马链球菌兽疫亚种烯醇化酶对小鼠肺泡巨噬细胞吞噬功能的影响   总被引：1，自引：1，他引：0  

陈楷文  华承薇  袁宸  潘飞  蔺辉星  范红结  马喆 《畜牧兽医学报》2020,51(10):2576-2583

旨在研究马链球菌兽疫亚种（Streptococcus equi ssp.zooepidemicus，SEZ）烯醇化酶（enolase，Eno）对小鼠肺泡巨噬细胞（RAW264.7）吞噬能力的影响。通过构建原核表达质粒获得重组烯醇化酶（rEno），采用台盼蓝活细胞计数法，判定在不同处理浓度和时间下rEno蛋白对RAW264.7细胞的细胞毒性。将rEno蛋白与RAW264.7细胞共孵育后，用SEZ作用于细胞并检测细胞吞菌数量，判断RAW264.7细胞对SEZ的吞噬活性。进一步通过活细胞稳定同位素标记技术（SILAC）和蛋白质谱分析技术（LC-MS/MS），筛选到RAW264.7细胞中可能与SEZ Eno存在相互作用的候选蛋白。结果发现，10 μg·mL^-1 rEno蛋白处理对RAW264.7细胞有明显的细胞毒性，且10 μg·mL^-1 rEno蛋白处理RAW264.7细胞2和4 h可显著抑制其对SEZ的吞噬作用（P<0.01、P<0.05）。初步筛选到RAW264.7细胞中动力蛋白激活蛋白亚单位蛋白（dynactin subunit protein 2，Dctn）、整合素α-M蛋白（integrin alpha-M）等17种可能与Eno发生互作的蛋白。本研究获得了rEno重组表达蛋白，发现rEno可减少RAW264.7细胞对SEZ的吞噬，互作蛋白的初步筛选也为进一步揭示Eno在SEZ抗吞噬中的作用机制奠定了基础。  相似文献   

Simultaneous Flow Cytometric Analysis of Phagocytosis and Oxidative Burst Activity in Equine Leukocytes     

Flaminio MJ  Rush BR  Davis EG  Hennessy K  Shuman W  Wilkerson MJ 《Veterinary research communications》2002,26(2):85-92

This paper describes a method for simultaneously measuring phagocytosis and oxidative burst activity in equine peripheral blood leukocytes by flow cytometry. Opsonized propidium iodide-labelled Staphylococcus aureus (PI-Sa) was used to measure the uptake of bacteria by equine phacocytes and the oxidative burst activity by oxidation of dihydrorhodamine 123. The requirements to achieve optimal activity of phagocytosis and oxidative burst are described. The advantage of the simultaneous technique is that it provides both independent and comparative values for phagocytosis and the oxidative burst, for the detection of impaired mechanisms of microbial destruction. Furthermore, the technique allows evaluation of opsonization activity in this context.  相似文献   

Neutrophil Phagocytosis Following Inoculation of Salmonella choleraesuis into Swine     

Stabel TJ  Fedorka-Cray PJ  Gray JT 《Veterinary research communications》2002,26(2):103-109

Neutrophils are an important mediator of host defence, especially in early stages of infection. A major function of neutrophils is the uptake and killing of invading microbes. Little is known about the effect of neutrophil activity on the pathogenesis and development of the carrier state in swine following infection with Salmonella choleraesuis. A human whole-blood microassay using flow cytometry was modified to measure the effect of S. choleraesuis infection in vivo on the rate of ingestion, or rate of uptake, of homologous bacteria by porcine neutrophils. Pigs were inoculated intranasally with 5–8×10⁸ CFU S. choleraesuis and blood was collected in heparinized tubes at –5, 0, 1, 2, 3 and 4 days post inoculation (PI). Heat-killed S. choleraesuis were labelled with fluorescein isothiocyanate and incubated for various times with diluted whole blood. Red blood cells were lysed, external non-phagocytized bacteria were quenched with a commercially available lysing solution, and fluorescence from internalized bacteria labelled with fluorescein isothiocyanate was detected by flow cytometry. The rate of uptake by neutrophils did not increase until 2 days PI and then remained elevated to 4 days PI. The minimal uptake of S. choleraesuis early after exposure to these organisms may provide an opportunity for the pathogen to colonize and/or replicate to levels that facilitate establishment of a carrier state or clinical infection in swine.  相似文献   

