减毒痘苗病毒天坛株的构建、致病性及免疫原性研究
|
摘要
痘苗病毒天坛株作为疫苗，在我国消灭天花的历史上居功至伟。近年来，痘苗病毒天坛株作为活病毒载体在预防艾滋病、狂犬病和流感等传染病方面取得了巨大成就。然而，颅内接种实验表明，痘苗病毒天坛株对小鼠的致死率较高，具有一定的神经毒性。临床接种痘苗病毒天坛株疫苗也可能导致少数接种者出现严重副作用，例如脑炎、结膜炎、心肌炎及全身性发痘等。因此，如何构建更加低毒、更加安全且高效的痘苗病毒天坛株疫苗或载体成为研究的热点。
通过同源重组和Cre-LoxP技术，定点敲除痘苗病毒基因组中的复制非必需基因，是构建减毒疫苗或载体的有效方法。基因重组作为基因工程的重要技术之一，包括同源重组、位点特异性重组以及转座重组。同源重组技术，又叫基因打靶技术，是目前研究痘苗病毒基因功能、构建减毒型痘苗病毒疫苗和载体的主要方法。该技术通过构建含有左右重组臂、启动子和筛选标记基因的穿梭载体，使病毒基因组与外源DNA序列发生重组，进而定点修饰或缺失基因组上某一目的基因，构建缺失型痘苗病毒。进而，通过体外和体内实验对缺失型痘苗病毒的生长动力学、病毒粒子表型、细胞间传播能力、毒力、宿主范围和免疫原性等特性进行系统分析。在构建缺失型毒株的过程中，通常需要引入筛选标记基因，用于病毒筛选。然而，筛选标记基因作为外源基因的存在不符合商业疫苗生物安全的要求，在研究过程中需要将其删除。Cre-LoxP系统作为一种特异性重组技术，能够定点将外源基因删除，广泛用于重组病毒、细菌和转基因动物等的研究。Cre重组酶能够特异性的识别LoxP位点（两个反向重复序列）和其间的间隔区域。当两个同向LoxP序列位于同一条DNA链上，Cre重组酶能有效切除位于两个LoxP序列之间的序列。
本研究基于传统痘苗病毒天坛株，分别构建了缺失TC7L-TK2L基因（15,262~25,450）和TA35R基因（138,881~139,570）的弱毒株vMVTT1、vMVTT2和vMVTT3。其中，TC7L-TK2L基因包括TC7L、TC6L、TC5L、TC4L、TC3L、TC2L、TC1L、TN1L、TN2L、TM1L、TM2L、TK1L和TK2L共13个片段。vMVTT1缺失了TC7L-TK2L基因，vMVTT2缺失了TA35R基因，vMVTT3同时缺失了TC7L-TK2L和TA35R基因。通过体内和体外实验对缺失型病毒的生长动力学、细胞间传播能力、病毒表型、毒力下降水平、宿主范围以及免疫原性进行了分析。首先，在BHK-21、Vero、MDCK、HeLa和PK15五种细胞中，通过结晶紫染色实验和MTT实验等多种方法分析了缺失型毒株（vMVTT1、vMVTT2和vMVTT3）和野生型VTT在细胞间传播能力、复制能力、病毒表型及细胞毒性和宿主范围之间的差异，并对缺失毒株的遗传稳定性进行了评估。然后，通过鼻内和颅内感染途径分别用vMVTT1、vMVTT2、vMVTT3和野生型VTT感染小鼠，通过统计小鼠体重下降程度和死亡情况分析缺失TC7L-TK2L基因和TA35R基因后天坛株痘苗病毒毒力下降水平；用缺失型毒株通过皮内途径感染家兔，观察家兔接种部位病灶变化大小和程度，进一步分析缺失型毒株对皮肤的损伤程度及其毒力下降水平。最后，分别用vMVTT1、vMVTT2、vMVTT3和野生型VTT接种BALB/c小鼠，检测血清中IL-2、IL-4、IL-10和IFN-γ的含量和特异性抗体水平，并进行攻毒保护实验，分析小鼠的免疫水平，进而评价vMVTT1、vMVTT2和vMVTT3的免疫原性。
结果表明，通过同源重组和Cre-LoxP系统，成功构建了缺失TC7L-TK2L基因和TA35R基因且不含外源基因的减毒痘苗病毒天坛株vMVTT1、vMVTT2和vMVTT3，均具有良好的遗传稳定性。电镜结果表明缺失TC7L-TK2L基因和TA35R基因不影响天坛株病毒粒子的形态发生。但是，对其生长动力学、细胞间传播能力、毒力和免疫原性有较大影响。与野毒相比较，vMVTT1、vMVTT2和vMVTT3在PK15、Vero、HeLa、MDCK和BHK-21细胞中的复制能力均下降，复制能力由大到小依次为VTT> vMVTT2> vMVTT1> vMVTT3。细胞与细胞间传播能力依次为VTT>vMVTT2>vMVTT1>vMVTT3，其中vMVTT3仅在BHK-21细胞上有一定的传播能力，在Vero、HeLa、MDCK和PK15细胞中传播能力较弱。MTT实验表明缺失型毒株对5种细胞的杀伤能力依次为VTT>vMVTT2> vMVTT1>vMVTT3。家兔皮肤致病力实验表明，VTT组对兔皮的损伤最大，毒性最强，vMVTT3组对兔皮基本无损伤，毒性最弱。而vMVTT1组和vMVTT2组对家兔皮肤均有致病力，且vMVTT2皮肤致病力较强。另外，痘斑形成的大小与感染缺失型病毒的滴度也有较大关系。通过鼻内和颅内接种途径感染小鼠，结果表明接种途径对病毒毒力的评估有较大影响。鼻内接种途径实验表明vMVTT1、vMVTT2和vMVTT3毒力明显下降，其中vMVTT3毒力下降约100倍。而颅内接种实验表明vMVTT2毒力下降80倍，vMVTT1和vMVTT3毒力下降3200倍。另外，细胞因子检测和中和试验结果表明缺失TC7L-TK2L基因和TA35R基因对天坛株痘苗病毒的免疫原性几乎没有影响，接种小鼠后没有显著降低其免疫应答水平，仍可诱导产生较高水平的IL-2、IL-4、IL-10和IFN-γ，并可刺激小鼠体内产生高水平中和抗体。攻毒保护试验结果表明vMVTT1、vMVTT2和vMVTT3作为疫苗对小鼠具有较好的保护作用，用高剂量（500×LD50）的野生型VTT攻击小鼠也没有出现死亡。综上所述，本研究所构建的三个缺失型毒株的毒力均有不同程度的下降，其中vMVTT3毒力下降最为明显且保持了较好的免疫原性，具有作为疫苗或载体的潜能。
本实验基于VTT，在不引进外源标记基因的条件下，同时缺失TC7L-TK2L和TA35R两段基因，并成功获得毒力下降且免疫原性较好的缺失型痘苗病毒天坛株，具备作为疫苗或载体的潜能，在各种传染性疾病的防治等领域具有较好的应用前景。
Widespread use of vaccinia virus as smallpox vaccine has eradicated variolavirus. Recently, vaccinia virus is a live recombinant vector against HIV, rabies andother infectious diseases that have been intensively studied. However, intracranialinoculation test show a high death rate of mice and neurotoxicity. Clinical vaccinationof vaccinia virus may also lead to severe side effects, such as myocarditis,encephalitis, conjunctivitis and systemic hair blain, etc. Therefore, how to build amore safe and efficient vaccinia virus vaccine strain is becoming a hot spot ofresearch.
Deletion of non-essential gene in vaccinia virus genome, is one of the effectivemethod to construct attenuated vaccine or vector by homologous recombination andCre-LoxP technology. Genetic recombination, as one of the important geneticengineering technology, includes the homologous recombination, site specificrestructuring and reorganization. Homologous recombination technology, also knownas gene targeting technology, is currently method in gene function research. Thetechnology of DNA restructuring was done by shuttle vector which containsrestructuring arm, promoter and EGFP. Attenuated vaccinia Tian Tan strain isconstructed by deletion of the target gene. Growth dynamics, virus particlesphenotype, intercellular communication ability, virulence, immunogenicity and hostrange features of vaccinia virus are analysised through in vitro and in vivo. Screeningmarker genes are used in the process of construction. However, screening markergenes are foreign genes which do not conform to the requirements of the commercialvaccines of biological safety. Thus, screening marker genes should be deleted in theprocess of research. Cre-loxP site-specific gene recombination system may removeexogenous gene, which has been used widely in the research of recombinant viruses,bacteria and transgenic animal. Cre recombinase can recognize LoxP site and remove the sequence when LoxP sequence located on the same DNA strand.
In the present study, we constructed modified VTT genome (vMVTT1, vMVTT2and vMVTT3) by deleting TC7L-TK2L or TA35R genes, which resulted in reducedvirulence. vMVTT1deletes TC7L-TK2L gene. vMVTT2deletes TA35R gene.vMVTT3deletes both TC7L-TK2L and TA35R genes. Here, we examine the mutantvirus which removed TC7L-TK2L (15,262-25,450) including TC7L, TC6L, TC5L,TC4L, TC3L, TC2L, TC1L, TN1L, TN2L, TM1L, TM2L, TK1L, and TK2L or asingle open reading frame TA35R (138,881-139,570) from vaccinia Tian Tan strain.Growth dynamics, virus particles phenotype, intercellular communication ability,virulence, immunogenicity and host range features of vaccinia virus are analysisedthrough in vitro and in vivo. First, the diffusing capacity and cytotoxicity ofvMVTT1、vMVTT2、vMVTT3and VTT in BHK-21、Vero、MDCK、HeLa and PK15cells were detected by crystal violet staining and MTT colorimetric assay. We alsoevaluate the genetic stability of the strains. Second, the body weight changes ofBALB/c mice were analyzed through intranasal inoculation, and the death rates ofBALB/c mice were analyzed by intracranial inoculation. Rabbits were test thevirulence of strains by intradermal inoculation. The immunogenicity of the vMVTT1,vMVTT2and vMVTT3was evaluated in BALB/c mice. IL-2, IL-4, IL-10, IFN-γ andVTT-specific antiboady was tested in peripheral blood. Protection from lethalchallenge was also tested.
The results showed that three attenuated strains (vMVTT1, vMVTT2andvMVTT3) deleting TC7L-TK2L or TA35R gene were successful constructed, andthey had well genetic stability. There were no significant differences in morphologyamong vMVTT1, vMVTT2, vMVTT3and VTT, suggesting that deletion of theTC7L-TK2L and TA35R genes did not affect the virus morphogenesis. However, ithas a great influence on the growth dynamics, intercellular communication ability,toxicity and immunogenicity. Replication was decreased in the cells compared toVTT (VTT> vMVTT2> vMVTT1> vMVTT3). Spread of cell-to-cell were changed(VTT> vMVTT2> vMVTT1> vMVTT3). vMVTT3spread in BHK-21cells. ButvMVTT3nearly could not spread in Vero, HeLa, MDCK and PK15cells. The mutantvMVTT3did not cause detectable ulceration when it was inoculated intradermally in rabbits. And skin virulence of vMVTT2was stronger than vMVTT1. Additionally,size of the spots also related to the titer of virus. Inoculation of virus was important tovirulence. The vMVTT3was found to be100-fold less virulent than VTT based onthe body weight loss of mice intranasally infected with either virus, and3200-fold lessvirulent based on the intracranial lethal infectious dose in mice. In addition, wedemonstrated mutants induced immune responses was equal to those induced by VTTimmunization. vMVTT3protect mice effectively from the challenge of a massivedose of VTT, suggesting that removal of TC7L-TK2L and TA35R genes fromvaccinia Tian Tan strain-based vaccines will keep their immunogenicity. In this study,the virulence of vMVTT1, vMVTT2and vMVTT3was decreased. The vMVTT3mutant was shown to be avirulent and immunogenic.
Base on VTT, TC7L-TK2L and TA35R genes are deleted without inserting ofEGFP. vMVTT3can be used as a safe viral vector or as platform of vaccines forinfectious diseases and cancer.
通过同源重组和Cre-LoxP技术，定点敲除痘苗病毒基因组中的复制非必需基因，是构建减毒疫苗或载体的有效方法。基因重组作为基因工程的重要技术之一，包括同源重组、位点特异性重组以及转座重组。同源重组技术，又叫基因打靶技术，是目前研究痘苗病毒基因功能、构建减毒型痘苗病毒疫苗和载体的主要方法。该技术通过构建含有左右重组臂、启动子和筛选标记基因的穿梭载体，使病毒基因组与外源DNA序列发生重组，进而定点修饰或缺失基因组上某一目的基因，构建缺失型痘苗病毒。进而，通过体外和体内实验对缺失型痘苗病毒的生长动力学、病毒粒子表型、细胞间传播能力、毒力、宿主范围和免疫原性等特性进行系统分析。在构建缺失型毒株的过程中，通常需要引入筛选标记基因，用于病毒筛选。然而，筛选标记基因作为外源基因的存在不符合商业疫苗生物安全的要求，在研究过程中需要将其删除。Cre-LoxP系统作为一种特异性重组技术，能够定点将外源基因删除，广泛用于重组病毒、细菌和转基因动物等的研究。Cre重组酶能够特异性的识别LoxP位点（两个反向重复序列）和其间的间隔区域。当两个同向LoxP序列位于同一条DNA链上，Cre重组酶能有效切除位于两个LoxP序列之间的序列。
本研究基于传统痘苗病毒天坛株，分别构建了缺失TC7L-TK2L基因（15,262~25,450）和TA35R基因（138,881~139,570）的弱毒株vMVTT1、vMVTT2和vMVTT3。其中，TC7L-TK2L基因包括TC7L、TC6L、TC5L、TC4L、TC3L、TC2L、TC1L、TN1L、TN2L、TM1L、TM2L、TK1L和TK2L共13个片段。vMVTT1缺失了TC7L-TK2L基因，vMVTT2缺失了TA35R基因，vMVTT3同时缺失了TC7L-TK2L和TA35R基因。通过体内和体外实验对缺失型病毒的生长动力学、细胞间传播能力、病毒表型、毒力下降水平、宿主范围以及免疫原性进行了分析。首先，在BHK-21、Vero、MDCK、HeLa和PK15五种细胞中，通过结晶紫染色实验和MTT实验等多种方法分析了缺失型毒株（vMVTT1、vMVTT2和vMVTT3）和野生型VTT在细胞间传播能力、复制能力、病毒表型及细胞毒性和宿主范围之间的差异，并对缺失毒株的遗传稳定性进行了评估。然后，通过鼻内和颅内感染途径分别用vMVTT1、vMVTT2、vMVTT3和野生型VTT感染小鼠，通过统计小鼠体重下降程度和死亡情况分析缺失TC7L-TK2L基因和TA35R基因后天坛株痘苗病毒毒力下降水平；用缺失型毒株通过皮内途径感染家兔，观察家兔接种部位病灶变化大小和程度，进一步分析缺失型毒株对皮肤的损伤程度及其毒力下降水平。最后，分别用vMVTT1、vMVTT2、vMVTT3和野生型VTT接种BALB/c小鼠，检测血清中IL-2、IL-4、IL-10和IFN-γ的含量和特异性抗体水平，并进行攻毒保护实验，分析小鼠的免疫水平，进而评价vMVTT1、vMVTT2和vMVTT3的免疫原性。
结果表明，通过同源重组和Cre-LoxP系统，成功构建了缺失TC7L-TK2L基因和TA35R基因且不含外源基因的减毒痘苗病毒天坛株vMVTT1、vMVTT2和vMVTT3，均具有良好的遗传稳定性。电镜结果表明缺失TC7L-TK2L基因和TA35R基因不影响天坛株病毒粒子的形态发生。但是，对其生长动力学、细胞间传播能力、毒力和免疫原性有较大影响。与野毒相比较，vMVTT1、vMVTT2和vMVTT3在PK15、Vero、HeLa、MDCK和BHK-21细胞中的复制能力均下降，复制能力由大到小依次为VTT> vMVTT2> vMVTT1> vMVTT3。细胞与细胞间传播能力依次为VTT>vMVTT2>vMVTT1>vMVTT3，其中vMVTT3仅在BHK-21细胞上有一定的传播能力，在Vero、HeLa、MDCK和PK15细胞中传播能力较弱。MTT实验表明缺失型毒株对5种细胞的杀伤能力依次为VTT>vMVTT2> vMVTT1>vMVTT3。家兔皮肤致病力实验表明，VTT组对兔皮的损伤最大，毒性最强，vMVTT3组对兔皮基本无损伤，毒性最弱。而vMVTT1组和vMVTT2组对家兔皮肤均有致病力，且vMVTT2皮肤致病力较强。另外，痘斑形成的大小与感染缺失型病毒的滴度也有较大关系。通过鼻内和颅内接种途径感染小鼠，结果表明接种途径对病毒毒力的评估有较大影响。鼻内接种途径实验表明vMVTT1、vMVTT2和vMVTT3毒力明显下降，其中vMVTT3毒力下降约100倍。而颅内接种实验表明vMVTT2毒力下降80倍，vMVTT1和vMVTT3毒力下降3200倍。另外，细胞因子检测和中和试验结果表明缺失TC7L-TK2L基因和TA35R基因对天坛株痘苗病毒的免疫原性几乎没有影响，接种小鼠后没有显著降低其免疫应答水平，仍可诱导产生较高水平的IL-2、IL-4、IL-10和IFN-γ，并可刺激小鼠体内产生高水平中和抗体。攻毒保护试验结果表明vMVTT1、vMVTT2和vMVTT3作为疫苗对小鼠具有较好的保护作用，用高剂量（500×LD50）的野生型VTT攻击小鼠也没有出现死亡。综上所述，本研究所构建的三个缺失型毒株的毒力均有不同程度的下降，其中vMVTT3毒力下降最为明显且保持了较好的免疫原性，具有作为疫苗或载体的潜能。
本实验基于VTT，在不引进外源标记基因的条件下，同时缺失TC7L-TK2L和TA35R两段基因，并成功获得毒力下降且免疫原性较好的缺失型痘苗病毒天坛株，具备作为疫苗或载体的潜能，在各种传染性疾病的防治等领域具有较好的应用前景。
Widespread use of vaccinia virus as smallpox vaccine has eradicated variolavirus. Recently, vaccinia virus is a live recombinant vector against HIV, rabies andother infectious diseases that have been intensively studied. However, intracranialinoculation test show a high death rate of mice and neurotoxicity. Clinical vaccinationof vaccinia virus may also lead to severe side effects, such as myocarditis,encephalitis, conjunctivitis and systemic hair blain, etc. Therefore, how to build amore safe and efficient vaccinia virus vaccine strain is becoming a hot spot ofresearch.
