Activation of granulocytes by direct interaction with dendritic cells

Granulocytes from human peripheral blood were co-cultured with conventional dendritic cells (cDC) or plasmacytoid DCs (pDC) to examine the effects of DCs on the activation or function of granulocytes. After co-culture of granulocytes with DCs, expression of the activation markers of granulocytes (CD63 and CD64) was up-regulated, and increased expression of CD50, the activation marker and ligand for CD209 (DC-SIGN) was also observed. The interaction of granulocytes with DCs was visualized as the cluster where DCs, especially cDCs, were surrounded by granulocytes to form a 'rosette'. After co-culture of granulocytes with cDCs, the secretion of elastase from granulocytes was enhanced significantly when examined cytohistochemically and by enzyme-linked immunosorbent assay. An increase in myeloperoxidase (another activation index of granulocytes) was also observed after co-culture with DCs. These findings suggest the functional and phenotypical activation of granulocytes by interaction with DCs. Furthermore, we examined the involvement of adhesion molecules in the granulocyte-DC interaction, and found that CD209 participates to some extent in this interaction.     

中性粒细胞与淋巴细胞比值和急性冠状动脉综合征的相关性及其对急性心肌梗死的诊断价值

目的 探讨中性粒细胞与淋巴细胞比值（NLR）与急性冠状动脉综合征（ACS）患者冠脉严重程度之间的关 系,并分析NLR对急性心肌梗死的诊断价值。方法 回顾性分析2016年11月—2017年3月于我院行冠状动脉造影 检查的259例ACS患者的临床资料,其中不稳定型心绞痛组（UA组）148例、急性非ST段抬高型心肌梗死组（NSTEMI组）46例、急性ST段抬高型心肌梗死组（STEMI组）65例。比较3组的白细胞计数（WBC）、中性粒细胞计数（N）、淋巴细胞计数（L）、NLR、高敏C反应蛋白（hs-CRP）、心肌酶谱、血脂、血糖、肝功能等指标的差异。根据冠状动脉造影结果将患者按照冠状动脉病变支数分为单支病变组（70例）、双支病变组（70例）和三支病变组（119例）,比较不同病变支数组间炎性细胞的差异。同时绘制NLR的受试者工作特征（ROC）曲线,评价NLR对急性心肌梗死的预测价值。结果 UA组、NSTEMI组及STEMI组间WBC、N、NLR、hs-CRP、cTnT、CK、CK-MB水平依次升高,STEMI组L值低于UA组,差异有统计学意义（P＜0.05）;WBC、NLR随冠状动脉病变支数的增加而呈上升趋势,但仅三支病变组与单支病变组间差异有统计学意义;NLR诊断急性心肌梗死的ROC曲线下面积（AUC）为0.865（95%CI：0.814~0.916）。当NLR=4.22时,其对急性心肌梗死的诊断效能最高,敏感度为72.1％,特异度为95.3％。结论 NLR水平与ACS患者病情严重程度呈正相关,并对诊断急性心肌梗死具有较高的诊断价值,可作为ACS患者病情的预测因素。     

Stimulatory response of neutrophils from periodontitis patients with periodontal pathogens

OBJECTIVE: Neutrophils play a crucial role in the defense of invading bacteria by releasing biologically active molecules. The response of peripheral blood neutrophils was studied in periodontitis-affected patients and in healthy controls towards stimulation to Porphyromonas gingivalis (Pg) and Actinobacillus actinomycetemcomitans (Aa) extracts. MATERIALS AND METHODS: Peripheral venous blood was drawn from 23 adult patients with moderate to advanced chronic periodontitis (probing depth >or=5 mm, attachment loss >or=3 mm), and 30 healthy volunteers. Neutrophil response followed by metalloproteinase-9 (MMP-9) and interleukin-8 (IL-8) secretion was assayed by zymography and enzyme-linked immunosorbent assay, respectively, on both whole blood and purified neutrophils. In addition to periodontal pathogen extracts, known stimulating agents were tested, such as Escherichia coli-lipopolysaccharide (LPS), phytohemagglutinin, and zymosan A. RESULTS: Neutrophil response, expressed as a secretion ratio under stimulated and non-stimulated conditions, measured in whole blood, showed no differences between periodontitis and healthy controls. Instead, in purified neutrophils from patients, MMP-9 exhibited a significantly higher secretion ratio with LPS and Pg (1.5- to 2-fold), whereas IL-8 showed a larger increase in secretion ratio (3- to 7-fold) in the presence of Pg, Aa, LPS, and zymosan A. CONCLUSION: Peripheral neutrophils of periodontitis-affected patients are more reactive as suggested by their significantly higher response toward periodontal pathogen extracts and other stimulating agents.     

