马立克病病毒超强毒株GX0101感染宿主部分病毒基因表达水平及致病阶段分析

doi:doi: 10.11843/j.issn.0366-6964.2016.08.014

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (8): 1635-1644.doi: doi: 10.11843/j.issn.0366-6964.2016.08.014

马立克病病毒超强毒株GX0101感染宿主部分病毒基因表达水平及致病阶段分析

马圣明^1，2，滕蔓²，余祖华¹，党露²，李会珍³，丁轲¹，邓瑞广²，罗俊^1，2*

（1.河南科技大学动物科技学院，动物疫病与公共安全重点实验室，洛阳 471003；2.河南省农业科学院动物免疫学重点实验室，农业部动物免疫学重点实验室，河南省动物免疫学重点实验室，郑州 450002；3.河南农业大学牧医工程学院，郑州450002）

收稿日期:2016-03-21 出版日期:2016-08-23 发布日期:2016-09-08
通讯作者: 罗俊，E-mail：luojun593@aliyun.com
作者简介:马圣明（1991-），男，河南罗山人，硕士生，主要从事动物病毒分子致病机制研究，Tel：0371-65756056，E-mail：mashengming66@163.com
基金资助:
国家自然科学基金（31372445）；国家重点研发计划（2016YFD0500802）

The Viral Gene Expression Profiles and Pathogenic Phases of the Disease Caused by Very Virulent MDV Strain GX0101

MA Sheng-ming ^1，2，TENG Man²，YU Zu-hua¹，DANG Lu ²，LI Hui-zhen³，DING Ke¹，DENG Rui-guang²，LUO Jun ^1，2*

(1.Key Laboratory of Animal Disease and Public Safety，College of Animal Science and Technology，
Henan University of Science and Technology，Luoyang 471003，China；2.Key Laboratory of Animal
Immunology of the Ministry of Agriculture，Henan Provincial Key Laboratory of Animal Immunology，
Henan Academy of Agricultural Sciences，Zhengzhou 450002，China；3.College of Animal Science
and Veterinary Medicine，Henan Agricultural University，Zhengzhou 450002，China)

Received:2016-03-21 Online:2016-08-23 Published:2016-09-08

摘要/Abstract

摘要：

旨在探讨马立克病病毒（MDV）超强毒株GX0101对宿主的感染过程及致病阶段，即“Cornell模型”，为进一步使用该毒株研究MDV的致病或致瘤机制奠定基础。用GX0101接种1日龄SPF鸡，建立了感染宿主的动物模型；然后用RT-qPCR分析10个具有代表性的MDV蛋白编码基因在感染不同时间点的动态基因转录水平。结果显示，GX0101感染发病鸡的临床症状及剖检病变与典型的马立克病一致；MDV编码的结构蛋白基因gB、gE和UL6、非结构蛋白基因UL42和UL52、立早期基因ICP4、水平传播相关基因UL13以及磷酸化蛋白基因pp38具有相似的转录水平和趋势，均在GX0101感染后7 d上升至第一个峰值，10 d降至低谷，14 d又逐渐升高并在21 d达到第二个峰值，30～60 d逐渐下降并处于较低的转录水平；MDV编码的原癌基因meq和毒力相关基因RLORF6在GX0101感染7 d即持续升高，并在14～60 d的试验周期内较长时间处于高转录水平。结合作者此前研究及本文结果分析，GX0101感染宿主的“Cornell模型”：早期增殖性-限制性感染期为1～7 d，潜伏感染期约为10 d前后，晚期溶细胞性感染及免疫抑制期发生于14～21 d，而T细胞转化及淋巴瘤形成阶段可能在14 d前后就已经开始。

关键词: 马立克病病毒, 超强毒株, GX0101, 实时荧光定量PCR, 基因表达, 致病阶段

Abstract:

