The purinergic P2×7 receptor is expressed on monocytes in Behçet's disease and is modulated by TNF‐α

The P2×7 receptor (P2×7r) is expressed in innate immune cells (e.g. monocyte/macrophages), playing a key role in IL‐1β release. Since innate immune activation and IL‐1β release seem to be implicated in Behçet's disease (BD), a systemic immune‐inflammatory disorder of unknown origin, we hypothesized that P2×7r is involved in the pathogenesis of the disease. Monocytes were isolated from 18 BD patients and 17 healthy matched controls. In BD monocytes, an increased P2×7r expression and Ca²⁺ permeability induced by the selective P2×7r agonist 2′‐3′‐O‐(4‐benzoylbenzoyl)ATP (BzATP) was observed. Moreover, IL‐1β release from LPS‐primed monocytes stimulated with BzATP was markedly higher in BD patients than in controls. TNF‐α‐incubated monocytes from healthy subjects almost reproduced the findings observed in BD patients, as demonstrated by the increase in P2×7r expression and BzATP‐induced Ca²⁺ intake. Our results provide evidence that in BD monocytes both the expression and function of the P2×7r are increased compared with healthy controls, as the possible result, at least in part, of a positive modulating effect of TNF‐α on the receptor. These data indicate P2×7r as a new potential therapeutic target for the control of BD, further supporting the rationale for the use of anti‐TNF‐α drugs in the treatment of the disease.     

外周血CD14~+CD16~+细胞与稳定型心绞痛患者冠状动脉硬化病变程度的相关性研究

目的:单核细胞在整个动脉硬化进程中至关重要,根据细胞表面CD14和CD16的差异表达,单核细胞可分为CD14+CD16-和CD14+CD16+两种亚群,本研究目的是探讨外周血单核细胞亚群与稳定型心绞痛患者冠状动脉硬化病变程度的相关性。方法:入选97例进行冠状动脉造影的稳定型心绞痛患者,根据造影结果分为无冠心病组(non-coronary artery disease,non-CAD)、单支病变组(single-vessel disease,SVD)及多支病变组(multiple-vessel disease,MVD),并对病变的严重程度进行Gensini评分。通过流式细胞术测定外周血两种单核细胞亚群数量。结果:外周血单核细胞CD14+CD16+亚群在MVD组的比例高于SVD组和non-CAD组(MVD:15.2±4.6;SVD:12.5±3.7,P0.001;non-CAD:7.5±2.5,P0.001),且与Gensini评分的相关性分析显示,CD14+CD16+亚群与冠状动脉病变严重程度呈正相关。结论:外周血CD14+CD16+细胞与稳定型心绞痛患者冠状动脉硬化病变程度相关。     

ATG16L1 and NOD2 polymorphisms enhance phagocytosis in monocytes of Crohn’s disease patients

AIM:To investigate if the presence of relevant genetic polymorphisms has effect on the effectual clearance of bacteria by monocytes and granulocytes in patients with Crohn’s disease(CD).METHODS:In this study,we assessed the differential responses in phagocytosis by measuring the phagocytic activity and the percentage of active phagocytic monocytes and granulocytes in inflammatory bowel disease patients as well as healthy controls.As both autophagy related like 1(ATG16L1)and immunityrelated guanosine triphosphatase gene are autophagy genes associated with CD and more recently nucleo-tide-binding ligomerization domain-containing protein2(NOD2)has been identified as a potent inducer of autophagy we genotyped the patients for these variants and correlated this to the phagocytic reaction.The genotyping was done with restriction fragment length polymorphisms analysis and the phagocytosis was determined with the pHrodo?Escherichia coli Bioparticles Phagocytosis kit for flowcytometry.RESULTS:In this study,we demonstrate that analysis of the monocyte and granulocyte populations of patients with CD and ulcerative colitis showed a comparable phagocytic activity(ratio of mean fluorescence intensity)between the patient groups and the healthy controls.CD patients show a significantly higher phagocytic capacity(ratio mean percentage of phagocytic cells)compared to healthy controls(51.91%±2.85%vs 37.67%±7.06%,P=0.05).The extend of disease was not of influence.However,variants of ATG16L1(WT:2.03±0.19 vs homozygoot variant:4.38±0.37,P<0.009)as well as NOD2(C-ins)(heterozygous variant:42.08±2.94 vs homozygous variant:75.58±4.34(P=0.05)are associated with the phagocytic activity in patients with CD.CONCLUSION:Monocytes of CD patients show enhanced phagocytosis associated with the presence of ATG16L1 and NOD2 variants.This could be part of the pathophysiological mechanism resulting in the disease.     

