摘要
目的:比较巨噬细胞源趋化因子(MDC)、补体片段C3d3、志贺毒素B亚单位(STxB)和小鼠β-防御素2(mBD2)分别与柯萨奇病毒B3(CVB3)VP1构建的融合基因疫苗及VP1基因疫苗对小鼠的免疫效果。方法:将雄性BALB/c小鼠随机分为6组,每组22只,分别于股四头肌注射pcDNA3、pcDNA3/VP1、pcDNA3/MDC-VP1、pcDNA3/VP1-C3d3、pcD-NA3/STxB-VP1和pcDNA3/mBD2-VP1,接种剂量为每次100μg/只,3周免疫1次,共3次。每次免疫后第14天内眦静脉取血,用微量中和试验滴定血清中和抗体滴度。第3次免疫后第21天,每组随机取3只小鼠,制备脾淋巴细胞,用CCK-8细胞计数法检测特异性CTL的杀伤活性;每组取3只小鼠以3LDLD50CVB3病毒攻击,第7天取血处死,检测血清病毒滴度;其余小鼠以5LDLD50的CVB3攻击,观察各组的生存情况。结果:除了pcDNA3对照组,其他各组的中和抗体滴度均随免疫次数的增加而提高(P<0.01),第3次免疫后,pcD-NA3/MDC-VP1、pcDNA3/VP1-C3d3和pcDNA3/mBD2-VP1组的抗体滴度明显高于pcDNA3/VP1组(P<0.01);pcDNA3/STxB-VP1和pcDNA3/mBD2-VP1组CTL杀伤活性显著高于其他各组(P<0.01)。致死量病毒攻击后,pcDNA3/MDC-VP1、pcDNA3/VP1-C3d3和pcDNA3/mBD2-VP1组小鼠血中病毒滴度显著低于其他组(P<0.01);pcDNA3/MDC-VP1和pcD-NA3/VP1-C3d3组小鼠生存率显著高于其他各组(P<0.05)。结论:pcDNA3/MDC-VP1和pcDNA3/VP1-C3d3能诱导出较强的体液免疫和细胞免疫,能有效降低血中病毒滴度,获得较高的小鼠生存率;整体效果优于其他组。
AIM：To compare the immunogenicity and protective effects on CVB3 infected mice of four DNA fusion vaccines coupling coxsackievirus B3（CVB3） VP1 with macrophage-derived chemokine（MDC）,C3d3,shiga toxin B subuit（STxB） and mouse beta-defensin-2（mBD2）,respectively.METHODS：BALB/c mice were divided into 6 groups randomly and inoculated in quadriceps at 3-week interval for 3 times with pcDNA3,pcDNA3/VP1,pcDNA3/MDC-VP1,pcDNA3/VP1-C3d3,pcDNA3/STxB-VP1 and pcDNA3/mBD2-VP1,respectively.Fourteen days after every inoculation,serum samples were collected and CVB3 specific neutralizing antibodies were determined.Three weeks after the last immunization,the mice were treated in three ways.First,the spleen cells were isolated from 3 mice of each group and specific CTL activities were tested.Second,3 mice of each group were further challenged with 3LDLD50 CVB3 and sacrificed 7 days later,and their blood viral titers were evaluated.Third,the rest mice of each group were subjected to intraperitoneal（i.p.） challenge with 5LDLD50 CVB3 and their survival was observed.RESULTS：The neutralizing antibodies against CVB3 were induced in pcDNA3/VP1,pcDNA3/MDC-VP1,pcDNA3/VP1-C3d3,pcDNA3/STxB-VP1 and pcDNA3/mBD2-VP1 groups,and antibody titers correlated with the number of injections（P〈0.01）.After three immunizations,the antibody titers in pcDNA3/MDC-VP1,pcDNA3/VP1-C3d3 and pcDNA3/mBD2-VP1 groups were higher than the ones in pcDNA3/VP1and pcDNA3/STxB-VP1 groups（P〈0.01）.The specific CTL activities in both pcDNA3/STxB-VP1 and pcDNA3/mBD2-VP1 groups were significantly stronger than those in the other groups（P〈0.01）.After CVB3 challenge,the blood viral titers in the pcDNA3/MDC-VP1,pcDNA3/VP1-C3d3 and pcDNA3/mBD2-VP1 groups were lower than those in the other groups（P〈0.01）,and the pcDNA3/MDC-VP1 and pcDNA3/VP1-C3d3 mice survived longer than the others（P〈0.05）.CONCLUSION：Both pcDNA3/MDC-VP1 and pcDNA3/VP1-C3d3 vaccines could induce stronger immune responses,resulting in higher survival rates and
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2010年第2期103-106,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
河北省科学技术研究与发展计划(08276101D-93)
河北省中医药管理局科学研究计划(2007118)
河北省卫生厅科技研究计划(08262)
