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1、THANK YOU,感谢聆听，批评指导,2020,Immunomodulatory effect of swine CCL20 chemokine in DNA vaccination against CSFV,猪CCL20趋化因子在抗猪瘟病毒的 DNA疫苗中的免疫调节作用,1.Abstract,The objective of this work was to explore whether a plasmid expressing CCL20 chemokine could improve the immune response against CSFV in co-administrat
2、ion with a DNA vaccine expressing the E2 protein. The immunization of pigs with the DNA vaccine formulation,that contains swine CCL20 chemokine, resulted in the homogenous induction of detectable levels of CSFV antibodies at 36 days after the rst injection,这项工作的目的是探讨CCL20 趋化因子质粒表达是否可以提高DNA 疫苗表达E2 蛋白
3、联合控制加强对猪瘟病毒的免疫应答。免疫猪的 DNA 疫苗配方包含猪 CCL20 趋化因子，结果在疫苗第一次注射后的第36 天检测到同源诱导的猪瘟病毒的抗体水平,兽医专业英语.ppt,Remarkably, immunized animals with E2 DNA vaccine in co-administration with the plasmid containing swine CCL20 developed high titers of neutralizing antibodies against homologous and heterologous（同源和异源）CSFV st
4、rains and were totally protected upon a lethal viral challenge (sterilizing protection,引人注意的是，E2 DNA 疫苗和含猪 CCL20 质粒共同作用下的免疫动物，在致死量病毒的作用（减毒保护）下产生对同源及异源猪瘟病毒株高效价的中和抗体，并使之得到完全保护。这证实 CCL20 具有增加抗体免疫反应的作用,兽医专业英语.ppt,2.Introduction,It is a small cytokine belonging to the CC chemokine family. It is strongly
5、chemotactic for lymphocytes and weakly attracts neutrophils（中性白细胞）. CCL20 is implicated in the formation and function of mucosal lymphoid tissues（粘膜淋巴组织） via chemoattraction of lymphocytes and dendritic cells towards the epithelial cells（上皮细胞） surrounding these tissues. CCL20 elicits its effects on
6、its target cells by binding and activating the chemokine receptor CCR6,What is CCL20,兽医专业英语.ppt,Classical swine fever virus is the causative agent（病原体） of classical swine fever (CSF), one of the most important swine infectious diseases. CSFV is an icosahedral(二十面的) RNA virus with (+) polarity（正极） an
7、d a lipid envelope（脂质包膜） that belongs to the Pestivirus Genus（瘟病毒属）of the Flaviviridae family（黄病毒科,Then,lets briefly know CSFV,兽医专业英语.ppt,3.Materials and methods,兽医专业英语.ppt,兽医专业英语.ppt,4.Results,Fig.1.Induction of E2 specic antibodies after immunization and CSFV challenge. Open bars indicate pigs inj
8、ected 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). Values are expressed as meanstandard deviations,兽医专业英语.ppt,Fig.2.Kinetics of specic CSFV IFN gamma-producing cells induced after immunization and
9、CSFV challenge.Open bars indicate pigs injected with pE2 and pCDNA3.1+(pigs 13).Black bars indicate pigs injected with plasmids pE2 and pCCL20 (pigs46).Grey bars indicate control pigs(pigs79).Specic CSFV IFN gamma-producing cells are expressed as spots per 5105PBMC.Values are expressed as mean stand
10、ard deviations,兽医专业英语.ppt,Fig.3.Lack of pyrexia upon CSFV challenge in pigs immunized with pE2 and pCCL20. Following CSFV challenge, rectal temperature was daily measured.Pyrexia was considered as rectal temperature 40C. Pigs inoculated with pE2 and pCDNA3.1+ (13). Pigs inoculated with pE2 and pCCL2
11、0 (46).Pigs injected with PBS (79,兽医专业英语.ppt,Fig.4.IFN alpha levels in serum at 3 and 7days post CSFV challenge.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
12、IFN-alpha are expressed as Units of IFN-alpha/mL of serum.Values are expressed as mean standard deviations,兽医专业英语.ppt,Lack of detectable CSFV RNA in animals co-administered pE2 and pCCL20,兽医专业英语.ppt,The protection conferred by pE2 and pCCL20 mixtureassociates with a better induction of neutralizing
13、antibodies upon challenge,兽医专业英语.ppt,5.Discussion,DNA immunization can be a useful tool for understanding the intrinsic immunological mechanisms involved in protection against a given pathogen or to map immunogenic protective determinants. Illustrating the versatility of DNA immunization, plasmid-ba
14、sed cytokines（细胞因子）,chemokines（趋化因子）, co-stimulatory molecules and enhancers of Dendritic Cells (DC) recruitment have been used as adjuvants to potentiate protective immune responses against different pathogens,兽医专业英语.ppt,For CSFV, we have shown that immunization with E2 DNA vaccine induced specic T
15、 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 insufcient prim
16、ing of the T and B lymphocytes after vaccination,兽医专业英语.ppt,Taken into account the feasibility of DNA immunization and its protective capacity against CSFV，we determined to use this methodology as a tool to analyze the potential of swine CCL20 chemokine to enhance the immune response elicited against E2 of CSFV in pigs. Here, we have 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 of antibodies