Deletion of non-essential gene in vaccinia virus genome, is one of the effectivemethod to construct attenuated vaccine or vector by homologous recombination andCre-LoxP technology. Genetic recombination, as one of the important geneticengineering technology, includes the homologous recombination, site specificrestructuring and reorganization. Homologous recombination technology, also knownas gene targeting technology, is currently method in gene function research. Thetechnology of DNA restructuring was done by shuttle vector which containsrestructuring arm, promoter and EGFP. Attenuated vaccinia Tian Tan strain isconstructed by deletion of the target gene. Growth dynamics, virus particlesphenotype, intercellular communication ability, virulence, immunogenicity and hostrange features of vaccinia virus are analysised through in vitro and in vivo. Screeningmarker genes are used in the process of construction. However, screening markergenes are foreign genes which do not conform to the requirements of the commercialvaccines of biological safety. Thus, screening marker genes should be deleted in theprocess of research. Cre-loxP site-specific gene recombination system may removeexogenous gene, which has been used widely in the research of recombinant viruses,bacteria and transgenic animal. Cre recombinase can recognize LoxP site and remove the sequence when LoxP sequence located on the same DNA strand.
In the present study, we constructed modified VTT genome (vMVTT1, vMVTT2and vMVTT3) by deleting TC7L-TK2L or TA35R genes, which resulted in reducedvirulence. vMVTT1deletes TC7L-TK2L gene. vMVTT2deletes TA35R gene.vMVTT3deletes both TC7L-TK2L and TA35R genes. Here, we examine the mutantvirus which removed TC7L-TK2L (15,262-25,450) including TC7L, TC6L, TC5L,TC4L, TC3L, TC2L, TC1L, TN1L, TN2L, TM1L, TM2L, TK1L, and TK2L or asingle open reading frame TA35R (138,881-139,570) from vaccinia Tian Tan strain.Growth dynamics, virus particles phenotype, intercellular communication ability,virulence, immunogenicity and host range features of vaccinia virus are analysisedthrough in vitro and in vivo. First, the diffusing capacity and cytotoxicity ofvMVTT1、vMVTT2、vMVTT3and VTT in BHK-21、Vero、MDCK、HeLa and PK15cells were detected by crystal violet staining and MTT colorimetric assay. We alsoevaluate the genetic stability of the strains. Second, the body weight changes ofBALB/c mice were analyzed through intranasal inoculation, and the death rates ofBALB/c mice were analyzed by intracranial inoculation. Rabbits were test thevirulence of strains by intradermal inoculation. The immunogenicity of the vMVTT1,vMVTT2and vMVTT3was evaluated in BALB/c mice. IL-2, IL-4, IL-10, IFN-γ andVTT-specific antiboady was tested in peripheral blood. Protection from lethalchallenge was also tested.
The results showed that three attenuated strains (vMVTT1, vMVTT2andvMVTT3) deleting TC7L-TK2L or TA35R gene were successful constructed, andthey had well genetic stability. There were no significant differences in morphologyamong vMVTT1, vMVTT2, vMVTT3and VTT, suggesting that deletion of theTC7L-TK2L and TA35R genes did not affect the virus morphogenesis. However, ithas a great influence on the growth dynamics, intercellular communication ability,toxicity and immunogenicity. Replication was decreased in the cells compared toVTT (VTT> vMVTT2> vMVTT1> vMVTT3). Spread of cell-to-cell were changed(VTT> vMVTT2> vMVTT1> vMVTT3). vMVTT3spread in BHK-21cells. ButvMVTT3nearly could not spread in Vero, HeLa, MDCK and PK15cells. The mutantvMVTT3did not cause detectable ulceration when it was inoculated intradermally in rabbits. And skin virulence of vMVTT2was stronger than vMVTT1. Additionally,size of the spots also related to the titer of virus. Inoculation of virus was important tovirulence. The vMVTT3was found to be100-fold less virulent than VTT based onthe body weight loss of mice intranasally infected with either virus, and3200-fold lessvirulent based on the intracranial lethal infectious dose in mice. In addition, wedemonstrated mutants induced immune responses was equal to those induced by VTTimmunization. vMVTT3protect mice effectively from the challenge of a massivedose of VTT, suggesting that removal of TC7L-TK2L and TA35R genes fromvaccinia Tian Tan strain-based vaccines will keep their immunogenicity. In this study,the virulence of vMVTT1, vMVTT2and vMVTT3was decreased. The vMVTT3mutant was shown to be avirulent and immunogenic.
Base on VTT, TC7L-TK2L and TA35R genes are deleted without inserting ofEGFP. vMVTT3can be used as a safe viral vector or as platform of vaccines forinfectious diseases and cancer.
引文
[1] MENDES EA, FONSECA FG, CASERIO BM, et al. Recombinant Vaccinesagainst T. gondii: Comparison between Homologous and Heterologous VaccinationProtocols Using Two Viral Vectors Expressing SAG1[J]. PloS one,2013,8(5):e63201.
[2] SEKIGUCHI S, KIMURA K, CHIYO T, et al. Immunization with arecombinant vaccinia virus that encodes nonstructural proteins of the hepatitis C virussuppresses viral protein levels in mouse liver[J]. PloS one,2012,7(12): e51656.
[3] FLYNN BJ, KASTENMULLER K, WILLE-REECE U, et al. Immunizationwith HIV Gag targeted to dendritic cells followed by recombinant New York vacciniavirus induces robust T-cell immunity in nonhuman primates[J]. Proceedings of theNational Academy of Sciences of the United States of America,2013,108(17):7131-7136.
[4] WOLLENBERG A, ENGLER R. Smallpox, vaccination and adverse reactionsto smallpox vaccine[J]. Current opinion in allergy and clinical immunology,2004,4(4):271-275.
[5] QIN L, LIANG M, EVANS DH. Genomic analysis of vaccinia virus strainTianTan provides new insights into the evolution and evolutionary relationshipsbetween Orthopoxviruses[J]. Virology, Available online19April2013.
[6] MOSS B. Genetically engineered poxviruses for recombinant gene expression,vaccination, and safety[J]. Proceedings of the National Academy of Sciences of theUnited States of America,1996,93(21):11341-11348.
[7] BOYD O, TURNER PC, MOYER RW, CONDIT RC, MOUSSATCHE N.The E6protein from vaccinia virus is required for the formation of immaturevirions[J]. Virology,2010,399(2):201-211.
[8] DOMS RW, BLUMENTHAL R, MOSS B. Fusion of intra-and extracellularforms of vaccinia virus with the cell membrane[J]. Journal of virology,1990,64(10):4884-4892.
[9] CUDMORE S, COSSART P, GRIFFITHS G, WAY M. Actin-based motility ofvaccinia virus[J]. Nature,1995,378(6557):636-638.
[10] DOCEUL V, HOLLINSHEAD M, VAN DER LINDEN L, SMITH GL.Repulsion of superinfecting virions: a mechanism for rapid virus spread[J]. Science(New York, NY,2010,327(5967):873-876.
[11] CARTER GC, LAW M, HOLLINSHEAD M, SMITH GL. Entry of thevaccinia virus intracellular mature virion and its interactions withglycosaminoglycans[J]. The Journal of general virology,2005,86(Pt5):1279-1290.
[12] VANDERPLASSCHEN A, HOLLINSHEAD M, SMITH GL. Intracellularand extracellular vaccinia virions enter cells by different mechanisms[J]. The Journalof general virology,1998,79(Pt4):877-887.
[13] HSIAO JC, CHUNG CS, CHANG W. Vaccinia virus envelope D8L proteinbinds to cell surface chondroitin sulfate and mediates the adsorption of intracellularmature virions to cells[J]. Journal of virology,1999,73(10):8750-8761.
[14] CHUNG CS, HSIAO JC, CHANG YS, CHANG W. A27L protein mediatesvaccinia virus interaction with cell surface heparan sulfate[J]. Journal of virology,1998,72(2):1577-1585.
[15] HSIAO JC, CHUNG CS, CHANG W. Cell surface proteoglycans arenecessary for A27L protein-mediated cell fusion: identification of the N-terminalregion of A27L protein as the glycosaminoglycan-binding domain[J]. Journal ofvirology,1998,72(10):8374-8379.
[16] RODRIGUEZ D, ESTEBAN M, RODRIGUEZ JR. Vaccinia virus A17Lgene product is essential for an early step in virion morphogenesis[J]. Journal ofvirology,1995,69(8):4640-4648.
[17] VAZQUEZ MI, RIVAS G, CREGUT D, SERRANO L, ESTEBAN M. Thevaccinia virus14-kilodalton (A27L) fusion protein forms a triple coiled-coil structureand interacts with the21-kilodalton (A17L) virus membrane protein through aC-terminal alpha-helix[J]. Journal of virology,1998,72(12):10126-10137.
[18] FOO CH, WHITBECK JC, PONCE-DE-LEON M, et al. The myristatemoiety and amino terminus of vaccinia virus l1constitute a bipartite functional regionneeded for entry[J]. Journal of virology,2012,86(10):5437-5451.
[19] CHAN WM, WARD BM. Increased interaction between vaccinia virusproteins A33and B5is detrimental to infectious extracellular enveloped virionproduction[J]. Journal of virology,2012,86(15):8232-8244.
[20] PEDERSEN K, SNIJDER EJ, SCHLEICH S, et al. Characterization ofvaccinia virus intracellular cores: implications for viral uncoating and corestructure[J]. Journal of virology,2000,74(8):3525-3536.
[21] ZHANG Y, KECK JG, MOSS B. Transcription of viral late genes isdependent on expression of the viral intermediate gene G8R in cells infected with aninducible conditional-lethal mutant vaccinia virus[J]. Journal of virology,1992,66(11):6470-6479.
[22] KECK JG, KOVACS GR, MOSS B. Overexpression, purification, and latetranscription factor activity of the17-kilodalton protein encoded by the vaccinia virusA1L gene[J]. Journal of virology,1993,67(10):5740-5748.
[23] HUBBS AE, WRIGHT CF. The A2L intermediate gene product is requiredfor in vitro transcription from a vaccinia virus late promoter[J]. Journal of virology,1996,70(1):327-331.
[24] LUTTGE BG, MOYER RW. Suppressors of a host range mutation in therabbitpox virus serpin SPI-1map to proteins essential for viral DNA replication[J].Journal of virology,2005,79(14):9168-9179.
[25] CURLEE KV, HONG JS, CLANCY JP, et al. Membrane transplantation tocorrect integral membrane protein defects[J]. Journal of molecular medicine (Berlin,Germany),2003,81(8):511-520.
[26] SODEIK B, KRIJNSE-LOCKER J. Assembly of vaccinia virus revisited: denovo membrane synthesis or acquisition from the host?[J]. Trends in microbiology,2002,10(1):15-24.
[27] RIETDORF J, PLOUBIDOU A, RECKMANN I, et al. Kinesin-dependentmovement on microtubules precedes actin-based motility of vaccinia virus[J]. Naturecell biology,2001,3(11):992-1000.
[28] CUDMORE S, BLASCO R, VINCENTELLI R, et al. A vaccinia virus coreprotein, p39, is membrane associated[J]. Journal of virology,1996,70(10):6909-6921.
[29] ZHU R, HUANG W, WANG W, et al. Comparison on virulence andimmunogenicity of two recombinant vaccinia vaccines, Tian Tan and Guang9strains,expressing the HIV-1envelope gene[J]. PloS one,2012,7(11): e48343.
[30] FANG Q, YANG L, ZHU W, et al. Host range, growth property, andvirulence of the smallpox vaccine: vaccinia virus Tian Tan strain[J]. Virology,2005,335(2):242-251.
[31] SUMNER RP, REN H, SMITH GL. Deletion of immunomodulator C6fromvaccinia virus strain Western Reserve enhances virus immunogenicity and vaccineefficacy[J]. The Journal of general virology,2013,94(Pt5):1121-1126.
[32] CLARK RH, KENYON JC, BARTLETT NW, TSCHARKE DC, SMITHGL. Deletion of gene A41L enhances vaccinia virus immunogenicity and vaccineefficacy[J]. The Journal of general virology,2006,87(Pt1):29-38.
[33] COTTINGHAM MG, ANDERSEN RF, SPENCER AJ, et al.Recombination-mediated genetic engineering of a bacterial artificial chromosomeclone of modified vaccinia virus Ankara (MVA)[J]. PloS one,2008,3(2): e1638.
[34] JENTARRA GM, HECK MC, YOUN JW, et al. Vaccinia viruses withmutations in the E3L gene as potential replication-competent, attenuated vaccines:scarification vaccination[J]. Vaccine,2008,26(23):2860-2872.
[35] STAIB C, KISLING S, ERFLE V, SUTTER G. Inactivation of the viralinterleukin1beta receptor improves CD8+T-cell memory responses elicited uponimmunization with modified vaccinia virus Ankara[J]. The Journal of general virology,2005,86(Pt7):1997-2006.
[36] VLIEGEN I, YANG G, HRUBY D, JORDAN R, NEYTS J. Deletion of thevaccinia virus F13L gene results in a highly attenuated virus that mounts a protectiveimmune response against subsequent vaccinia virus challenge[J]. Antiviral research,2012,93(1):160-166.
[37] ALCAMI A, SMITH GL. Vaccinia, cowpox, and camelpox viruses encodesoluble gamma interferon receptors with novel broad species specificity[J]. Journal ofvirology,1995,69(8):4633-4639.
[38] VERARDI PH, JONES LA, AZIZ FH, AHMAD S, YILMA TD. Vacciniavirus vectors with an inactivated gamma interferon receptor homolog gene (B8R) areattenuated In vivo without a concomitant reduction in immunogenicity[J]. Journal ofvirology,2001,75(1):11-18.
[39] DENES B, GRIDLEY DS, FODOR N, et al. Attenuation of a vaccine strainof vaccinia virus via inactivation of interferon viroceptor[J]. The journal of genemedicine,2006,8(7):814-823.
[40] LEGRAND FA, VERARDI PH, JONES LA, et al. Induction of potenthumoral and cell-mediated immune responses by attenuated vaccinia virus vectorswith deleted serpin genes[J]. J Virol,2004,78(6):2770-2779.
[41] JACKSON SS, ILYINSKII P, PHILIPPON V, et al. Role of genes thatmodulate host immune responses in the immunogenicity and pathogenicity ofvaccinia virus[J]. Journal of virology,2005,79(10):6554-6559.
[42] SYMONS JA, ADAMS E, TSCHARKE DC, et al. The vaccinia virus C12Lprotein inhibits mouse IL-18and promotes virus virulence in the murine intranasalmodel[J]. The Journal of general virology,2002,83(Pt11):2833-2844.
[43] READING PC, SMITH GL. Vaccinia virus interleukin-18-binding proteinpromotes virulence by reducing gamma interferon production and natural killer andT-cell activity[J]. Journal of virology,2003,77(18):9960-9968.
