Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (19): 4096-4101.doi: 10.3864/j.issn.0578-1752.2011.19.021

• VETERINARY SCIENCE • Previous Articles     Next Articles

Co-expression of MIC-5 Gene of Eimeria stiedai and Rabbit IL-15 in Eucaryotic Cell Line

 MENG  Qing-Ling, QIAO  Jun, CAI  Xue-Peng, TIAN  Guang-Fu, YAN  Hong-Bin, LUO  Xue-Nong   

  1. 1.中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室
    2.石河子大学动物科技学院
  • Received:2010-03-29 Online:2011-10-01 Published:2011-08-23

Abstract: 【Objective】 The aim of this study is to construct a recombinant co-expression vector expressing Eimeria stiedai microneme protein 5 (MIC-5) and rabbit IL-15 in eukaryotic cells, which may lay a foundation for further study immune regulatory DNA vaccine against infection of Eimeria stiedai. 【Method】 E. stiedai MIC-5 gene and rabbit's IL-15 gene were respectively cloned into plasmid pBudce4.1 to construct recombinant vector pB-MIC-5-IL-15 for co-expression, and then the recombinant plasmid pB-MIC-5-IL-15 was transfected into BHK-21 cell line for expression. The RT-PCR and indirect fluorescent immunoassay were used to detect the transcription and expression of interesting genes in BHK-21 cell line, respectively. 【Result】 The results showed that E. stiedai MIC-5 gene and rabbit's IL-15 gene  were successfully transcripted. And interesting genes of E.stiedai MIC-5 and rabbit's IL-15 were successfully expressed at 36, 48 and 72 hour post-transfection, respectively. 【Conclusion】 The recombinant co-expression vector was successfully constructed, which can express E. stiedai MIC-5 and rabbit IL-15 gene in eukaryotic cells.

Key words:

[1]Carruthers V B, Tomley F M. Microneme proteins in apicomplexans. Subcellular Biochemistry, 2008, 47:33-45.

[2]Tomley F M, Clarke L E, Kawazoe U, Dijkema R, Kok J J. Sequence of the gene encoding an immunodominant microneme protein of Eimeria tenella. Molecular and Biochemical Parasitology, 1991, 49 (2): 277-288.

[3]Tomley F M, Bumstead J M, Billington K J, Dunn P P. Molecular cloning and characterization of a novel acidic microneme prtein (Etmic-2) from the apicomplexan protozoan parasite,Eimeria tenella. Molecular and Biochemical Parasitology, 1996, 79(2): 195-206.

[4]Tomley F M, Billington K J, Bumstead J M, Clark J D, Monaghan P. EtMIC4: a microneme protein from Eimeria tenella that contains tandem arrays of epidermal growth factor-like repeats and thrombospondin type-I repeats. International Journal for Parasitology, 2001, 31(12): 1303-1310.

[5]Brown P J, Billington K J, Bumstead J M, Clark J D, Tomley F M. A microneme protein from Eimeria tenella with homology to the Apple domains of coagulation factor XI and plasma pre-kallikrein. Molecular and Biochemical Parasitology, 2000, 107(1): 91-102.

[6]Labbe M, de Venevelles P, Girard-Misguich F, Bourdieu C, Guillaume A, Péry P. Eimeria tenella microneme protein EtMIC3: identification, localisation and role in host cell infection. Molecular & Biochemical Parasitology, 2005, 140(1): 43-53.

[7]Lai L, Simpson P, Bumstead J, Tomley F, Matthews S. Complete NMR assignments for the second microneme adhesive repeat (MAR) domain from Eimeria tenella microneme protein EtMIC3. Biomolecular NMR Assignments, 2009, 3(2):175-177.

[8]Periz J, Ryan R, Blake D P, Tomley F M. Eimeria tenella microneme protein EtMIC4: capture of the full-length transcribed sequence and comparison with other microneme proteins. Parasitology Research, 2009,104(3):717-21.

[9]Periz J, Gill A C, Hunt L, Brown P, Tomley F M. The microneme proteins EtMIC4 and EtMIC5 of Eimeria tenella form a novel, ultra-high molecular mass protein complex that binds target host cells. Journal of Biological Chemistry, 2007,282(23):16891-16898.

[10]Zuo J, Stohlman S A, Parra G I, Bergmann C C. IL-15 independent maintenance of virus-specific CD8+ T cells in the CNS during chronic infection. Journal of Neuroimmunology, 2009, 207(1): 32-38.

