獸醫(yī)專業(yè)英語學(xué)習(xí)教案

1、會計(jì)學(xué)1獸醫(yī)專業(yè)英語獸醫(yī)專業(yè)英語第一頁，編輯于星期三：十七點(diǎn) 三十四分。vaccine formulation,that contains swine CCL20 chemokine, resulted in the homogenous induction of detectablelevels of CSFV antibodies at 36 days after the first injection. 這項(xiàng)工作的目的是探討這項(xiàng)工作的目的是探討CCL20 CCL20 趨化因子質(zhì)粒表達(dá)是否可以提高趨化因子質(zhì)粒表達(dá)是否可以提高DNA DNA 疫苗表達(dá)疫苗表達(dá)E2 E2 蛋白聯(lián)合控制加強(qiáng)對豬瘟

2、病毒的免疫應(yīng)答。免疫豬的蛋白聯(lián)合控制加強(qiáng)對豬瘟病毒的免疫應(yīng)答。免疫豬的 DNA DNA 疫苗配方包含豬疫苗配方包含豬 CCL20 CCL20 趨化因子，結(jié)果在疫苗第一次注射后的趨化因子，結(jié)果在疫苗第一次注射后的第第36 36 天檢測到同源誘導(dǎo)的豬瘟病毒的抗體水平。天檢測到同源誘導(dǎo)的豬瘟病毒的抗體水平。第1頁/共29頁第二頁，編輯于星期三：十七點(diǎn) 三十四分。 Remarkably, immunized animals with E2 DNA vaccine in co-administration with the plasmid containing swine CCL20 developed

3、 high titers of neutralizing antibodies against homologous and heterologous（同源和異源）CSFV strains and were totally protected upon a lethal viral challenge (sterilizing protection). 引人注意的是，引人注意的是，E2 DNA E2 DNA 疫苗和含豬疫苗和含豬 CCL20 CCL20 質(zhì)粒共同質(zhì)粒共同作用下的免疫動物，在致作用下的免疫動物，在致死量死量病毒的病毒的作用作用（減毒保護(hù)）下（減毒保護(hù)）下產(chǎn)生對同源及異源豬瘟病毒株

4、高效價(jià)的中和抗體產(chǎn)生對同源及異源豬瘟病毒株高效價(jià)的中和抗體，并并使之使之得到得到完全保護(hù)。這證實(shí)完全保護(hù)。這證實(shí) CCL20 CCL20 具有增加抗體免疫反應(yīng)的作用。具有增加抗體免疫反應(yīng)的作用。第2頁/共29頁第三頁，編輯于星期三：十七點(diǎn) 三十四分。via chemoattraction of lymphocytes and dendritic cells towards the epithelial cells（上皮細(xì)胞） surrounding these tissues. CCL20 elicits its effects on its target cells by binding a

5、nd activating the chemokine receptor CCR6.What is CCL20？第3頁/共29頁第四頁，編輯于星期三：十七點(diǎn) 三十四分。virus with (+) polarity（正極）and a lipid envelope（脂質(zhì)包膜）that belongs to the Pestivirus Genus（瘟病毒屬瘟病毒屬）of the Flaviviridae family（黃病毒科）.Then,lets briefly know CSFV.第4頁/共29頁第五頁，編輯于星期三：十七點(diǎn) 三十四分。Cells and viruesConstruction

6、 of the plasmid expressing full-length CCL20 chemokineDNA immunization of pigsCSFV challendge第5頁/共29頁第六頁，編輯于星期三：十七點(diǎn) 三十四分。Isolation of porcine PBMC and Elispot assay fordetection CSFV-specific IFN-gamma producing cellsAntibody detectionRT-PCR real timeELISA for IFN-alpha and IFN-gamma detection from

7、serum samples第6頁/共29頁第七頁，編輯于星期三：十七點(diǎn) 三十四分。Fig.1.Induction of E2 specific antibodies after immunization and CSFV challenge. Open bars indicate pigs injected with pE2 and pCDNA3.1+ (pigs 13).Black bars indicate pigs injected with plasmids pE2 and pCCL20 (pigs 46).Grey bars indicate control pigs (pigs79

8、). Values are expressed as meanstandard deviations.第7頁/共29頁第八頁，編輯于星期三：十七點(diǎn) 三十四分。Fig.2.Kinetics of specific CSFV IFN gamma-producing cells induced after immunization and CSFV challenge.Open bars indicate pigs injected with pE2 and pCDNA3.1+(pigs 13).Black bars indicate pigs injected with plasmids pE2

9、and pCCL20 (pigs46).Grey bars indicate control pigs(pigs79).Specific CSFV IFN gamma-producing cells are expressed as spots per 5105PBMC.Values are expressed as mean standard deviations第8頁/共29頁第九頁，編輯于星期三：十七點(diǎn) 三十四分。 of pyrexia upon CSFV challenge in pigs immunized with pE2 and pCCL20. Following CSFV ch

