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1.
《中国兽医学报》2017,(6):1007-1011
为研究猪瘟(CSF)、猪繁殖与呼吸综合征(PRRS)免疫猪肺门淋巴结中淋巴细胞对PRRSV N类特异性肽刺激的应答特点,试验从猪肺门淋巴结中分离T淋巴细胞,分别用PRRSV N类特异性肽进行刺激,待细胞培养至24,48h时,离心取细胞上清液,检测上清液中细胞因子TGF-β1、IL-10、IL-12分泌含量的变化。结果显示,PRRSV N类特异性肽能够抑制TGF-β1、IL-10的分泌,表明N1、N2肽具有减弱免疫抑制,促进抗炎性细胞因子分泌的作用;此外IL-12的分泌量也减少,且有逐渐恢复平衡的趋势,提示PRRSV N类特异性肽可能具有双面性,有待于进一步研究。本试验为PRRSV合成肽疫苗及疫苗佐剂的研制提供理论依据。  相似文献   

2.
《中国兽医学报》2016,(4):549-552
本试验探讨PRRSV特异性肽-N对CSFV弱毒疫苗免疫猪骨髓细胞因子分泌量的影响。选择已免疫CSFV弱毒疫苗的猪为实验动物,提取四肢长骨骨髓,进行T淋巴细胞的分离培养。分别在培养后的24、72 h,用PRRSV特异性肽N1、N2对所培养细胞进行刺激,再进行培养12 h,经离心方法得到细胞上清液,最终检测上清液中细胞因子IFN-γ、IL-12、IL-10的分泌含量变化。结果显示,PRRSV特异性肽-N对于免疫猪骨髓中IFN-γ的分泌影响较小,能够抑制IL-12的分泌,不利于机体免疫反应,对IL-10的分泌影响复杂,需进一步研究。  相似文献   

3.
应用PRRSV-GP5特异性肽刺激CSF、PRRS免疫猪外周血淋巴细胞,采用ELISA方法测定不同时间点淋巴细胞分泌IFN-γ及IL-10的水平,分析PRRSV-GP5特异性肽刺激后,淋巴细胞培养上清中细胞因子的变化规律。结果显示,CSFV高抗PRRSV低抗体组于PRRSV免疫14d,肽刺激组中IFN-γ水平显著高于对照组(P<0.05);其他组均差异不显著(P>0.05)。PRRSV免疫14d,细胞培养24,72h后加PRRSV-GP5肽刺激,各组中试验组与对照组相比IL-10水平明显降低(P<0.01);PRRSV免疫28d,各组中肽刺激组与空白组相比,IL-10水平均无显著性差异(P>0.05)。结果表明,PRRSV-GP5肽在PRRSV免疫早期可以促进IFN-γ的分泌,并抑制IL-10的产生,但在免疫后期对淋巴细胞分泌的调节作用还需进一步研究。  相似文献   

4.
试验从猪骨髓中分离T淋巴细胞,待细胞培养达到相应时间点时,分别用PRRSV GP5类特异性肽进行刺激,继而检测细胞因子IL-10、IL-12以及IFN-γ分泌量的变化情况。结果显示:在PRRSV GP5类特异性肽刺激后,骨髓中IL-10分泌量减少,表明G1、G2肽具有减弱免疫抑制,促进抗炎性细胞因子产生的作用;此外,IL-12的分泌量也减少,提示猪的免疫保护作用减弱,表明PRRSV GP5类特异性肽可能具有双面性;而IFN-γ的分泌量变化不稳定,有待于进一步研究。  相似文献   

