You Position: Home > Paper
Generation and immunogenicity of a recombinant replication incompetent adenovirus expressing human rotavirus NSP4 protein
( views:0, downloads:0 )
- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 9
- DOI:
- 10.3969/j.issn.1009-3079.2006.09.004
- Key Word:
- 轮状病毒;NSP4;腺病毒载体;免疫
Abstract： 目的:研究以非复制型重组腺病毒表达人轮状病毒NSP4基因作为新型轮状病毒基因疫苗的可行性.方法:用AdEasy系统构建表达NSP4蛋白的重组腺病毒,并通过形态学观察、PCR、RT-PCR、Southern blot和Western blot等对重组腺病毒进行鉴定.通过滴鼻途径免疫♀小鼠,对免疫后母鼠生产的乳鼠(4日龄)用SA11株轮状病毒灌胃攻击,观察腹泻的产生情况并进行腹泻评级.结果:获得了重组腺病毒rvAdEasyNSP4.Southern blot表明,在重组腺病毒中确有特异性的NSP4基因整合,Western blot证明,目的蛋白得到表达.初次免疫小鼠后血清IgG抗体的滴度可达1:1 000,但阳转率仅为28.5%;再次免疫后,血清抗体阳转率达到了85.7%;第3次免疫后,血清抗体阳转率达到100%.血清IgA阳转率可以达到71.4%.实验组乳鼠攻毒后腹泻的百分率和腹泻评分(反应腹泻严重程度)均较对照组低.结论:腺病毒载体可有效表达轮状病毒非结构蛋白NSP4并可有效诱导小鼠免疫反应,这些结果的取得为新型轮状病毒疫苗的研制提供了依据.
- [1]Zhang MD.,Zeng CQY.,Morris AP.,etc.A functional NSP4 enterotoxin peptide secreted from rotavirus-infected cells[J].2000,74(24).
- [2]Borgan MA,Mori Y,Ito N,etc.Antigenic analysis of nonstructural protein (NSP) 4 of group A avian rotavirus strain PO-13.[J].2003,47(9).
- [3]Murphy TV,Gargiullo PM,Massoudi MS,etc.Intussusception among infants given an oral rotavirus vaccine.[J].2001,344(8).
- [4]Oka T,Horie Y,Suzuki Y,etc.Diarrhea induction by rotavirus NSP4 in the homologous mouse model system.[J].1999,262(2).
- [5]J M, Ball,P, Tian,C Q, Zeng,etc.Age-dependent diarrhea induced by a rotaviral nonstructural glycoprotein.[J].1996,272.
- [6]West K,Hayasaka D,Ennis FA,etc.Recombinant adenovirus vector vaccine induces stronger cytotoxic T-cell responses than recombinant vaccinia virus vector, plasmid DNA, or a combination of these.[J].2005,18(4).
- [7]N, Wolins,J, Lozier,T L, Eggerman,etc.Intravenous administration of replication-incompetent adenovirus to rhesus monkeys induces thrombocytopenia by increasing in vivo platelet clearance.[J].2003,123(5).
- [8]Rodrigo C,Grimprel E,Giaquinto C,etc.Nosocomial rotavirus infection in European countries: a review of the epidemiology, severity and economic burden of hospital-acquired rotavirus disease.[J].2006,25(1 Suppl.).
- [9]Iosef C,Chang KO,Saif LJ,etc.Systemic and intestinal antibody responses to NSP4 enterotoxin of Wa human rotavirus in a gnotobiotic pig model of human rotavirus disease.[J].2002,68(1).
- [10]Dong Y,Saif LJ,Zeng CQ,etc.Mutations in rotavirus nonstructural glycoprotein NSP4 are associated with altered virus virulence.[J].1998,72(5).
- [11]Chung I,Deisseroth AB.Recombinant adenoviral vector containing tumor-specific L-plastin promoter fused to cytosine deaminase gene as a transcription unit: generation and functional test.[J].2004,27(6).
- [12]Magnusson KE.,Mirazimi A.,Svensson L..A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum[J].2003,84(4).
- [13]Angel J,Franco MA,Greenberg HB.Immunity and correlates of protection for rotavirus vaccines[J].2006,24(15).
- [14]Browne EP.,Taylor JA.,Bellamy AR..Membrane-destabilizing activity of rotavirus NSP4 is mediated by a membrane-proximal amphipathic domain[J].2000,81(8).
- [15]Berkhout B,Havenga M,Kaspers J,etc.Novel replication-incompetent vector derived from adenovirus type 11 (ad11) for vaccination and gene therapy: low seroprevalence and non-cross-reactivity with ad5.[J].2004,78(23).
- [16]Lu X,Nguyen DC,Barratt Boyes SM,etc.Protection of mice and poultry from lethal H5N1 avian influenza virus through adenovirus-based immunization.[J].2006,80(4).
- [17]Huang H,Schroeder F,McPherson T,etc.Interaction(s) of rotavirus non-structural protein 4 (NSP4) C-terminal peptides with model membranes[J].2004,380(Pt.3).
- [18]Patton JT,Silvestri LS,Tortorici MA,etc.Rotavirus glycoprotein NSP4 is a modulator of viral transcription in the infected cell[J].2005,79(24).
- [19]Ertl HC,Zhou D.Therapeutic potential of adenovirus as a vaccine vector for chronic virus infections.[J].2006,6(1).
- [20]Leung AK,Kellner JD,Davies HD.Rotavirus gastroenteritis.[J].2005,22(5).
- [21]Vesikari T,Verstraeten T,Mrukowicz J,etc.Burden of rotavirus disease in European Union countries.[J].2006,25(1 Suppl.).