10、allenge, rectal temperature was daily measured.Pyrexia was considered as rectal temperature 40 C. Pigs inoculated with pE2 and pCDNA3.1+ (13). Pigs inoculated with pE2 and pCCL20 (46).Pigs injected with PBS (79).第9頁/共29頁第十頁，編輯于星期三：十七點(diǎn) 三十四分。Fig.4.IFN alpha levels in serum at 3 and 7days post CSFV cha

11、llenge.Open bars indicate pigs injected with pE2 and pCDNA3.1+(pigs13).Black bars indicate pigs injected with plasmids pE2 and pCCL20 (pigs46).Grey bars indicate control pigs (pigs 79).Levels of IFN-alpha are expressed as Units of IFN-alpha/mL of serum.Values are expressed as mean standard deviation

12、s.第10頁/共29頁第十一頁，編輯于星期三：十七點(diǎn) 三十四分。Lack of detectable CSFV RNA in animals co-administered pE2 and pCCL20第11頁/共29頁第十二頁，編輯于星期三：十七點(diǎn) 三十四分。The protection conferred by pE2 and pCCL20 mixtureassociates with a better induction of neutralizing antibodies upon challenge第12頁/共29頁第十三頁，編輯于星期三：十七點(diǎn) 三十四分。cytokines（細(xì)胞因

13、子）,chemokines（趨化因子）, co-stimulatory molecules and enhancers of Dendritic Cells (DC) recruitment have been used as adjuvants to potentiate protective immune responses against different pathogens.第13頁/共29頁第十四頁，編輯于星期三：十七點(diǎn) 三十四分。 For CSFV, we have shown that immunization with E2 DNA vaccine induced speci

14、fic T cell responses in the absence of neutralizing antibodies before viral challenge.Nevertheless, some vaccinated animals showed a delay in the induction of neutralizing antibodies and they did not achieve enough clinical protection after CSFV challenge.This response was explained by an insufficie

15、nt priming of the T and B lymphocytes after vaccination.第14頁/共29頁第十五頁，編輯于星期三：十七點(diǎn) 三十四分。shown that co-injection of pCCL20 with pE2 in domestic pigs results in an enhancement of the humoral response elicited, as indicated by the induction ofantibodies against E2 glycoprotein in all immunized pigs.第15頁/

16、共29頁第十六頁，編輯于星期三：十七點(diǎn) 三十四分。pCCL20 in comparison with the pE2 injected pigs.Remarkably, pigs immunized with pE2 and pCCL20 elicited,at days 7 and 13 p.c., high levels of neutralizing antibodies. Also, these antibodies neutralized a heterologous（異種的）CSFV strain with high titers, suggesting that co-expre

17、ssion of CCL20expands the antigenic spectrum of the antibodies induced by E2.在pE2組的免疫后第36天，抗E2特定抗體的平均值顯示的是陰性。經(jīng)過CSFV的感染第3天，對比只免疫pE 2的動物，混合免疫pE2+pCCL20組，E2抗體的平均值顯示陽性。值得注意的是用pE2和pCCL20進(jìn)行誘導(dǎo)的免疫豬在攻毒后第7天、第13天，有高水平中和抗體。當(dāng)然，這些抗體以一個(gè)高滴度中和異源CSFV株，意味著聯(lián)合表達(dá)的CCL20擴(kuò)大了由E2誘導(dǎo)的抗體的抗原圖譜。第16頁/共29頁第十七頁，編輯于星期三：十七點(diǎn) 三十四分。CCL20

18、to enhance the T helper cell response associated with the induction of neutralizing antibodiesagainst CSFV in pigs is suggested. 最近的研究表明CCL20趨化因子可以增強(qiáng)經(jīng)過DNA疫苗免疫的小鼠體液應(yīng)答。我們的結(jié)果證實(shí)了這樣的一個(gè)小鼠模型實(shí)驗(yàn)。與此同時(shí)，揭示了CCL20趨化因子增強(qiáng)T細(xì)胞應(yīng)答和誘導(dǎo)豬中抗CSFV中和抗體的產(chǎn)生相關(guān)的能力。第17頁/共29頁第十八頁，編輯于星期三：十七點(diǎn) 三十四分。detected in the sera. 這里的實(shí)驗(yàn)報(bào)告了致死性病毒感染

