equal number of healthy participants to obtain their peripheral blood mononuclear cells (PBMCs). qRT-PCR was performed to estimate the expression of interferon-stimulated genes (ISGs) including CXCL10, OAS3, OAS1, MX1, IFIT3, IFIT2, IFI6 and AIM2. Receiver-operating characteristic (ROC) analysis was adapted to diagnostic value of these genes to distinguish healthy controls and patients with SS.ResultsISGs including CXCL10, OAS3, OAS1, MX1, IFIT3, IFIT2, IFI6 and AIM2 were all upregulated in PBMCs of patients with SS. Area under the ROC curve (AUC) of the 8 ISGs were all more than 0.5. IFIT3 exhibited the highest diagnostic value, followed by AIM2, IFIT2 and CXCL10, according to the Yoden Index.ConclusionISGs including CXCL10, OAS3, OAS1, MX1, IFIT3, IFIT2, IFI6 and AIM2 were upregulated in patients with SS and they have diagnostic value for syphilis.  相似文献   

17.
Widespread metabolic changes associated with injury facilitate the delivery of nutrients to the immune system. The effect of specific nutrients administered by the enteral route on the immune response in trauma victims is not well understood. The purpose of this study was to examine whether the synthesis of proinflammatory cytokines (tumor necrosis factor-alpha [TNF-alpha], interleukin-1 beta [IL-1 beta], and interleukin-6 [IL-6]) by peripheral blood mononuclear cells (PBMCs) are influenced by the nature of the dietary fat in critically injured trauma victims. We measured plasma TNF-alpha, IL-1 beta, and IL-6 and their release stimulated by phytohemagglutinin (PHA) and endotoxin (lipopolysaccharide, LPS) from PBMCs of 13 severely injured (injury severity score = 30 +/- 2) patients once within 48-60 h after injury and then after 7 d of enteral feeding (1.5 g protein[P].kg-1.d-1). Group I (n = 6) received diet A (Crucial) and group II (n = 7) received diet B (Impact). The plasma levels of TNF-alpha and IL-1 beta in trauma patients are not significantly different from those in healthy volunteers, but plasma IL-6 levels are significantly increased (five times) in severely injured patients. Stimulation of TNF-alpha and IL-1 beta secretion by LPS and PHA were significantly higher in patients than in control subjects; in contrast, there was no stimulation of IL-6 because of trauma or nutritional support by either of the diets. Stimulation of IL-1 beta by LPS was normalized by Crucial but was further enhanced by Impact. The higher fat content in Crucial may contribute in part to the apparent immunomodulation. Crucial seems to be a better choice in correcting the nutritional deficiency.  相似文献   

18.
目的 观察重症脓毒症患者外周血单核细胞(PBMC)表面膜CD14(mCD14)、人类白细胞抗原( HLA)-DR及炎性因子的表达及意义.方法 选取重症脓毒症患者35例(病例组)和健康志愿者15例(对照组).于入院后第1、3、5天检测其PBMC表面mCD14、HLA-DR表达,血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-10浓度及急性生理学和慢性健康状况Ⅱ评分(APACHEⅡ)、全身性感染相关器官功能衰竭评分(SOFA评分).结果 病例组患者PBMC表面mCD14、HLA-DR表达分别为(2.61±1.59)%、( 10.25±5.35)%,明显低于对照组的(5.57±1.53)%、(59.28±14.76)%,血清TNF-α、IL-10浓度分别为(96.66±45.38)、( 149.79±77.15) ng/L,明显高于对照组的(0.12±0.00)、(5.67±2.16) ng/L,差异均有统计学意义(P< 0.05或<0.01).病例组患者死亡10例,存活25例,28d病死率28.6%(10/35),死亡患者与存活患者PBMC表面mCD14、HLA-DR表达及SOFA评分、APACHEⅡ在入院后第1、3天比较差异均无统计学意义(P>0.05),第5天存活患者PBMC表面mCD14、HLA-DR表达明显高于死亡患者[(5.12±2.03)%比(2.75±0.67)%;(35.12±9.29)%比(13.06±5.87)% ](P< 0.01或<0.05),SOFA评分、APACHEⅡ明显低于死亡患者[(4.48±1.71)分比(10.70±3.16)分;(9.36±5.57)分比(25.60±10.88)分](P<0.01),而两者血清TNF-α、IL-10浓度在入院后第1、3、5天比较差异均无统计学意义(P>0.05).结论 重症脓毒症患者PBMC表面mCD14、HLA-DR表达与患者预后密切相关,血清TNF-α、IL-10浓度在入院后5d内的动态变化不能反映患者疾病的演变和预后.  相似文献   

19.
20.
在急性乙肝和慢性乙肝患者的外周血单个核细胞都有不同程度的乙型肝炎病毒感染。外周血单个核细胞不但可以从外周血摄取乙型肝炎病毒 ,而且乙型肝炎病毒可在其内复制、转录、翻译并能释放病毒颗粒 ,具有一定的感染性。乙型肝炎病毒感染外周血单个核细胞可逃避免疫反应 ,导致病情的隐匿化。当外周血单个核细胞所处的外环境改变时 ,可导致其再次激活 ,引起肝移植术后乙肝复发、乙肝的输血传播及母婴传播  相似文献   

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