BACKGROUND & AIMS: :Methods to determine cytokine protein content in samples of interest, such as enzyme-linked immunosorbent assay (ELISA), are often labor-intensive and costly. Furthermore, because ELISA requires relatively large sample volumes and protein concentrations, it is difficult using this technique to determine protein content for multiple cytokines from individual samples. Recently, Luminex has developed an open source hardware platform combining flow cytometry- and bead-based antibody capture that is capable of detecting multiple analytes from a single sample. In the present study we employed the Luminex 200 platform to determine the cytokine protein content in discrete brain regions of C57BL/6J mice. In spike-and-recovery experiments, known concentrations of murine recombinant interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)alpha were added either singly or as a mixture of all three to whole brain homogenates containing known quantities of total protein. Spiked samples were assayed for either a single cytokine or for multiple cytokines using 1-plex or 3-plex assay kits, respectively. In whole mouse brain homogenate we recovered between 81% and 103% of the recombinant cytokines. We then injected C57BL/6J mice intraperitoneally with bacterial lipopolysaccharide (LPS) and sacrificed them 4h later. We detected in samples taken from LPS-stimulated mice 4- to 870-fold increases in serum or spleen cytokine protein, and 1.5- to 16-fold increases in cytokine protein in discrete brain regions, relative to protein content in samples obtained from vehicle-treated animals. These results indicate that multiple cytokines may be reliably assayed from discrete regions of mouse brain using a single sample.

背景与目标： : 确定目标样品中细胞因子蛋白含量的方法，例如酶联免疫吸附测定 (ELISA)，通常是劳动密集型且昂贵的。此外，由于ELISA需要相对较大的样品体积和蛋白质浓度，因此使用该技术很难确定来自单个样品的多种细胞因子的蛋白质含量。最近，Luminex开发了一种结合流式细胞术和基于珠子的抗体捕获的开源硬件平台，该平台能够从单个样品中检测多种分析物。在本研究中，我们使用Luminex 200平台来确定C57BL/6J小鼠离散脑区域中的细胞因子蛋白含量。在穗和恢复实验中，已知浓度的鼠重组白细胞介素 (IL)-1β，IL-6，并且将肿瘤坏死因子 (TNF) α 单独或作为所有三种混合物添加到包含已知量的总蛋白的全脑匀浆中。使用1-plex或3-plex检测试剂盒对加标样品进行单一细胞因子或多种细胞因子的测定，分别。在整个小鼠脑匀浆中，我们回收了81% 至103% 的重组细胞因子。然后，我们向C57BL/6J小鼠腹腔注射细菌脂多糖 (LPS)，并在4小时后将其处死。我们在从LPS刺激的小鼠身上采集的样本中检测到血清或脾脏增加4至870倍细胞因子蛋白，相对于从媒介物处理的动物获得的样品中的蛋白质含量，离散脑区域中的细胞因子蛋白增加1.5至16倍这些结果表明，使用单个样品可以从小鼠脑的离散区域可靠地测定多种细胞因子。

BACKGROUND & AIMS: :Rheumatoid arthritis (RA) is characterized by proliferation of synoviocytes that produce proinflammatory cytokines, which are implicated in the pathogenesis of RA. When human fibroblast-like synoviocytes line MH7A was treated with cigarette smoke condensate (CSC), either mainstream or sidestream, expression levels of interleukin (IL)-1alpha, IL-1beta, IL-6, IL-8, and CYP1A1 mRNA were upregulated in both time- and dose-dependent manners. The upregulatory effects of CSC on these cytokines were not significantly inhibited by alpha-naphthoflavone, an aryl hydrocarbon receptor (AhR) antagonist, suggesting that the effects of CSC were independent of AhR. Cycloheximide treatment indicated that the augmenting effect of CSC on IL-1alpha, IL-1beta and IL-8, but not IL-6 and CYP1A1, mRNA expression requires de novo protein synthesis. CSC also induced cytokines at protein levels and further augmented the effects of tumor necrosis factor alpha on induction of these cytokines at both mRNA and protein levels. These results support the epidemiological studies indicating a strong association between heavy cigarette smoking and pathogenesis of RA.

背景与目标： : 类风湿关节炎 (RA) 的特征是滑膜细胞的增殖，产生促炎细胞因子，这与RA的发病机理有关。当人类成纤维细胞样滑膜细胞系MH7A用香烟烟雾冷凝物 (CSC) 处理时，无论是主流还是侧流，白介素 (IL)-1α，IL-1beta，IL-6，IL-8和cyp1a1mrna的表达水平均随时间和剂量而增加。芳基烃受体 (AhR) 拮抗剂 α-萘黄酮并未显着抑制CSC对这些细胞因子的上调作用，这表明CSC的作用与AhR无关。环己酰亚胺处理表明，CSC对IL-1alpha，IL-1beta和IL-8的增强作用，但对IL-6和CYP1A1，mRNA表达不需要从头合成蛋白质。CSC还在蛋白质水平上诱导了细胞因子，并进一步增强了肿瘤坏死因子 α 在mRNA和蛋白质水平上诱导这些细胞因子的作用。这些结果支持流行病学研究，表明大量吸烟与RA的发病机理之间存在很强的关联。