[44] DAI K, LIU Y, LIU M, et al. Pathogenicity and immunogenicity ofrecombinant Tiantan Vaccinia Virus with deleted C12L and A53R genes[J]. Vaccine,2008,26(39):5062-5071.
[45] ZHU W, FANG Q, ZHUANG K, et al. The attenuation of vaccinia Tian Tanstrain by the removal of the viral M1L-K2L genes[J]. Journal of virological methods,2007,144(1-2):17-26.
[46] KIENY MP, LATHE R, DRILLIEN R, et al. Expression of rabies virusglycoprotein from a recombinant vaccinia virus[J]. Nature,1984,312(5990):163-166.
[47] RUPPRECHT CE, BLASS L, SMITH K, et al. Human infection due torecombinant vaccinia-rabies glycoprotein virus[J]. The New England journal ofmedicine,2001,345(8):582-586.
[48]毛春生，金宁一，李季枝. A基因侧翼重组痘苗病毒的生物学特性研究[J].河南农业大学学报,1997,31(4):314.
[49] BULLER RM, CHAKRABARTI S, COOPER JA, TWARDZIK DR, MOSSB. Deletion of the vaccinia virus growth factor gene reduces virus virulence[J].Journal of virology,1988,62(3):866-874.
[50] LEE MS, ROOS JM, MCGUIGAN LC, et al. Molecular attenuation ofvaccinia virus: mutant generation and animal characterization[J]. Journal of virology,1992,66(5):2617-2630.
[51] LANGLAND JO, JACOBS BL. The role of the PKR-inhibitory genes, E3Land K3L, in determining vaccinia virus host range[J]. Virology,2002,299(1):133-141.
[52] CHANG HW, URIBE LH, JACOBS BL. Rescue of vaccinia virus lackingthe E3L gene by mutants of E3L[J]. Journal of virology,1995,69(10):6605-6608.
[53] VIJAYSRI S, JENTARRA G, HECK MC, et al. Vaccinia viruses withmutations in the E3L gene as potential replication-competent, attenuated vaccines:intra-nasal vaccination[J]. Vaccine,2008,26(5):664-676.
[54] LANGLAND JO, KASH JC, CARTER V, et al. Suppression ofproinflammatory signal transduction and gene expression by the dual nucleic acidbinding domains of the vaccinia virus E3L proteins[J]. Journal of virology,2006,80(20):10083-10095.
[55] CHANG PY, POGO BG. Reintroduction of gene(s) into an attenuateddeletion mutant of vaccinia virus strain IHD-W[J]. Microbial pathogenesis,1993,15(5):347-357.
[56] LANGLAND JO, JACOBS BL. Inhibition of PKR by vaccinia virus: role ofthe N-and C-terminal domains of E3L[J]. Virology,2004,324(2):419-429.
[57] HOLZER GW, FALKNER FG. Construction of a vaccinia virus deficient inthe essential DNA repair enzyme uracil DNA glycosylase by a complementing cellline[J]. Journal of virology,1997,71(7):4997-5002.
[58] HOLZER GW, REMP G, ANTOINE G, et al. Highly efficient induction ofprotective immunity by a vaccinia virus vector defective in late gene expression[J].Journal of virology,1999,73(6):4536-4542.
[59] WHITE SD, JACOBS BL. The amino terminus of the vaccinia virus E3protein is necessary to inhibit the interferon response[J]. Journal of virology,2012,86(10):5895-5904.
[60] GOEBEL SJ, JOHNSON GP, PERKUS ME, et al. The complete DNAsequence of vaccinia virus[J]. Virology,1990,179(1):247-266,517-263.
[61] ANTOINE G, SCHEIFLINGER F, DORNER F, FALKNER FG. Thecomplete genomic sequence of the modified vaccinia Ankara strain: comparison withother orthopoxviruses[J]. Virology,1998,244(2):365-396.
[62] UPTON C, SLACK S, HUNTER AL, EHLERS A, ROPER RL. Poxvirusorthologous clusters: toward defining the minimum essential poxvirus genome[J].Journal of virology,2003,77(13):7590-7600.
[63] SHCHELKUNOV SN, TOTMENIN AV, SANDAKHCHIEV LS. Analysis ofthe nucleotide sequence of23.8kbp from the left terminus of the genome of variolamajor virus strain India-1967[J]. Virus research,1996,40(2):169-183.
[64] MASSUNG RF, LIU LI, QI J, et al. Analysis of the complete genome ofsmallpox variola major virus strain Bangladesh-1975[J]. Virology,1994,201(2):215-240.
[65] SHCHELKUNOV SN, TOTMENIN AV, LOPAREV VN, et al. Alastrimsmallpox variola minor virus genome DNA sequences[J]. Virology,2000,266(2):361-386.
[66] ROPER RL. Characterization of the vaccinia virus A35R protein and its rolein virulence[J]. Journal of virology,2006,80(1):306-313.
[67] REHM KE, JONES GJ, TRIPP AA, METCALF MW, ROPER RL. Thepoxvirus A35protein is an immunoregulator[J]. J Virol,2010,84(1):418-425.
[68] ROPER RL. Characterization of the Vaccinia Virus A35R Protein and ItsRole in Virulence[J]. JOURNAL OF VIROLOGY,2006,80(1):306–313.
[69] STACK J, HAGA IR, SCHRODER M, et al. Vaccinia virus protein A46Rtargets multiple Toll-like-interleukin-1receptor adaptors and contributes tovirulence[J]. The Journal of experimental medicine,2005,201(6):1007-1018.
[70] FERRIER-REMBERT A, DRILLIEN R, TOURNIER JN, GARIN D,CRANCE JM. Short-and long-term immunogenicity and protection induced bynon-replicating smallpox vaccine candidates in mice and comparison with thetraditional1st generation vaccine[J]. Vaccine,2008,26(14):1794-1804.
[71] STITTELAAR KJ, VAN AMERONGEN G, KONDOVA I, et al. Modifiedvaccinia virus Ankara protects macaques against respiratory challenge withmonkeypox virus[J]. J Virol,2005,79(12):7845-7851.
[72] BELYAKOV IM, EARL P, DZUTSEV A, et al. Shared modes of protectionagainst poxvirus infection by attenuated and conventional smallpox vaccine viruses[J].Proceedings of the National Academy of Sciences of the United States of America,2003,100(16):9458-9463.
[73] BLANCHARD TJ, ALCAMI A, ANDREA P, SMITH GL. Modifiedvaccinia virus Ankara undergoes limited replication in human cells and lacks severalimmunomodulatory proteins: implications for use as a human vaccine[J]. The Journalof general virology,1998,79(Pt5):1159-1167.
[74] HOWLES S, GUIMARAES-WALKER A, YANG H, et al. Vaccination witha modified vaccinia virus Ankara (MVA)-vectored HIV-1immunogen induces modestvector-specific T cell responses in human subjects[J]. Vaccine,2010,28(45):7306-7312.
[75] EARL PL, COTTER C, MOSS B, et al. Design and evaluation of multi-gene,multi-clade HIV-1MVA vaccines[J]. Vaccine,2009,27(42):5885-5895.
[76] GUO W, CLEVELAND B, DAVENPORT T, LEE K, HU SL. Purification ofrecombinant vaccinia virus-expressed monomeric HIV-1gp120to apparenthomogeneity[J]. Protein expression and purification,2013,90(1):34-39.
[77] REHM KE, ROPER RL. Deletion of the A35gene from Modified VacciniaVirus Ankara increases immunogenicity and isotype switching[J]. Vaccine,2011,29(17):3276-3283.
[78] LI P, WANG N, ZHOU D, et al. Disruption of MHC class II-restrictedantigen presentation by vaccinia virus[J]. J Immunol,2005,175(10):6481-6488.
[79] JIN B, WANG RY, QIU Q, et al. Induction of potent cellular immuneresponse in mice by hepatitis C virus NS3protein with double-stranded RNA[J].Immunology,2007,122(1):15-27.
[80] SHARMA DP, RAMSAY AJ, MAGUIRE DJ, ROLPH MS, RAMSHAW IA.Interleukin-4mediates down regulation of antiviral cytokine expression and cytotoxicT-lymphocyte responses and exacerbates vaccinia virus infection in vivo[J]. Journal ofvirology,1996,70(10):7103-7107.
[81] REHM KE, CONNOR RF, JONES GJ, YIMBU K, ROPER RL. Vacciniavirus A35R inhibits MHC class II antigen presentation[J]. Virology,2010,397(1):176-186.
[82] JONES T. IMVAMUNE, an attenuated modified vaccinia Ankara virusvaccine for smallpox infection[J]. Current opinion in molecular therapeutics,2008,10(4):407-417.
[83] BACKES S, SPERLING KM, ZWILLING J, et al. Viral host-range factorC7or K1is essential for modified vaccinia virus Ankara late gene expression inhuman and murine cells, irrespective of their capacity to inhibit protein kinaseR-mediated phosphorylation of eukaryotic translation initiation factor2alpha[J]. TheJournal of general virology,2010,91(Pt2):470-482.
[84] ASSARSSON E, GREENBAUM JA, SUNDSTROM M, et al. Kineticanalysis of a complete poxvirus transcriptome reveals an immediate-early class ofgenes[J]. Proceedings of the National Academy of Sciences of the United States ofAmerica,2008,105(6):2140-2145.
[85] KECK JG, BALDICK CJ, JR., MOSS B. Role of DNA replication invaccinia virus gene expression: a naked template is required for transcription of threelate trans-activator genes[J]. Cell,1990,61(5):801-809.
[86] PERKUS ME, GOEBEL SJ, DAVIS SW, et al. Vaccinia virus host rangegenes[J]. Virology,1990,179(1):276-286.
[87] HSIAO JC, CHUNG CS, DRILLIEN R, CHANG W. The cowpox virus hostrange gene, CP77, affects phosphorylation of eIF2alpha and vaccinia viral translationin apoptotic HeLa cells[J]. Virology,2004,329(1):199-212.
[88] NAJERA JL, GOMEZ CE, DOMINGO-GIL E, GHERARDI MM,ESTEBAN M. Cellular and biochemical differences between two attenuated poxvirusvaccine candidates (MVA and NYVAC) and role of the C7L gene[J]. Journal ofvirology,2006,80(12):6033-6047.
[89] GOMEZ CE, NAJERA JL, KRUPA M, ESTEBAN M. The poxvirus vectorsMVA and NYVAC as gene delivery systems for vaccination against infectiousdiseases and cancer[J]. Current gene therapy,2008,8(2):97-120.
[90] SUTTER G, RAMSEY-EWING A, ROSALES R, MOSS B. Stableexpression of the vaccinia virus K1L gene in rabbit cells complements the host rangedefect of a vaccinia virus mutant[J]. Journal of virology,1994,68(7):4109-4116.
[91] GARCIA MA, MEURS EF, ESTEBAN M. The dsRNA protein kinase PKR:virus and cell control[J]. Biochimie,2007,89(6-7):799-811.
[92] DAVIES MV, CHANG HW, JACOBS BL, KAUFMAN RJ. The E3L andK3L vaccinia virus gene products stimulate translation through inhibition of thedouble-stranded RNA-dependent protein kinase by different mechanisms[J]. Journalof virology,1993,67(3):1688-1692.
[93] MENG X, CHAO J, XIANG Y. Identification from diverse mammalianpoxviruses of host-range regulatory genes functioning equivalently to vaccinia virusC7L[J]. Virology,2008,372(2):372-383.
[94] MENG X, JIANG C, ARSENIO J, et al. Vaccinia virus K1L and C7L inhibitantiviral activities induced by type I interferons[J]. Journal of virology,2009,83(20):10627-10636.
[95] UNGER B, NICHOLS RJ, STANITSA ES, TRAKTMAN P. Functionalcharacterization of the vaccinia virus I5protein[J]. Virology journal,2008,5:148.
[96] CHIU WL, LIN CL, YANG MH, TZOU DL, CHANG W. Vaccinia virus4c(A26L) protein on intracellular mature virus binds to the extracellular cellular matrixlaminin[J]. Journal of virology,2007,81(5):2149-2157.
[97] CHING YC, CHUNG CS, HUANG CY, et al. Disulfide bond formation atthe C termini of vaccinia virus A26and A27proteins does not require viral redoxenzymes and suppresses glycosaminoglycan-mediated cell fusion[J]. Journal ofvirology,2009,83(13):6464-6476.
[98] CHANG SJ, CHANG YX, IZMAILYAN R, TANG YL, CHANG W.Vaccinia virus A25and A26proteins are fusion suppressors for mature virions anddetermine strain-specific virus entry pathways into HeLa, CHO-K1, and L cells[J]. JVirol,2010,84(17):8422-8432.
[99] HOU YT, YANG XK, HU YW. Variation in the HindIII restriction fragmentsof DNA from the Chinese Tian Tan strain of vaccinia virus[J]. The Journal of generalvirology,1985,66(Pt8):1819-1823.
[100] SMITH CA, HU FQ, SMITH TD, et al. Cowpox virus genome encodes asecond soluble homologue of cellular TNF receptors, distinct from CrmB, that bindsTNF but not LT alpha[J]. Virology,1996,223(1):132-147.
[101] SCHREIBER M, RAJARATHNAM K, MCFADDEN G. Myxoma virus T2protein, a tumor necrosis factor (TNF) receptor homolog, is secreted as a monomerand dimer that each bind rabbit TNFalpha, but the dimer is a more potent TNFinhibitor[J]. The Journal of biological chemistry,1996,271(23):13333-13341.
[102] MOLOUGHNEY JG, MONKEN CE, TAO H, et al. Vaccinia virus leads toATG12-ATG3conjugation and deficiency in autophagosome formation[J]. Autophagy,2011,7(12):1434-1447.
[103] CHEN RA, RYZHAKOV G, COORAY S, RANDOW F, SMITH GL.Inhibition of IkappaB kinase by vaccinia virus virulence factor B14[J]. PLoSpathogens,2008,4(2): e22.
[104] BENFIELD CT, MANSUR DS, MCCOY LE, et al. Mapping the IkappaBkinase beta (IKKbeta)-binding interface of the B14protein, a vaccinia virus inhibitorof IKKbeta-mediated activation of nuclear factor kappaB[J]. The Journal of biologicalchemistry,2011,286(23):20727-20735.
[105] YU E, ZHAI D, JIN C, et al. Structural determinants of caspase-9inhibitionby the vaccinia virus protein, F1L[J]. The Journal of biological chemistry,2011,286(35):30748-30758.
[106] GERLIC M, FAUSTIN B, POSTIGO A, et al. Vaccinia virus F1L proteinpromotes virulence by inhibiting inflammasome activation[J]. Proceedings of theNational Academy of Sciences of the United States of America,2013,110(19):7808-7813.
[107] BENFIELD CT, REN H, LUCAS SJ, BAHSOUN B, SMITH GL. Vacciniavirus protein K7is a virulence factor that alters the acute immune response toinfection[J]. The Journal of general virology, Online in5April2013
[108] KIM MK, BREITBACH CJ, MOON A, et al. Oncolytic andimmunotherapeutic vaccinia induces antibody-mediated complement-dependentcancer cell lysis in humans[J]. Science translational medicine,2013,5(185):185ra163.
[109] MOORE ZS, SEWARD JF, LANE JM. Smallpox[J]. Lancet,2006,367(9508):425-435.
[110] WHO recommendations concerning the distribution, handling and synthesisof variola virus DNA, May2008[J]. Releve epidemiologique hebdomadaire/Sectiond'hygiene du Secretariat de la Societe des Nations=Weekly epidemiological record/Health Section of the Secretariat of the League of Nations,2008,83(44):393-395.
[111] BREMAN JG, ARITA I, FENNER F. Preventing the return of smallpox[J].The New England journal of medicine,2003,348(5):463-466.
[112] LANE JM, MILLAR JD. Risks of smallpox vaccination complications inthe United States[J]. American journal of epidemiology,1971,93(4):238-240.
[113] LANE JM, MILLAR JD, NEFF JM. Smallpox and smallpox vaccinationpolicy[J]. Annual review of medicine,1971,22:251-272.
[114] NELL P, KOHL KS, GRAHAM PL, et al. Progressive vaccinia as anadverse event following exposure to vaccinia virus: case definition and guidelines ofdata collection, analysis, and presentation of immunization safety data[J]. Vaccine,2007,25(31):5735-5744.
[115] VELLOZZI C, LANE JM, AVERHOFF F, et al. Generalized vaccinia,progressive vaccinia, and eczema vaccinatum are rare following smallpox (vaccinia)vaccination: United States surveillance,2003[J]. Clin Infect Dis,2005,41(5):689-697.
[116] KEMPER AR, DAVIS MM, FREED GL. Expected adverse events in amass smallpox vaccination campaign[J]. Eff Clin Pract,2002,5(2):84-90.