[11]Dubois S P, Waldmann T A, Muller J R. Survival adjustment of mature dendritic cells by IL-15. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(24): 8662-8667.

[12]Yajima T, Nishimura H, Sad S, Shen H, Kuwano H, Yoshikai Y. A novel role of IL-15 in early activation of memory CD8+ CTL after reinfection. Journal of Immunology, 2005, 174(6): 3590-3597.

[13]Diab A, Cohen A D, Alpdogan O, Perales M A. IL-15: targeting CD8+ T cells for immunotherapy. Cytotherapy, 2005, 7(1): 23-35.

[14]Rückert R, Brandt K, Bulanova E, Mirghomizadeh F, Paus R, Bulfone-Paus S. Dendritic cell-derived IL-15 controls the induction of CD8 T cell immune responses. European Journal of Immunology, 2003, 33(12): 3493-3503.

[15]Becknell B, Caligiuri M A. Interleukin-2, interleukin-15, and their roles in human natural killer cells. Advances in Immunology, 2005, 86: 209-239.

[16]McInnes I B, Gracie J A. Interleukin-15: a new cytokine target for the treatment of inflammatory diseases. Current Opinion in Pharmacology, 2004, 4(4): 392-397.

[17]Dann S M, Wang H C, Gambarin K J, Actor J K, Robinson P, Lewis D E, Caillat-Zucman S, White A C Jr. Interleukin-15 activates human natural killer cells to clear the intestinal protozoan cryptosporidium. Journal of Infectious Diseases, 2005, 192(7): 1294-1302.

[18]Ing R, Gros P, Stevenson M M. Interleukin-15 enhances innate and adaptive immune reponses to blood-stage malaria infection in mice. Infection and Immunity, 2005, 73(5): 3172-3177.

 [19]孟庆玲, 才学鹏, 乔 军, 骆学农,景志忠. 中国白兔IL-15基因的克隆、序列分析及其表达研究. 中国兽医学报, 2008, 28(5): 518-522.

Meng Q L, Cai X P, Qiao J, Luo X N, Jing Z Z. Cloning, sequence analysis of Chinese white rabbit IL-15 gene and its expression in E.coli. Chinese Journal of Veterinary Science, 2008, 28(5): 518-522. (in Chinese)

[20]孟庆玲. 减毒鼠伤寒沙门氏菌介导的斯氏艾美耳球虫DNA疫苗研究[D]. 兰州:甘肃农业大学博士学位论文,2008:35-50.

Meng Q L. An Attenuated Salmonella Typhimurium DNA Vaccine Against Eimeria Stiedai[D]. Lanzhou: Gansu Agricultural University, 2008:35-50. (in Chinese)

[21]Gravisaco M J, Mongini C, Alvarez E, Ruybal P, Escalada A, Sanchez-Lockhart M, Hajos S, Waldner C. IL-2, IL-10,  IL-15 and TNF are key regulators of murine T-cell lymphoma growth. International Journal of Molecular Medicine, 2003, 12(4): 627-632.

[22]Ozdemir, Savasan S. Combinational IL-2/IL-15 induction does not further enhance IL-15-induced lymphokine-activated killer cell cytotoxicity against human leukemia/lymphoma cells. Clinical Immunology, 2005, 115: 240-249.

[23]王世若,王兴龙,韩文瑜. 现代动物免疫学. 长春:吉林科学技术出版社,2001: 174-182.

Wang S R, Wang X L, Han W Y. Moderm Immunology of Animal. Jilin: Jilin Science and Technology Press, 2001: 174-182. (in Chinese)

[24]Calarota S A, Dai A, Trocio J N, Weiner D B, Lori F, Lisziewicz J. IL-15 as memory T-cell adjuvant for topical HIV-1 DermaVir vaccine. Vaccine, 2008, 26(40): 5188-5195.

[25]Gursel M, Gregoriadis G. Interleukin-15 acts as an immunological co-adjuvant for liposomal antigen in vivo. Immunology Letters, 1997, 55(3): 161-165.

[26]Wongi M, Lillehoj H S, Burnside J, Weining K C, Staeheli P, Zhu J J. Adjuvant effects of IL-1β, IL-2, IL-8, IL-15, IFN-α, IFN-γ TGF-β4 and lymphotactin on DNA vaccination against Eimeria acervulina. Vaccine, 2002, 20: 267-274.
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