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排序方式: 共有122条查询结果,搜索用时 156 毫秒
1.
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
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
2.
Differences in uptake and killing of pathogenic and non‐pathogenic bacteria by haemocyte subpopulations of penaeid shrimp,Litopenaeus vannamei, (Boone) 下载免费PDF全文
V V Tuan J J Dantas‐Lima K V Thuong W Li K Grauwet P Bossier H J Nauwynck 《Journal of fish diseases》2016,39(2):163-174
Phagocytosis is an important function of both invertebrate and vertebrate blood cells. In this study, the phagocytic activity of haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei, (Boone), against pathogenic and non‐pathogenic particles was investigated in vitro. The haemocytes of penaeid shrimp were firstly separated by centrifugation on a continuous density gradient of iodixanol into four fractions with five subpopulations (sub), of which sub 1 (hyalinocytes) and sub 4 (semi‐granulocytes) have the main function in phagocytosis of both pathogenic and non‐pathogenic bacteria as well as fluorescent polystyrene beads. It was found that these haemocyte subpopulations engulfed virulent Vibrio campbellii and Vibrio harveyi at a higher rate than non‐virulent Escherichia coli and polystyrene beads. When these bacteria were mixed with shrimp haemocyte subpopulations and incubated for 180 min, the percentage of viable intracellular V. campbellii (25.5 ± 6.0%) recovered was significantly higher than the percentage recovered from V. harveyi (13.5 ± 1.1%). No viable intracellular E. coli was observed in this study. In contrast to V. harveyi and E. coli, V. campbellii containing endosomes did not acidify in time. Incubation of haemocyte subpopulations with the most virulent V. campbellii strain resulted in a significant drop in haemocyte viability (41.4 ± 6.3% in sub 1 and 30.2 ± 15.1% in sub 4) after 180 min post‐inoculation in comparison with the less virulent V. harveyi (84.1 ± 5.6% in sub 1 and 83.4 ± 4.1% in sub 4) and non‐virulent E. coli (92.7 ± 2.8% in sub 1 and 92.3 ± 5.6% in sub 4) and polystyrene beads (91.9 ± 1.6% in sub 1 and 84.4 ± 3.4% in sub 4). These findings may be a valuable tool for monitoring shrimp health and immunological studies. 相似文献
3.
Phenotypical parameters as a tool to evaluate the immunostimulatory effects of laminarin in Oncorhynchus mykiss 下载免费PDF全文
Byron Morales‐Lange Jörn Bethke Paulina Schmitt Luis Mercado 《Aquaculture Research》2015,46(11):2707-2715
Laminarin is a β‐glucan from the brown algae Laminaria digitata (J.V. Lamour), which can activate innate immune responses. With the aim of developing a strategy to evaluate specific phenotypical parameters of the effects of laminarin in trout innate immunity, we (i) fed fish with laminarin‐supplemented diet (0.2 g kg?1 day?1) for 21 days and (ii) treated fish with a single dose of intraperitoneal injected laminarin (0.08 g kg?1 fish). The evaluation of cellular and humoral immune parameters was established at phenotypic level by the phagocytic activity of headkidney macrophages and detection of inflammatory cytokines in head kidney and gill tissue by indirect ELISA. Results showed that both delivery methods of laminarin produce an increase in the phagocytic activity in head kidney macrophages and a significant increase in the production of TNFα and IL‐8 in gill tissue at day 21. Additionally, some of these parameters were significantly correlated (P < 0.025), which places them as new potential combined markers to detect activation of trout defense mechanisms by laminarin. These results highlight the importance of developing new protocols to quantitate immune parameters, in order to evaluate immunostimulants in fish farming. 相似文献
4.
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
5.
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
6.
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
7.
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
8.
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
9.
Immunostimulant effects of nucleotide isolated from yeast RNA on carp, Cyprinus carpio L. 总被引:2,自引:0,他引:2
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
10.