19、后與這里的實(shí)驗(yàn)報(bào)告了致死性病毒感染后與OIE ManualOIE Manual（國（國際獸醫(yī)手冊）指示的相似，在經(jīng)過際獸醫(yī)手冊）指示的相似，在經(jīng)過pE2+pE2+pCCLpCCL2020免疫的動物免疫的動物產(chǎn)生的中和抗體與經(jīng)得起的完全保護(hù)相關(guān)。這些動物產(chǎn)生的中和抗體與經(jīng)得起的完全保護(hù)相關(guān)。這些動物既既沒有疾病癥狀沒有疾病癥狀，在血清中，在血清中也檢測不到也檢測不到RNARNA病毒病毒。第18頁/共29頁第十九頁，編輯于星期三：十七點(diǎn) 三十四分。相反，只有經(jīng)過相反，只有經(jīng)過pE2pE2免疫的豬在感染后第免疫的豬在感染后第7 7天有較低水平的中天有較低水平的中和抗體。這個(gè)滴度的減少可以與感染后第和

20、抗體。這個(gè)滴度的減少可以與感染后第3 3和第和第7 7天的動物天的動物發(fā)熱，輕微癥狀，病理發(fā)現(xiàn)和血清中病毒的測定相聯(lián)系。發(fā)熱，輕微癥狀，病理發(fā)現(xiàn)和血清中病毒的測定相聯(lián)系。第19頁/共29頁第二十頁，編輯于星期三：十七點(diǎn) 三十四分。detected by RT-PCR real timeclinical signs and the virus load（病毒載量）detected in these animals were similar to previously described for pigs inoculated（接種）with empty plasmid(pCDNA3.1+) in

21、 similar experiments.第20頁/共29頁第二十一頁，編輯于星期三：十七點(diǎn) 三十四分。and pCCL20 speeds up the induction of the specific humoral response. Likely,the effect of CCL20 on vaccination was related to theinduction of recruitment of APC（抗原提呈細(xì)胞） that further enhance cooperation and activation of T and B cells, generating a

22、more potent humoral response after vaccination.只接種只接種pE2pE2的動物產(chǎn)生特定的動物產(chǎn)生特定-IFN-IFN產(chǎn)生細(xì)胞，但是在注射后第產(chǎn)生細(xì)胞，但是在注射后第3636天幾天幾乎沒有可測定的特定抗乎沒有可測定的特定抗E2E2抗體的產(chǎn)生。這些結(jié)果表明聯(lián)合控制的抗體的產(chǎn)生。這些結(jié)果表明聯(lián)合控制的pE2pE2和和pCCL20pCCL20加快誘導(dǎo)特殊體液應(yīng)答。同樣，加快誘導(dǎo)特殊體液應(yīng)答。同樣，CCL20CCL20在疫苗的效果與在疫苗的效果與APCAPC聚集誘導(dǎo)相關(guān)，進(jìn)一步加強(qiáng)聯(lián)合操縱和聚集誘導(dǎo)相關(guān)，進(jìn)一步加強(qiáng)聯(lián)合操縱和T T、B B細(xì)胞的活性，免疫后產(chǎn)

23、細(xì)胞的活性，免疫后產(chǎn)生更強(qiáng)潛在的體液應(yīng)答生更強(qiáng)潛在的體液應(yīng)答。第21頁/共29頁第二十二頁，編輯于星期三：十七點(diǎn) 三十四分。早期報(bào)道表明經(jīng)過早期報(bào)道表明經(jīng)過E2-DNAE2-DNA疫苗免疫之后，疫苗免疫之后，-IFN-IFN表達(dá)表達(dá)T T細(xì)胞的細(xì)胞的特定誘導(dǎo)。同樣，特定誘導(dǎo)。同樣，CSFVCSFV特特定定-IFN-IFN產(chǎn)生細(xì)胞在產(chǎn)生細(xì)胞在病毒病毒感染前感染前與注射質(zhì)粒組與注射質(zhì)粒組頻數(shù)頻數(shù)相似。感染后包括相似。感染后包括-IFN-IFN誘導(dǎo)誘導(dǎo)控制控制病毒復(fù)制可病毒復(fù)制可以解釋質(zhì)粒注射組以解釋質(zhì)粒注射組（pE2pE2組）組）在感染后第在感染后第3 3天觀察到的天觀察到的CSFVCSFV特定特

24、定-IFN-IFN產(chǎn)生細(xì)胞產(chǎn)生細(xì)胞的應(yīng)答的減少的應(yīng)答的減少。第22頁/共29頁第二十三頁，編輯于星期三：十七點(diǎn) 三十四分。in blood to tissues to halt viral replication there. On the otherhand, these results could be related to the levels of neutralizing antibodies found at 7 days p.c. in animals immunized with pE2 and pCCL20, compared to those of animals immu

25、nized with pE2 alone.第23頁/共29頁第二十四頁，編輯于星期三：十七點(diǎn) 三十四分。found by Elispot（酶聯(lián)免疫斑點(diǎn)技術(shù)）.Probably the number of IFN-gamma producing cells may not be high enough to produce detectable levels in blood.These results corroborate previous reports where no correlation was found between the number of IFN-gamma-producing cells and the cytokine levels in serum measured by ELISA .第24頁/共29頁第二十五頁，編輯于星期三：十七點(diǎn) 三十四分。at