摘要
为提高犬瘟热病毒(canine distemper virus,CDV)重组质粒的免疫应答水平,合成4条CpG-寡脱氧核苷酸(CpG-oligodeoxynueleotides,CpG-ODN);通过体外淋巴细胞增殖试验确定1条刺激活性最显著的CpG-ODN,序列为5-′TCGTCGTTTTGTCGTTTTGTCGTT-3′。将30只健康毕格犬随机均分为2大组,每大组再分为5小组,分别以不同的剂量肌肉注射CDV附着蛋白(H)重组质粒pcDNA-H和选定的CpG-ODN。在免疫后的第2、4、6周分别测定各组动物的抗CDV中和抗体和γ-干扰素水平。结果表明选定的CpG-ODN对提高抗CDV中和抗体效价和干扰素水平具有一定作用,且与对照组差异显著(P<0.05);但不同剂量的质粒pcDNA-H和CpG-ODN的效果不同,其中以30μg/只的CpG-ODN和50μg/只的pcDNA-H的剂量组合效果最佳。以选定剂量的CpG-ODN和pcDNA-H免疫5只健康毕格犬,同时以质粒pcDNA-H与生理盐水作为对照;4周后所有动物均接种CDV组织匀浆毒。结果发现试验组动物均未出现犬瘟热临床症状,且攻毒后第3周CDV检测为阴性,而pcDNA-H对照组和生理盐水对照组动物的CDV检测阳性率为2/5和5/5;这表明CpG能够明显促进重组质粒pcDNA-H的免疫保护作用。
To enhance the immunocompetence of recombinant plasmid against canine distemper virus, four CpG-oligodeoxynueleotides （CpG-ODN） were designed and used in this research. Following lymphocytes proliferation test determined by 3 H-TdR incorporation, one CpG-ODN was selected with optimistic immunostimulatory activity which sequence is 5＇-TCGTCGTTTT- GTCGTTTTGTCGTT-3＇. 30 healthy Beagle dogs were assigned into two groups and each group was further divided into 5 subgroups. All the subgroups were tested with different dose of CpG ODN and recombinant plasmid pcDNA-H （which contain the haemagglutinin gene of CDV） respectively. On the 2nd, 4th and 6th week after immunization, the neutralization antibody specific to CDV and the level of gama-interferon （IFN-γ） were determined, and the data showed that the CpG-ODN could elicit a better antibody response to CDV and increase the level of IFN-γ. Fur thermore, the effect of different levels of CpG-ODN were dissimilar on improving immune response, while the optimistic CpG-ODN level is 30μg with 50μg pcDNA-H for each animal. Five Beagle dogs were inoculated with optimistic level of CpG-ODN and pcDNA-H, while another 10 dogs were only injected with pcDNA-H or normal saline water as control, all the animals were challenged with CDV 4 weeks post immunization, and the data showed that there were no Canine Distemper clinical symptom and no CDV infection in the first group, while 2（40%） and 5（100%） animals were infected with CDV in the pcDNA-H and normal saline water control group respectively.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2008年第12期1737-1742,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
江苏省高校自然科学研究计划(04KJB230161)