[117] CONO J, CASEY CG, BELL DM. Smallpox vaccination and adversereactions. Guidance for clinicians[J]. MMWR Recomm Rep,2003,52(RR-4):1-28.
[118] F. FENNER DAH, I. ARITA, Z. JEZEK, AND I. D. LADNYI,. Smallpoxand Its Eradication. World Health Organization: Geneva, Switzerland1988.
[119] GREENBERG RN, KENNEDY JS. ACAM2000: a newly licensed cellculture-based live vaccinia smallpox vaccine[J]. Expert opinion on investigationaldrugs,2008,17(4):555-564.
[120] JACOBS BL, LANGLAND JO, KIBLER KV, et al. Vaccinia virus vaccines:past, present and future[J]. Antiviral research,2009,84(1):1-13.
[121] DOWNIE AW, MC CK. The antibody response in man following infectionwith viruses of the pox group. III. Antibody response in smallpox[J]. The Journal ofhygiene,1958,56(4):479-487.
[122] BELSHE RB, NEWMAN FK, FREY SE, et al. Dose-dependentneutralizing-antibody responses to vaccinia[J]. The Journal of infectious diseases,2004,189(3):493-497.
[123] COUCH RB, WINOKUR P, EDWARDS KM, et al. Reducing the dose ofsmallpox vaccine reduces vaccine-associated morbidity without reducing vaccinationsuccess rates or immune responses[J]. The Journal of infectious diseases,2007,195(6):826-832.
[124] MARRIOTT KA, PARKINSON CV, MOREFIELD SI, et al. Clonalvaccinia virus grown in cell culture fully protects monkeys from lethal monkeypoxchallenge[J]. Vaccine,2008,26(4):581-588.
[125] ARTENSTEIN AW, JOHNSON C, MARBURY TC, et al. A novel, cellculture-derived smallpox vaccine in vaccinia-naive adults[J]. Vaccine,2005,23(25):3301-3309.
[126] OSBORNE JD, DA SILVA M, FRACE AM, et al. Genomic differences ofVaccinia virus clones from Dryvax smallpox vaccine: the Dryvax-like ACAM2000and the mouse neurovirulent Clone-3[J]. Vaccine,2007,25(52):8807-8832.
[127] JANG HC, KIM CJ, KIM KH, et al. A randomized, double-blind,controlled clinical trial to evaluate the efficacy and safety of CJ-50300, a newlydeveloped cell culture-derived smallpox vaccine, in healthy volunteers[J]. Vaccine,2010,28(36):5845-5849.
[128] WELTZIN R, LIU J, PUGACHEV KV, et al. Clonal vaccinia virus grownin cell culture as a new smallpox vaccine[J]. Nature medicine,2003,9(9):1125-1130.
[129] KENNER J, CAMERON F, EMPIG C, JOBES DV, GURWITH M.LC16m8: an attenuated smallpox vaccine[J]. Vaccine,2006,24(47-48):7009-7022.
[130] SAITO T, FUJII T, KANATANI Y, et al. Clinical and immunologicalresponse to attenuated tissue-cultured smallpox vaccine LC16m8[J]. Jama,2009,301(10):1025-1033.
[131] VOLLMAR J, ARNDTZ N, ECKL KM, et al. Safety and immunogenicityof IMVAMUNE, a promising candidate as a third generation smallpox vaccine[J].Vaccine,2006,24(12):2065-2070.
[132] MEISINGER-HENSCHEL C, SCHMIDT M, LUKASSEN S, et al.Genomic sequence of chorioallantois vaccinia virus Ankara, the ancestor of modifiedvaccinia virus Ankara[J]. The Journal of general virology,2007,88(Pt12):3249-3259.
[133] WILCK MB, SEAMAN MS, BADEN LR, et al. Safety andimmunogenicity of modified vaccinia Ankara (ACAM3000): effect of dose and routeof administration[J]. The Journal of infectious diseases,2010,201(9):1361-1370.
[134] SPEERS WC, WESLEY RB, NEFF JM, GOLDSTEIN J, LOURIE B.Evaluation of two kinds of smallpox vaccine: CVI-78and calf lymph vaccine. II.Clinical and serologic observations of response to revaccination with calf lymphvaccine[J]. Pediatric research,1975,9(8):628-632.
[135] MONATH TP, CALDWELL JR, MUNDT W, et al. ACAM2000clonalVero cell culture vaccinia virus (New York City Board of Health strain)--asecond-generation smallpox vaccine for biological defense[J]. Int J Infect Dis,2004,8Suppl2: S31-44.
[136] HATCH GJ, GRAHAM VA, BEWLEY KR, et al. Assessment of theprotective effect of IMVAMUNE(R) and ACAM2000(R) vaccines against aerosolisedMonkeypox virus in cynomolgus macaques[J]. J Virol, on line May2013.
[137] FREY SE, NEWMAN FK, KENNEDY JS, et al. Comparison of the safetyand immunogenicity of ACAM1000, ACAM2000and Dryvax in healthyvaccinia-naive adults[J]. Vaccine,2009,27(10):1637-1644.
[138] GREENBERG RN, KENNEDY JS, CLANTON DJ, et al. Safety andimmunogenicity of new cell-cultured smallpox vaccine compared with calf-lymphderived vaccine: a blind, single-centre, randomised controlled trial[J]. Lancet,2005,365(9457):398-409.
[139] POLAND GA, GRABENSTEIN JD, NEFF JM. The US smallpoxvaccination program: a review of a large modern era smallpox vaccinationimplementation program[J]. Vaccine,2005,23(17-18):2078-2081.
[140] BEACHKOFSKY TM, CARRIZALES SC, BIDINGER JJ, et al. Adverseevents following smallpox vaccination with ACAM2000in a military population[J].Archives of dermatology,2010,146(6):656-661.
[141] SARKAR JK, MITRA AC, MUKHERJEE MK. The minimum protectivelevel of antibodies in smallpox[J]. Bulletin of the World Health Organization,1975,52(3):307-311.
[142] FREY SE, COUCH RB, TACKET CO, et al. Clinical responses toundiluted and diluted smallpox vaccine[J]. The New England journal of medicine,2002,346(17):1265-1274.
[143] MCCLAIN DJ, HARRISON S, YEAGER CL, et al. Immunologicresponses to vaccinia vaccines administered by different parenteral routes[J]. TheJournal of infectious diseases,1997,175(4):756-763.
[144] KIM SH, CHOI SJ, PARK WB, et al. Detailed kinetics of immuneresponses to a new cell culture-derived smallpox vaccine in vaccinia-naive adults[J].Vaccine,2007,25(33):6287-6291.
[145] MESEDA CA, MAYER AE, KUMAR A, et al. Comparative evaluation ofthe immune responses and protection engendered by LC16m8and Dryvax smallpoxvaccines in a mouse model[J]. Clin Vaccine Immunol,2009,16(9):1261-1271.
[146] KEMPE CH, FULGINITI V, MINAMITANI M, SHINEFIELD H.Smallpox vaccination of eczema patients with a strain of attenuated live vaccinia(CVI-78)[J]. Pediatrics,1968,42(6):980-985.
[147] WESLEY RB, SPEERS WC, NEFF JM, RUBEN FL, LOURIE B.Evaluation of two kinds of smallpox vaccine: CVI-78and calf lymph vaccine. I.Clinical and serologic response to primary vaccination[J]. Pediatric research,1975,9(8):624-628.
[148] MEYER H, SUTTER G, MAYR A. Mapping of deletions in the genome ofthe highly attenuated vaccinia virus MVA and their influence on virulence[J]. TheJournal of general virology,1991,72(Pt5):1031-1038.
[149] STITTELAAR KJ, OSTERHAUS AD. MVA: a cuckoo in the vaccinenest?[J]. Vaccine,2001,19(27): V-VI.
[150] KENNEDY JS, GREENBERG RN. IMVAMUNE: modified vacciniaAnkara strain as an attenuated smallpox vaccine[J]. Expert review of vaccines,2009,8(1):13-24.
[151] FREY SE, NEWMAN FK, KENNEDY JS, et al. Clinical and immunologicresponses to multiple doses of IMVAMUNE (Modified Vaccinia Ankara) followed byDryvax challenge[J]. Vaccine,2007,25(51):8562-8573.
[152] PARRINO J, MCCURDY LH, LARKIN BD, et al. Safety, immunogenicityand efficacy of modified vaccinia Ankara (MVA) against Dryvax challenge invaccinia-naive and vaccinia-immune individuals[J]. Vaccine,2007,25(8):1513-1525.
[153] PERKUS ME, TARTAGLIA J, PAOLETTI E. Poxvirus-based vaccinecandidates for cancer, AIDS, and other infectious diseases[J]. Journal of leukocytebiology,1995,58(1):1-13.
[154] TYKODI SS, THOMPSON JA. Development of modified vacciniaAnkara-5T4as specific immunotherapy for advanced human cancer[J]. Expertopinion on biological therapy,2008,8(12):1947-1953.
[155] SEAMAN MS, WILCK MB, BADEN LR, et al. Effect of vaccination withmodified vaccinia Ankara (ACAM3000) on subsequent challenge with Dryvax[J].The Journal of infectious diseases,2010,201(9):1353-1360.
[156] MACK TM, NOBLE J, JR., THOMAS DB. A prospective study of serumantibody and protection against smallpox[J]. The American journal of tropicalmedicine and hygiene,1972,21(2):214-218.
[157] RAMANATHAN VD, KUMAR M, MAHALINGAM J, et al. A Phase1study to evaluate the safety and immunogenicity of a recombinant HIV type1subtypeC-modified vaccinia Ankara virus vaccine candidate in Indian volunteers[J]. AIDSresearch and human retroviruses,2009,25(11):1107-1116.
[158] CURRIER JR, NGAUY V, DE SOUZA MS, et al. Phase I safety andimmunogenicity evaluation of MVA-CMDR, a multigenic, recombinant modifiedvaccinia Ankara-HIV-1vaccine candidate[J]. PloS one,2010,5(11): e13983.
[159] MWAU M, CEBERE I, SUTTON J, et al. A human immunodeficiencyvirus1(HIV-1) clade A vaccine in clinical trials: stimulation of HIV-specific T-cellresponses by DNA and recombinant modified vaccinia virus Ankara (MVA) vaccinesin humans[J]. The Journal of general virology,2004,85(Pt4):911-919.
[160] MCCORMACK S, STOHR W, BARBER T, et al. EV02: a Phase I trial tocompare the safety and immunogenicity of HIV DNA-C prime-NYVAC-C boost toNYVAC-C alone[J]. Vaccine,2008,26(25):3162-3174.
[161] AMATO RJ, HAWKINS RE, KAUFMAN HL, et al. Vaccination ofmetastatic renal cancer patients with MVA-5T4: a randomized, double-blind,placebo-controlled phase III study[J]. Clin Cancer Res,2010,16(22):5539-5547.
[162] GARCIA-HERNANDEZ E, GONZALEZ-SANCHEZ JL,ANDRADE-MANZANO A, et al. Regression of papilloma high-grade lesions (CIN2and CIN3) is stimulated by therapeutic vaccination with MVA E2recombinantvaccine[J]. Cancer gene therapy,2006,13(6):592-597.
[163] ARLEN PM, PAZDUR M, SKARUPA L, RAUCKHORST M, GULLEYJL. A randomized phase II study of docetaxel alone or in combination withPANVAC-V (vaccinia) and PANVAC-F (fowlpox) in patients with metastatic breastcancer (NCI05-C-0229)[J]. Clinical breast cancer,2006,7(2):176-179.
[164] ZEMP FJ, CORREDOR JC, LUN X, MURUVE DA, FORSYTH PA.Oncolytic viruses as experimental treatments for malignant gliomas: using a scourgeto treat a devil[J]. Cytokine&growth factor reviews,2010,21(2-3):103-117.
[165] PARK BH, HWANG T, LIU TC, et al. Use of a targeted oncolytic poxvirus,JX-594, in patients with refractory primary or metastatic liver cancer: a phase I trial[J].The lancet oncology,2008,9(6):533-542.
[166] FENNER FH, DA.; ARITA, I.; JEZEK, Z.; LADNYI, I. Smallpox and itsEradication. World Health Organization. Geneva, Switzerland1988.
[167] PANICALI D, PAOLETTI E. Construction of poxviruses as cloning vectors:insertion of the thymidine kinase gene from herpes simplex virus into the DNA ofinfectious vaccinia virus[J]. Proceedings of the National Academy of Sciences of theUnited States of America,1982,79(16):4927-4931.
[168] JOHNSTON JB, MCFADDEN G. Technical knockout: understandingpoxvirus pathogenesis by selectively deleting viral immunomodulatory genes[J].Cellular microbiology,2004,6(8):695-705.
[169] PASTORET PP, BROCHIER B. The development and use of avaccinia-rabies recombinant oral vaccine for the control of wildlife rabies; a linkbetween Jenner and Pasteur[J]. Epidemiology and infection,1996,116(3):235-240.
[170] HANLON CA, NIEZGODA M, HAMIR AN, et al. First North Americanfield release of a vaccinia-rabies glycoprotein recombinant virus[J]. Journal ofwildlife diseases,1998,34(2):228-239.
[171] SLATE D, ALGEO TP, NELSON KM, et al. Oral rabies vaccination innorth america: opportunities, complexities, and challenges[J]. PLoS neglected tropicaldiseases,2009,3(12): e549.
[172] KANTOFF PW, SCHUETZ TJ, BLUMENSTEIN BA, et al. Overallsurvival analysis of a phase II randomized controlled trial of a Poxviral-basedPSA-targeted immunotherapy in metastatic castration-resistant prostate cancer[J]. JClin Oncol,2010,28(7):1099-1105.
[173] SUTTER G, MOSS B. Nonreplicating vaccinia vector efficiently expressesrecombinant genes[J]. Proceedings of the National Academy of Sciences of the UnitedStates of America,1992,89(22):10847-10851.
[174] AMATO RJ.5T4-modified vaccinia Ankara: progress in tumor-associatedantigen-based immunotherapy[J]. Expert opinion on biological therapy,2010,10(2):281-287.
[175] TARTAGLIA J, PERKUS ME, TAYLOR J, et al. NYVAC: a highlyattenuated strain of vaccinia virus[J]. Virology,1992,188(1):217-232.
[176] OCKENHOUSE CF, SUN PF, LANAR DE, et al. Phase I/IIa safety,immunogenicity, and efficacy trial of NYVAC-Pf7, a pox-vectored, multiantigen,multistage vaccine candidate for Plasmodium falciparum malaria[J]. The Journal ofinfectious diseases,1998,177(6):1664-1673.
[177] BART PA, GOODALL R, BARBER T, et al. EV01: a phase I trial inhealthy HIV negative volunteers to evaluate a clade C HIV vaccine, NYVAC-Cundertaken by the EuroVacc Consortium[J]. Vaccine,2008,26(25):3153-3161.
[178] SHEPPARD NC, BATES AC, SATTENTAU QJ. A functional human IgMresponse to HIV-1Env after immunization with NYVAC HIV C[J]. AIDS (London,England),2007,21(4):524-527.
[179] GOMEZ CE, NAJERA JL, JIMENEZ EP, et al. Head-to-head comparisonon the immunogenicity of two HIV/AIDS vaccine candidates based on the attenuatedpoxvirus strains MVA and NYVAC co-expressing in a single locus the HIV-1BX08gp120and HIV-1(IIIB) Gag-Pol-Nef proteins of clade B[J]. Vaccine,2007,25(15):2863-2885.
[180] NAJERA JL, GOMEZ CE, GARCIA-ARRIAZA J, SORZANO CO,ESTEBAN M. Insertion of vaccinia virus C7L host range gene into NYVAC-Bgenome potentiates immune responses against HIV-1antigens[J]. PloS one,2010,5(6): e11406.
[181] WHO. Global eradication of smallpox: WHO Global Commission for theCertification od Smallpox Eradication.[J]. J Med Assoc Thai,1979,62(8):461.
[182]娄元梅,阮力,朱既明.痘苗病毒毒力相关基因的研究进展[J].病毒学报,1996,12(1):88-96.
[183] PERDIGUERO B, ESTEBAN M. The interferon system and vaccinia virusevasion mechanisms[J]. J Interferon Cytokine Res,2009,29(9):581-598.
[184] GIRGIS NM, DEHAVEN BC, XIAO Y, et al. The Vaccinia viruscomplement control protein modulates adaptive immune responses during infection[J].J Virol,2011,85(6):2547-2556.
[185] RAMSEY-EWING A, MOSS B. Restriction of vaccinia virus replication inCHO cells occurs at the stage of viral intermediate protein synthesis[J]. Virology,1995,206(2):984-993.