馥感啉口服液联合脾氨肽治疗小儿反复呼吸道感染的临床研究

目的探讨馥感啉口服液联合脾氨肽治疗小儿反复呼吸道感染的临床疗效。方法选取2015年9月—2016年3月在郑州市第三人民院治疗的反复呼吸道感染患儿98例,随机分为对照组(49例)和治疗组(49例)。对照组口服脾氨肽口服冻干粉,2 mg/次,1次/d;治疗组在对照组基础上口服馥感啉口服液,10 mL/次,1~3岁患儿3次/d,4~6岁4次/d,7~12岁5次/d。两组患儿均经过3个月治疗。观察两组患儿临床疗效,比较治疗前后两组患儿临床症状改善时间、血清学指标、白细胞分类和白三烯D4(LTD4)水平。结果治疗后,对照组临床总有效率为81.63%,显著低于治疗组的95.92%。两组比较差异具有统计学意义(P0.05)。治疗后,治疗组发热、咳嗽和肺部啰音消失时间均显著短于对照组(P0.05)。治疗后,两组血清IL-1β、IL-6、和肿瘤坏死因子-α(TNF-α)水平均显著降低,胰岛素1号增长因子(IGF-1)、干扰素-γ(IFN-γ)和25-OH维生素D_3[25-(OH)D_3]水平均显著升高,同组比较差异具有统计学意义(P0.05);且治疗组上述血清学指标改善程度明显好于对照组(P0.05)。治疗后,两组鼻咽分泌物中中性粒细胞、嗜酸性粒细胞和LTD4水平均显著降低,单核巨噬细胞显著升高,同组比较差异具有统计学意义(P0.05);且治疗组上述指标改善后水平显著优于对照组(P0.05)。结论馥感啉口服液联合脾氨肽治疗小儿反复呼吸道感染可有效改善临床症状,降低炎症水平,具有一定的临床推广应用价值。     

Proteolysis of the endothelial cell protein C receptor by neutrophil proteinase 3

BACKGROUND: The endothelial cell protein C receptor (EPCR) presents protein C to the thrombin:thrombomodulin complex on the endothelium of large vessels, and enhances the generation of activated protein C (APC) and activation of protease-activated receptor-1. A previous report has demonstrated binding of soluble (s) EPCR to activated neutrophils via surface proteinase 3 (PR3). METHODS: We now report further characterization of this interaction. Activated neutrophils and purified PR3 both decrease endothelial cell (EC) surface EPCR, suggestive of its proteolysis. RESULTS: When added to purified recombinant sEPCR, PR3 produced multiple cleavages, with early products including 20 kDa N-terminal and C-terminal (after Lys(176)) fragments. The binding of active site blocked PR3 to sEPCR was studied by surface plasmon resonance. Estimates of the K(D) of 18.5-102 nM were obtained with heterogeneous binding, suggestive of more than a single interaction site. CONCLUSIONS: This work demonstrates PR3 binding to and proteolysis of EPCR and suggests a mechanism by which anticoagulant and cell protective pathways can be down-regulated during inflammation.     

Measuring calprotectin in plasma and blood with a fully automated turbidimetric assay

Calprotectin in plasma and blood might prove to be a useful biomarker of inflammation and infection; however, automated methods for analysing the concentration of calprotectin in those materials are lacking. We have validated a fully automated turbidimetric method and present health-related reference limits. Calprotectin was measured by Siemens Advia XPT with the Bühlmann fCAL® turbo test (Bühlmann Laboratories AG, Schönenbuch, Switzerland), a particle enhanced turbidimetric immunoassay for quantification of calprotectin in fecal extracts. Plasma and serum samples were analysed directly, while whole blood was first extracted with M-PER® Mammalian Protein Extraction Reagent (ThermoFisher) and diluted with B-CAL-EX (Bühlmann). We studied analytical imprecision, estimated health-related reference limits and examined the correlation between neutrophil-calprotectin (blood-calprotectin adjusted for plasma-calprotectin) and the neutrophil count. The intermediate (‘day-to-day’) coefficient of variation was 3.5 and 1.0% for heparin-plasma-calprotectin at 0.52?mg/L and 3.53?mg/L, respectively, and 4.9% for heparin-blood-calprotectin at 50.2?mg/L. Health-related reference limits were 0.470–3.02?mg/L for calprotectin in heparin-plasma, 50.8–182?mg/L for calprotectin in heparin-blood, 0.534–2.41% for the ratio between them and 24.7–33.3?pg for the mean amount of calprotectin per neutrophil. Compared to heparin-plasma, calprotectin concentrations were significantly lower in EDTA-plasma and higher in serum (p?<?.05). Correlation between neutrophil-calprotectin and the neutrophil count was excellent. We have shown that the Bühlmann fCAL® turbo test can be used to measure calprotectin in plasma and blood.     