BACKGROUND & AIMS: :Asthma is a triad of intermittent airway obstruction, bronchial smooth muscle cell hyperreactivity to bronchoconstrictors, and chronic bronchial inflammation. From an aetiological standpoint, asthma is a heterogeneous disease, but often appears as a form of immediate hypersensitivity. Many patients with asthma have other manifestations of atopy, such as rhinitis or eczema. Even among non-atopic patients with asthma, the pathophysiology of airway constriction is similar, raising the hypothesis that alternative mechanisms of mast cell degranulation may underlie the disease. The primary inflammatory lesion of asthma consists of accumulation of CD4(+) T helper type 2 (TH2) lymphocytes and eosinophils in the airway mucosa. TH2 cells orchestrate the asthmatic inflammation through the secretion of a series of cytokines, particularly interleukin 4 (IL-4), IL-13, IL-5, and IL-9. IL-4 is the major factor regulating IgE production by B cells, and is required for optimal TH2 differentiation. However, blocking IL-4 is not sufficient to inhibit the development of asthma in experimental models. In contrast, inhibition of IL-13, another TH2 cytokine whose signal transduction pathway overlaps with that of IL-4, completely blocks airway hyperreactivity in mouse asthma models. IL-5 is a key factor for eosinophilia and could therefore be responsible for some of the tissue damage seen in chronic asthma. IL-9 has pleiotropic activities on allergic mediators such as mast cells, eosinophils, B cells and epithelial cells, and might be a good target for therapeutic interventions. Finally, chemokines, which can be produced by many cell types from inflamed lungs, play a major role in recruiting the mediators of asthmatic inflammation. Genetic studies have demonstrated that multiple genes are involved in asthma. Several genome wide screens point to chromosome 5q31--33 as a major susceptibility locus for asthma and high IgE values. This region includes a cluster of cytokine genes, and genes encoding IL-3, IL-4, IL-5, IL-9, IL-13, granulocyte macrophage colony stimulating factor, and the beta chain of IL-12. Interestingly, for some of these cytokines, a linkage was also established between asthma and their receptor. Another susceptibility locus has been mapped on chromosome 12 in a region that contains other potential candidate cytokine genes, including the gene encoding interferon gamma, the prototypical TH1 cytokine with inhibitory activities for TH2 lymphocytes. Taken together, both experimental and genetic studies point to TH2 cytokines, such as IL-4, IL-13, IL-5, and IL-9, as important targets for therapeutic applications in patients with asthma.

背景与目标： : 哮喘是间歇性气道阻塞，支气管平滑肌细胞对支气管收缩剂的高反应性和慢性支气管炎症的三联征。从病因学的角度来看，哮喘是一种异质性疾病，但通常以即刻超敏反应的形式出现。许多哮喘患者有特应性的其他表现，如鼻炎或湿疹。即使在非特应性哮喘患者中，气道收缩的病理生理也相似，这提出了肥大细胞脱颗粒的替代机制可能是该疾病的基础的假设。哮喘的主要炎性病变包括气道粘膜中CD4 () T辅助2型 (TH2) 淋巴细胞和嗜酸性粒细胞的积累。TH2细胞通过分泌一系列细胞因子，特别是白介素4 (IL-4)，IL-13，IL-5和IL-9来协调哮喘炎症。IL-4是调节b细胞产生IgE的主要因子，是最佳TH2分化所必需的。然而，在实验模型中，阻断IL-4不足以抑制哮喘的发展。相反，抑制IL-13 (另一种TH2细胞因子的信号转导途径与IL-4的信号转导途径重叠) 完全阻断了小鼠哮喘模型中的气道高反应性。IL-5是嗜酸性粒细胞增多的关键因素，因此可能是慢性哮喘中某些组织损伤的原因。IL-9对肥大细胞，嗜酸性粒细胞，b细胞和上皮细胞等过敏介质具有多效性，可能是治疗干预的良好靶标。最后，趋化因子可以由发炎的肺部产生的许多细胞类型产生，在募集哮喘炎症介质中起主要作用。遗传学研究表明，多种基因与哮喘有关。几个基因组宽筛选显示5q31 -- 33号染色体是哮喘和高IgE值的主要易感位点。该区域包括细胞因子基因簇，以及编码IL-3，IL-4，IL-5，IL-9，IL-13，粒细胞巨噬细胞集落刺激因子和IL-12 β 链的基因。有趣的是，对于其中一些细胞因子，哮喘和它们的受体之间也建立了联系。另一个易感性基因座已定位在12号染色体上，该区域包含其他潜在的候选细胞因子基因，包括编码干扰素 γ 的基因，干扰素 γ 是对TH2淋巴细胞具有抑制活性的原型TH1细胞因子。总之，实验和遗传学研究都指出，TH2细胞因子 (例如IL-4，IL-13，IL-5和IL-9) 是哮喘患者治疗应用的重要靶标。