[186] HINTHONG O, JIN XL, SHISLER JL. Characterization of wild-type andmutant vaccinia virus M2L proteins' abilities to localize to the endoplasmic reticulumand to inhibit NF-kappaB activation during infection[J]. Virology,2008,373(2):248-262.
[187] WITTEK R, MULLER HK, WYLER R. Length heterogeneity in the DNAof vaccinia virus is eliminated on cloning the virus[J]. FEBS letters,1978,90(1):41-46.
[188] BULLER RM, SMITH GL, CREMER K, NOTKINS AL, MOSS B.Decreased virulence of recombinant vaccinia virus expression vectors is associatedwith a thymidine kinase-negative phenotype[J]. Nature,1985,317(6040):813-815.
[189] KOTWAL GJ, MOSS B. Analysis of a large cluster of nonessential genesdeleted from a vaccinia virus terminal transposition mutant[J]. Virology,1988,167(2):524-537.
[190] BARTLETT N, SYMONS JA, TSCHARKE DC, SMITH GL. The vacciniavirus N1L protein is an intracellular homodimer that promotes virulence[J]. TheJournal of general virology,2002,83(Pt8):1965-1976.
[191] KOTWAL GJ, HUGIN AW, MOSS B. Mapping and insertionalmutagenesis of a vaccinia virus gene encoding a13,800-Da secreted protein[J].Virology,1989,171(2):579-587.
[192] AOYAGI M, ZHAI D, JIN C, et al. Vaccinia virus N1L protein resembles aB cell lymphoma-2(Bcl-2) family protein[J]. Protein Sci,2007,16(1):118-124.
[193] SEET BT, JOHNSTON JB, BRUNETTI CR, et al. Poxviruses and immuneevasion[J]. Annual review of immunology,2003,21:377-423.
[194] TAYLOR JM, BARRY M. Near death experiences: poxvirus regulation ofapoptotic death[J]. Virology,2006,344(1):139-150.
[195] GONZALEZ JM, ESTEBAN M. A poxvirus Bcl-2-like gene familyinvolved in regulation of host immune response: sequence similarity and evolutionaryhistory[J]. Virology journal,2010,7:59.
[196] BOWIE A, KISS-TOTH E, SYMONS JA, et al. A46R and A52R fromvaccinia virus are antagonists of host IL-1and toll-like receptor signaling[J].Proceedings of the National Academy of Sciences of the United States of America,2000,97(18):10162-10167.
[197] GRAHAM SC, BAHAR MW, COORAY S, et al. Vaccinia virus proteinsA52and B14Share a Bcl-2-like fold but have evolved to inhibit NF-kappaB ratherthan apoptosis[J]. PLoS pathogens,2008,4(8): e1000128.
[198] ZHANG L, VILLA NY, RAHMAN MM, et al. Analysis of vacciniavirus-host protein-protein interactions: validations of yeast two-hybrid screenings[J].Journal of proteome research,2009,8(9):4311-4318.
[199] OSEROFF C, PETERS B, PASQUETTO V, et al. Dissociation betweenepitope hierarchy and immunoprevalence in CD8responses to vaccinia virus westernreserve[J]. J Immunol,2008,180(11):7193-7202.
[200] STETSON DB, MEDZHITOV R. Type I interferons in host defense[J].Immunity,2006,25(3):373-381.
[201] SADLER AJ, WILLIAMS BR. Interferon-inducible antiviral effectors[J].Nature reviews,2008,8(7):559-568.
[202] UNTERHOLZNER L, SUMNER RP, BARAN M, et al. Vaccinia virusprotein C6is a virulence factor that binds TBK-1adaptor proteins and inhibitsactivation of IRF3and IRF7[J]. PLoS pathogens,2011,7(9): e1002247.
[203] GARCIA-ARRIAZA J, NAJERA JL, GOMEZ CE, et al. A candidateHIV/AIDS vaccine (MVA-B) lacking vaccinia virus gene C6L enhances memoryHIV-1-specific T-cell responses[J]. PloS one,2011,6(8): e24244.
[204] HAGA IR, BOWIE AG. Evasion of innate immunity by vaccinia virus[J].Parasitology,2005,130Suppl: S11-25.
[205] BILLINGS B, SMITH SA, ZHANG Z, LAHIRI DK, KOTWAL GJ. Lackof N1L gene expression results in a significant decrease of vaccinia virus replicationin mouse brain[J]. Annals of the New York Academy of Sciences,2004,1030:297-302.
[206] SMITH KA, STALLARD V, ROOS JM, et al. Host range selection ofvaccinia recombinants containing insertions of foreign genes into non-codingsequences[J]. Vaccine,1993,11(1):43-53.
[207] SHCHELKUNOV S, TOTMENIN A, KOLOSOVA I. Species-specificdifferences in organization of orthopoxvirus kelch-like proteins[J]. Virus genes,2002,24(2):157-162.
[208] PIRES DE MIRANDA M, READING PC, TSCHARKE DC, MURPHY BJ,SMITH GL. The vaccinia virus kelch-like protein C2L affects calcium-independentadhesion to the extracellular matrix and inflammation in a murine intradermalmodel[J]. The Journal of general virology,2003,84(Pt9):2459-2471.
[209] SHIMSHEK DR, KIM J, HUBNER MR, et al. Codon-improved Crerecombinase (iCre) expression in the mouse[J]. Genesis,2002,32(1):19-26.
[210] POMERANTSEV AP, SITARAMAN R, GALLOWAY CR, KIVOVICH V,LEPPLA SH. Genome engineering in Bacillus anthracis using Cre recombinase[J].Infection and immunity,2006,74(1):682-693.
[211]贺淹才.简明基因工程原理[M].科学出版社:北京2005.
[212]邹艳,阮力,娄元梅,朱诚.痘苗病毒天坛株非必需区TA25R…TA36L的两步重组表达载体的构建[J].病毒学报,1998,14(2):97-103.
[213] SCHEIFLINGER F, DORNER F, FALKNER FG. Transient markerstabilisation: a general procedure to construct marker-free recombinant vacciniavirus[J]. Archives of virology,1998,143(3):467-474.
[214]周小云,刘颖,饶力群,王芙,戴凯凡等.适用于疫苗株筛选的痘苗病毒载体的构建[J].病毒学报,2004,20(4):315-321.
[215]房有荣,阎辉.以GFP-Zeocin为筛选标志的重组痘苗病毒载体质粒的构建.《2011年浙江省医学会医学病毒学分会、医学微生物与免疫学分会学术年会论文汇编》2011.
[216] CUBITT AB, HEIM R, ADAMS SR, et al. Understanding, improving andusing green fluorescent proteins[J]. Trends in biochemical sciences,1995,20(11):448-455.
[217] HIKICHI M, KIDOKORO M, HARAGUCHI T, et al. MicroRNAregulation of glycoprotein B5R in oncolytic vaccinia virus reduces viral pathogenicitywithout impairing its antitumor efficacy[J]. Mol Ther,2011,19(6):1107-1115.
[218] FAHY AS, CLARK RH, GLYDE EF, SMITH GL. Vaccinia virus proteinC16acts intracellularly to modulate the host response and promote virulence[J]. TheJournal of general virology,2008,89(Pt10):2377-2387.
[219] MOSS B. Replicating and host-restricted non-replicating vaccinia virusvectors for vaccine development[J]. Developments in biological standardization,1994,82:55-63.
[220] KUMAR S, BOYLE DB. A poxvirus bidirectional promoter element withearly/late and late functions[J]. Virology,1990,179(1):151-158.
[221] BOYLE DB. Quantitative assessment of poxvirus promoters in fowlpoxand vaccinia virus recombinants[J]. Virus genes,1992,6(3):281-290.
[222]朱自满,孙念绪,张超,吴淼.应用Cre/loxP系统构建含人PTEN基因的重组腺病毒载体[J].消化外科,2005,4(2):124-127.
[223]王艳春,姜娜,展德文,邱炎等. Cre-LoxP系统在炭疽芽胞杆菌基因敲除中的应用及eag基因的敲除[J].生物化学与生物物理进展,2009:1-10.
[224] HOESS RH, ZIESE M, STERNBERG N. P1site-specific recombination:nucleotide sequence of the recombining sites[J]. Proceedings of the NationalAcademy of Sciences of the United States of America,1982,79(11):3398-3402.
[225] KOTWAL GJ, MOSS B. Vaccinia virus encodes a secretory polypeptidestructurally related to complement control proteins[J]. Nature,1988,335(6186):176-178.
[226] GEDEY R, JIN XL, HINTHONG O, SHISLER JL. Poxviral regulation ofthe host NF-kappaB response: the vaccinia virus M2L protein inhibits induction ofNF-kappaB activation via an ERK2pathway in virus-infected human embryonickidney cells[J]. Journal of virology,2006,80(17):8676-8685.
[227]殷震,刘景华.动物病毒学（第二版）.北京：科学出版社1997.
[228] REED LJ, MUENCH, H. A simple method of estimating fifty percentendpoints[J]. Am J Hyg,1938,27:493–497.
[229] FULGINITI VA, PAPIER A, LANE JM, NEFF JM, HENDERSON DA.Smallpox vaccination: a review, part II. Adverse events[J]. Clin Infect Dis,2003,37(2):251-271.
[230] GURVICH EB, VILESOVA IS. Vaccinia virus in postvaccinalencephalitis[J]. Acta virologica,1983,27(2):154-159.
[231] HENDERSON DA MB. Smallpox and vacciniaIn: Plotkin SA, OrensteinWA, editors. Vaccines. Philadelphia: W.B. Saunders Company.1999:74-97.
[232] KUTINOVA L, LUDVIKOVA V, SIMONOVA V, et al. Search for optimalparent for recombinant vaccinia virus vaccines. Study of three vaccinia virus vaccinalstrains and several virus lines derived from them[J]. Vaccine,1995,13(5):487-493.
[233] CARROLL MW, MOSS B. Host range and cytopathogenicity of the highlyattenuated MVA strain of vaccinia virus: propagation and generation of recombinantviruses in a nonhuman mammalian cell line[J]. Virology,1997,238(2):198-211.
[234] YU W, FANG Q, ZHU W, et al. One time intranasal vaccination with amodified vaccinia Tiantan strain MVTT(ZCI) protects animals against pathogenicviral challenge[J]. Vaccine,2010,28(9):2088-2096.
[235] TURNER PC, MOYER RW. Orthopoxvirus fusion inhibitor glycoproteinSPI-3(open reading frame K2L) contains motifs characteristic of serine proteinaseinhibitors that are not required for control of cell fusion[J]. J Virol,1995,69(10):5978-5987.
[236] LUDVIKOVA V, KUTINOVA L, SIMONOVA V, OTAVOVA M.Evaluation of various virulence tests with low virulence vaccinia virus in mice[J].Biologicals,1994,22(2):187-190.
[237] LI G, CHEN N, FENG Z, et al. Genomic sequence and analysis of avaccinia virus isolate from a patient with a smallpox vaccine-related complication[J].Virology journal,2006,3:88.
[238]董树林.天坛株痘苗毒种创始人齐长庆[J].微生物学及免疫学进展,2009,37(3):1-3.
[239]金奇,陈南海,陈淑霞,黄静,冯泽华等.痘苗病毒天坛株全基因组结构特点的分析[J].中国科学(C辑),1997,27(6):563-567.
[240] MARIES VAN DEN BROEK MFB, GABRIELE KO¨HLER, MARIJKEBARNER. IL-4and IL-10Antagonize IL-12-Mediated Protection Against AcuteVaccinia Virus Infection with a Limited Role of IFN-g and Nitric Oxide Synthetase2[J]. The Journal of Immunology,2000,164:371–378.
[241] BORN TL, MORRISON LA, ESTEBAN DJ, et al. A poxvirus protein thatbinds to and inactivates IL-18, and inhibits NK cell response[J]. J Immunol,2000,164(6):3246-3254.
[242] ANDERSON KP, LIE YS, LOW MA, FENNIE EH. Effects of tumornecrosis factor-alpha treatment on mortality in murine cytomegalovirus-infectedmice[J]. Antiviral research,1993,21(4):343-355.
[243] ZINKERNAGEL RM. Immunology taught by viruses[J]. Science (NewYork, NY,1996,271(5246):173-178.
[244] MALONEY G, SCHRODER M, BOWIE AG. Vaccinia virus protein A52Ractivates p38mitogen-activated protein kinase and potentiateslipopolysaccharide-induced interleukin-10[J]. The Journal of biological chemistry,2005,280(35):30838-30844.
[245] KURILLA MG, SWAMINATHAN S, WELSH RM, KIEFF E,BRUTKIEWICZ RR. Effects of virally expressed interleukin-10on vaccinia virusinfection in mice[J]. Journal of virology,1993,67(12):7623-7628.
[246] VAN DEN BROEK M, BACHMANN MF, KOHLER G, et al. IL-4andIL-10antagonize IL-12-mediated protection against acute vaccinia virus infectionwith a limited role of IFN-gamma and nitric oxide synthetase2[J]. J Immunol,2000,164(1):371-378.
[247] IAN A. RAMSHAW MEA, SUSAN M. PHILLIPS, DAVID B. BOYL.Recovery of immunodeficient mice from a vaccinia virus/IL-2recombinantinfection[J]. Nature,1987,329:545-546.
[248] KIDOKORO M, TASHIRO M, SHIDA H. Genetically stable and fullyeffective smallpox vaccine strain constructed from highly attenuated vacciniaLC16m8[J]. Proceedings of the National Academy of Sciences of the United States ofAmerica,2005,102(11):4152-4157.
[249] PHELPS AL, GATES AJ, HILLIER M, EASTAUGH L, ULAETO DO.Comparative efficacy of modified vaccinia Ankara (MVA) as a potential replacementsmallpox vaccine[J]. Vaccine,2007,25(1):34-42.
[250] BERHANU A, WILSON RL, KIRKWOOD-WATTS DL, et al. Vaccinationof BALB/c mice with Escherichia coli-expressed vaccinia virus proteins A27L, B5R,and D8L protects mice from lethal vaccinia virus challenge[J]. Journal of virology,2008,82(7):3517-3529.
[2] SEKIGUCHI S, KIMURA K, CHIYO T, et al. Immunization with arecombinant vaccinia virus that encodes nonstructural proteins of the hepatitis C virussuppresses viral protein levels in mouse liver[J]. PloS one,2012,7(12): e51656.
[3] FLYNN BJ, KASTENMULLER K, WILLE-REECE U, et al. Immunizationwith HIV Gag targeted to dendritic cells followed by recombinant New York vacciniavirus induces robust T-cell immunity in nonhuman primates[J]. Proceedings of theNational Academy of Sciences of the United States of America,2013,108(17):7131-7136.
[4] WOLLENBERG A, ENGLER R. Smallpox, vaccination and adverse reactionsto smallpox vaccine[J]. Current opinion in allergy and clinical immunology,2004,4(4):271-275.
[5] QIN L, LIANG M, EVANS DH. Genomic analysis of vaccinia virus strainTianTan provides new insights into the evolution and evolutionary relationshipsbetween Orthopoxviruses[J]. Virology, Available online19April2013.
[6] MOSS B. Genetically engineered poxviruses for recombinant gene expression,vaccination, and safety[J]. Proceedings of the National Academy of Sciences of theUnited States of America,1996,93(21):11341-11348.
[7] BOYD O, TURNER PC, MOYER RW, CONDIT RC, MOUSSATCHE N.The E6protein from vaccinia virus is required for the formation of immaturevirions[J]. Virology,2010,399(2):201-211.
[8] DOMS RW, BLUMENTHAL R, MOSS B. Fusion of intra-and extracellularforms of vaccinia virus with the cell membrane[J]. Journal of virology,1990,64(10):4884-4892.
[9] CUDMORE S, COSSART P, GRIFFITHS G, WAY M. Actin-based motility ofvaccinia virus[J]. Nature,1995,378(6557):636-638.
[10] DOCEUL V, HOLLINSHEAD M, VAN DER LINDEN L, SMITH GL.Repulsion of superinfecting virions: a mechanism for rapid virus spread[J]. Science(New York, NY,2010,327(5967):873-876.
[11] CARTER GC, LAW M, HOLLINSHEAD M, SMITH GL. Entry of thevaccinia virus intracellular mature virion and its interactions withglycosaminoglycans[J]. The Journal of general virology,2005,86(Pt5):1279-1290.
[12] VANDERPLASSCHEN A, HOLLINSHEAD M, SMITH GL. Intracellularand extracellular vaccinia virions enter cells by different mechanisms[J]. The Journalof general virology,1998,79(Pt4):877-887.
[13] HSIAO JC, CHUNG CS, CHANG W. Vaccinia virus envelope D8L proteinbinds to cell surface chondroitin sulfate and mediates the adsorption of intracellularmature virions to cells[J]. Journal of virology,1999,73(10):8750-8761.