尼罗罗非鱼鼠李糖凝集素 (RBL-1)在非特异性细胞防御病原菌感染中的功能     

木亮亮  白皓  李嘉东  邱丽  尹晓雪  叶剑敏 《水产学报》2022,46(5):730-740

为了探究鼠李糖凝集素（Rhamnose-binding lectin, RBL）在硬骨鱼非特异性细胞防御病原菌感染中的作用,本研究以尼罗罗非鱼为研究模型,首先通过分离头肾单核/巨噬细胞进行体外菌应激实验,发现在罗非鱼两种重要的致病菌-无乳链球菌（Streptococcus agalactiae）和嗜水气单胞菌（Aeromonas hydrophila）应激后,尼罗罗非鱼L-鼠李糖凝集素1（OnRBL-1）的表达量显著上调。然后,通过荧光定量PCR（qRT-PCR）检测发现OnRBL-1重组蛋白能够调节病原菌诱导细胞炎症因子的表达,包括显著抑制菌诱导的IL-6、IL-8和TNF-α的表达,和促进IL-10和TGF-β的表达。此外,通过流式细胞术检测证实OnRBL-1具有促进单核/巨噬细胞的吞噬作用,同时增强呼吸爆发水平和上调活性氧的释放。以上研究结果表明,OnRBL-1在罗非鱼单核/巨噬细胞非特异性细胞防御中发挥重要的调节作用。本研究为探讨RBL-1在硬骨鱼宿主防御病原菌感染中的功能提供了参考,并有助于完善和丰富硬骨鱼类RBL的功能和在抗菌免疫应答中的基础理论体系,具有重要的科学意义。  相似文献   

The effect of various stresses,corticosteroids and adrenergic agents on phagocytosis in the rainbow trout Oncorhynchus mykiss     

Yuwaraj K. Narnaware  Bridget I. Baker  Mike G. Tomlinson 《Fish physiology and biochemistry》1994,13(1):31-40

The effect of acute and chronic stress on the phagocytic activity of putative macrophages from the rainbow trout. Oncorhynchus mykiss has been assessed, using an in vitro phagocytic index, in which the average number of engulfed yeast cells in a population of phagocytes is determined. An injection stress given under light anaesthesia, or a longer noise stress combined with confinement, both significantly reduced, within 3 h, the level of phagocytic activity of macrophages from the spleen and pronephros. Daily injection stress over six days had a lesser effect on the proportion of phagocytically active cells even though plasma cortisol levels were equally raised. Daily dexamethasone injection depressed the proportion of phagocytically active cells more than saline injection. In these in vivo experiments, it was not possible to determine whether stress and steroids depressed the phagocytic activity of individual macrophages or caused the active macrophages to migrate out of the spleen and pronephros. Administration of cortisol (200 nM) to trout macrophages in vitro failed to depress phagocytic activity within a 3h period but both α- and β-adrenergic agonists (10 μM) were usually depressive. It is proposed that the autonomic nervous system may be an early regulator of macrophage phagocytosis following stress and that corticosteroids only exert their suppressive effect on macrophage activity in the longer term. To whom reprint requests should be addressed.  相似文献   

Immunostimulant effects of nucleotide isolated from yeast RNA on carp, Cyprinus carpio L.   总被引：2，自引：0，他引：2  

M Sakai  K Taniguchi  K Mamoto  H Ogawa  & M Tabata 《Journal of fish diseases》2001,24(8):433-438