5.
应用PRRSV-GP5特异性肽刺激CSF、PRRS免疫猪外周血淋巴细胞,采用EI。IsA方法测定不同时间点淋巴细胞分泌IFN-7及IL-10的水平,分析PRRSv_GP5特异性肽刺激后,淋巴细胞培养上清中细胞因子的变化规律。结果显示,CSFV高抗PRRSV低抗体组于PRRSV免疫14d,肽刺激组中IFN-7水平显著高于对照组(P〈0.05);其他组均差异不显著(P〉0.05)。PRRSV免疫14d,细胞培养24,72h后加PRRSV—GP5肽刺激,各组中试验组与对照组相比IL-10水平明显降低(P〈0.01);PRRSV免疫28d,各组中肽刺激组与空白组相比,IL-10水平均无显著性差异(P〉0.05)。结果表明,PRRSV-GP5肽在PRRSV免疫早期可以促进IFN-7的分泌,并抑制IL-10的产生,但在免疫后期对淋巴细胞分泌的调节作用还需进-步研究。  相似文献   

6.
生物矿化技术可以增强口蹄疫(foot-and-mouth, FMD)病毒样颗粒(virus-like particles, VLPs)的耐热性,但并无试验数据支撑矿化FMD VLPs用于临床的可行性。本研究旨在评价矿化FMD VLPs疫苗在牛和猪上的免疫原性。用VLPs和矿化VLPs免疫猪,采用液相阻断ELISA(LPBE)检测猪血清中的特异性抗体滴度,并经酶联免疫斑点试验(ELISpot)检测免疫猪外周血淋巴细胞(PBLs)经体外刺激后分泌的特异性IFN-γ。用矿化A型VLPs、矿化O型VLPs、矿化的A型和O型二价VLPs分别免疫牛,采用LPBE、中和试验、夹心ELISA检测血清中的特异性抗体、中和抗体以及IL-4和IFN-γ的含量。结果显示,VLPs与矿化VLPs免疫猪的血清抗体水平无显著差异,相对于VLPs,矿化VLPs免疫猪的PBLs分泌的IFN-γ水平高于VLPs免疫猪(P<0.05);3种疫苗免疫牛血清中IL-4和IFN-γ水平均高于PBS组,矿化二价疫苗与相应的矿化单价疫苗免疫牛的血清特异性抗体和中和抗体效价均无显著差异。综上所述,矿化VLPs对牛和猪均具有良好的...  相似文献   

7.
选择已成功免疫猪瘟(CSF)和猪繁殖与呼吸综合征(PRRS)弱毒疫苗的仔猪作为实验动物,提取其骨髓、胸腺、肺门淋巴结和脾脏,进行T淋巴细胞的分离培养,然后用PRRSV特异性肽GP5-1、GP5-2对培养的细胞进行刺激,再培养至24,48h时间点时,离心取细胞上清液,检测上清液中TGF-β1的分泌含量的变化。结果显示,PRRSV GP5类特异性肽刺激作用后,CSF、PRRS免疫猪组织淋巴细胞中TGF-β1的分泌量减少,表明GP5-1、GP5-2肽具有减弱免疫抑制,恢复宿主抗病毒免疫应答的作用。本试验为PRRSV合成肽疫苗及疫苗佐剂的研制提供了理论依据。  相似文献   

8.
不同CSF免疫状态下猪PRRS易感性及IFN-γ分泌细胞应答   总被引:1,自引:0,他引:1  
采用酶联免疫斑点检测技术(ELISpot)检测自然状态下猪外周血单核细胞(PBMC)中分泌IFN-γ的细胞数,并用带T细胞表位的猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)特异性小分子多肽刺激培养的PBMC,观察IFN-γ的分泌变化。结果显示,猪瘟病毒(Classical fever virus,CSFV)抗体阳性组中感染PRRSV比率小于CSFV抗体阴性组。CSFV抗体阳性猪PBMC中IFN-γ分泌细胞数量均高于CS—FV抗体阴性组,CSFV抗体阴性且受PRRSV感染猪的PBMC对PRRSV多肽刺激不应答。结果表明,对CSFV疫苗应答好的猪对PRRSV感染有一定的抵抗,其细胞免疫处于活动状态,提示2种传染病的免疫应答机理有部分相关性。  相似文献   