[14] CHUNG CS, HSIAO JC, CHANG YS, CHANG W. A27L protein mediatesvaccinia virus interaction with cell surface heparan sulfate[J]. Journal of virology,1998,72(2):1577-1585.
[15] HSIAO JC, CHUNG CS, CHANG W. Cell surface proteoglycans arenecessary for A27L protein-mediated cell fusion: identification of the N-terminalregion of A27L protein as the glycosaminoglycan-binding domain[J]. Journal ofvirology,1998,72(10):8374-8379.
[16] RODRIGUEZ D, ESTEBAN M, RODRIGUEZ JR. Vaccinia virus A17Lgene product is essential for an early step in virion morphogenesis[J]. Journal ofvirology,1995,69(8):4640-4648.
[17] VAZQUEZ MI, RIVAS G, CREGUT D, SERRANO L, ESTEBAN M. Thevaccinia virus14-kilodalton (A27L) fusion protein forms a triple coiled-coil structureand interacts with the21-kilodalton (A17L) virus membrane protein through aC-terminal alpha-helix[J]. Journal of virology,1998,72(12):10126-10137.
[18] FOO CH, WHITBECK JC, PONCE-DE-LEON M, et al. The myristatemoiety and amino terminus of vaccinia virus l1constitute a bipartite functional regionneeded for entry[J]. Journal of virology,2012,86(10):5437-5451.
[19] CHAN WM, WARD BM. Increased interaction between vaccinia virusproteins A33and B5is detrimental to infectious extracellular enveloped virionproduction[J]. Journal of virology,2012,86(15):8232-8244.
[20] PEDERSEN K, SNIJDER EJ, SCHLEICH S, et al. Characterization ofvaccinia virus intracellular cores: implications for viral uncoating and corestructure[J]. Journal of virology,2000,74(8):3525-3536.
[21] ZHANG Y, KECK JG, MOSS B. Transcription of viral late genes isdependent on expression of the viral intermediate gene G8R in cells infected with aninducible conditional-lethal mutant vaccinia virus[J]. Journal of virology,1992,66(11):6470-6479.
[22] KECK JG, KOVACS GR, MOSS B. Overexpression, purification, and latetranscription factor activity of the17-kilodalton protein encoded by the vaccinia virusA1L gene[J]. Journal of virology,1993,67(10):5740-5748.
[23] HUBBS AE, WRIGHT CF. The A2L intermediate gene product is requiredfor in vitro transcription from a vaccinia virus late promoter[J]. Journal of virology,1996,70(1):327-331.
[24] LUTTGE BG, MOYER RW. Suppressors of a host range mutation in therabbitpox virus serpin SPI-1map to proteins essential for viral DNA replication[J].Journal of virology,2005,79(14):9168-9179.
[25] CURLEE KV, HONG JS, CLANCY JP, et al. Membrane transplantation tocorrect integral membrane protein defects[J]. Journal of molecular medicine (Berlin,Germany),2003,81(8):511-520.
[26] SODEIK B, KRIJNSE-LOCKER J. Assembly of vaccinia virus revisited: denovo membrane synthesis or acquisition from the host?[J]. Trends in microbiology,2002,10(1):15-24.
[27] RIETDORF J, PLOUBIDOU A, RECKMANN I, et al. Kinesin-dependentmovement on microtubules precedes actin-based motility of vaccinia virus[J]. Naturecell biology,2001,3(11):992-1000.
[28] CUDMORE S, BLASCO R, VINCENTELLI R, et al. A vaccinia virus coreprotein, p39, is membrane associated[J]. Journal of virology,1996,70(10):6909-6921.
[29] ZHU R, HUANG W, WANG W, et al. Comparison on virulence andimmunogenicity of two recombinant vaccinia vaccines, Tian Tan and Guang9strains,expressing the HIV-1envelope gene[J]. PloS one,2012,7(11): e48343.
[30] FANG Q, YANG L, ZHU W, et al. Host range, growth property, andvirulence of the smallpox vaccine: vaccinia virus Tian Tan strain[J]. Virology,2005,335(2):242-251.
[31] SUMNER RP, REN H, SMITH GL. Deletion of immunomodulator C6fromvaccinia virus strain Western Reserve enhances virus immunogenicity and vaccineefficacy[J]. The Journal of general virology,2013,94(Pt5):1121-1126.
[32] CLARK RH, KENYON JC, BARTLETT NW, TSCHARKE DC, SMITHGL. Deletion of gene A41L enhances vaccinia virus immunogenicity and vaccineefficacy[J]. The Journal of general virology,2006,87(Pt1):29-38.
[33] COTTINGHAM MG, ANDERSEN RF, SPENCER AJ, et al.Recombination-mediated genetic engineering of a bacterial artificial chromosomeclone of modified vaccinia virus Ankara (MVA)[J]. PloS one,2008,3(2): e1638.
[34] JENTARRA GM, HECK MC, YOUN JW, et al. Vaccinia viruses withmutations in the E3L gene as potential replication-competent, attenuated vaccines:scarification vaccination[J]. Vaccine,2008,26(23):2860-2872.
[35] STAIB C, KISLING S, ERFLE V, SUTTER G. Inactivation of the viralinterleukin1beta receptor improves CD8+T-cell memory responses elicited uponimmunization with modified vaccinia virus Ankara[J]. The Journal of general virology,2005,86(Pt7):1997-2006.
[36] VLIEGEN I, YANG G, HRUBY D, JORDAN R, NEYTS J. Deletion of thevaccinia virus F13L gene results in a highly attenuated virus that mounts a protectiveimmune response against subsequent vaccinia virus challenge[J]. Antiviral research,2012,93(1):160-166.
[37] ALCAMI A, SMITH GL. Vaccinia, cowpox, and camelpox viruses encodesoluble gamma interferon receptors with novel broad species specificity[J]. Journal ofvirology,1995,69(8):4633-4639.
[38] VERARDI PH, JONES LA, AZIZ FH, AHMAD S, YILMA TD. Vacciniavirus vectors with an inactivated gamma interferon receptor homolog gene (B8R) areattenuated In vivo without a concomitant reduction in immunogenicity[J]. Journal ofvirology,2001,75(1):11-18.
[39] DENES B, GRIDLEY DS, FODOR N, et al. Attenuation of a vaccine strainof vaccinia virus via inactivation of interferon viroceptor[J]. The journal of genemedicine,2006,8(7):814-823.
[40] LEGRAND FA, VERARDI PH, JONES LA, et al. Induction of potenthumoral and cell-mediated immune responses by attenuated vaccinia virus vectorswith deleted serpin genes[J]. J Virol,2004,78(6):2770-2779.
[41] JACKSON SS, ILYINSKII P, PHILIPPON V, et al. Role of genes thatmodulate host immune responses in the immunogenicity and pathogenicity ofvaccinia virus[J]. Journal of virology,2005,79(10):6554-6559.
[42] SYMONS JA, ADAMS E, TSCHARKE DC, et al. The vaccinia virus C12Lprotein inhibits mouse IL-18and promotes virus virulence in the murine intranasalmodel[J]. The Journal of general virology,2002,83(Pt11):2833-2844.
[43] READING PC, SMITH GL. Vaccinia virus interleukin-18-binding proteinpromotes virulence by reducing gamma interferon production and natural killer andT-cell activity[J]. Journal of virology,2003,77(18):9960-9968.
[44] DAI K, LIU Y, LIU M, et al. Pathogenicity and immunogenicity ofrecombinant Tiantan Vaccinia Virus with deleted C12L and A53R genes[J]. Vaccine,2008,26(39):5062-5071.
[45] ZHU W, FANG Q, ZHUANG K, et al. The attenuation of vaccinia Tian Tanstrain by the removal of the viral M1L-K2L genes[J]. Journal of virological methods,2007,144(1-2):17-26.
[46] KIENY MP, LATHE R, DRILLIEN R, et al. Expression of rabies virusglycoprotein from a recombinant vaccinia virus[J]. Nature,1984,312(5990):163-166.
[47] RUPPRECHT CE, BLASS L, SMITH K, et al. Human infection due torecombinant vaccinia-rabies glycoprotein virus[J]. The New England journal ofmedicine,2001,345(8):582-586.
[48]毛春生，金宁一，李季枝. A基因侧翼重组痘苗病毒的生物学特性研究[J].河南农业大学学报,1997,31(4):314.
[49] BULLER RM, CHAKRABARTI S, COOPER JA, TWARDZIK DR, MOSSB. Deletion of the vaccinia virus growth factor gene reduces virus virulence[J].Journal of virology,1988,62(3):866-874.
[50] LEE MS, ROOS JM, MCGUIGAN LC, et al. Molecular attenuation ofvaccinia virus: mutant generation and animal characterization[J]. Journal of virology,1992,66(5):2617-2630.
[51] LANGLAND JO, JACOBS BL. The role of the PKR-inhibitory genes, E3Land K3L, in determining vaccinia virus host range[J]. Virology,2002,299(1):133-141.
[52] CHANG HW, URIBE LH, JACOBS BL. Rescue of vaccinia virus lackingthe E3L gene by mutants of E3L[J]. Journal of virology,1995,69(10):6605-6608.
[53] VIJAYSRI S, JENTARRA G, HECK MC, et al. Vaccinia viruses withmutations in the E3L gene as potential replication-competent, attenuated vaccines:intra-nasal vaccination[J]. Vaccine,2008,26(5):664-676.
[54] LANGLAND JO, KASH JC, CARTER V, et al. Suppression ofproinflammatory signal transduction and gene expression by the dual nucleic acidbinding domains of the vaccinia virus E3L proteins[J]. Journal of virology,2006,80(20):10083-10095.
[55] CHANG PY, POGO BG. Reintroduction of gene(s) into an attenuateddeletion mutant of vaccinia virus strain IHD-W[J]. Microbial pathogenesis,1993,15(5):347-357.
[56] LANGLAND JO, JACOBS BL. Inhibition of PKR by vaccinia virus: role ofthe N-and C-terminal domains of E3L[J]. Virology,2004,324(2):419-429.
[57] HOLZER GW, FALKNER FG. Construction of a vaccinia virus deficient inthe essential DNA repair enzyme uracil DNA glycosylase by a complementing cellline[J]. Journal of virology,1997,71(7):4997-5002.
[58] HOLZER GW, REMP G, ANTOINE G, et al. Highly efficient induction ofprotective immunity by a vaccinia virus vector defective in late gene expression[J].Journal of virology,1999,73(6):4536-4542.
[59] WHITE SD, JACOBS BL. The amino terminus of the vaccinia virus E3protein is necessary to inhibit the interferon response[J]. Journal of virology,2012,86(10):5895-5904.
[60] GOEBEL SJ, JOHNSON GP, PERKUS ME, et al. The complete DNAsequence of vaccinia virus[J]. Virology,1990,179(1):247-266,517-263.
[61] ANTOINE G, SCHEIFLINGER F, DORNER F, FALKNER FG. Thecomplete genomic sequence of the modified vaccinia Ankara strain: comparison withother orthopoxviruses[J]. Virology,1998,244(2):365-396.
[62] UPTON C, SLACK S, HUNTER AL, EHLERS A, ROPER RL. Poxvirusorthologous clusters: toward defining the minimum essential poxvirus genome[J].Journal of virology,2003,77(13):7590-7600.
[63] SHCHELKUNOV SN, TOTMENIN AV, SANDAKHCHIEV LS. Analysis ofthe nucleotide sequence of23.8kbp from the left terminus of the genome of variolamajor virus strain India-1967[J]. Virus research,1996,40(2):169-183.
[64] MASSUNG RF, LIU LI, QI J, et al. Analysis of the complete genome ofsmallpox variola major virus strain Bangladesh-1975[J]. Virology,1994,201(2):215-240.
[65] SHCHELKUNOV SN, TOTMENIN AV, LOPAREV VN, et al. Alastrimsmallpox variola minor virus genome DNA sequences[J]. Virology,2000,266(2):361-386.
[66] ROPER RL. Characterization of the vaccinia virus A35R protein and its rolein virulence[J]. Journal of virology,2006,80(1):306-313.
[67] REHM KE, JONES GJ, TRIPP AA, METCALF MW, ROPER RL. Thepoxvirus A35protein is an immunoregulator[J]. J Virol,2010,84(1):418-425.
[68] ROPER RL. Characterization of the Vaccinia Virus A35R Protein and ItsRole in Virulence[J]. JOURNAL OF VIROLOGY,2006,80(1):306–313.
[69] STACK J, HAGA IR, SCHRODER M, et al. Vaccinia virus protein A46Rtargets multiple Toll-like-interleukin-1receptor adaptors and contributes tovirulence[J]. The Journal of experimental medicine,2005,201(6):1007-1018.
[70] FERRIER-REMBERT A, DRILLIEN R, TOURNIER JN, GARIN D,CRANCE JM. Short-and long-term immunogenicity and protection induced bynon-replicating smallpox vaccine candidates in mice and comparison with thetraditional1st generation vaccine[J]. Vaccine,2008,26(14):1794-1804.
[71] STITTELAAR KJ, VAN AMERONGEN G, KONDOVA I, et al. Modifiedvaccinia virus Ankara protects macaques against respiratory challenge withmonkeypox virus[J]. J Virol,2005,79(12):7845-7851.
[72] BELYAKOV IM, EARL P, DZUTSEV A, et al. Shared modes of protectionagainst poxvirus infection by attenuated and conventional smallpox vaccine viruses[J].Proceedings of the National Academy of Sciences of the United States of America,2003,100(16):9458-9463.
[73] BLANCHARD TJ, ALCAMI A, ANDREA P, SMITH GL. Modifiedvaccinia virus Ankara undergoes limited replication in human cells and lacks severalimmunomodulatory proteins: implications for use as a human vaccine[J]. The Journalof general virology,1998,79(Pt5):1159-1167.
[74] HOWLES S, GUIMARAES-WALKER A, YANG H, et al. Vaccination witha modified vaccinia virus Ankara (MVA)-vectored HIV-1immunogen induces modestvector-specific T cell responses in human subjects[J]. Vaccine,2010,28(45):7306-7312.
[75] EARL PL, COTTER C, MOSS B, et al. Design and evaluation of multi-gene,multi-clade HIV-1MVA vaccines[J]. Vaccine,2009,27(42):5885-5895.
[76] GUO W, CLEVELAND B, DAVENPORT T, LEE K, HU SL. Purification ofrecombinant vaccinia virus-expressed monomeric HIV-1gp120to apparenthomogeneity[J]. Protein expression and purification,2013,90(1):34-39.
[77] REHM KE, ROPER RL. Deletion of the A35gene from Modified VacciniaVirus Ankara increases immunogenicity and isotype switching[J]. Vaccine,2011,29(17):3276-3283.
[78] LI P, WANG N, ZHOU D, et al. Disruption of MHC class II-restrictedantigen presentation by vaccinia virus[J]. J Immunol,2005,175(10):6481-6488.
[79] JIN B, WANG RY, QIU Q, et al. Induction of potent cellular immuneresponse in mice by hepatitis C virus NS3protein with double-stranded RNA[J].Immunology,2007,122(1):15-27.
[80] SHARMA DP, RAMSAY AJ, MAGUIRE DJ, ROLPH MS, RAMSHAW IA.Interleukin-4mediates down regulation of antiviral cytokine expression and cytotoxicT-lymphocyte responses and exacerbates vaccinia virus infection in vivo[J]. Journal ofvirology,1996,70(10):7103-7107.
[81] REHM KE, CONNOR RF, JONES GJ, YIMBU K, ROPER RL. Vacciniavirus A35R inhibits MHC class II antigen presentation[J]. Virology,2010,397(1):176-186.
[82] JONES T. IMVAMUNE, an attenuated modified vaccinia Ankara virusvaccine for smallpox infection[J]. Current opinion in molecular therapeutics,2008,10(4):407-417.
[83] BACKES S, SPERLING KM, ZWILLING J, et al. Viral host-range factorC7or K1is essential for modified vaccinia virus Ankara late gene expression inhuman and murine cells, irrespective of their capacity to inhibit protein kinaseR-mediated phosphorylation of eukaryotic translation initiation factor2alpha[J]. TheJournal of general virology,2010,91(Pt2):470-482.
[84] ASSARSSON E, GREENBAUM JA, SUNDSTROM M, et al. Kineticanalysis of a complete poxvirus transcriptome reveals an immediate-early class ofgenes[J]. Proceedings of the National Academy of Sciences of the United States ofAmerica,2008,105(6):2140-2145.
[85] KECK JG, BALDICK CJ, JR., MOSS B. Role of DNA replication invaccinia virus gene expression: a naked template is required for transcription of threelate trans-activator genes[J]. Cell,1990,61(5):801-809.
[86] PERKUS ME, GOEBEL SJ, DAVIS SW, et al. Vaccinia virus host rangegenes[J]. Virology,1990,179(1):276-286.