Nucleotides from yeast RNA were evaluated for their ability to enhance the non-specific immune responses in carp, Cyprinus carpio . Oral administration of nucleotides to fish daily for 3 days resulted in enhanced responses of phagocytic and nitroblue tetrazolium (NBT) activities in kidney phagocytic cells. This activation of kidney cells was observed for at least 10 days post-treatment. The serum complement and lysozyme activities also increased in fish treated with nucleotides. Furthermore, the number of Aeromonas hydrophila in nucleotide-treated fish significantly decreased in the blood, kidney and liver after intraperitoneal injection. Thus yeast nucleotides appear to enhance non-specific immune responses in fish.  相似文献   

The effects of vitamin E on non-specific immune response of the juvenile soft-shelled turtle <Emphasis Type="Italic">Pelodiscus sinensis</Emphasis>   总被引：1，自引：0，他引：1  

Xian-Qing?Zhou  Cui-Juan?NiuEmail author  Ru-Yong?Sun 《Fisheries Science》2005,71(3):612-617

ABSTRACT:   This study was conducted to determine the effect of dietary vitamin E supplements on non-specific immune responses in juvenile soft-shelled turtles. Turtles were fed diets supplemented with vitamin E at 0 (control), 50, 250, 500, 1000 and 5000 mg/kg, respectively, for 4 weeks. Results showed that blood cell phagocytosis and serum bactericidal activity were significantly improved in turtles from two diets (250 and 500 mg/kg vitamin E supplementation), while had no significant improvement in turtles from the three remaining diets (50, 1000 and 5000 mg/kg vitamin E supplementation) when compared to turtles from the control group. Serum bacteriolytic activity in turtles from diets with 50, 250, 500 and 1000 mg/kg vitamin E supplementation were higher than activity levels quantified for the control group, but no significant difference was observed between the 5000 mg/kg vitamin E supplementation diet and the control diet. The results suggest that vitamin E has an upper and lower threshold for improving non-specific immune function in soft-shelled turtles, and the optimal supplementation may be between 250 and 500 mg/kg.  相似文献   

异黄酮植物雌激素对小鼠免疫功能的影响   总被引：39，自引：0，他引：39  

张荣庆  韩正康 《南京农业大学学报》1993,16(2):64-68

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Cellular Defence Reaction of American Cockroach , Periplaneta americana to Metarhizium anisopliae Isolate CQMa102     

ZHONG Wei  YIN You ping 《保鲜与加工》2003,(6):89-92100

Injection of conidia solution of Metarhizium anisopliae isolate CQMa102 into the hemocoel of Periplaneta americana caused significant changes of the total hemocytes of the cockroach. P. americana eliminated large amount of the spores efficiently by phagocytosis and nodulation.The experiment shows that the immune system of the cockroach was hardly injured after injected with high dose spores, and the higher dose of spores was injected,the higher mortality of the insect was. However, low dose injection strengthened the immune ability of the insect in certain degree. The saline slightly induced the immune reaction of the insect.  相似文献   

不同鹿胎及胎盘制剂对老年雄性大鼠免疫机能影响的研究   总被引：4，自引：1，他引：3  

韩广金  杨桂芹  邹兴淮 《畜牧与兽医》2006,38(1):8-10

研究了鹿胎及胎盘制剂对老年雄性大鼠免疫器官指数、血清球蛋白含量和巨噬细胞吞噬率及吞噬指数的影响。结果表明,鹿胎及胎盘制剂对老年雄性大鼠胸腺和脾脏指数影响不显著(P>0.05);但提高了老年雄性大鼠血清球蛋白含量,其中鹿胎制剂Ⅰ号组最高,比对照组高出62.64%,与其他各组相比有显著差异(P<0.05);鹿胎制剂Ⅱ号组和人胎盘粉处理组大鼠巨噬细胞吞噬率和吞噬指数与对照组相比有极显著提高(P<0.01),说明鹿胎及胎盘制剂对老年雄性大鼠具有提高免疫机能的作用。  相似文献