罗婷,李虹娟,王霄,段志青,李萍,王海漩,胡凝珠,胡云章
LUO Ting,LI Hong-juan,WANG Xiao,DUAN Zhi-qing,LI Ping,WANG Hai-xuan,HU Ning-zhu,HU Yun-zhang

• 1:中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室
• 2:昆明医科大学生物所

摘要(Abstract)：

目的探讨甲磺酸去铁胺(deferoxamine,DFO)佐剂对甲型肝炎病毒(hepatitis A virus antigen,HAV)抗原诱导小鼠体液免疫应答的影响。方法将不同剂量的DFO(0.6、1、4、8 mg)佐剂分别与HAV抗原18 EU混合,免疫ICR小鼠,并设生理盐水对照组(生理盐水200μl)、HAV抗原对照组(HAV 18 EU)、铝佐剂对照组(铝佐剂300μg+HAV抗原18 EU)和复合佐剂组(HAV抗原18 EU+DFO 1 mg+铝佐剂150μg),均经腹部皮下多点注射,0.2 ml/只,共免疫1次。分别于免疫后第4、8、12、16周,采用间接ELISA法检测小鼠血清中抗-HAV IgG水平,并进行安全性试验。结果除生理盐水对照组外,其他各组小鼠血清在各时间点均能检测到抗-HAV IgG;除DFO 0.6 mg组外,抗体滴度均在第8周达峰值;不同剂量DFO组的抗体水平均明显高于HAV抗原对照组,其中DFO 4 mg和8 mg组在8~16周时抗体水平显著高于HAV抗原对照组,且差异有统计学意义(P<0.05),但与铝佐剂对照组比较,差异无统计学意义(P>0.05);在8~16周时,同一时间DFO 4 mg、DFO 8 mg和复合佐剂组的抗体水平较高,并能维持较长时间,至第16周时仍能达到与铝佐剂对照组相当的水平。结论 DFO佐剂能显著增强HAV抗原诱导小鼠的体液免疫应答,有望开发为一种可替代铝佐剂的新型佐剂。
Objective To investigate the effect of deferoxamine(DFO)on humoral immune response induced by hepatitis A virus(HAV)antigen in mice. Methods ICR mice were immunized with 18 EU HAV antigen combined with DFO at various dosages(0. 6,1,4 and 8 mg)respectively by s. c. injection in several sites at a volume of 0. 2 ml,using 200 μl of physiological saline,18 EU HAV antigen alone and 300 μg aluminum adjuvant + 18 EU HAV antigen and complex adjuvant(18 EU HAV antigen + 1 mg DFO + 150 μg aluminum adjuvant)as controls. Serum samples were collected at weeks 4,8,12 and 16 after immunization,and determined for anti-HAV IgG levels by indirect ELISA. Meanwhile,safety test was performed. Results Anti-HAV IgGs were detected in sera of mice at various time points in various groups except physiological saline control group. The IgG titers in various groups except DFO 0. 6 mg group reached peak values at weeks 8 after immunization. The anti-HAV IgG levels of mice after immunization with HAV antigen combined with DFO adjuvant at various dosages were higher,of which those in DFO 4 and 8 mg groups at weeks 8 ~ 16 were significantly higher,than that in HAV antigen control group(P < 0. 05),which showed no significant difference with that in aluminum adjuvant control group(P < 0. 05). The IgG levels in DFO 4 mg,DFO 8 mg and complex adjuvant control groups at the same time points at weeks 8 ~ 16 were relatively high and persistent,which were equal to that in aluminum adjuvant control group until week 16. Conclusion DFO enhanced the humoral immune response induced by HAV in mice significantly,which might be developed as a novel adjuvant instead of aluminum adjuvant.

