The roles of a ribosomal protein S19 polymer in a mouse model of carrageenan-induced acute pleurisy

C5-deficient mice usually present moderate neutrophil activation during the initiation phase of acute inflammation. Conversely, C5a receptor (C5aR)-deficient mice show unusually excessive activation of neutrophils. We identified the ribosomal protein S19 (RP S19) polymer, which is cross-linked at Lys122 and Gln137 by transglutaminases in apoptotic neutrophils, as a second C5aR ligand during the resolution phase of acute inflammation. The RP S19 polymer promotes apoptosis via the neutrophil C5aR and phagocytosis via the macrophage C5aR. To confirm the roles of the RP S19 polymer, we employed a carrageenan-induced acute pleurisy mouse model using C57BL/6J mice with a knock-in of the Gln137Glu mutant RP S19 gene and replaced the RP S19 polymer with either an S-tagged C5a/RP S19 recombinant protein or the RP S19^122–145 peptide monomer and dimer (as functional C5aR agonists/antagonists) and the RP S19^122–145 peptide trimer (as a functional C5aR antagonist). Neutrophils and macrophages were still present in the thoracic cavities of the knock-in mice at 24 h and 7 days after carrageenan injection, respectively. Knock-in mice showed structural organization and severe hemorrhaging from the surrounding small vessels of the alveolar walls in the lung parenchyma. In contrast to the RP S19^122–145 peptide monomer and trimer, the simultaneous presence of S-tagged C5a/RP S19 and the RP S19^122–145 peptide dimer completely improved the physiological and pathological acute inflammatory cues. The RP S19 polymer, especially the dimer, appears to play a role at the resolution phase of carrageenan-induced acute pleurisy in C57BL/6J model mice.     

过敏性紫癜患者中性粒细胞及其IgA Fc受体对血管内皮细胞凋亡的影响及机制

目的 研究中性粒细胞及其IgA Fc受体(CD89)在过敏性紫癜(HSP)发病中的作用和对血管内皮细胞的损伤效应及调控机制.方法 入组30例急性期HSP患儿及9例健康对照儿童,分离外周血中性粒细胞;Jacalin亲和层析分离HSP患者血清IgA后,采用聚丙烯葡聚糖凝胶纯化IgA.实时定量PCR(qPCR)、Western印迹分别检测中性粒细胞CD89 mRNA和蛋白表达,流式细胞仪分析中性粒细胞活化标志分子CD11b的表达.分别将HSP患儿中性粒细胞、健康对照中性粒细胞与血管内皮细胞HUVEC共培养,以HSP患儿血清分离的IgA (HSP IgA)、单体IgA(mIgA)、磷酸盐缓冲液(空白对照组)分别处理,流式细胞仪检测HUVEC细胞凋亡,ELISA检测上清液中白细胞介素8(IL-8)、肿瘤坏死因子α(TNF-α)水平.结果 HSP患儿组中性粒细胞CD89 mRNA表达与健康对照组差异无统计学意义(P=0.98),但蛋白表达(0.60±0.16)低于健康对照组(0.83±0.24,P=0.03).HSP患者中性粒细胞CD11b表达(1 880.25±388.29)显著高于健康对照组(1 109.25±364.25,P＜0.01).与HSP患者中性粒细胞共培养组HUVEC凋亡率(37.44％±5.49％)高于与健康对照中性粒细胞共培养组(17.14％±4.45％,P＜ 0.01).HSP IgA处理组HUVEC凋亡率明显高于mIgA处理组(均P＜0.01)和空白对照组(P＜0.01),且上清液中IL-8、TNF-α浓度明显高于mIgA处理组(均P＜0.01)和空白对照组(P值分别为0.01、0.02).结论 HSP患者外周血中性粒细胞活化,并可诱导血管内皮细胞凋亡.而且HSP IgA能促进中性粒细胞介导的血管内皮细胞凋亡和炎症因子IL-8、TNF-α分泌,而mIgA则可能具有一定抑制效应.     

