应用吞噬性能选育矮小鸡的抗病群体     

麻慧  韩红兵  宁中华  连正兴 《中国农业科学》2016,49(23):4628-4637

【目的】单核巨噬细胞在免疫系统中发挥着重要的作用,试验研究了对单核巨噬细胞吞噬性能的选择影响矮小鸡G1代抗传染性支气管炎病毒(IBV)的能力。【方法】在矮小鸡290日龄时,检测了G0代500只个体(母鸡400只,公鸡100只)的吞噬指数(PI),并根据PI的大小分为强吞噬力组(HPIG)和弱吞噬力组(LPIG)。建立2×2交配组合:HPIG♂×HPIG♀(强公强母组),LPIG♂×HPIG♀(弱公强母组),HPIG♂×LPIG♀(强公弱母组),LPIG♂×LPIG♀(弱公弱母组)。随机选择400只1日龄G1代雏鸡(每组100只,公母各半),其中360只采用滴鼻方式人工接种含IBV M41病毒的鸡胚尿囊液,40只作为对照,连续观察14d,记录死亡数,制作石蜡切片进行H.E.染色,并通过红细胞凝集抑制试验(HI)测定15 d时存活鸡只的抗体效价。G1代鸡20周龄时,选择母鸡12只,其中强组母鸡后代6只,弱组母鸡后代6只,以病毒模拟物Poly I:C刺激体外培养的单核巨噬细胞,采用荧光定量PCR的方法测定细胞因子及主要组织相容性复合体(MHC)m RNA的表达水平。【结果】G0代不同个体对异源红细胞的吞噬能力差异显著,通过测定G0代个体的吞噬指数,建立交配组合,孵育得到G1代鸡。对G1代鸡的IBV攻毒试验的结果为强公强母组后代的死亡率(33.3±0.05)%显著低于弱公弱母组(55.6±0.05)%,其他两个组合后代的死亡率介于上述值之间,弱公强母组为(43.3±0.05)%,强公弱母组为(47.8±0.05)%;母鸡对后代的影响大于公鸡,强母组后代的死亡率(38.3±0.04)%显著低于弱母组(51.7±0.04)%。攻毒组在接种IBV M41病毒3 d后表现出咳嗽、呼吸困难、食欲减退、精神沉郁等临床症状,对病死鸡的气管及肾脏的H.E.染色结果可见典型的病灶,气管上皮细胞坏死脱落,肾小管上皮细胞发生空泡变性等,而对照组均无临床症状及组织病变。对198只攻毒后存活个体抗体滴度的测定结果显示强组母鸡后代抗体滴度(8.45±0.07)显著高于弱组母鸡后代的抗体滴度(8.10±0.08)。利用病毒模拟物Poly I:C刺激体外培养的单核巨噬细胞2 h后,强吞噬力个体(强组母鸡后代)细胞因子IFNγ和IL-1β的表达量分别是弱吞噬力个体(弱组母鸡后代)的5.14倍(P0.05)和2.41倍(P0.05)。强吞噬力个体MHCⅠ的表达量显著高于弱吞噬力个体,而MHCⅡ的表达量差异不显著。【结论】通过测定体外培养的单核巨噬细胞的吞噬性能,按照吞噬指数的高低建立交配组合,强吞噬力母鸡后代的攻毒死亡率显著低于弱吞噬力母鸡后代,且其抗体滴度、细胞因子(IFNγ、IL-1β)和MHCⅠ的表达量显著高于弱吞噬力母鸡后代,说明强吞噬力母鸡后代具有较强的抗IBV的能力。因此单核巨噬细胞的吞噬能力可以作为培育抗IBV品系的一种指标。  相似文献   

聚酰胺-胺对大鼠腹腔巨噬细胞吞噬功能的影响          下载免费PDF全文

林检生  杨奔  陈安  吴丹  胡兆勇  喻嵘  谭劲 《勤云标准版测试》2017,37(7):726-729

目的 研究聚酰胺-胺（poly-amidoamine,PAMAM）对大鼠腹腔巨噬细胞吞噬功能的影响。方法 采用CCK-8试剂盒检测PAMAM对巨噬细胞的细胞毒性。通过巨噬细胞吞噬印度墨汁实验和吞噬鸡红细胞实验,观察PAMAM对巨噬细胞吞噬功能的影响。采用扫描电子显微镜（SEM）观察巨噬细胞吞噬鸡红细胞的形貌。结果 0.1-10 mg/mL的G5 PAMAM-NH2和10 mg/mL的G4.5 PAMAM-COOH对巨噬细胞有毒性（P<0.05）。0.01 mg/mL的G5 PAMAM-NH2、G4.5 PAMAM-COOH、G5 PAMAM-OH对巨噬细胞均无毒性（P>0.05）。0.01 mg/mL的PAMAM对巨噬细胞吞噬鸡红细胞形貌、吞噬率和吞噬指数均无影响（P>0.05）。结论 PAMAM的浓度在0.01 mg/mL范围内,未观察到其对大鼠腹腔巨噬细胞吞噬功能有影响。  相似文献   