BACKGROUND & AIMS: :Regular exercise is a good non-pharmacological treatment of metabolic syndrome in that it improves obesity, diabetes, and inflammation. The 72 kDa extracellular heat shock protein (eHsp72) is released during exercise, thus stimulating the inflammatory responses. The aim of the present work was to evaluate the effect of regular exercise on the eHsp72-induced release of IL-1β, IL-6, and TNFα by macrophages from genetically obese Zucker rats (fa/fa) (ObZ), using lean Zucker (LZ) rats (Fa/fa) to provide reference values. ObZ presented a higher plasma concentration of eHsp72 than LZ, and exercise increased that concentration. In response to eHsp72, the macrophages from ObZ released less IL-1β and TNFα, but more IL-6, than macrophages from LZ. While eHsp72 stimulated the release of IL-1β, TNFα, and IL-6 in the macrophages from healthy LZ (with respect to the constitutive release), it inhibited the release of IL-1β and IL-6 in macrophages from ObZ. The habitual exercise improved the release of inflammatory cytokines by macrophages from ObZ in response to eHsp72 (it increased IL-1β and TNFα, and decreased IL-6), tending to values closer to those determined in healthy LZ. A deregulated macrophage inflammatory and stress response induced by eHsp72 underlies MS, and this is improved by habitual exercise.

背景与目标： : 定期运动是代谢综合征的一种很好的非药物治疗方法，因为它可以改善肥胖，糖尿病和炎症。在运动过程中释放72 kDa的细胞外热休克蛋白 (eHsp72)，从而刺激炎症反应。本工作的目的是评估定期运动对遗传肥胖Zucker大鼠 (fa/fa) (ObZ) 巨噬细胞eHsp72-induced释放IL-1β，IL-6和tnf α 的影响，使用精益Zucker (LZ) 大鼠 (Fa/fa) 提供参考值。ObZ的eHsp72血浆浓度高于LZ，运动增加了该浓度。响应eHsp72，来自ObZ的巨噬细胞比来自LZ的巨噬细胞释放更少的IL-1β 和tnf α，但更IL-6。虽然eHsp72刺激了健康LZ中巨噬细胞中IL-1β，tnf α 和IL-6的释放 (相对于组成型释放)，但它抑制了ObZ中巨噬细胞中IL-1β 和IL-6的释放。习惯性运动改善了巨噬细胞对eHsp72的响应从ObZ释放的炎性细胞因子 (它增加了IL-1β 和tnf α，并降低了IL-6)，趋向于接近健康LZ中确定的值。由eHsp72引起的巨噬细胞炎症和应激反应失调是MS的基础，而习惯性运动可以改善这种反应。

BACKGROUND & AIMS: :Th17-related cytokines (IL-17, IL-23, and IL-26) and receptors (IL-17R and IL-23R) were evaluated in MS patients under immunomodulatory IFN-β1 therapy during a 2year follow-up. Before the initiation of treatment, no significant difference was found in cytokine or receptor expression between controls and MS patients. Of the three cytokines evaluated, IL-26 was the highest in the patients' sera. The amount of IL-17 and CD13(+)IL-17R(+) cells was steadily decreased whereas IL-23 and IL-26 levels were gradually increased with IFN-β1 therapy. The patients in progressive phase had very high levels of IL-17. Th17-associated parameters should be considered in the immunomodulatory IFN-β1 therapy of MS.

背景与目标： : 在2年的随访中，在免疫调节IFN-β1治疗的MS患者中评估了Th17-related细胞因子 (IL-17，IL-23和IL-26) 和受体 (IL-17R和IL-23R)。在开始治疗之前，对照组和MS患者之间的细胞因子或受体表达没有发现显着差异。在评估的三种细胞因子中，IL-26在患者血清中最高。IFN-β1治疗后，IL-17和CD13(+)IL-17R(+) 细胞的数量稳步下降，而IL-23和IL-26水平逐渐上升。进展期患者的IL-17水平非常高。在MS的免疫调节IFN-β1治疗中应考虑Th17-associated参数。

BACKGROUND & AIMS: :The ability to induce several cytokines relevant to tuberculosis (TNF-α, IL-1β, IL-6, IL-12p40 and IL-23) by cord factor (trehalose dimycolate) from Mycobacterium alvei CR-21(T) and Mycobacterium brumae CR-270(T) was studied in the cell lines RAW 264.7 and THP-1, and compared to the ability of cord factor from Mycobacterium tuberculosis H37Rv, where this glycolipid appears to be implicated in the pathogenesis of tuberculosis. Details of the fine structure of these molecules were obtained by NMR and MS. The mycoloyl residues were identified as α and (ω-1)-methoxy in M. alvei CR-21(T) and α in M. brumae CR-270(T); in both cases they were di-unsaturated instead of cyclopropanated as found in M. tuberculosis. In RAW 264.7 cells, cord factors from M. alvei CR-21(T), M. brumae CR-270(T) and M. tuberculosis differed in their ability to stimulate IL-6, the higher levels corresponding to the cord factor from M. tuberculosis. In THP-1 cells, a similar overall profile of cytokines was found for M. alvei CR-21(T) and M. brumae CR-270(T), with high proportions of IL-1β and TNF-α, and different from M. tuberculosis, where IL-6 and IL-12p40 prevailed. The data obtained indicate that cord factors from the atypical mycobacteria M. alvei CR-21(T) and M. brumae CR-270(T) stimulated the secretion of several pro-inflammatory cytokines, although there were some differences with those of M. tuberculosis H37Rv. This finding seems to be due to their particular mycoloyl substituents and could be of interest when considering the potential adjuvanticity of these molecules.