The present work was performed to study the infection phases of Marek’s disease (MD)，namely ‘Cornell Model’，caused by the very virulent (vv) Marek’s disease virus (MDV) strain GX0101.For the first step，one-day-old specific pathogen free (SPF) chickens were challenged by GX0101 to establish the animal model of MD.Then，the kinetic transrption levels of 10 representative MDV protein-coding genes at different days post-infection (dpi) were determined by real-time quantitative PCR (RT-qPCR).The results showed that the onset of clinical symptoms and autopsy lesions of GX0101-infected birds was consistent with the typical MD cases.Some of the MDV protein-coding genes，including the structural protein-coding genes of gB，gE and UL6，the nonstructural protein-coding genes of UL42 and UL52，the immediate early infected cell protein 4 gene (ICP4)，the horizontal transmission associated gene of UL13 and the 38 kD phosphorylated protein gene (pp38)，demonstrated a similar transcription pattern，which increased to a first peak at 7 dpi，fell to a bottom level at 10 dpi，increased again at 14 dpi and reached to a second peak at 21 dpi，and then gradually declined to low levels from 30 to 60 dpi.However，both of the unique MDV oncogene meq and virulence related gene RLORF6 represented a persistent increasing transcription trend from 7 dpi，and then kept at a higher transcription level from 14 to 60 dpi.Our previous and present data indicates that the ‘Cornell Model’ of MD caused by GX0101 are as blow：the early cytolytic phase occurs at 1-7 dpi，the latent phase establishes at about 10 dpi and the late cytolytic and immunosuppressive phase appears at 14-21 dpi，while the T cell transformation and lymphomagenesis possibly starts at about 14 dpi.

Key words: Marek’s disease virus, vvMDV, GX0101, RT-qPCR, gene expression, pathogenic phase

中图分类号:

S852.659.1

马圣明，滕蔓，余祖华，党露，李会珍，丁轲，邓瑞广，罗俊. 马立克病病毒超强毒株GX0101感染宿主部分病毒基因表达水平及致病阶段分析[J]. 畜牧兽医学报, 2016, 47(8): 1635-1644.

MA Sheng-ming，TENG Man，YU Zu-hua，DANG Lu，LI Hui-zhen，DING Ke，DENG Rui-guang，LUO Jun. The Viral Gene Expression Profiles and Pathogenic Phases of the Disease Caused by Very Virulent MDV Strain GX0101[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(8): 1635-1644.