华蟾毒精对小鼠巨噬细胞功能的影响     

孟显华  于洋  郝璐 《西北农林科技大学学报(自然科学版)》2016,44(8):61-65

【目的】阐明华蟾毒精(CBG)对巨噬细胞功能的影响,为CBG的开发和利用提供理论依据。【方法】利用Balb/c小鼠制备与纯化腹腔巨噬细胞,采用不同质量浓度(0.5,1.0,1.5,2.0mg/L)CBG作用于巨噬细胞,采用MTT法检测CBG对巨噬细胞吞噬金黄色葡萄球菌的影响;采用Griess法检测CBG对巨噬细胞分泌NO的影响;采用ELISA法检测CBG对巨噬细胞分泌IL-6、TNF-α和GM-CSF的影响。【结果】CBG质量浓度为1.0,1.5和2.0mg/L组均能极显著提高巨噬细胞的吞噬功能;CBG质量浓度为0.5 mg/L组能显著促进巨噬细胞NO的分泌,而1.0,1.5和2.0mg/L CBG组均能极显著促进巨噬细胞NO的分泌;CBG质量浓度为1.0,1.5和2.0mg/L组均能极显著促进巨噬细胞IL-6的分泌;1.0mg/L CBG组能显著促进巨噬细胞TNF-α的分泌,1.5和2.0mg/L CBG组均能极显著促进巨噬细胞TNF-α的分泌;CBG质量浓度为0.5,1.0,1.5和2.0mg/L组均能极显著促进巨噬细胞GM-CSF的分泌。【结论】CBG能提高巨噬细胞的吞噬功能和分泌功能,说明CBG具有良好的抗炎、抗肿瘤和提高免疫等作用。  相似文献   

What is your diagnosis? Purple granules within neutrophils of a dog          下载免费PDF全文

Francisco O. Conrado  Rose E. Raskin 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2017,46(4):639-640

  相似文献   

Flow cytometric characterization of bovine blood neutrophil phagocytosis of fluorescent bacteria and zymosan particles   总被引：1，自引：0，他引：1  

A M Saad  M Hageltorn 《Acta veterinaria Scandinavica》1985,26(3):289-307

A Flow Cytometric method for the evaluation of the phagocytic capacity of bovine blood neutrophils is described. The neutrophils were isolated from bovine blood by a one step discontinuous gradient of Percoll. By this technique of isolation, 90 ± 2.8 % (mean ± s) of the granulocytes in the whole blood were recovered.Isolated neutrophils were incubated with FITC labeled S. aureus or zymosan particles in a ratio of 1:20 and 1:10, respectively, and a final serum concentration of 10 %. Phagocytosis was terminated after 15 min and the number of extracellular bacteria or zymosan particles and the percentage of phagocytic granulocytes were registered by Flow Cytometry (FCM). FCM and microscopic studies revealed that eosinophils play a minor role in the phagocytosis of bacteria. The neutrophils were the main population of the granulocytes which were actively phagocytic. Variation among cows in the ability of their blood neutrophils to phagocytize bacteria was evident.  相似文献   

Phagocytic plasmacytoma in a dog     

Yearley JH  Stanton C  Olivry T  Dean GA 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2007,36(3):293-296

A 4-year-old neutered male Golden Retriever was presented to the oncology service of the North Carolina State University Veterinary Teaching Hospital for staging of a histiocytic sarcoma of the left forelimb, diagnosed on the basis of biopsies submitted by the referring veterinarian. Cytologic assessment of aspirates of 2 splenic nodules identified on ultrasonographic examination of the abdomen revealed a highly phagocytic population of neoplastic round cells morphologically suggestive of plasma cells. Histologic assessment of the forelimb mass after amputation of the limb revealed a neoplastic round cell population demonstrating extensive cytophagia and erythrophagia. Immunohistochemical analysis of the tumor population revealed it to be negative for BLA.36 with sporadic positivity for lysozyme and CD79a. Immunofluorescent evaluation revealed weak tumor cell positivity for immunoglobulin (Ig) A and IgM, but extensive strong positivity for IgG, confirming the plasma cell origin of the tumor. Although extensive phagocytic activity may strongly suggest histiocytic origin, plasma cell origin must also be considered among the differential diagnoses for phagocytic round cell tumors.  相似文献   

Phagocytic and oxidative burst activity of phagocytic cells in peripheral blood and uterine washings in cows with clinical endometritis before and after intrauterine use of cephapirin and methisoprinol          下载免费PDF全文

Piotr Brodzki  Urszula Lisiecka  Adam Brodzki  Renata Pyz‐Łukasik  Leszek Krakowski 《Animal Science Journal》2018,89(10):1389-1397