背景与目标： : 在细胞系原始264.7和THP-1中研究了脐带因子 (海藻糖二CR-270) 诱导几种与结核病相关的细胞因子 (TNF-α，IL-1β，IL-6，IL-12p40和IL-23) 的能力，与结核分枝杆菌H37Rv的脐带因子的能力相比，该糖脂似乎与结核病的发病机理有关。通过NMR和MS获得了这些分子的精细结构的详细信息。菌酰基残基在CR-21(T) 中被鉴定为 α 和 (ω-1)-甲氧基，而在CR-270(T) 中被鉴定为 α; 在这两种情况下，它们都是二不饱和的，而不是在结核分枝杆菌中发现的环丙烷。在原始264.7细胞中，来自肺泡CR-21(T)，布鲁氏菌CR-270(T) 和结核分枝杆菌的脐带因子在刺激IL-6的能力上有所不同，较高水平对应于结核分枝杆菌的脐带因子。在THP-1细胞中，发现肺泡支原体CR-21(T) 和CR-270支原体 (T) 具有相似的细胞因子总体特征，IL-1β 和TNF-α 比例很高，与结核支原体不同，IL-6和IL-12p40盛行。获得的数据表明，来自非典型分枝杆菌CR-21(T) 和布鲁氏菌CR-270(T) 的脐带因子刺激了几种促炎细胞因子的分泌，尽管与结核分枝杆菌H37Rv有一些差异。这一发现似乎是由于它们特定的mycoloyl取代基，并且在考虑这些分子的潜在可调性时可能会引起关注。

BACKGROUND & AIMS: :Preeclampsia (PE), a specific syndrome of pregnancy, can be classified into early and late onset, depending on whether clinical manifestations occur before or after 34 weeks' gestation. We determined whether plasma concentrations of Hsp60 and Hsp70 were related to circulating cytokine levels, as well as kidney and liver functions, in early- and late-onset PE. Two hundred and thirty-seven preeclamptic women (95 with early- and 142 with late-onset PE) were evaluated. Plasma levels of Hsp60, Hsp70, and their specific antibodies, tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1, IL-10, IL-12, and soluble TNF-α-receptor I (sTNFRI) concentrations, were determined by enzyme-linked immunosorbent assay (ELISA). Concentrations of Hsp70, TNF-α, IL-1β, IL-12, and sTNFRI were significantly elevated in patients with early-onset PE compared with women with late-onset PE; IL-10 levels were significantly lower in the early-onset PE group. Concentrations of urea, uric acid, proteinuria, glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), and lactate dehydrogenase (LDH) were also significantly higher in early-onset PE. The percentage of infants with intrauterine growth restriction was also significantly higher in women with early-onset PE. There were positive correlations between Hsp70 levels and TNF-α, TNFRI, IL-1β, IL-12, GOT, GPT, LDH, and uric acid concentrations in early-onset PE group. Thus, early-onset PE was associated with greater maternal and fetal impairment. There are differences in pathophysiology between early- and late-onset PE, highlighting by the difference in Hsp70 levels.

背景与目标： : 先兆子痫 (PE) 是一种特定的妊娠综合征，可分为早发和晚发，具体取决于临床表现是在妊娠34周之前还是之后发生。我们确定了早期和晚期PE中Hsp60和Hsp70的血浆浓度是否与循环细胞因子水平以及肾脏和肝脏功能有关。评估了二百三十七名先兆子痫妇女 (95例早发性PE和142例晚发性PE)。通过酶联免疫吸附测定 (ELISA) 测定血浆Hsp60，Hsp70及其特异性抗体，肿瘤坏死因子-α (TNF-α)，白介素 (IL)-1，IL-10，IL-12和可溶性TNF-α 受体I (sTNFRI) 的浓度。与晚发PE患者相比，早发PE患者的Hsp70，TNF-α，IL-1β，IL-12和sTNFRI浓度显着升高; 早发PE组的IL-10水平显着降低。早发性PE的尿素，尿酸，蛋白尿，谷氨酸草酰乙酸转氨酶 (GOT)，谷氨酸丙酮酸转氨酶 (GPT) 和乳酸脱氢酶 (LDH) 的浓度也显着较高。早发性PE的女性中，宫内生长受限的婴儿百分比也显着更高。早发性PE组Hsp70水平与TNF-α，TNFRI，IL-1β，IL-12，GOT，GPT，LDH和尿酸浓度呈正相关。因此，早发性PE与更大的母体和胎儿损害有关。早发性PE和晚发性PE之间的病理生理差异，突出表现为Hsp70水平的差异。

BACKGROUND & AIMS: :Leptin is a hormone secreted by adipocytes. Besides controlling appetite and body weight, it has been suggested that leptin plays a role in inflammation and hemopoiesis. In this study we demonstrate that the pro-inflammatory/hemopoietic cytokines, IL-1beta, IL-6, TNF-alpha, and interferon-gamma, significantly inhibit gene expression and secretion of leptin by bone marrow adipocytes. These findings are in agreement with the data recently obtained from non-medullary adipose tissues. Within the bone marrow environment, leptin regulation by these pleiotropic cytokines could contribute to controlling the proliferation and differentiation of hemopoietic precursors as well as the maturation of stromal cells.

