• 中国中文核心期刊
  • 中国科学引文数据库核心期刊
  • 中国科技核心期刊
  • 中国高校百佳科技期刊
高级检索

重组HSP65-6P277乳酸乳球菌疫苗对链脲佐菌素诱导的1型糖尿病小鼠的降糖作用

吴洁, 刘晓锐, 杨雪, 侯景, 徐茂磊, 申丽丽, 黄东成, 刘坤锋

吴洁, 刘晓锐, 杨雪, 侯景, 徐茂磊, 申丽丽, 黄东成, 刘坤锋. 重组HSP65-6P277乳酸乳球菌疫苗对链脲佐菌素诱导的1型糖尿病小鼠的降糖作用[J]. 中国药科大学学报, 2014, 45(1): 106-110. DOI: 10.11665/j.issn.1000-5048.20140120
引用本文: 吴洁, 刘晓锐, 杨雪, 侯景, 徐茂磊, 申丽丽, 黄东成, 刘坤锋. 重组HSP65-6P277乳酸乳球菌疫苗对链脲佐菌素诱导的1型糖尿病小鼠的降糖作用[J]. 中国药科大学学报, 2014, 45(1): 106-110. DOI: 10.11665/j.issn.1000-5048.20140120
WU Jie, LIU Xiaorui, YANG Xue, HOU Jing, XU Maolei, SHEN Lili, HUANG Dongcheng, LIU Kunfeng. Hypoglycemic effect of Lactococcus lactis vaccine containing HSP65-6P277 on streptozotocin-induced type 1 diabetic mice[J]. Journal of China Pharmaceutical University, 2014, 45(1): 106-110. DOI: 10.11665/j.issn.1000-5048.20140120
Citation: WU Jie, LIU Xiaorui, YANG Xue, HOU Jing, XU Maolei, SHEN Lili, HUANG Dongcheng, LIU Kunfeng. Hypoglycemic effect of Lactococcus lactis vaccine containing HSP65-6P277 on streptozotocin-induced type 1 diabetic mice[J]. Journal of China Pharmaceutical University, 2014, 45(1): 106-110. DOI: 10.11665/j.issn.1000-5048.20140120

重组HSP65-6P277乳酸乳球菌疫苗对链脲佐菌素诱导的1型糖尿病小鼠的降糖作用

基金项目: 国家自然科学基金资助项目(No.81172973,31270985);中央高校基本科研业务费研究生专项资助项目(No.JKY2011076);江苏高校优势学科建设工程资助项目

Hypoglycemic effect of Lactococcus lactis vaccine containing HSP65-6P277 on streptozotocin-induced type 1 diabetic mice

