K型和D型CpG寡脱氧核苷酸对猪外周血单个核细胞免疫刺激活性的比较研究  

Difference between Immunostimulation Activity of K and D Types of CpG Oligodeoxynu-cleotides in Peripheral Blood Mononuclear Cells of Porcine

作　　者：朱鸿飞[1] 李光富[2] 朱建中[2] 卢春[2] 邵国清[3] 张兆松[2] 

机构地区：[1]南京农业大学生物制品工程技术中心,江苏南京210095 [2]南京医科大学医学分子生物学研究所,江苏南京210029 [3]江苏省农业科学院兽医研究所,江苏南京210095

出　　处：《南京医科大学学报（自然科学版）》2003年第6期542-544,553,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家高技术研究发展计划资助项目(863计划;2001AA243071)

摘　　要：目的:比较K、D两型CpG寡脱氧核苷酸(CpG 0DN)对猪外周血单个核细胞(PBMC)免疫刺激活性的差异.方法:用人工合成的@K型和D型两类CpG 0DN和不含CpG基元的对照0DN D48分别刺激已分离的猪PBMC,3H-TdR掺入法测定每分闪烁次数(cpm)值,计算刺激指数(SI).双抗体夹心ELISA法测定活化细胞上清中IFN-γ和IL-2表达水平.结果:D和K型CpGODN对猪PBMC增殖均具有较强刺激作用,K型的增殖效应高于D型.含'GTCGTT'基元的2006作用尤为显著,但不能有效刺激猪PBMC分泌IFN-γ和IL-2.D型刺激增殖效应低于K型,但能有效地促进猪PBMC分泌IFN-γ和IL-2.结论:K型和D型CpG 0DN在刺激猪PBMC增殖及其分泌IFN-γ、IL-2的水平有所不同.Objective: To investigate the difference between K and D types of CpG oligodeoxynucleotides(ODN) immunostimulation activity in peripheral blood mononuclear cells(PBMC)of porcine. Methods: K and D types of CpG ODNs 2006,20061,2016,D19.D28, D25,and D48 without CpG motif as control were synthesized based on previous research data. PBMC of Porcine was isolated and further added to the above synthesized CpG ODNs, respectively. On proliferation assay in vitro, the incorporation of 3H-thymidine was determined to calculate stimulation index(SI), and a solid phase sandwich ELISA was performanced to evaluate IFN-r and IL-2 levels from the supernatant of CpG-activated cells. Results: Both D and K types of CpG ODN could strong stimulation activity in PBMC of porcine according to the SI value. Moreover, stimulation activity of K type was stronger than that of D, especially displayed in CpG ODN 2006 which contains 'GTCGTT' motif. However, the amounts of both IFN-'/ and IL-2 from D type of CpG ODN-activated cells was dramatically increased than that of K type CpG ODN. Conclusion: There are differences between immunostimulation activity and expression of cy-tokines IFN-r and IL-2 secreted by CpG ODN-activated PBMC of porcine between K and D types.

关 键 词：CPG寡脱氧核苷酸 CPG基元 IFN—γ IL—2 猪 

分 类 号：Q74[生物学—分子生物学]