背景与目标： : 瘦素是一种由脂肪细胞分泌的激素。除了控制食欲和体重外，还表明瘦素在炎症和造血中起作用。在这项研究中，我们证明了促炎/造血细胞因子IL-1beta，IL-6，TNF-α 和干扰素-γ 显着抑制骨髓脂肪细胞的基因表达和瘦素分泌。这些发现与最近从非髓脂肪组织获得的数据一致。在骨髓环境中，这些多效性细胞因子对瘦素的调节可能有助于控制造血前体的增殖和分化以及基质细胞的成熟。

BACKGROUND & AIMS: :The galactoside-binding protein galectin-3 is increasingly recognized as an important player in cancer development, progression, and metastasis via its interactions with various galactoside-terminated glycans. We have shown previously that circulating galectin-3, which is increased up to 30-fold in cancer patients, promotes blood-borne metastasis in an animal cancer model. This effect is partly attributable to the interaction of galectin-3 with unknown receptor(s) on vascular endothelial cells and causes endothelial secretion of several metastasis-promoting cytokines. Here we sought to identify the galectin-3-binding molecule(s) on the endothelial cell surface responsible for the galectin-3-mediated cytokine secretion. Using two different galectin-3 affinity purification processes, we extracted four cell membrane glycoproteins, CD146/melanoma cell adhesion molecule (MCAM)/MUC18, CD31/platelet endothelial cell adhesion molecule-1 (PECAM-1), CD144/VE-cadherin, and CD106/Endoglin, from vascular endothelial cells. CD146 was the major galectin-3-binding ligand and strongly co-localized with galectin-3 on endothelial cell surfaces treated with exogenous galectin-3. Moreover, galectin-3 bound to N-linked glycans on CD146 and induced CD146 dimerization and subsequent activation of AKT signaling. siRNA-mediated suppression of CD146 expression completely abolished the galectin-3-induced secretion of IL-6 and G-CSF cytokines from the endothelial cells. Thus, CD146/MCAM is the functional galectin-3-binding ligand on endothelial cell surfaces responsible for galectin-3-induced secretion of metastasis-promoting cytokines. We conclude that CD146/MCAM interactions with circulating galectin-3 may have an important influence on cancer progression and metastasis.

背景与目标： : 半乳糖苷结合蛋白galectin-3通过与各种半乳糖苷末端聚糖的相互作用，越来越被认为是癌症发展，进展和转移的重要参与者。我们先前已经证明，在动物癌症模型中，循环galectin-3在癌症患者中增加了30倍，可促进血源性转移。这种作用部分归因于galectin-3与血管内皮细胞上未知受体的相互作用，并引起内皮分泌几种促进转移的细胞因子。在这里，我们试图鉴定内皮细胞表面负责galectin-3-mediated细胞因子分泌的galectin-3-binding分子。使用两种不同的galectin-3亲和纯化过程，我们从血管内皮细胞中提取了四种细胞膜糖蛋白，CD146/黑色素瘤细胞粘附分子 (MCAM)/MUC18，CD31/血小板内皮细胞粘附分子-1 (PECAM-1)，CD144/VE-钙粘蛋白和CD106/Endoglin。CD146是主要的galectin-3-binding配体，与galectin-3强烈共定位在用外源galectin-3处理的内皮细胞表面上。此外，galectin-3与CD146上的N-连接聚糖结合并诱导CD146二聚化和随后AKT信号传导的激活。siRNA介导的CD146表达抑制完全消除了内皮细胞galectin-3-induced分泌IL-6和g-csf细胞因子。因此，CD146/MCAM是内皮细胞表面的功能性galectin-3-binding配体，负责galectin-3-induced分泌促进转移的细胞因子。我们得出结论，CD146/MCAM与循环galectin-3的相互作用可能对癌症的进展和转移有重要影响。

BACKGROUND & AIMS: :Clinical features suggest differences in immune response among periodontitis forms, albeit a large number of cytokines and chemokines remain to be evaluated. The saliva is an available source of mediators and its analysis would be valuable in order to understand pathophysiological differences. The objective of this study was analyze chemokines/cytokines profile in whole saliva of individuals with severe periodontitis (Stage III) presenting moderate [Grade B; GB] or rapid progression rate with a localized incisor-molar pattern [Grade C; GC/IMP]. A case-control study was designed for each periodontitis group. GB (n = 9) and GC/IMP (n = 7) patients and their healthy controls (C-GB, n = 9 and C-GC, n = 7) were evaluated. Non-stimulated saliva samples were assessed by a multiplex assay for a total of 40 cytokines, C-C and C-X-C motif chemokines. GC/IMP group presented higher levels of CCL17 and CCL27 (p = 0.04, FDR > 0.05), and lower levels of CCL2 (p = 0.04, FDR > 0.05) and CCL25 (p = 0.006, FDR < 0.05) when compared to its control. GB patients had higher levels of IL-6, IL-1β (p = 0.04, FDR > 0.05), and elevated pro-inflammatory (TNF-α,IL-1β,INF-γ,IL-6, IL-16): anti-inflammatory (IL-2, IL-4, IL-10) ratio (p = 0.01, FDR < 0.05) compared to its control [p-values by Mann-Whitney test, and False Discovery Rate (FDR) by Benjamini-Hochburg corrections]. CCL-chemokines and cytokines contributed to differences between GC/C-GC and GB/C-GB, respectively (p < 0.05, PERMANOVA test). These preliminary data revealed that each periodontitis phenotype presented distinct immune profiles differentially expressed in saliva compared to their related controls, suggesting differences in the etiopathogenesis of GB and GC/IMP.