  • 摘要: 探讨表达HSP65-6P277蛋白的乳酸乳球菌活载体疫苗对链脲佐菌素(STZ)诱导的1型糖尿病小鼠的降糖作用及可能的作用机制。采用多次小剂量腹腔注射STZ建立1型糖尿病模型。将诱导型表达菌株Lactococcus lactis NZ9000 pCYT∶HSP65-6P277和组成型表达菌株L.lactis NZ9000 pHJ∶HSP65-6P277及无关蛋白对照菌株L.lactis NZ9000 pCYT∶Nuc灌胃1型糖尿病小鼠(2×109CFU,200 μL),每天灌胃1次,连续7 d,之后每周灌胃1次,持续16周。每月从小鼠眼眶静脉丛取血1次,血糖检测仪检测STZ造模小鼠血糖水平并称体重。4个月后将小鼠处死,取小鼠脾脏做脾细胞增殖实验,采用ELISA试剂盒分别检测脾细胞上清液中的IL-10和IFN-γ细胞因子水平以及血清IgG抗体水平。结果显示,与对照组相比,pCYT∶HSP65-6P277组和pHJ∶HSP65-6P277组可以一定程度上降低STZ造模小鼠血糖水平(P<;0.05),并能维持其正常体重稳定,降低脾细胞针对HSP65和P277的增殖(P<;0.05),脾细胞上清液中IFN-γ细胞因子水平明显降低(P<;0.05),血清IgG抗体水平无明显差异(P >;0.05)。
    Abstract: To study the hypoglycemic effect and mechanisms of vaccine HSP65-6P277 protein on type 1 diabetic mice induced by streptozotocin(STZ), Lactococcus lactis live vector was used to express the vaccine protein. The mouse model of type 1 diabetes mellitus were established by intraperitoneal injections of multiple low dose STZ(MLD-STZ)and divided into three groups. The inducible expression strain L. lactis NZ9000 pCYT ∶HSP65-6P277, constitutive expression strain L. lactis NZ9000 pHJ ∶HSP65-6P277 and control strains L. lactis NZ9000 pCYT ∶Nuc were respectively immunized to diabetic mice orally once daily(2×109 CFU, 200 μL)in the first week and once every week in the following weeks lasting for 16 weeks. Fasting blood glucose levels were measured once a month on blood samples taken from orbital venous plexus. Four months later, the mice were sacrificed. Spleen cell proliferation experiment was performed. IL-10 and IFN-γ cytokine levels of spleen cells and serum IgG levels were assayed using ELISA kits. The results showed that pCYT ∶HSP65-6P277 group and pHJ ∶HSP65-6P277 group have low blood glucose(P < 0. 05), and maintain normal body weight, compared with control group(pCYT ∶Nuc). On the other hand, cytokine IFN-γ levels were significantly lower(P < 0. 05)in spleen cells supernatant and spleen cell proliferation was reduced against HSP65 and P277(P< 0. 05), but serum IgG levels were not significantly different(P> 0. 05).
  • [1] Nicholas D,Odumosu O,Langridge WH.Autoantigen based vaccines for type 1 diabetes[J]. Discov Med,2011, 11(59):293-301.
    [2] Raz I,Elias D,Avron A,et al.Beta-cell function in new-onset type 1 diabetes and tyimmunomodulation with a heat-shock protein peptide(DiaPep277):a randomised,double-blind,phase II trial[J].Lancet,2001, 358(9 295):1 749-1 753.
    [3] EliasI D,Cohen R.Peptide therapy for dia NOD mice[J].Lancet,1994,343(8 899):704-706.
    [4] Cohen R,Elias D.The hsp60 peptide betes in p277 arrests the autoimmune diabetes induced by the toxin streptozotocin[J].Diabetes,1996,45(9):1 168-1 177.
    [5] Jin L,Wang Y,Xiong Q,et al.Long-lasting specific antibodies against P277 induced by mucosaladministration of P277 repeat sequences carried by Hsp65 in the absence of adjuvant[J].Vaccine,2007,25(11):2 043-2 050.
    [6] Liang J,Aihua Z,Yu W,et al.HSP65 serves as an immunogenic carrier for a diabetogenic peptide P277 inducing anti-inflammatory immune response in NOD mice by nasal administration[J]. Vaccine,2010, 28(19):3 312-3 317.
    [7] Chen QM, Wu GJ, Wu J, et al. Purification and stability research of mycobacterial heat shock protein 65 and its fusion protein Hsp65-6P277[J]. Pharm Biotechnol(药物生物技术),2007,14(4):249-254.
    [8] Huibregtse IL,Marietta EV,Rashtak S, et al.Induction of antigen-specific tolerance by oral administration of lactococcus lactis delivered immunodominant DQ8-restricted gliadin peptide in sensitized nonobese diabetic Abo Dq8 transgenic mice[J].J Immunol,2009, 183(4):2 390-2 396.
    [9] Bermúdez-Humarán LG.Lactococcus lactis as a live vector for mucosal delivery of therapeutic proteins[J]. Hum Vaccin,2009, 5(4):264-267.
    [10] Sun W,Wang L,Zhang Z,et al.Intramuscular transfer of naked calcitonin gene-related peptide gene prevents autoimmune diabetes induced bymultiple low-dose streptozotocin in C57BL mice[J]. Eur J Immunol,2003, 33(1):233-242.
      
    [11] Csorba TR,Lyon AW,Hollenberg MD.Autoimmunity and the pathogenesis of type 1 diabetes[J]. Crit Rev Clin Lab Sci,2010,47(2):51-71.
    [11] Morello E,Bermúdez-Humarán LG,Llull D,et al.Lactococcus lactis an efficient cell factory for recombinant protein production and secretion[J]. J Mol Microbiol Biotech,2008, 14(1/2/3):48-58.
    [12] Takiishi T,Korf H,Van Belle TL,et al.Reversal of autoimmune diabetes by restoration of antigen-specific tolerance using genetically modified Lactococcus lactis in mice[J].J Clin Invest,2012,122(5):1 717-1 725.
    [13] Rezende RM,Oliveira RP,Medeiros SR,et al.Hsp65-producing lactococcus lactis prevents experimental autoimmune encephalomyelitis in mice by inducing CD4+LAP+regulatory T cells[J].J Autoimmun,2012,40:45-57.
计量
  • 文章访问数:  1340
  • HTML全文浏览量:  1
  • PDF下载量:  1820
  • 被引次数: 0
出版历程
  • 刊出日期:  2014-02-24

目录

    /

    返回文章
    返回
    x 关闭 永久关闭