Decreased phagocytic function in neutrophils and monocytes from peripheral blood in periodontal disease

Phagocytosis by neutrophils and monocytes constitutes the main defense mechanism against bacterial challenges in periodontitis. Phagocytosis by neutrophils has already been evaluated, whereas phagocytic function of monocytes has hardly been addressed so far.

Objectives

The aim of this study was to assess phagocytosis by neutrophils and monocytes in periodontitis.

Material and Methods

The sample included 30 subjects with severe periodontitis and 27 control subjects without periodontal disease. The phagocytic index (PhI) was calculated as the mean number of adhered/ingested Saccharomyces cerevisiae per phagocytozing monocyte or neutrophil multiplied by the percentage of phagocytes involved in phagocytosis.

Results

A significant reduction in phagocyte functions was observed in individuals with periodontitis. The median of PhI of neutrophils using non-sensitized S. cerevisiae was 3 for the control group, and 1.5 for the periodontitis group (p=0.01, Mann-Whitney test). The median of PhI of monocytes with non-sensitized S. cerevisiae was 26.13 for the control group, and 13.23 for the periodontitis group (p=0.03, Mann Whitney test). The median of PhI of monocytes assessed with sensitized S. cerevisiae was 97.92 for the control group and 60.1 for the periodontitis group (p=0.005, t-test).

Conclusion

The data demonstrated a reduction in the function of phagocytes, suggesting a decrease in immune defenses in periodontitis.     

超高分子聚乙烯颗粒刺激单核细胞高表达CD147/EMMPRIN

目的探讨不同浓度磨损颗粒对单核细胞表达细胞外基质金属蛋白酶诱导剂（CD147/EMMPRIN）的影响。方法根据颗粒/细胞比值实验分为0、1、10、100、500、1000共6组,将不同浓度的超高分子聚乙烯（UHMWPE）颗粒与人单核细胞株THP-1共培养24h,台盼蓝染色检测THP-1细胞存活率,利用流式细胞术、Real-time PCR和Western blot检测THP-1细胞CD147/EMMPRIN的表达。结果加入UHMWPE颗粒前及共培养24h后THP-1细胞存活率均大于90%。流式细胞术、Real-timePCR和Westernblot结果均显示加入UHMWPE颗粒后,CD147/EMMPRIN表达较对照组（即颗粒/细胞比值为0）增加,当颗粒/细胞比值在1-100范围时,CD147/EMMPRIN表达与颗粒浓度正相关,颗粒/细胞比值大于或等于100时,CD147/EMMPRIN稳定保持在高表达水平,不随颗粒浓度升高而变化。结论 UHMWPE颗粒刺激单核细胞高表达CD147/EMMPRIN,并呈浓度依赖性。     

手足口病患儿外周血单核细胞亚群的研究

目的：探讨未经治疗和随访1、3、5 d手足口患儿外周血单核细胞亚群的变化。方法流式细胞仪检测104例未经治疗手足口患儿（包括阴性患儿62例,EV71阳性患儿37例,CVA16阳性患儿5例）和随访1、3、5 d的患儿外周血单核细胞CD14highCD16-亚群、CD14highCD16＋亚群和CD14lowCD16＋亚群。结果与健康儿童组相比,手足口患儿CD14highCD16＋单核细胞亚群所占总单核细胞比例明显升高（t =4.092,P ＜0.001）;CD14highCD16-亚群比例明显降低。阴性、EV71型、CVA16型患儿之间CD14highCD16-亚群、CD14highCD16＋亚群和CD14lowCD16＋亚群差异均无统计学意义。结论未经治疗的手足口患儿外周血单核细胞亚群的变化可能与EV71及CVA16病毒的持续感染有关,与病毒种类相关性不明显。     