背景与目标： : 临床特征表明牙周炎形式之间的免疫反应存在差异，尽管仍有大量细胞因子和趋化因子有待评估。唾液是介质的可用来源，其分析对于了解病理生理差异将很有价值。这项研究的目的是分析重度牙周炎 (III期) 个体的整个唾液中的趋化因子/细胞因子谱，这些个体表现出中度 [B级; GB] 或具有局部切牙-磨牙模式的快速进展速度 [C级; GC/IMP]。针对每个牙周炎组设计了病例对照研究。评估了GB (n = 9) 和GC/IMP (n = 7) 患者及其健康对照 (C-GB，n = 9和C-GC，n = 7)。通过多重分析评估了未刺激的唾液样品，共40种细胞因子，c-c和c-x-c基序趋化因子。与对照相比，GC/IMP组表现出较高水平的CCL17和CCL27 (p = 0.04，FDR> 0.05) 和较低水平的CCL2 (p = 0.04，FDR> 0.05) 和CCL25 (p = 0.006，FDR <0.05)。GB患者的IL-6，IL-1β 水平较高 (p = 0.04，FDR> 0.05)，促炎性 (TNF-α，IL-1β，INF-γ，IL-6，IL-16): 抗炎 (IL-2，IL-4，IL-10) 比率升高 (p = 0.01，FDR <0.05) 与其对照 [通过Mann-Whitney检验的p值和通过本杰明尼-霍奇堡校正的错误发现率 (FDR)] 相比。CCL-趋化因子和细胞因子分别导致GC/c-gc和GB/c-gb之间的差异 (p <0.05，PERMANOVA测试)。这些初步数据显示，与相关对照相比，每种牙周炎表型在唾液中表现出不同的免疫谱，表明GB和GC/IMP的发病机理存在差异。

BACKGROUND & AIMS: :Plasticin-L1 (GLVNGLLSSVLGGGQGGGGLLGGIL) is a conformationally flexible glycine/leucine-rich peptide originally isolated from norepinephrine-stimulated skin secretions of the South-American Santa Fe frog Leptodactylus laticeps (Leptodactylidae). A nuclear magnetic resonance/molecular dynamics characterization of plasticin-L1 in the presence of dodecylphosphocholine (DPC) and DPC/sodium dodecylsulphate micelles as membrane-mimetic models showed that the peptide has affinity for both neutral and anionic membranes. The peptide adopts a stable helical conformation at the N-terminal region and a more disordered helix at the C-terminal region, separated by an unstructured loop wherein the highest number of glycines is localized. In both micelle environments, plasticin-L1 slowly inserts between the detergent head groups but always remains localized at the micelle/water interface. Plasticin-L1 lacks direct antimicrobial activity but stimulates cytokine production by macrophages. Incubation with plasticin-L1 (20 μg/mL) significantly (P < 0.05) increased the production of the proinflammatory cytokines IL-1β, IL-12, IL-23, and TNF-α from unstimulated peritoneal macrophages from both C57BL/6 and BALB/C mice. The peptide also increased IL-6 production by unstimulated (P < 0.01) and lipopolysaccharide-stimulated (P < 0.01) macrophages, whereas the effects on production of the anti-inflammatory cytokine IL-10 were not significant. These findings suggest that plasticin-L1 may play an immunomodulatory role in vivo by stimulating cytokine production from frog skin macrophages in response to microbial pathogens. This peptide may represent a template for the design of peptides with therapeutic applications as immunostimulatory agents.

背景与目标： : Plasticin-L1 (glvngllssvlgggggqgggggllggil) 是一种构象灵活的甘氨酸/亮氨酸肽，最初是从南美圣达菲青蛙leptodactyleps (Leptodactylidae) 的去甲肾上腺素刺激的皮肤分泌物中分离出来的。在十二烷基磷酸胆碱 (DPC) 和DPC/十二烷基硫酸钠胶束作为膜模拟模型的存在下，plasticin-L1的核磁共振/分子动力学表征表明，该肽对中性和阴离子膜均具有亲和力。肽在N端区域采用稳定的螺旋构象，在C端区域采用更无序的螺旋，由非结构化环隔开，其中甘氨酸的数量最多。在两种胶束环境中，plasticin-L1缓慢地插入洗涤剂头基团之间，但始终保持在胶束/水界面处。Plasticin-L1缺乏直接的抗微生物活性，但刺激巨噬细胞产生细胞因子。与plasticin-L1 (20 μ g/mL) 孵育显着 (P < 0.05) 增加了来自C57BL/6和BALB/C小鼠的未刺激腹膜巨噬细胞的促炎细胞因子IL-1β，IL-12，IL-23和TNF-α 的产生。该肽还增加了未刺激 (P < 0.01) 和脂多糖刺激 (P < 0.01) 的巨噬细胞产生的IL-6，而对抗炎细胞因子IL-10产生的影响并不显著。这些发现表明，plasticin-L1可能通过刺激青蛙皮肤巨噬细胞对微生物病原体的反应产生细胞因子而在体内发挥免疫调节作用。该肽可以代表设计具有作为免疫刺激剂的治疗应用的肽的模板。