参考文献

［1］SAIF Y M.禽病学［M］.苏敬良，高福，索勋，译.12版.北京：中国农业出版社，2012：129-151.
SAIF Y M.Avain disease［M］.SU J L，GAO F，SUO X，translated.12ed.Beijing：China Agriculture Press，2012：129-151.(in Chinese)
［2］DAVISON A J，EBERLE R，EHLERS B，et al.The order Herpesvirales［J］.Arch Virol，2009，154(1)：171-177.
［3］WITTER R L.Increased virulence of Marek’s disease virus field isolates［J］.Avian Dis，1997，41(1)：149-163.
［4］TENG L Q，WEI P，SONG Z B，et al.Molecular epidemiological investigation of Marek’s disease virus from Guangxi，China［J］.Arch Virol，2011，156(2)：203-206.
［5］TIAN M，ZHAO Y，LIN Y，et al.Comparative analysis of oncogenic genes revealed unique evolutionary features of field Marek’s disease virus prevalent in recent years in China［J］.Virol J，2011，8：121.
［6］ZHANG Y P，LIU C J，ZHANG F，et al.Sequence analysis of the Meq gene in the predominant Marek’s disease virus strains isolated in China during 2006-2008［J］.Virus Genes，2011，43(3)：353-357.
［7］YU Z H，TENG M，LUO J，et al.Molecular characteristics and evolutionary analysis of field Marek’s disease virus prevalent in vaccinated chicken flocks in recent years in China［J］.Virus Genes，2013，47(2)：282-291.
［8］ZHANG Y P，LV H C，BAO K Y，et al.Molecular and pathogenicity characterization of Gallid herpesvirus 2 newly isolated in China from 2009 to 2013［J］.Virus Genes，2016，52(1)：51-60.
［9］CUI N，SU S，SUN P，et al.Isolation and pathogenic analysis of virulent Marek’s disease virus field strain in China［J］.Poult Sci，2016，pii：pew073.
［10］CALNEK B W.Pathogenesis of Marek’s disease virus infection［J］.Curr Top Microbiol Immunol，2001，255：25-55.
［11］BAIGENT S J，DAVISON F，NAIR V.Marek’s disease virus：biology and life cycle，in Marek’s Disease［M］.An Evolving Problem.Elsevier Academic Press，Oxford，United Kingdom，2004：62-77.
［12］HU X，QIN A，XU W，et al.Transcriptional analysis of host responses to Marek’s disease virus infection in chicken thymus［J］.Intervirology，2015，58(2)：95-105.
［13］ZHANG Z，CUI Z.Isolation of recombinant field strains of Marek’s disease virus integrated with reticuloendotheliosis virus genome fragments［J］.Sci China C Life Sci，2005，48(1)：81-88.
［14］CUI Z，ZHUANG G，XU X，et al.Molecular and biological characterization of a Marek’s disease virus field strain with reticuloendotheliosis virus LTR insert［J］.Virus Genes，2010，40(2)：236-243.
［15］SUN A J，LAWRENCE P，ZHAO Y G，et al.A BAC clone of MDV strain GX0101 with REV-LTR integration retained its pathogenicity［J］.Chin Sci Bull，2009，54(15)：2641-2647.
［16］SUN A J，XU X Y，PETHERBRIDGE L，et al.Functional evaluation of the role of reticuloendotheliosis virus long terminal repeat (LTR) integrated into the genome of a field strain of Marek’s disease virus［J］.Virology，2010，397(2)：270-276.
［17］SU S，CUI N，SUN A，et al.Sequence analysis of the whole genome of a recombinant Marek’s disease virus strain，GX0101，with a reticuloendotheliosis virus LTR insert［J］.Arch Virol，2013，158(9)：2007-2014.
［18］SU S，CUI N，CUI Z，et al.Complete genome sequence of a recombinant Marek’s disease virus field strain with one reticuloendotheliosis virus long terminal repeat insert［J］.J Virol，2012，86(24)：13818-13819.
［19］LUO J，SUN A J，TENG M，et al.Expression profiles of microRNAs encoded by the oncogenic Marek’s disease virus reveal two distinct expression patterns in vivo during different phases of disease［J］.J Gen Virol，2011，92(Pt 3)：608-620.
［20］YU Z H，TENG M，SUN A J，et al.Virus-encoded miR-155 ortholog is an important potential regulator but not essential for the development of lymphomas induced by very virulent Marek’s disease virus［J］.Virology，2014，448，55-64.
［21］TENG M，YU Z H，SUN A J，et al.The significance of the individual Meq-clustered miRNAs of Marek’s disease virus in oncogenesis［J］.J Gen Virol，2015，96(Pt 3)：637-649.
［22］ZHAO P，LI X J，TENG M，et al.In vivo expression patterns of microRNAs of Gallid herpesvirus 2 (GaHV-2) during the virus life cycle and development of Marek’s disease lymphomas［J］.Virus Genes，2015，50(2)：245-252.
［23］CHI J Q，TENG M，YU Z H，et al.Marek’s disease virus-encoded analog of microRNA-155 activates the oncogene c-Myc by targeting LTBP1 and suppressing the TGF-β signaling pathway［J］.Virology，2015，476：72-84.
［24］CHI X J，LU Y X，ZHAO P，et al.Interaction domain of glycoproteins gB and gH of Marek’s disease virus and identification of an antiviral peptide with dual functions［J］.PLoS One，2013，8(2)：e54761.
［25］SCHUMACHER D，TISCHER B K，REDDY S M，et al.Glycoproteins E and I of Marek’s disease virus serotype 1 are essential for virus growth in cultured cells［J］.J Virol，2001，75(23)：11307-11318.
［26］LZUMIYA Y，JANG H K，CAI J S，et al.Identification and sequence analysis of the Marek’s disease virus serotype 2 gene homologous to the herpes simplex virus type 1 UL52 protein［J］.J Vet Med Sci，1999，61(6)：683-687.
［27］KATO K，LZUMIYA Y，TOHYA Y，et al.Identification and characterization of Marek’s disease virus serotype 1 (MDV1) ICP22 gene product：MDV1 ICP22 transactivates the MDV1 ICP27 promoter synergistically with MDV1 ICP4［J］.Vet Microbiol，2002，85(4)：305-313.
［28］JAROSINSKI K W，OSTERRIEDER N.Further analysis of Marek’s disease virus horizontal transmission confirms that U(L)44 (gC) and U(L)13 protein kinase activity are essential，while U(S)2 is nonessential［J］.J Virol，2010，84(15)：7911-7916.
［29］GIMENO I M，WITTER R L，HUNT H D，et al.The pp38 gene of Marek’s disease virus (MDV) is necessary for cytolytic infection of B cells and maintenance of the transformed state but not for cytolytic infection of the feather follicle epithelium and horizontal spread of MDV［J］.J Virol，2005，79(7)：4545-4549.
［30］LI Y，SUN S，SU S，et al.Deletion of the Meq gene significantly decreases immunosuppression in chickens caused by pathogenic Marek’s disease virus［J］.Virol J，2011，8：2.
［31］SPATZ S J，SILVA R F.Polymorphisms in the repeat long regions of oncogenic and attenuated pathotypes of Marek’s disease virus 1［J］.Virus Genes，2007，35(1)：41-53.
［32］崔治中.禽反转录病毒与DNA病毒间的基因重组及其流行病学意义［J］.病毒学报，2006，22(2)：150-154.
CUI Z Z.The epidemiological significance of the genetic recombination between avian retroviruses and DNA virus［J］.Chinese Journal of Virology，2006，22(2)：150-154.(in Chinese)