Serum soluble CD163 levels in patients with influenza-associated encephalopathy

Background: Influenza-associated encephalopathy (IE) is a serious complication during influenza viral infection. Common clinical symptoms of IE include seizures and progressive coma with high-grade fever. We previously reported that hypercytokinemia and monocyte/macrophage activation may play an important role in the pathogenesis of IE. CD163 is a scavenger receptor for hemoglobin–haptoglobin complexes and is expressed by monocytes/macrophages. Proteolytic cleavage of monocyte-bound CD163 by matrix metalloproteinases releases soluble CD163 (sCD163). However, there have been no reports regarding serum sCD163 levels in IE patients. Methods: We measured serum levels of sCD163 as a marker of monocyte/macrophage activation in IE patients with poor outcomes, those without neurological sequelae, influenza patients without IE, and control subjects. Results: Serum sCD163 levels were significantly higher in IE patients with poor outcomes than in those without neurological sequelae. In particular, sCD163 levels in cases of death were significantly higher than those in other cases. Conclusions: Our results suggest that monocyte/macrophage activation is related to the pathogenesis of severe IE.     

Monocyte activity is linked with abdominal aortic aneurysm diameter

Background

Systemic inflammation and increased matrix metalloproteinase (MMP) cause elastin degradation leading to abdominal aortic aneurysm (AAA) expansion. Several prospective studies report that statin therapy can reduce AAA expansion through anti-inflammation. We hypothesize that monocyte activity plays a pivotal role in this AAA development and this study examines patient peripheral blood monocyte cell adhesion, transendothelial migration, and MMP concentrations between AAA and non-AAA patients.

Materials and methods

Peripheral blood was collected and monocytes isolated from control (n = 15) and AAA (n = 13) patients. Monocyte adhesion, transmigration, and permeability assays were assessed. Luminex assays determined MMP-9 and tissue inhibitor of metalloproteinase-4 (TIMP-4) concentrations from cell culture supernatant and patient serum.

Results

AAA patient monocytes showed increased adhesion to the endothelium relative fluorescence units (RFU, 0.33 ± 0.17) versus controls (RFU, 0.13 ± 0.04; P = 0.005). Monocyte transmigration was also increased in AAA patients (RFU, 0.33 ± 0.11) compared with controls (RFU, 0.25 ± 0.04, P = 0.01). Greater numbers of adhesive (R² = 0.66) and transmigratory (R² = 0.86) monocytes were directly proportional to the AAA diameter. Significantly higher serum levels of MMP-9 (2149.14 ± 947 pg/mL) were found in AAA patients compared with controls (1189.2 ± 293; P = 0.01). TIMP-4 concentrations were significantly lower in AAA patients (826.7 ± 100 pg/mL) compared with controls (1233 ± 222 pg/mL; P = 0.02). Cell culture supernatant concentrations of MMP and TIMP from cocultures were higher than monocyte-only cultures.

Conclusions

Monocytes from AAA patients have greater adhesion and transmigration through the endothelium in vitro, leading to elevated MMP-9 levels and the appropriate decrease in TIMP-4 levels. The ability to modulate monocyte activity may lead to novel medical therapies to decrease AAA expansion.     

白细胞吸附疗法在重症炎症性肠病中的应用进展

炎症性肠病(IBD)是一种以反复发作的、多种免疫因子参与的慢性肠道炎症,其发病率在逐年增加。其发病机制可能与机体免疫反应、肠道炎性活动、肠道功能紊乱、精神状态、遗传等因素有关。目前主要的治疗方式以营养疗法、药物治疗、单克隆抗体相关的生物治疗、免疫抑制剂疗法为主,因发病机制仍不十分明确,这些疗法都不能完全有效地控制炎症的发展。近年来研究发现,血液中的白细胞(中性白细胞和单核细胞)的激活在IBD的发生与发展中起着至关重要的作用,因此,有效地清除血液中激活的白细胞,中断炎症反应链,可以达到控制疾病进展的目的。许多研究者将白细胞免疫吸附疗法应用于对重症炎症性肠病(SIBD)病人的治疗,并取得了一定的效果。这是对目前IBD治疗的补充,也是一种研究思路,但其吸附效果、疗效、不良反应等还有待进一步研究。