人参皂苷Re抑制心肌缺血再灌注损伤中性粒细胞浸润和髓过氧化物酶活性的研究

目的 :观察人参皂苷Re对心肌缺血再灌注中性粒细胞 (PMNs)浸润及髓过氧化物酶 (MPO)活性的影响 ,探讨人参皂苷Re对心肌缺血再灌注损伤的防治作用。方法 :结扎SD大鼠左冠状动脉前降支 (LAD) ,建立大鼠缺血再灌注动物模型 ;苏木精 伊红染色光镜检测心肌组织中PMNs浸润的数量 ;MPO法测定心肌MPO活性。结果 :①心肌缺血再灌注后 ,光镜下假手术组未发现PMNs ,缺血再灌注组和人参皂苷Re治疗组 (简称Re治疗组 )PMNs浸润明显增加。缺血再灌注组PMNs数为 (8.16 6 7± 1.14 72 )个 /视野 ;Re治疗组为 (4 .5 0 0 0±1.0 4 88)个 /视野 ;两者与假手术组比较差异有统计学意义 (P <0 .0 1) ;Re治疗组与缺血再灌注组相比 ,PMNs浸润明显减少 ,差异有统计学意义 (P <0 .0 1)。②缺血再灌注组、Re治疗组MPO活性均较假手术组明显增加(P <0 .0 1) ,Re治疗组与缺血再灌注组相比MPO活性明显降低 (P <0 .0 1)。结论 :心肌缺血再灌注诱导PMNs的浸润和MPO活性的增加 ,人参皂苷Re可显著抑制心肌缺血再灌注PMNs浸润和MPO活性     

Erythrophagocytosis by dysplastic neutrophils in chronic myelomonocytic leukemia and subsequent transformation to acute myeloid leukemia

Erythrophagocytosis by neutrophils is a rare phenomenon in myeloid malignancies, and its clinicopathologic significance is not fully understood. We report a unique case of erythrophagocytosis by dysplastic neutrophils in chronic myelomonocytic leukemia (CMML) and subsequent transformation to acute myeloid leukemia (AML). Review of multiple marrow samples, both pretreatment and post-treatment, demonstrated a correlation between percentage of dysplastic neutrophils and degree of erythrophagocytosis. Erythrophagocytosis was observed only in dysplastic forms of neutrophils. Post-transplant marrows with engraftment of donor cells showed no neutrophilic dysplasia or erythrophagocytosis. Possible mechanisms of neutrophilic erythrophagocytosis in myeloid malignancies are discussed.     

Role of neutrophils and mast cells in acute indomethacin-induced small bowel injury in the rat

The aim of this study was to determine whether the injury to the rat jejunum during the first 4.5h exposure to indomethacin is due to an influx of neutrophils or degranulation of resident mast cells. Indomethacin and vehicle both caused changes in villous morphology (length, width, etc.) while only indomethacin injured the small bowel, as indicated by increased histological lesion score and⁵¹Cr-ethylene diamine tetraacetate (EDTA) flux across the intestinal epithelium. Immunohistochemical staining showed the same small increase in neutrophil density (predominantly in the submucosa) following exposure to vehicle as following exposure to indomethacin. Chronic oral administration of indomethacin for 48 h did cause increased tissue neutrophil density compared to that in vehicle-fed controls. Mast cell depletion (using dexamethasone) did not alter either the indomethacin-induced increase in⁵¹Cr-EDTA clearance or the increase in neutrophil density caused by the vehicle and by indomethacin. However, the lesion score following exposure to indomethacin was significantly lower in mast-cell-depleted animals than in control animals. We conclude that the acute phase of indomethacin-induced intestinal injury is not associated with neutrophil influx. Increased neutrophils seen after chronic indomethacin may result from injury rather than be causative. Mast cells appear to exacerbate the initial stages of indomethacin-induced intestinal injury.     

Role of neutrophil-mediated inflammation in aspirin-induced gastric mucosal injury

The objectives of this study were to determine the roles of neutrophil-endothelial cell interactions and oxygen-derived free radicals in the pathogenesis of aspirin-induced gastric mucosal injury in rats. Oral administration of acidified aspirin (200 mg/kg) resulted in linear hemorrhagic erosions and an increase in myeloperoxidase activity, an index of neutrophil infiltration, in the gastric mucosa. Aspirin-induced gastric damage and the increase in myeloperoxidase activity were significantly inhibited by the injection of anti-CD11a, anti-CD11b, anti-intercellular adhesion molecule-1 monoclonal antibodies, and the combination of superoxide dismutase and catalase, which are scavengers of active oxygen species. These results suggest that neutrophil-endothelial adhesive interactions, which occur via CD11a/CD18-and CD11b/CD18-dependent interactions with intercellular adhesion molecule-1, and oxygen-derived free radicals produced by neutrophils are implicated in the production of aspirin-induced gastric mucosal injury.