9.
用猪瘟疫苗和高致病性猪繁殖与呼吸综合征弱毒疫苗对猪进行免疫,在免疫后的14d和28d采集外周血液,分析特异性抗体表达量和外周血T淋巴细胞表型的变化,并用猪繁殖与呼吸综合征病毒(PRRSV)特异性肽对淋巴细胞进行刺激。结果显示,在PRRSV免疫后14d,机体PRRSV抗体水平较低,其中CSFV低抗组中CD4+细胞百分数明显降低,CD4+/CD8+细胞的比值也明显降低;用PRRSV特异性肽刺激淋巴细胞24h后,CSFV低抗组与高抗组中CD3+、CD8+细胞的百分数都明显升高,CD4+细胞的百分数及CD4+/CD8+细胞的比值都明显降低。在PRRSV免疫后28d,CSFV抗体和PRRSV抗体的产生都比较高,CD3+细胞、CD4+T淋巴细胞百分数及CD4+/CD8+细胞的比值也明显升高,用肽刺激以后,CSFV高抗组中的CD3+细胞百分数降低,CSFV低抗组中的细胞百分数升高,CD4+/CD8+细胞的比值在所有组中均下降。结果表明,PRRSV特异性肽在PRRSV免疫早期可以使CD3+细胞的百分数升高,CD4+/CD8+细胞的比值降低,其详细的机理还有待进一步的探究。  相似文献   

10.
为了研究猪繁殖与呼吸综合征病毒(PRRSV)GP5重组蛋白(rGP5)对猪外周血单个核细胞(PBMC)功能的影响,本研究选取PRRSV-PC疫苗株中的ORF5基因片段,对其进行原核表达并纯化,结果显示rGP5以可溶性和包涵体两种形式表达,分子质量约为30 ku;将该纯化蛋白免疫大耳白兔,按常规方法制备抗血清并进行酶联免疫吸附试验(ELISA),结果显示,纯化后的rGP5可以与PRRSV阳性血清特异性结合;制备的兔抗rGP5多克隆抗体效价高达1.64 000,可用于后续试验。无菌采集PRRSV阴性猪外周血并分离PBMC,加入终浓度为10μg/mL的rGP5体外培养后,采用间接免疫荧光试验(IFA)检测重组蛋白与PBMC的结合,结果显示rGP5与PBMC共培养1 h后,PBMC出现红色荧光,表明r GP5可以结合PBMC。进一步采用不同浓度的rGP5体外刺激猪PBMC,利用ELISA方法检测淋巴细胞中IL-10、IFN-γ、TGF-β1及TNF-α的分泌水平,同时采用CCK-8细胞增殖检测试剂盒测定rGP5对PBMC增殖的影响,以及不同浓度的rGP5刺激PBMC分泌NO的情况,结果显示,不同浓度的rGP5均显著促进PBMC细胞因子IL-10、IFN-γ和TNF-α的分泌(p0.05);同时rGP5能够促进PBMC增殖(p0.05),也能增加NO分泌(p0.05),上述结果表明GP5是PRRSV重要的抗原,发挥着重要的免疫作用。本实验首次研究了rGP5对猪PBMC免疫功能的影响,揭示PRRSV影响宿主免疫反应的具体机制,有助于阐明PRRSV与宿主细胞相互作用的分子基础,为进一步研究其免疫保护性提供了实验依据。  相似文献   

11.
12.
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。  相似文献   

13.
采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。  相似文献   

14.
本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。  相似文献   

15.
REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.  相似文献   

16.
用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10~0.82μg/g。表明:蜂蜜应视为天然富硒营养品。  相似文献   

17.
乳酸杆菌益生作用机制的研究进展   总被引:2,自引:0,他引:2  
乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和 Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。  相似文献   

18.
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.  相似文献   

19.
乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制.  相似文献   

20.
OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis.  相似文献   

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