BACKGROUND & AIMS: :Exposure to Trypanosoma cruzi parasites induces monocytes and macrophages to produce various endogenous mediators, including prostaglandins and cytokines. To clarify the involvement of monocytes as an important source of inflammatory mediators in Chagas disease patients, we evaluated PBMC before and after depletion of adherent cells (monocytes) from patients with indeterminate (IND) and cardiac (CARD) clinical forms and from non-infected individuals (NI). We demonstrated that after the partial depletion of adherent cells, production of PGE2 was slightly decreased in patients with Chagas disease. Inhibition of the cells by indomethacin increased the proliferation in PBMC cells from patients after antigen stimulation. Pro-inflammatory cytokines as IL-2 and IFN-γ also had a greater decrease after partial depletion of adherent cells in both clinical forms of Chagas disease. IL-10 and IL-5 levels were also reduced after partial depletion of adherent cells both in IND and CARD patients. In addition, we evaluated the APC potential of B cells and observed that the MHCII and CD80 molecules had an increased expression after partial depletion of most monocytes in all groups. Thus, inflammatory mediators produced by monocytes seem to be important to modulate immune responses in Chagas disease by regulating the processes of inflammation and antigen presentation.

背景与目标： : 暴露于克氏锥虫寄生虫会诱导单核细胞和巨噬细胞产生各种内源性介质，包括前列腺素和细胞因子。为了阐明单核细胞作为Chagas病患者炎症介质的重要来源的参与，我们评估了来自不确定 (IND) 和心脏 (CARD) 临床形式的患者以及未感染的患者的粘附细胞 (单核细胞) 耗竭前后的PBMC。个体 (NI)。我们证明，在部分耗尽贴壁细胞后，恰加氏病患者PGE2的产生略有减少。消炎痛对细胞的抑制作用增加了抗原刺激后患者PBMC细胞的增殖。在两种临床形式的恰加氏病中，粘附细胞部分耗尽后，促炎细胞因子如IL-2和IFN-γ 也有更大的降低。IND和CARD患者的粘附细胞部分耗竭后，IL-10和IL-5水平也降低。此外，我们评估了b细胞的APC潜力，并观察到在所有组中大多数单核细胞部分耗竭后，MHCII和CD80分子的表达增加。因此，单核细胞产生的炎症介质似乎对于通过调节炎症和抗原呈递过程来调节恰加人病的免疫反应很重要。

BACKGROUND & AIMS: BACKGROUND:The balance between inflammatory and anti-inflammatory responses of the immune system has been demonstrated to determine the fate of transplanted allografts. Here we analyzed CD19+CD24hiCD38hi immature transitional regulatory B (TRB) cells, as well as the gene and protein levels of interleukin (IL)-10 and transforming growth factor (TGF)-β in the three separate groups, include of stable transplanted subjects, chronic antibody-mediated rejection (cAMR) patients, and healthy individuals. METHOD:Peripheral blood mononuclear cells (PBMCs) from stable subjects (n = 36), cAMR patients (n = 36) and healthy controls (n = 18) were isolated. Flowcytometry was performed for CD19, CD24, and CD38 surface markers. ELISA and quantitative real-time PCR were performed for IL-10 and TGF-β cytokines. RESULT:The percentages of immature TRB cells were significantly decrease in cAMR patients (0.98%) versus stable recipients (2.81%) and healthy subjects (4.03%) (P = 0.001 and P < 0.001, respectively). Total lymphocytes, circulating B cells, memory and mature subsets of B cells did not show any significant difference between the groups. TGF-β mRNA was 3-fold upregulated in the cAMR group compared to stable patients (P < 0.001.), but without significant alteration at the protein level. Also, long-term survival renal transplant recipients had a higher protein but not mRNA levels of IL-10 than short-term survival renal transplant recipients. CONCLUSION:It seems that immature TRB cell subpopulation might be a crucial regulator of immune system response and plays an important role in determining the transplantation outcome. Furthermore, immunosuppressive IL-10 and TGF-β cytokines might act as a double sword and can exhibit either pathogenic or protective effects against allograft.

背景与目标：

BACKGROUND & AIMS: :Recent seroepidemiological and immunohistochemical studies have demonstrated an association between microbial infections and atherosclerosis. However, the mechanisms underlying this association are widely unknown. In the present study, arterial specimens obtained at autopsy after sudden death were analyzed concerning (1) the presence of Chlamydia pneumoniae, cytomegalovirus, herpes simplex virus, and Helicobacter pylori; (2) the expression of secretory group IIA phospholipase A(2) (sPLA(2)-IIA) and of proinflammatory cytokines; and (3) the stage of atherosclerosis. Genomic DNA of microbial pathogens was determined by the polymerase chain reaction technique. The expression of sPLA(2)-IIA was studied immunohistochemically by using monoclonal antibodies against human sPLA(2)-IIA. Transcripts specific for sPLA(2)-IIA, interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma were identified by reverse transcription-polymerase chain reaction. In 18 of 102 analyzed specimens, DNA of microbial pathogens was found. Thirteen sections were positive for C pneumoniae, whereas 2 specimens were positive either for cytomegalovirus or for herpes simplex virus. One section contained genomic DNA of all 3 pathogens simultaneously. None of the analyzed tissues exhibited nucleic acids specific for H pylori. In addition to macrophage infiltrates, the presence of microbial DNA was closely associated with the occurrence of transcripts specific for proinflammatory cytokines and sPLA(2)-IIA. Pathogens as well as sPLA(2)-IIA and cytokines were found to be present not only in advanced but also in early stages of atherosclerosis. In tissues negative for sPLA(2)-IIA and cytokine expression, none of the pathogens could be identified. Because macrophages exposed to phospholipase A(2)-treated lipoproteins are transformed into foam cells in vitro, the results of this study suggest an alternative mechanism by which microbial infections may act in a proatherogenic fashion in vessel walls.

