Construction of a recombinant adenovirus overexpressing rat TIPE2 gene

doi:10.3969/j.issn.2095-4344.2014.11.017

Abstract

Abstract:

BACKGROUND: Tumor necrosis factor-α-induced protein-8 like-2 (TIPE2), an anti-inflammatory protein, through the T cell receptor (TCR) and TOLL-like receptor signaling pathway, implements negative regulation of adaptive immunity and innate immunity, and thus effectively maintains the stable internal environment of the body.

OBJECTIVE: To construct a recombinant adenovirus that can overexpress rat TIPE2 gene.

METHODS: TIPE2 cDNA target gene was amplified from rat’s lymphocytes using RT-PCR, cloned into shuttle plasmid pShuttle-clontech, and then subcloned into artificial adenovirus vector AdC68. Hereafter, HEK 293 cells were transfected to generate a recombinant adenovirus. HEK293A cells were infected using this recombinant adenovirus, and then TIPE2 gene level was tested by western blot method.

RESULTS AND CONCLUSION: Based on results of PCR, digestion identification and sequencing, the obtained cDNA was the coding sequence region of TIPE2. Western blot findings showed that the recombinant adenovirus could overexpress TIPE2 gene. These findings indicate that the recombinant adenovirus is constructed successfully and can express TIPE2 gene stably.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: adenoviridae, gene expression, transfection

CLC Number:

R394.2

Zhang You-bin, Yu Yun-sheng, Shen Zhen-ya, Zhou Dong-ming. Construction of a recombinant adenovirus overexpressing rat TIPE2 gene[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(11): 1743-1748.

Figures/Tables 4

References

[1] Zhang G, Zhang W, Lou Y,et al.TIPE2 deficiency accelerates neointima formation by downregulating smooth muscle cell differentiation. Cell Cycle.2013；12(3): 501-510.

[2] Laliberté B, Wilson AM, Nafisi H, et al.TNFAIP8: a new effector for Galpha(i) coupling to reduce cell death and induce cell transformation. J Cell Physiol, 2010. 225(3):865-874.

[3] Freundt EC, Bidere N, Lenardo MJ.A different TIPE of immune homeostasis. Cell, 2008. 133(3): 401-402.

[4] Lou Y, Liu S.The TIPE (TNFAIP8) family in inflammation, immunity, and cancer. Mol Immunol, 2011. 49(1-2):4-7.

[5] Zhang X, Wang J, Fan C, et al.Crystal structure of TIPE2 provides insights into immune homeostasis. Nat Struct Mol Biol, 2009. 16(1): 89-90.

[6] Capone S, D'Alise AM, Ammendola V, et al.Development of chimpanzee adenoviruses as vaccine vectors: challenges and successes emerging from clinical trials. Expert Rev Vaccines. 2013;12(4):379-393.

[7] Hoskote A, Carter C, Rees P, et al. Acute right ventricular failure after pediatric cardiac transplant: predictors and long-term outcome in current era of transplantation medicine. J Thorac Cardiovasc Surg. 2010;139(1):146-153

[8] 张仁福.心脏移植现状及进展[J].医学研究杂志,2008,37(7):4-6.

[9] 胡盛寿,王春生,董念国，等.心脏移植的多中心研究[J].中华器官移植杂志,2012,33(5):264-266.

[10] 肖长波,娄勇,张毅强，等.原位心脏移植3例的围术期治疗[J].中国实用医刊,2013,40(18):57-59.

[11] Sun H, Gong S, Carmody RJ, et al.TIPE2, a negative regulator of innate and adaptive immunity that maintains immune homeostasis. Cell.2008；133(3):415-426.

[12] 蒋丽娜,姚咏明.肿瘤坏死因子α诱导蛋白8家族的研究进展[J].中华损伤与修复杂志:电子版,2011,06(2)：268-278.

[13] 栾樱译,姚咏明,董宁,等.肿瘤坏死因子-α诱导蛋白-8样分子2对小鼠调节性T细胞免疫调节功能的影响[J].中华创伤杂志,2012, 28(4):316-321.

[14] 朱玉楠.免疫负调控因子TIPE2与核蛋白PCNP双向调节在白血病细胞中的生物学作用[D]. 开封:河南大学,2013.

[15] 栾樱译.TIPE2在CD4+CD25+调节性T细胞中的表达及其对免疫抑制功能的影响[D].广州:南方医科大学,2012.

[16] 宋立军.免疫负调控基因IL-37和TIPE2在SLE患者的表达及其意义[D].济南:山东大学,2012.

[17] Sun H, Zhuang G, Chai L, et al.TIPE2 controls innate immunity to RNA by targeting the phosphatidylinositol 3-kinase-Rac pathway. J Immunol.2012;189(6): 2768-2773.

[18] 石春燕,何琼,杨晶晶，等.TIPE2过表达质粒转染大鼠佐剂型关节炎滑膜成纤维样细胞及分析[J].中国免疫学杂志,2013,29(12)：1240-1243,1253.

[19] 章桂忠.Ⅰ.TIPE2负性调控VSMCs功能抗动脉粥样硬化的机制;Ⅱ.TIPE1蛋白在小鼠组织及TIPE1、TIPE2 mRNA在细胞系的表达谱检测[D]. 济南:山东大学,2012.

[20] 刘瑞敏,孙颖川|,张爱红,等.TIPE2在类风湿关节炎发病中的表达及其意义[J].中华微生物学和免疫学杂志,2013,33(3)： 212-215.

[21] 张虎.TIPE2在白血病细胞中的表达及其功能研究[D].开封:河南大学,2012.

[22] 朱江木.肌细胞感染后TIPE2与PCNP分子在JNK1通路中的作用[D].开封:河南大学,2012.

[23] 孙颖川.类风湿性关节炎中PCNP和TIPE2的表达及意义[D].开封:河南大学,2012.

[24] 徐凯,朱其明,冯建鹏.免疫抑制剂的研究进展[J].医学综述,2012, 18(14):2177-2180.

[25] 黄金沐，池惠琼，张忠阳．他克莫司的研究概况[J]. 海峡药学，2010,22(11):148-150．

[26] 郭斌,范明,陈伟高．FK506的临床研究与应用进展[J]．实用临床医学,2002,3(3):114-115.

[27] González-Vílchez F, Vázquez de Prada JA, Paniagua MJ, et al.Rejection after conversion to a proliferation signal inhibitor in chronic heart transplantation. Clin Transplant. 2013;27(6): E649-58.

[28] McCoy K, Tatsis N, Korioth-Schmitz B, et al.Effect of preexisting immunity to adenovirus human serotype 5 antigens on the immune responses of nonhuman primates to vaccine regimens based on human- or chimpanzee-derived adenovirus vectors. J Virol. 2007;81(12): 6594-6604.

[29] Zhou D, Wu TL, Emmer KL, et al.Hexon-modified recombinant E1-deleted adenovirus vectors as dual specificity vaccine carriers for influenza virus. Mol Ther. 2013;21(3): 696-706.

[30] Tatsis N, Lasaro MO, Lin SW, et al. Adenovirus vector-induced immune responses in nonhuman primates: responses to prime boost regimens. J Immunol. 2009; 182(10):6587-6599.

[31] Li H, Tuyishime S, Wu TL, et al. Adeno-associated virus vectors serotype 2 induce prolonged proliferation of capsid-specific CD8+ T cells in mice. Mol Ther. 2011;19(3): 536-546.