[87] HSIAO JC, CHUNG CS, DRILLIEN R, CHANG W. The cowpox virus hostrange gene, CP77, affects phosphorylation of eIF2alpha and vaccinia viral translationin apoptotic HeLa cells[J]. Virology,2004,329(1):199-212.
[88] NAJERA JL, GOMEZ CE, DOMINGO-GIL E, GHERARDI MM,ESTEBAN M. Cellular and biochemical differences between two attenuated poxvirusvaccine candidates (MVA and NYVAC) and role of the C7L gene[J]. Journal ofvirology,2006,80(12):6033-6047.
[89] GOMEZ CE, NAJERA JL, KRUPA M, ESTEBAN M. The poxvirus vectorsMVA and NYVAC as gene delivery systems for vaccination against infectiousdiseases and cancer[J]. Current gene therapy,2008,8(2):97-120.
[90] SUTTER G, RAMSEY-EWING A, ROSALES R, MOSS B. Stableexpression of the vaccinia virus K1L gene in rabbit cells complements the host rangedefect of a vaccinia virus mutant[J]. Journal of virology,1994,68(7):4109-4116.
[91] GARCIA MA, MEURS EF, ESTEBAN M. The dsRNA protein kinase PKR:virus and cell control[J]. Biochimie,2007,89(6-7):799-811.
[92] DAVIES MV, CHANG HW, JACOBS BL, KAUFMAN RJ. The E3L andK3L vaccinia virus gene products stimulate translation through inhibition of thedouble-stranded RNA-dependent protein kinase by different mechanisms[J]. Journalof virology,1993,67(3):1688-1692.
[93] MENG X, CHAO J, XIANG Y. Identification from diverse mammalianpoxviruses of host-range regulatory genes functioning equivalently to vaccinia virusC7L[J]. Virology,2008,372(2):372-383.
[94] MENG X, JIANG C, ARSENIO J, et al. Vaccinia virus K1L and C7L inhibitantiviral activities induced by type I interferons[J]. Journal of virology,2009,83(20):10627-10636.
[95] UNGER B, NICHOLS RJ, STANITSA ES, TRAKTMAN P. Functionalcharacterization of the vaccinia virus I5protein[J]. Virology journal,2008,5:148.
[96] CHIU WL, LIN CL, YANG MH, TZOU DL, CHANG W. Vaccinia virus4c(A26L) protein on intracellular mature virus binds to the extracellular cellular matrixlaminin[J]. Journal of virology,2007,81(5):2149-2157.
[97] CHING YC, CHUNG CS, HUANG CY, et al. Disulfide bond formation atthe C termini of vaccinia virus A26and A27proteins does not require viral redoxenzymes and suppresses glycosaminoglycan-mediated cell fusion[J]. Journal ofvirology,2009,83(13):6464-6476.
[98] CHANG SJ, CHANG YX, IZMAILYAN R, TANG YL, CHANG W.Vaccinia virus A25and A26proteins are fusion suppressors for mature virions anddetermine strain-specific virus entry pathways into HeLa, CHO-K1, and L cells[J]. JVirol,2010,84(17):8422-8432.
[99] HOU YT, YANG XK, HU YW. Variation in the HindIII restriction fragmentsof DNA from the Chinese Tian Tan strain of vaccinia virus[J]. The Journal of generalvirology,1985,66(Pt8):1819-1823.
[100] SMITH CA, HU FQ, SMITH TD, et al. Cowpox virus genome encodes asecond soluble homologue of cellular TNF receptors, distinct from CrmB, that bindsTNF but not LT alpha[J]. Virology,1996,223(1):132-147.
[101] SCHREIBER M, RAJARATHNAM K, MCFADDEN G. Myxoma virus T2protein, a tumor necrosis factor (TNF) receptor homolog, is secreted as a monomerand dimer that each bind rabbit TNFalpha, but the dimer is a more potent TNFinhibitor[J]. The Journal of biological chemistry,1996,271(23):13333-13341.
[102] MOLOUGHNEY JG, MONKEN CE, TAO H, et al. Vaccinia virus leads toATG12-ATG3conjugation and deficiency in autophagosome formation[J]. Autophagy,2011,7(12):1434-1447.
[103] CHEN RA, RYZHAKOV G, COORAY S, RANDOW F, SMITH GL.Inhibition of IkappaB kinase by vaccinia virus virulence factor B14[J]. PLoSpathogens,2008,4(2): e22.
[104] BENFIELD CT, MANSUR DS, MCCOY LE, et al. Mapping the IkappaBkinase beta (IKKbeta)-binding interface of the B14protein, a vaccinia virus inhibitorof IKKbeta-mediated activation of nuclear factor kappaB[J]. The Journal of biologicalchemistry,2011,286(23):20727-20735.
[105] YU E, ZHAI D, JIN C, et al. Structural determinants of caspase-9inhibitionby the vaccinia virus protein, F1L[J]. The Journal of biological chemistry,2011,286(35):30748-30758.
[106] GERLIC M, FAUSTIN B, POSTIGO A, et al. Vaccinia virus F1L proteinpromotes virulence by inhibiting inflammasome activation[J]. Proceedings of theNational Academy of Sciences of the United States of America,2013,110(19):7808-7813.
[107] BENFIELD CT, REN H, LUCAS SJ, BAHSOUN B, SMITH GL. Vacciniavirus protein K7is a virulence factor that alters the acute immune response toinfection[J]. The Journal of general virology, Online in5April2013
[108] KIM MK, BREITBACH CJ, MOON A, et al. Oncolytic andimmunotherapeutic vaccinia induces antibody-mediated complement-dependentcancer cell lysis in humans[J]. Science translational medicine,2013,5(185):185ra163.
[109] MOORE ZS, SEWARD JF, LANE JM. Smallpox[J]. Lancet,2006,367(9508):425-435.
[110] WHO recommendations concerning the distribution, handling and synthesisof variola virus DNA, May2008[J]. Releve epidemiologique hebdomadaire/Sectiond'hygiene du Secretariat de la Societe des Nations=Weekly epidemiological record/Health Section of the Secretariat of the League of Nations,2008,83(44):393-395.
[111] BREMAN JG, ARITA I, FENNER F. Preventing the return of smallpox[J].The New England journal of medicine,2003,348(5):463-466.
[112] LANE JM, MILLAR JD. Risks of smallpox vaccination complications inthe United States[J]. American journal of epidemiology,1971,93(4):238-240.
[113] LANE JM, MILLAR JD, NEFF JM. Smallpox and smallpox vaccinationpolicy[J]. Annual review of medicine,1971,22:251-272.
[114] NELL P, KOHL KS, GRAHAM PL, et al. Progressive vaccinia as anadverse event following exposure to vaccinia virus: case definition and guidelines ofdata collection, analysis, and presentation of immunization safety data[J]. Vaccine,2007,25(31):5735-5744.
[115] VELLOZZI C, LANE JM, AVERHOFF F, et al. Generalized vaccinia,progressive vaccinia, and eczema vaccinatum are rare following smallpox (vaccinia)vaccination: United States surveillance,2003[J]. Clin Infect Dis,2005,41(5):689-697.
[116] KEMPER AR, DAVIS MM, FREED GL. Expected adverse events in amass smallpox vaccination campaign[J]. Eff Clin Pract,2002,5(2):84-90.
[117] CONO J, CASEY CG, BELL DM. Smallpox vaccination and adversereactions. Guidance for clinicians[J]. MMWR Recomm Rep,2003,52(RR-4):1-28.
[118] F. FENNER DAH, I. ARITA, Z. JEZEK, AND I. D. LADNYI,. Smallpoxand Its Eradication. World Health Organization: Geneva, Switzerland1988.
[119] GREENBERG RN, KENNEDY JS. ACAM2000: a newly licensed cellculture-based live vaccinia smallpox vaccine[J]. Expert opinion on investigationaldrugs,2008,17(4):555-564.
[120] JACOBS BL, LANGLAND JO, KIBLER KV, et al. Vaccinia virus vaccines:past, present and future[J]. Antiviral research,2009,84(1):1-13.
[121] DOWNIE AW, MC CK. The antibody response in man following infectionwith viruses of the pox group. III. Antibody response in smallpox[J]. The Journal ofhygiene,1958,56(4):479-487.
[122] BELSHE RB, NEWMAN FK, FREY SE, et al. Dose-dependentneutralizing-antibody responses to vaccinia[J]. The Journal of infectious diseases,2004,189(3):493-497.
[123] COUCH RB, WINOKUR P, EDWARDS KM, et al. Reducing the dose ofsmallpox vaccine reduces vaccine-associated morbidity without reducing vaccinationsuccess rates or immune responses[J]. The Journal of infectious diseases,2007,195(6):826-832.
[124] MARRIOTT KA, PARKINSON CV, MOREFIELD SI, et al. Clonalvaccinia virus grown in cell culture fully protects monkeys from lethal monkeypoxchallenge[J]. Vaccine,2008,26(4):581-588.
[125] ARTENSTEIN AW, JOHNSON C, MARBURY TC, et al. A novel, cellculture-derived smallpox vaccine in vaccinia-naive adults[J]. Vaccine,2005,23(25):3301-3309.
[126] OSBORNE JD, DA SILVA M, FRACE AM, et al. Genomic differences ofVaccinia virus clones from Dryvax smallpox vaccine: the Dryvax-like ACAM2000and the mouse neurovirulent Clone-3[J]. Vaccine,2007,25(52):8807-8832.
[127] JANG HC, KIM CJ, KIM KH, et al. A randomized, double-blind,controlled clinical trial to evaluate the efficacy and safety of CJ-50300, a newlydeveloped cell culture-derived smallpox vaccine, in healthy volunteers[J]. Vaccine,2010,28(36):5845-5849.
[128] WELTZIN R, LIU J, PUGACHEV KV, et al. Clonal vaccinia virus grownin cell culture as a new smallpox vaccine[J]. Nature medicine,2003,9(9):1125-1130.
[129] KENNER J, CAMERON F, EMPIG C, JOBES DV, GURWITH M.LC16m8: an attenuated smallpox vaccine[J]. Vaccine,2006,24(47-48):7009-7022.
[130] SAITO T, FUJII T, KANATANI Y, et al. Clinical and immunologicalresponse to attenuated tissue-cultured smallpox vaccine LC16m8[J]. Jama,2009,301(10):1025-1033.
[131] VOLLMAR J, ARNDTZ N, ECKL KM, et al. Safety and immunogenicityof IMVAMUNE, a promising candidate as a third generation smallpox vaccine[J].Vaccine,2006,24(12):2065-2070.
[132] MEISINGER-HENSCHEL C, SCHMIDT M, LUKASSEN S, et al.Genomic sequence of chorioallantois vaccinia virus Ankara, the ancestor of modifiedvaccinia virus Ankara[J]. The Journal of general virology,2007,88(Pt12):3249-3259.
[133] WILCK MB, SEAMAN MS, BADEN LR, et al. Safety andimmunogenicity of modified vaccinia Ankara (ACAM3000): effect of dose and routeof administration[J]. The Journal of infectious diseases,2010,201(9):1361-1370.
[134] SPEERS WC, WESLEY RB, NEFF JM, GOLDSTEIN J, LOURIE B.Evaluation of two kinds of smallpox vaccine: CVI-78and calf lymph vaccine. II.Clinical and serologic observations of response to revaccination with calf lymphvaccine[J]. Pediatric research,1975,9(8):628-632.
[135] MONATH TP, CALDWELL JR, MUNDT W, et al. ACAM2000clonalVero cell culture vaccinia virus (New York City Board of Health strain)--asecond-generation smallpox vaccine for biological defense[J]. Int J Infect Dis,2004,8Suppl2: S31-44.
[136] HATCH GJ, GRAHAM VA, BEWLEY KR, et al. Assessment of theprotective effect of IMVAMUNE(R) and ACAM2000(R) vaccines against aerosolisedMonkeypox virus in cynomolgus macaques[J]. J Virol, on line May2013.
[137] FREY SE, NEWMAN FK, KENNEDY JS, et al. Comparison of the safetyand immunogenicity of ACAM1000, ACAM2000and Dryvax in healthyvaccinia-naive adults[J]. Vaccine,2009,27(10):1637-1644.
[138] GREENBERG RN, KENNEDY JS, CLANTON DJ, et al. Safety andimmunogenicity of new cell-cultured smallpox vaccine compared with calf-lymphderived vaccine: a blind, single-centre, randomised controlled trial[J]. Lancet,2005,365(9457):398-409.
[139] POLAND GA, GRABENSTEIN JD, NEFF JM. The US smallpoxvaccination program: a review of a large modern era smallpox vaccinationimplementation program[J]. Vaccine,2005,23(17-18):2078-2081.
[140] BEACHKOFSKY TM, CARRIZALES SC, BIDINGER JJ, et al. Adverseevents following smallpox vaccination with ACAM2000in a military population[J].Archives of dermatology,2010,146(6):656-661.
[141] SARKAR JK, MITRA AC, MUKHERJEE MK. The minimum protectivelevel of antibodies in smallpox[J]. Bulletin of the World Health Organization,1975,52(3):307-311.
[142] FREY SE, COUCH RB, TACKET CO, et al. Clinical responses toundiluted and diluted smallpox vaccine[J]. The New England journal of medicine,2002,346(17):1265-1274.
[143] MCCLAIN DJ, HARRISON S, YEAGER CL, et al. Immunologicresponses to vaccinia vaccines administered by different parenteral routes[J]. TheJournal of infectious diseases,1997,175(4):756-763.
[144] KIM SH, CHOI SJ, PARK WB, et al. Detailed kinetics of immuneresponses to a new cell culture-derived smallpox vaccine in vaccinia-naive adults[J].Vaccine,2007,25(33):6287-6291.
[145] MESEDA CA, MAYER AE, KUMAR A, et al. Comparative evaluation ofthe immune responses and protection engendered by LC16m8and Dryvax smallpoxvaccines in a mouse model[J]. Clin Vaccine Immunol,2009,16(9):1261-1271.
[146] KEMPE CH, FULGINITI V, MINAMITANI M, SHINEFIELD H.Smallpox vaccination of eczema patients with a strain of attenuated live vaccinia(CVI-78)[J]. Pediatrics,1968,42(6):980-985.
[147] WESLEY RB, SPEERS WC, NEFF JM, RUBEN FL, LOURIE B.Evaluation of two kinds of smallpox vaccine: CVI-78and calf lymph vaccine. I.Clinical and serologic response to primary vaccination[J]. Pediatric research,1975,9(8):624-628.
[148] MEYER H, SUTTER G, MAYR A. Mapping of deletions in the genome ofthe highly attenuated vaccinia virus MVA and their influence on virulence[J]. TheJournal of general virology,1991,72(Pt5):1031-1038.
[149] STITTELAAR KJ, OSTERHAUS AD. MVA: a cuckoo in the vaccinenest?[J]. Vaccine,2001,19(27): V-VI.
[150] KENNEDY JS, GREENBERG RN. IMVAMUNE: modified vacciniaAnkara strain as an attenuated smallpox vaccine[J]. Expert review of vaccines,2009,8(1):13-24.
[151] FREY SE, NEWMAN FK, KENNEDY JS, et al. Clinical and immunologicresponses to multiple doses of IMVAMUNE (Modified Vaccinia Ankara) followed byDryvax challenge[J]. Vaccine,2007,25(51):8562-8573.
[152] PARRINO J, MCCURDY LH, LARKIN BD, et al. Safety, immunogenicityand efficacy of modified vaccinia Ankara (MVA) against Dryvax challenge invaccinia-naive and vaccinia-immune individuals[J]. Vaccine,2007,25(8):1513-1525.
[153] PERKUS ME, TARTAGLIA J, PAOLETTI E. Poxvirus-based vaccinecandidates for cancer, AIDS, and other infectious diseases[J]. Journal of leukocytebiology,1995,58(1):1-13.
[154] TYKODI SS, THOMPSON JA. Development of modified vacciniaAnkara-5T4as specific immunotherapy for advanced human cancer[J]. Expertopinion on biological therapy,2008,8(12):1947-1953.
[155] SEAMAN MS, WILCK MB, BADEN LR, et al. Effect of vaccination withmodified vaccinia Ankara (ACAM3000) on subsequent challenge with Dryvax[J].The Journal of infectious diseases,2010,201(9):1353-1360.
[156] MACK TM, NOBLE J, JR., THOMAS DB. A prospective study of serumantibody and protection against smallpox[J]. The American journal of tropicalmedicine and hygiene,1972,21(2):214-218.
[157] RAMANATHAN VD, KUMAR M, MAHALINGAM J, et al. A Phase1study to evaluate the safety and immunogenicity of a recombinant HIV type1subtypeC-modified vaccinia Ankara virus vaccine candidate in Indian volunteers[J]. AIDSresearch and human retroviruses,2009,25(11):1107-1116.