IL‐24 protects against Salmonella typhimurium infection by stimulating early neutrophil Th1 cytokine production,which in turn activates CD8+ T cells

Salmonella are important intracellular pathogens in humans and other animal hosts. IL‐24 is a novel tumour suppressor and can mediate induction of Th1‐type cytokines from PBMC. However, the immunological consequences of this cytokine during intracellular pathogen infection in vivo remain unclear. In the present study, we used a virulent S. typhimurium C5 infected mouse model of typhoid fever to demonstrate that administration of exogenous IL‐24 had a protective effect against the bacteria. The IL‐24 glycosylation site mutant, in contrast, showed a decreased protective effect. Furthermore, the protective effect of IL‐24 was abrogated in IFN‐γ KO mice. More importantly, we demonstrated that IL‐24 predominately stimulated neutrophils to produce IFN‐γ and IL‐12, subsequently activating CD8⁺ T cells both in vivo and in vitro. In addition, IL‐24 could induce neutrophils to produce NO. These data indicate that the neutrophils activated by IL‐24 may play important roles in host defence against Salmonella infection in vivo. Our findings support the development of a novel cytokine immunotherapy against Salmonella.     

不同剂量胆红素对内毒素致大鼠急性肺损伤的保护作用

目的探讨不同剂量的胆红素对内毒素致急性肺损伤(ALI)的保护作用及其可能机制。方法将雄性Wistar大鼠(200~250 g)40只,按随机数字表法分为正常对照组、ALI动物模型组、大剂量胆红素干预组和小剂量胆红素干预组。观察肺组织形态学变化,检测肺组织肺系数(LI)、支气管肺泡灌洗液(BALF)中白细胞(WBC)计数和中性粒细胞(PMN)百分比、蛋白质含量(Pr)、肺泡通透指数(LPI);测定肺组织匀浆中超氧化物歧化酶(SOD)、脂质过氧化产物丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)含量变化。结果①ALI模型组LI、BALF中WBC计数、PMN百分比和Pr及LPI均显著高于生理盐水对照组(P均<0.01)。大剂量胆红素干预组LI,BALF中WBC计数和LPI均显著低于ALI模型组(P均<0.01),PMN百分比和Pr也明显低于ALI模型组(P均<0.05),且与生理盐水对照组差异无统计学意义。小剂量胆红素组LI、BALF中PMN百分比、Pr和LPI均显著低于ALI模型组(P均<0.05)。②ALI模型组MDA含量显著高于生理盐水对照组(P<0.01),SOD和GSH-Px水平却明显低于生理盐水对照组(P<0.01)。大剂量胆红素干预组肺匀浆中MDA含量明显低于ALI模型组(P<0.01),而肺匀浆中的SOD活性和GSH-Px含量显著高于ALI模型组(P<0.01,P<0.05)且与生理盐水对照组差异无统计学意义。小剂量胆红素干预组MDA含量明显高于正常对照组(P<0.05),而SOD活性和GSH-Px含量显著低于正常对照组(分别P<0.05、P<0.01)且与ALI模型组差异无统计学意义。结论胆红素通过其抗氧化作用对抗大鼠ALI并且存在剂量依赖性。     

长效局麻药对中性粒细胞呼吸爆发的抑制作用

目的观察几种长效局麻药对多形核中性粒细胞(PMN)释放氧自由基的抑制作用。方法选取近2周未用过任何药物的健康志愿者12人,抽取静脉血经分离得PMN悬液。将每人的血样分成6组:Ⅰ、Ⅱ组分别为单纯的反应混合物和参考样品,含有不同局麻药(浓度均为1×10-4)的Ⅲ~Ⅵ组为实验组。以fMLP或PMA激活反应,用细胞色素C减少法测定活化PMN产生的细胞外O2-含量的变化。Ⅰ,Ⅱ两组光密度之差作为对照值,各实验组吸光度与Ⅱ组吸光度之差为依赖于O2-的细胞色素C减少值,以反映PMN的O2-释放。结果未加局麻药的PMN悬液O2-的释放量为9.3±4.7nmol/106个细胞。而含有罗哌卡因、S(-)布比卡因、布比卡因和地卡因的PMN悬液O2-的释放量分别是对照值的77.5%、75.8%、88.3%和61.3%(P<0.01)。结论地卡因、罗哌卡因、布比卡因和S(-)布比卡因对活化的PMN氧自由基的释放有不同程度的抑制作用,不同局麻药之间的差异可能与药物的立体异构和脂溶性有关。     

