摘要
目的 改良新型B3型柯萨奇病毒 (CVB3)DNA疫苗chitosan pcDNA3 VP1,增强其预防CVB3性心肌炎的效果。方法 合成含融内体多肽和多聚赖氨酸的“载体多肽”HApLys16 ,与pcDNA3 VP1、脱乙酰壳多糖 (chitosan)依次复合制备chitosan pcDNA3 VP1 HApLys16疫苗 ,以含 2 0 0 μgDNA剂量疫苗隔周滴鼻免疫小鼠 ,3周后以 3×半数致死剂量 (LD50 )CVB3感染小鼠致病毒性心肌炎 ,检测感染后体重、体重 /心脏重比、心肌HE染色和心脏外观评价病毒性心肌炎预防效果。结果 (1)chitosan pcDNA3 VP1 HApLys16疫苗免疫组黏膜sIgA分泌增高。 (2 )病毒感染后体重显示 :pcDNA3组减重 4 .5 0 % ,chitosan pcDNA3 VP1组和chitosan pcDNA3 VP1 HApLys16组分别增重 1 75 %和 5 .4 4 %。 (3)感染后体重 /心脏重比值显示 :chitosan pcDNA3 VP1 HAplys16组比值最大 ,为 193.77,病毒性心肌炎的体征最轻。 (4)感染后心肌病理显示 :pcDNA3组 10 0 %发生严重病毒性心肌炎 ;chitosan pcDNA3 VP1组发病率 16 .7% ,程度轻 ;chitosan pcDNA3 VP1 HApLys16组不发病 ,心肌完全正常。 (5 )心脏外观显示 :pcDNA3组心脏增大有疤痕状白斑 ;chitosan pcDNA3 VP1组心脏基本正常 ;chitosan pcDNA3 VP1 HApLys16组心脏正常 ,提示无病毒性心肌炎发生。
Objective To enhance the effect of chitosan-pcDNA3-VP1 DNA vaccine to prevent onset of coxsackievirus B3(CVB3)-induced myocarditis. Methods HA-pLys16 “carrier peptide” containing “endosome fusogenic peptide” and poly-Lysine was synthesized and complexed with DNA and chitosan sequentially, resulting chitosan-pcDNA3-VP1-HApLys16 vaccine. It was administrated to BALB/c mice with a total dose of 200 μg pcDNA3-VP1. Mice were infected with 3(LD 50 CVB3 3 weeks later to induce myocarditis. Loss of body weight, body/heart weight ratio, outlook and HE staining of heart tissues was observed and compared as an indicative of severity of myocarditis. Results chitosan-pcDNA3-VP1-HApLys16 vaccine intranasal immunization enhanced secretion of mucosal sIgA. After 3×LD 50 CVB3 infection, pcDNA3 immunized mice groups lost their 5.75% weight following CVB3 infection, while chitosan-pcDNA3-VP1 and chitosan-pcDNA3-VP1-HApLys16 immunized mice increased their body weight by 1.75% and 5.44%. Body/heart weight ratio of chitosan-pcDNA3-VP1-HAplys16 group reached 193.77 suggesting lightened myocarditis. No myonecrosis or infiltrating immune cells existed in heart of chitosan-pcDNA3-VP1-HAplys16 immunized mice, and 1/6 of incidence and very light myocarditis were seen in chitosan-pcDNA3-VP1 immunized mice. While in pcDNA3 immunized mice 6/6 incidence and extremely severe myocarditis were observed. Heart from pcDNA3-immunized mice was bigger than normal ones and had a lot of white stripes, which were not observed in that of chitosan-pcDNA3-VP1 and chitosan-pcDNA3-VP1-HApLys16 immunized mice. Conclusion Novel chitosan-pcDNA3-VP1-HApLys16 vaccine containing “endosome fusogenic peptide” could more effectively prevent CVB3-induced myocarditis than chitosan-pcDNA3-VP1 vaccine.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2004年第4期334-338,共5页
National Medical Journal of China
基金
国家杰出青年科学基金资助项目 (3 992 5 0 3 1)
国家重点基础研究发展计划基金资助项目(2 0 0 1CB5 10 0 0 6)