The aim of the study was to evaluate phagocytic and killing activity of phagocytic cells in blood and uterine flush of cows with endometritis before and after intrauterine (i.u.) administration of cephapirin and methisoprinol. The research was carried out on 28 cows with clinical endometritis. Animals were divided into four groups, each composed of seven cows, depending on the i.u. treatment used: Group A—cephapirin; Group B—methisoprinol; Group C—cephapirin and methisoprinol at the same time; and a control group—without medication. Using flow cytometry technique, the phagocytic activity of granulocytes and monocytes was identified, as well as the oxidative burst activity of neutrophils in the peripheral blood and uterine washings. Summarizing the results of the research, i.u. infusion of cephapirin caused a reduction in the phagocytic and killing activity of phagocytes. The i.u. use of methisoprinol increased phagocytic and killing activity of phagocytes in the uterus. Administering both listed substances simultaneously showed a decrease in phagocytosis, presumably due to the dominating inhibitor effect of the antibiotic. However, also an increase of mean fluorescence intensity was observed, presumably caused by the methisoprinol. Intrauterine use of immunostimulatory substances, can improve the effectiveness of the treatment of endometritis in cows.  相似文献   

布鲁氏菌P39基因的克隆、原核表达及其糖基化修饰     

胡祥坤  卢天成  刘思阳  付玉芹  王茗宇  郑晗旭  王岩  王秀然 《中国畜牧兽医》2018,45(11):2996-3002

为探究糖基化修饰对布鲁氏菌P39蛋白免疫原性的影响,本试验对布鲁氏菌P39蛋白进行了表达和纯化,并对表达的蛋白进行了甘露六糖修饰。参照GenBank公布的布鲁氏菌P39基因序列设计引物,从布鲁氏菌16M基因组中克隆P39基因片段并连接至pMD19-T载体,转化大肠杆菌DH5α感受态细胞,提取阳性菌株质粒进行酶切等鉴定,鉴定正确后构建重组质粒pGEX-6P-1-P39,对重组蛋白进行诱异表达与条件优化。利用SDS-PAGE和Western blotting对诱导表达的目的蛋白进行分析,将鉴定正确的蛋白经GST亲和层析柱纯化。通过EDC/NHS法对纯化的P39蛋白进行甘露六糖修饰,研究甘露六糖修饰后目的蛋白对巨噬细胞吞噬的影响。结果显示,本试验成功克隆了片段大小为1 206 bp的目的基因,构建了pGEX-6P-1-P39原核表达载体,在大肠杆菌中成功表达了P39蛋白,该蛋白主要以可溶性形式存在。Western blotting结果显示,在约65 ku处有特异性条带。纯化后获得目的蛋白大小为43 ku,成功对目的蛋白P39进行了糖基化修饰,得到修饰产物与蛋白的摩尔比为2.3∶1。此外,糖基化修饰可显著提前蛋白激活小鼠巨噬细胞的吞噬作用时间。本试验结果可为研究甘露六糖修饰影响P39蛋白免疫原性的机制提供参考。  相似文献   

Combined phagocytosis-nitroblue tetrazolium reduction test in bovine neutrophils     

Valinoti JM  Zinkl JG  Jain NC 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》1988,17(4):99-104

An assay was developed for the simultaneous evaluation of phagocytosis and oxidative metabolism of bovine blood neutrophils. Phagocytosis was evaluated by using opsonized zymosan, and oxidative metabolism was evaluated by nitroblue tetrazolium (NBT) reduction. Normal bovine neutrophils exhibited moderate variation in ability to phagocytize zymosan, but little variation in ability to reduce NBT. The subcellular location of NBT reduction to formazan was determined by electron microscopy. Electron dense formazan precipitate was observed along the inner membrane of phagosomes enclosing zymosan particles and radiating from the membrane toward the center of the phagosome.  相似文献   

Cell cannibalism by malignant neoplastic cells: three cases in dogs and a literature review          下载免费PDF全文

Antonio Meléndez‐Lazo  Paola Cazzini  Melinda Camus  Georgina Doria‐Torra  Alberto Jesús Marco Valle  Laia Solano‐Gallego  Josep Pastor 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2015,44(2):287-294

Cell cannibalism refers to the engulfment of cells by nonprofessional phagocytic cells. Studies in human medicine have demonstrated a relationship between the presence of cell cannibalism by neoplastic cells and a poor outcome, and have shown a positive correlation with the presence of metastasis at the time of diagnosis. The biologic significance of cell cannibalism is unknown, but it is proposed that it may represent a novel mechanism of tumor immune evasion as a survival strategy in cases of unfavorable microenvironmental conditions. This report describes clinical and morphologic features of 3 cases of dogs with malignant neoplasia in which the presence of cellular cannibalism was observed in cytologic and histologic specimens. In the 1^st case, a dog with a primary tonsillar squamous cell carcinoma with metastasis to retropharyngeal lymph nodes had neoplastic epithelial cells engulfing neutrophils noted in cytologic examination of the lymph nodes. In the 2^nd case, neoplastic epithelial cells were seen engulfing each other in fine‐needle aspirates from a primary mammary carcinoma with lung metastasis. In the 3^rd case, poorly differentiated neoplastic mast cells from a recurrent, metastatic grade III mast cell tumor were observed cannibalizing eosinophils. A brief review of the literature describing known cell‐into‐cell relationships and the possible biologic significance and mechanisms involved in this phenomenon is provided. The relationship between cell cannibalism and distant metastasis should be explored in further studies, as it may prove to be a criterion of malignancy, as it is proposed in human medicine.  相似文献