Clinical symptoms and neutropenia: the balance of neutrophil development, functional activity, and cell death

Neutrophilic granulocytes form the major type of leukocytes with counts ranging from about 1500–5000 cells/μl of blood under normal conditions. Neutrophils protect our body against bacterial and fungal infections. For this purpose, these cells are equipped with a machinery to sense the site of an infection and, upon local extravasation, rapidly move towards the site with invading micro-organisms, to ingest and kill them. As will be described, for proper functioning of this line of defence, a number of prerequisites have to be fulfilled. The quantitative defects are diagnosed more often and easier than the mere qualitative phagocytic defects. Nonetheless, functional defects may accompany neutropenia. These functional defects are seen in severe congenital neutropenia of which the gene defect has recently been elucidated, as well as in the more complex and syndromal forms of neutropenia such as Shwachman syndrome or the metabolic disease glycogen storage disease type 1b (non-a). The background of functional neutrophil defects is briefly reviewed. Published online: 13 September 2002     

A Study of Changes in Uterine Leucocytes During Early Pregnancy in the Mouse-vole Interspesific Pregnancies

Mouse and vole embryos were allogeneically and xenogeneically transferred into pseudopregnant CD.1 and immunodeficient (seid)female mice,and we investigated the distribution of uterine leucocytes cells in the implantation sites on days 5,6,and 7 of pregnancy. Maerophages were evenly distributed in the endometrium on days 5-7.Neutrophils were rarely seen on days 5-7,but lymphocytes were found throughout the endometrium,often in groups associated with glands or the luminal epithelium.The number of uNK cells increased markedly at the mesometrial uriangle and the outer decidual area in the CD-1 uteri containing vole embryos;by contrast,seid uteri having vole embryos showed almost the same number as those having mouse embryos.Mast cells were present in large numbers at the myometrium,but rarely in the decidua in all types of pregnant uteri.Cells at the myometrium were more numerous in xenogeneic than in allogeneic transfer.Maay mast cells appeared in the inner decidua where xenogeneically transferred vole embryos were dead and aborted.These results suggest the possibility that uterine leucocytes mediate various immunological events in the mouse-vole interspesific pregnancies.     

四氧嘧啶糖尿病大鼠外周血嗜中性粒细胞吞噬功能改变

本文报道四氧嘧啶诱导的糖尿病大鼠外周血嗜中性粒细胞对中华墨汁颗粒的吞噬功能测定,结果显示:(1)糖尿病大鼠嗜中性粒细胞吞噬率和吞噬指数明显降低(P<0.01);(2)体外试验观察到在高葡萄糖浓度下,正常大鼠嗜中性粒细胞吞噬率和吞噬指数显著下降,并与葡萄糖浓度呈负相关关系(P<0.01),但相关关系不密切。高血糖可能是糖尿病状态下粒细胞吞噬功能受损的原因之一     

Modification of neutrophil chemotactic responsiveness during various inflammatory reactions

Various inflammatory reactions have been induced in order to examine the chemotactic response of polymorphonuclear leucocytes (PMN) collected under various experimental conditions. Cells were harvested from the pleural cavity of rats after the induction of three acute non specific inflammatory reactions and two immune reactions. The results obtained with two techniques of chemotactic assessment (agarose assay and Boyden chamber technique) demonstrated a variation of chemotactic response depending on the cell source and the chemoattractants used. Using agarose assay, we distinguished locomotor reactivity of PMN harvested after immune inflammatory reactions from that of PMN harvested after non immune inflammatory reactions. Chemokinetic and chemotactic responses to various chemoattractants were inhibited in the first case and not affected in the second. Using the Boyden chamber technique, inhibition of random or oriented migration of PMN harvested after immune inflammatory reactions after the injection of a non antigenic irritant such as calcium pyrophosphate crystals (CaPP) was also observed.