关键词(KeyWords)： 甲磺酸去铁胺;佐剂;甲型肝炎病毒抗原;体液免疫
Deferoxamine;Adjuvant;Hepatitis A virus antigen;Humoral immunity

Abstract:

Keywords:

基金项目(Foundation): 国家高技术研究发展计划(863计划“新型疫苗佐剂的研发及其应用研究”(2012AA02A406);; 云南省创新团队“中国医学科学院医学生物学研究所新型疫苗佐剂应用研究省创新团队”(2011CI140)

作者(Author): 罗婷,李虹娟,王霄,段志青,李萍,王海漩,胡凝珠,胡云章
LUO Ting,LI Hong-juan,WANG Xiao,DUAN Zhi-qing,LI Ping,WANG Hai-xuan,HU Ning-zhu,HU Yun-zhang

DOI: 10.13200/j.cnki.cjb.000246

参考文献(References)：

• [1]Li FX,Xia QM,Xu ZX,et al.Effects of desferrioxamine on HIF-1αexpression in rat lungs and on the pulmonary structure in rats subjected to hypoxia at simulated high altitude[J].Pract Med J,2008,24(17):2948-2951.(in Chinese)李福祥,夏前明,徐朝霞,等.去铁胺对大鼠肺组织低氧诱导因子-1表达及对模拟高原低氧大鼠肺组织结构的影响[J].实用医学杂志,2008,24(17):2948-2951.
• [2]Long FY,Yin GB,Liu ZM,et al.Expressions of hypoxiainducible factor-1α,twist and E-cadherin in human breast cancer tissue and their significances[J].Chin J Biologicals,2012,25(7):884-887.(in Chinese)龙方懿,印国兵,刘智敏,等.低氧诱导因子-1α、锌指转录抑制因子Twist和钙黏蛋白E在人乳腺癌组织中的表达及其意义[J].中国生物制品学杂志,2012,25(7):884-887.
• [3]Zhang W,Wu Y,Yan Q,et al.Deferoxamine enhances cell migration an d invasion through promotion of HIF-1alpha expression and epithelial-mesenchymal transition in colorectal cancer[J].Oncol Rep,2014,31(1):111-116.
• [4]Malyshev LLU,Kruglov SV,Liamina SV.Hypoxia,inflammation and phenotypic plasticity of macrophages:the central role of HIF-1 and NFkappaB[J].Patol Fiziol Eksp Ter,2012,(3):42-50.
• [5]Okumura CY,Hollands A,Tran DN,et al.A new pharmacological agent(AKB-4924)stabilizes hypoxia inducible factor-1(HIF-1)and increases skin innate defenses against bacterial infection[J].J Mol Med(Berl),2012,90(9):1079-1089.
• [6]Vecchi S,Bufali S,Skibinski DA,et al.Aluminum adjuvant dose guidelines in vaccine formulation for preclinical evaluations[J].J Pharm Sci,2012,101(1):17-20.
• [7]Scholz CC,Taylor CT.Targeting the HIF pathway in inflammation and immunity[J].Curr Opin Pharmacol,2013,13(4):646-653.
• [8]Gale DP,Maxwell PH.The role of HIF in immunity[J].Int J Biochem Cell Biol,2010,42(4):486-494.
• [9]Wobben R,Hüsecken Y,Lodewick C,et al.Role of hypoxia inducible factor-1αfor interferon synthesis in mouse dendritic cells[J].Biol Chem,2013,394(4):495-505.
• [10]Chen LZ,Chen JY,Wu M,et al.Different doses of deferoxamine clinical research for the treatment of blood transfusion dependency iron overload[J].Chin Clin Doctors,2010,38(5):35-37.(in Chinese)陈玲珍,陈嘉榆,巫进明,等.不同剂量去铁胺治疗输血依赖性铁过载的临床研究[J].中国临床医生,2010,38(5):35-37.
• [11]Miao X,Liu X,Yue Q,et al.Deferoxamine suppresses microglia activation and protects against secondary neural injury after intracerebral hemorrhage in rats[J].Nan Fang Yi Ke Da Xue Xue Bao,2012,32(7):970-975.