背景与目标： : 最近的血清流行病学和免疫组织化学研究表明，微生物感染与动脉粥样硬化之间存在关联。但是，这种关联的潜在机制尚不清楚。在本研究中，对猝死后尸检获得的动脉标本进行了分析 :( 1) 肺炎衣原体，巨细胞病毒，单纯疱疹病毒和幽门螺杆菌的存在; (2) 分泌型IIA磷脂酶A(2) (sPLA(2)-IIA) 和促炎细胞因子的表达; (3) 动脉粥样硬化的阶段。通过聚合酶链反应技术确定微生物病原体的基因组DNA。通过使用针对人sPLA(2)-IIA的单克隆抗体，免疫组织化学研究了sPLA(2)-IIA的表达。通过逆转录聚合酶链反应鉴定sPLA(2)-IIA，interleukin-1beta，肿瘤坏死因子-α 和干扰素-γ 的特异性转录本。在102个分析标本中的18个中，发现了微生物病原体的DNA。13个切片对肺炎C呈阳性，而2个标本对巨细胞病毒或单纯疱疹病毒呈阳性。一个部分同时包含所有3种病原体的基因组DNA。分析的组织均未显示出对幽门螺杆菌特异性的核酸。除了巨噬细胞浸润外，微生物DNA的存在与促炎细胞因子和sPLA(2)-IIA特异性转录本的发生密切相关。发现病原体以及sPLA(2)-IIA和细胞因子不仅存在于动脉粥样硬化的晚期，而且还存在于动脉粥样硬化的早期。在sPLA(2)-IIA和细胞因子表达阴性的组织中，无法鉴定出任何病原体。由于暴露于磷脂酶A(2) 处理的脂蛋白的巨噬细胞在体外转化为泡沫细胞，因此这项研究的结果表明，微生物感染可能以动脉粥样硬化的方式在血管壁中起作用的另一种机制。

BACKGROUND & AIMS: :Bacterial products [lipopolysaccharide (LPS) with Gram-negative bacteria and toxins, superantigens or cell wall fragments with Gram-positive bacteria] are the main activators of the septic shock cascade. These molecules interact with monocytes, macrophages and endothelial cells to produce inflammatory cytokines [tumour necrosis factor (TNF) and interleukins 1 and 6], and may activate other harmful pathways such as the coagulation system, complement cascade and lipid mediators. As a therapeutic strategy, antibodies directed against LPS have been well studied, although, on the whole, the clinical results have been disappointing. Other possible interventions that have not yet been tested clinically include natural intracellular antibacterial proteins (e.g. bacterial permeability-increasing protein) and high density lipoprotein (responsible for detoxifying LPS in the body). The stimulation pathway of responsive cells by bacterial products is also another possible target for intervention. Compounds under investigation include soluble CD14 and antibodies directed against CD14 or LPS binding protein. Antibodies directed against the cytokines are another option. Anti-TNF antibodies are currently being investigated, but conclusive evidence of their activity is still lacking. Soluble receptors (e.g. interleukin-1 receptor antagonist, or soluble TNF receptor) are another possibility; one soluble TNF receptor is still undergoing clinical investigation.

背景与目标： : 细菌产物 [具有革兰氏阴性细菌和毒素的脂多糖 (LPS)，具有革兰氏阳性细菌的超抗原或细胞壁碎片] 是败血性休克级联的主要激活剂。这些分子与单核细胞，巨噬细胞和内皮细胞相互作用，产生炎性细胞因子 [肿瘤坏死因子 (TNF) 和白介素1和6]，并可能激活其他有害途径，例如凝血系统，补体级联反应和脂质介质。作为一种治疗策略，针对LPS的抗体已经得到了很好的研究，尽管总的来说，临床结果令人失望。尚未进行临床测试的其他可能的干预措施包括天然细胞内抗菌蛋白 (例如细菌通透性增加蛋白) 和高密度脂蛋白 (负责体内LPS的解毒)。细菌产物对应答细胞的刺激途径也是另一个可能的干预目标。正在研究的化合物包括可溶性CD14和针对CD14或LPS结合蛋白的抗体。针对细胞因子的抗体是另一种选择。目前正在研究抗TNF抗体，但仍缺乏其活性的确凿证据。可溶性受体 (例如interleukin-1受体拮抗剂或可溶性TNF受体) 是另一种可能性; 一种可溶性TNF受体仍在进行临床研究。