[1]	宋慧子, 栾兆进, 王兆琛, 杜炜, 赵勇超, 张家新. 永生化绵羊附睾上皮细胞系的建立及其生物学特性分析[J]. 畜牧兽医学报, 2019, 50(9): 1822-1831.
[2]	李华振, 狄冉, 刘秋月, 胡文萍, 王翔宇, 马琳, 刘武军, 储明星. 类视黄醇X受体基因调控动物季节性发情和发情周期的研究进展[J]. 畜牧兽医学报, 2019, 50(8): 1525-1535.
[3]	李丽敏, 邹云婧, 郝雪飘, 赵兴华, 李睿文, 孙继国, 袁万哲. 脑心肌炎病毒感染神经细胞的基因表达谱分析[J]. 畜牧兽医学报, 2019, 50(8): 1709-1714.
[4]	李华振, 狄冉, 刘秋月, 胡文萍, 王翔宇, 马琳, 刘武军, 储明星. 类视黄醇X受体基因调控动物季节性发情和发情周期的研究进展[J]. 畜牧兽医学报, 2019, 50(8): 1525-1535.
[5]	李丽敏, 邹云婧, 郝雪飘, 赵兴华, 李睿文, 孙继国, 袁万哲. 脑心肌炎病毒感染神经细胞的基因表达谱分析[J]. 畜牧兽医学报, 2019, 50(8): 1709-1714.
[6]	刘豪丽, 滕蔓, 李会珍, 余祖华, 刘金玲, 丁轲, 张改平, 罗俊. 马立克病病毒编码的miR-M11-5p宿主靶基因的筛选与鉴定[J]. 畜牧兽医学报, 2019, 50(5): 1026-1038.
[7]	刘豪丽, 滕蔓, 李会珍, 余祖华, 刘金玲, 丁轲, 张改平, 罗俊. 马立克病病毒编码的miR-M11-5p宿主靶基因的筛选与鉴定[J]. 畜牧兽医学报, 2019, 50(5): 1026-1038.
[8]	占思远, 丁雪, 仲涛, 王林杰, 李利, 张红平. IGF1R和IGF2R在南江黄羊不同组织和肌细胞中的表达模式比较[J]. 畜牧兽医学报, 2019, 50(4): 701-711.
[9]	王波, 刁志成, 马勇, 李发弟, 曲扬华, 徐晨晨, 卢晓楠, 罗海玲. 日粮添加油脂和出生类型对羔羊肌肉胶原蛋白特性及相关基因表达的影响[J]. 畜牧兽医学报, 2019, 50(4): 771-780.
[10]	占思远, 丁雪, 仲涛, 王林杰, 李利, 张红平. IGF1R和IGF2R在南江黄羊不同组织和肌细胞中的表达模式比较[J]. 畜牧兽医学报, 2019, 50(4): 701-711.
[11]	王波, 刁志成, 马勇, 李发弟, 曲扬华, 徐晨晨, 卢晓楠, 罗海玲. 日粮添加油脂和出生类型对羔羊肌肉胶原蛋白特性及相关基因表达的影响[J]. 畜牧兽医学报, 2019, 50(4): 771-780.
[12]	王惠影, 郭保地, 何大乾, 龚绍明, 刘毅, 李光全, 徐琪, 陈国宏. 饲粮蛋氨酸对鹅再生羽绒品质、血清指标及基因mRNA表达量的影响[J]. 畜牧兽医学报, 2019, 50(2): 323-331.
[13]	王惠影, 郭保地, 何大乾, 龚绍明, 刘毅, 李光全, 徐琪, 陈国宏. 饲粮蛋氨酸对鹅再生羽绒品质、血清指标及基因mRNA表达量的影响[J]. 畜牧兽医学报, 2019, 50(2): 323-331.
[14]	胡会龙, 魏小兵, 王秋霞, 欧长波, 闫艺婷, 吕玉琼, 刘兴友. 发酵饲料中乳酸菌含量检测的EMA-qPCR方法[J]. 畜牧兽医学报, 2019, 50(10): 2166-2170.
[15]	张秋旭, 张润厚, 史晓雪, 李刚, 高爱琴. 添加不同形式亚麻油对肉羊生产性能、肉品质、脂肪酸含量及脂代谢相关酶基因mRNA表达量的影响[J]. 畜牧兽医学报, 2018, 49(9): 1928-1939.

马立克病病毒超强毒株GX0101感染宿主部分病毒基因表达水平及致病阶段分析

The Viral Gene Expression Profiles and Pathogenic Phases of the Disease Caused by Very Virulent MDV Strain GX0101

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价