[158] CURRIER JR, NGAUY V, DE SOUZA MS, et al. Phase I safety andimmunogenicity evaluation of MVA-CMDR, a multigenic, recombinant modifiedvaccinia Ankara-HIV-1vaccine candidate[J]. PloS one,2010,5(11): e13983.
[159] MWAU M, CEBERE I, SUTTON J, et al. A human immunodeficiencyvirus1(HIV-1) clade A vaccine in clinical trials: stimulation of HIV-specific T-cellresponses by DNA and recombinant modified vaccinia virus Ankara (MVA) vaccinesin humans[J]. The Journal of general virology,2004,85(Pt4):911-919.
[160] MCCORMACK S, STOHR W, BARBER T, et al. EV02: a Phase I trial tocompare the safety and immunogenicity of HIV DNA-C prime-NYVAC-C boost toNYVAC-C alone[J]. Vaccine,2008,26(25):3162-3174.
[161] AMATO RJ, HAWKINS RE, KAUFMAN HL, et al. Vaccination ofmetastatic renal cancer patients with MVA-5T4: a randomized, double-blind,placebo-controlled phase III study[J]. Clin Cancer Res,2010,16(22):5539-5547.
[162] GARCIA-HERNANDEZ E, GONZALEZ-SANCHEZ JL,ANDRADE-MANZANO A, et al. Regression of papilloma high-grade lesions (CIN2and CIN3) is stimulated by therapeutic vaccination with MVA E2recombinantvaccine[J]. Cancer gene therapy,2006,13(6):592-597.
[163] ARLEN PM, PAZDUR M, SKARUPA L, RAUCKHORST M, GULLEYJL. A randomized phase II study of docetaxel alone or in combination withPANVAC-V (vaccinia) and PANVAC-F (fowlpox) in patients with metastatic breastcancer (NCI05-C-0229)[J]. Clinical breast cancer,2006,7(2):176-179.
[164] ZEMP FJ, CORREDOR JC, LUN X, MURUVE DA, FORSYTH PA.Oncolytic viruses as experimental treatments for malignant gliomas: using a scourgeto treat a devil[J]. Cytokine&growth factor reviews,2010,21(2-3):103-117.
[165] PARK BH, HWANG T, LIU TC, et al. Use of a targeted oncolytic poxvirus,JX-594, in patients with refractory primary or metastatic liver cancer: a phase I trial[J].The lancet oncology,2008,9(6):533-542.
[166] FENNER FH, DA.; ARITA, I.; JEZEK, Z.; LADNYI, I. Smallpox and itsEradication. World Health Organization. Geneva, Switzerland1988.
[167] PANICALI D, PAOLETTI E. Construction of poxviruses as cloning vectors:insertion of the thymidine kinase gene from herpes simplex virus into the DNA ofinfectious vaccinia virus[J]. Proceedings of the National Academy of Sciences of theUnited States of America,1982,79(16):4927-4931.
[168] JOHNSTON JB, MCFADDEN G. Technical knockout: understandingpoxvirus pathogenesis by selectively deleting viral immunomodulatory genes[J].Cellular microbiology,2004,6(8):695-705.
[169] PASTORET PP, BROCHIER B. The development and use of avaccinia-rabies recombinant oral vaccine for the control of wildlife rabies; a linkbetween Jenner and Pasteur[J]. Epidemiology and infection,1996,116(3):235-240.
[170] HANLON CA, NIEZGODA M, HAMIR AN, et al. First North Americanfield release of a vaccinia-rabies glycoprotein recombinant virus[J]. Journal ofwildlife diseases,1998,34(2):228-239.
[171] SLATE D, ALGEO TP, NELSON KM, et al. Oral rabies vaccination innorth america: opportunities, complexities, and challenges[J]. PLoS neglected tropicaldiseases,2009,3(12): e549.
[172] KANTOFF PW, SCHUETZ TJ, BLUMENSTEIN BA, et al. Overallsurvival analysis of a phase II randomized controlled trial of a Poxviral-basedPSA-targeted immunotherapy in metastatic castration-resistant prostate cancer[J]. JClin Oncol,2010,28(7):1099-1105.
[173] SUTTER G, MOSS B. Nonreplicating vaccinia vector efficiently expressesrecombinant genes[J]. Proceedings of the National Academy of Sciences of the UnitedStates of America,1992,89(22):10847-10851.
[174] AMATO RJ.5T4-modified vaccinia Ankara: progress in tumor-associatedantigen-based immunotherapy[J]. Expert opinion on biological therapy,2010,10(2):281-287.
[175] TARTAGLIA J, PERKUS ME, TAYLOR J, et al. NYVAC: a highlyattenuated strain of vaccinia virus[J]. Virology,1992,188(1):217-232.
[176] OCKENHOUSE CF, SUN PF, LANAR DE, et al. Phase I/IIa safety,immunogenicity, and efficacy trial of NYVAC-Pf7, a pox-vectored, multiantigen,multistage vaccine candidate for Plasmodium falciparum malaria[J]. The Journal ofinfectious diseases,1998,177(6):1664-1673.
[177] BART PA, GOODALL R, BARBER T, et al. EV01: a phase I trial inhealthy HIV negative volunteers to evaluate a clade C HIV vaccine, NYVAC-Cundertaken by the EuroVacc Consortium[J]. Vaccine,2008,26(25):3153-3161.
[178] SHEPPARD NC, BATES AC, SATTENTAU QJ. A functional human IgMresponse to HIV-1Env after immunization with NYVAC HIV C[J]. AIDS (London,England),2007,21(4):524-527.
[179] GOMEZ CE, NAJERA JL, JIMENEZ EP, et al. Head-to-head comparisonon the immunogenicity of two HIV/AIDS vaccine candidates based on the attenuatedpoxvirus strains MVA and NYVAC co-expressing in a single locus the HIV-1BX08gp120and HIV-1(IIIB) Gag-Pol-Nef proteins of clade B[J]. Vaccine,2007,25(15):2863-2885.
[180] NAJERA JL, GOMEZ CE, GARCIA-ARRIAZA J, SORZANO CO,ESTEBAN M. Insertion of vaccinia virus C7L host range gene into NYVAC-Bgenome potentiates immune responses against HIV-1antigens[J]. PloS one,2010,5(6): e11406.
[181] WHO. Global eradication of smallpox: WHO Global Commission for theCertification od Smallpox Eradication.[J]. J Med Assoc Thai,1979,62(8):461.
[182]娄元梅,阮力,朱既明.痘苗病毒毒力相关基因的研究进展[J].病毒学报,1996,12(1):88-96.
[183] PERDIGUERO B, ESTEBAN M. The interferon system and vaccinia virusevasion mechanisms[J]. J Interferon Cytokine Res,2009,29(9):581-598.
[184] GIRGIS NM, DEHAVEN BC, XIAO Y, et al. The Vaccinia viruscomplement control protein modulates adaptive immune responses during infection[J].J Virol,2011,85(6):2547-2556.
[185] RAMSEY-EWING A, MOSS B. Restriction of vaccinia virus replication inCHO cells occurs at the stage of viral intermediate protein synthesis[J]. Virology,1995,206(2):984-993.
[186] HINTHONG O, JIN XL, SHISLER JL. Characterization of wild-type andmutant vaccinia virus M2L proteins' abilities to localize to the endoplasmic reticulumand to inhibit NF-kappaB activation during infection[J]. Virology,2008,373(2):248-262.
[187] WITTEK R, MULLER HK, WYLER R. Length heterogeneity in the DNAof vaccinia virus is eliminated on cloning the virus[J]. FEBS letters,1978,90(1):41-46.
[188] BULLER RM, SMITH GL, CREMER K, NOTKINS AL, MOSS B.Decreased virulence of recombinant vaccinia virus expression vectors is associatedwith a thymidine kinase-negative phenotype[J]. Nature,1985,317(6040):813-815.
[189] KOTWAL GJ, MOSS B. Analysis of a large cluster of nonessential genesdeleted from a vaccinia virus terminal transposition mutant[J]. Virology,1988,167(2):524-537.
[190] BARTLETT N, SYMONS JA, TSCHARKE DC, SMITH GL. The vacciniavirus N1L protein is an intracellular homodimer that promotes virulence[J]. TheJournal of general virology,2002,83(Pt8):1965-1976.
[191] KOTWAL GJ, HUGIN AW, MOSS B. Mapping and insertionalmutagenesis of a vaccinia virus gene encoding a13,800-Da secreted protein[J].Virology,1989,171(2):579-587.
[192] AOYAGI M, ZHAI D, JIN C, et al. Vaccinia virus N1L protein resembles aB cell lymphoma-2(Bcl-2) family protein[J]. Protein Sci,2007,16(1):118-124.
[193] SEET BT, JOHNSTON JB, BRUNETTI CR, et al. Poxviruses and immuneevasion[J]. Annual review of immunology,2003,21:377-423.
[194] TAYLOR JM, BARRY M. Near death experiences: poxvirus regulation ofapoptotic death[J]. Virology,2006,344(1):139-150.
[195] GONZALEZ JM, ESTEBAN M. A poxvirus Bcl-2-like gene familyinvolved in regulation of host immune response: sequence similarity and evolutionaryhistory[J]. Virology journal,2010,7:59.
[196] BOWIE A, KISS-TOTH E, SYMONS JA, et al. A46R and A52R fromvaccinia virus are antagonists of host IL-1and toll-like receptor signaling[J].Proceedings of the National Academy of Sciences of the United States of America,2000,97(18):10162-10167.
[197] GRAHAM SC, BAHAR MW, COORAY S, et al. Vaccinia virus proteinsA52and B14Share a Bcl-2-like fold but have evolved to inhibit NF-kappaB ratherthan apoptosis[J]. PLoS pathogens,2008,4(8): e1000128.
[198] ZHANG L, VILLA NY, RAHMAN MM, et al. Analysis of vacciniavirus-host protein-protein interactions: validations of yeast two-hybrid screenings[J].Journal of proteome research,2009,8(9):4311-4318.
[199] OSEROFF C, PETERS B, PASQUETTO V, et al. Dissociation betweenepitope hierarchy and immunoprevalence in CD8responses to vaccinia virus westernreserve[J]. J Immunol,2008,180(11):7193-7202.
[200] STETSON DB, MEDZHITOV R. Type I interferons in host defense[J].Immunity,2006,25(3):373-381.
[201] SADLER AJ, WILLIAMS BR. Interferon-inducible antiviral effectors[J].Nature reviews,2008,8(7):559-568.
[202] UNTERHOLZNER L, SUMNER RP, BARAN M, et al. Vaccinia virusprotein C6is a virulence factor that binds TBK-1adaptor proteins and inhibitsactivation of IRF3and IRF7[J]. PLoS pathogens,2011,7(9): e1002247.
[203] GARCIA-ARRIAZA J, NAJERA JL, GOMEZ CE, et al. A candidateHIV/AIDS vaccine (MVA-B) lacking vaccinia virus gene C6L enhances memoryHIV-1-specific T-cell responses[J]. PloS one,2011,6(8): e24244.
[204] HAGA IR, BOWIE AG. Evasion of innate immunity by vaccinia virus[J].Parasitology,2005,130Suppl: S11-25.
[205] BILLINGS B, SMITH SA, ZHANG Z, LAHIRI DK, KOTWAL GJ. Lackof N1L gene expression results in a significant decrease of vaccinia virus replicationin mouse brain[J]. Annals of the New York Academy of Sciences,2004,1030:297-302.
[206] SMITH KA, STALLARD V, ROOS JM, et al. Host range selection ofvaccinia recombinants containing insertions of foreign genes into non-codingsequences[J]. Vaccine,1993,11(1):43-53.
[207] SHCHELKUNOV S, TOTMENIN A, KOLOSOVA I. Species-specificdifferences in organization of orthopoxvirus kelch-like proteins[J]. Virus genes,2002,24(2):157-162.
[208] PIRES DE MIRANDA M, READING PC, TSCHARKE DC, MURPHY BJ,SMITH GL. The vaccinia virus kelch-like protein C2L affects calcium-independentadhesion to the extracellular matrix and inflammation in a murine intradermalmodel[J]. The Journal of general virology,2003,84(Pt9):2459-2471.
[209] SHIMSHEK DR, KIM J, HUBNER MR, et al. Codon-improved Crerecombinase (iCre) expression in the mouse[J]. Genesis,2002,32(1):19-26.
[210] POMERANTSEV AP, SITARAMAN R, GALLOWAY CR, KIVOVICH V,LEPPLA SH. Genome engineering in Bacillus anthracis using Cre recombinase[J].Infection and immunity,2006,74(1):682-693.
[211]贺淹才.简明基因工程原理[M].科学出版社:北京2005.
[212]邹艳,阮力,娄元梅,朱诚.痘苗病毒天坛株非必需区TA25R…TA36L的两步重组表达载体的构建[J].病毒学报,1998,14(2):97-103.
[213] SCHEIFLINGER F, DORNER F, FALKNER FG. Transient markerstabilisation: a general procedure to construct marker-free recombinant vacciniavirus[J]. Archives of virology,1998,143(3):467-474.
[214]周小云,刘颖,饶力群,王芙,戴凯凡等.适用于疫苗株筛选的痘苗病毒载体的构建[J].病毒学报,2004,20(4):315-321.
[215]房有荣,阎辉.以GFP-Zeocin为筛选标志的重组痘苗病毒载体质粒的构建.《2011年浙江省医学会医学病毒学分会、医学微生物与免疫学分会学术年会论文汇编》2011.
[216] CUBITT AB, HEIM R, ADAMS SR, et al. Understanding, improving andusing green fluorescent proteins[J]. Trends in biochemical sciences,1995,20(11):448-455.
[217] HIKICHI M, KIDOKORO M, HARAGUCHI T, et al. MicroRNAregulation of glycoprotein B5R in oncolytic vaccinia virus reduces viral pathogenicitywithout impairing its antitumor efficacy[J]. Mol Ther,2011,19(6):1107-1115.
[218] FAHY AS, CLARK RH, GLYDE EF, SMITH GL. Vaccinia virus proteinC16acts intracellularly to modulate the host response and promote virulence[J]. TheJournal of general virology,2008,89(Pt10):2377-2387.
[219] MOSS B. Replicating and host-restricted non-replicating vaccinia virusvectors for vaccine development[J]. Developments in biological standardization,1994,82:55-63.
[220] KUMAR S, BOYLE DB. A poxvirus bidirectional promoter element withearly/late and late functions[J]. Virology,1990,179(1):151-158.
[221] BOYLE DB. Quantitative assessment of poxvirus promoters in fowlpoxand vaccinia virus recombinants[J]. Virus genes,1992,6(3):281-290.
[222]朱自满,孙念绪,张超,吴淼.应用Cre/loxP系统构建含人PTEN基因的重组腺病毒载体[J].消化外科,2005,4(2):124-127.
[223]王艳春,姜娜,展德文,邱炎等. Cre-LoxP系统在炭疽芽胞杆菌基因敲除中的应用及eag基因的敲除[J].生物化学与生物物理进展,2009:1-10.
[224] HOESS RH, ZIESE M, STERNBERG N. P1site-specific recombination:nucleotide sequence of the recombining sites[J]. Proceedings of the NationalAcademy of Sciences of the United States of America,1982,79(11):3398-3402.
[225] KOTWAL GJ, MOSS B. Vaccinia virus encodes a secretory polypeptidestructurally related to complement control proteins[J]. Nature,1988,335(6186):176-178.
[226] GEDEY R, JIN XL, HINTHONG O, SHISLER JL. Poxviral regulation ofthe host NF-kappaB response: the vaccinia virus M2L protein inhibits induction ofNF-kappaB activation via an ERK2pathway in virus-infected human embryonickidney cells[J]. Journal of virology,2006,80(17):8676-8685.
[227]殷震,刘景华.动物病毒学（第二版）.北京：科学出版社1997.
[228] REED LJ, MUENCH, H. A simple method of estimating fifty percentendpoints[J]. Am J Hyg,1938,27:493–497.
[229] FULGINITI VA, PAPIER A, LANE JM, NEFF JM, HENDERSON DA.Smallpox vaccination: a review, part II. Adverse events[J]. Clin Infect Dis,2003,37(2):251-271.
[230] GURVICH EB, VILESOVA IS. Vaccinia virus in postvaccinalencephalitis[J]. Acta virologica,1983,27(2):154-159.
[231] HENDERSON DA MB. Smallpox and vacciniaIn: Plotkin SA, OrensteinWA, editors. Vaccines. Philadelphia: W.B. Saunders Company.1999:74-97.
[232] KUTINOVA L, LUDVIKOVA V, SIMONOVA V, et al. Search for optimalparent for recombinant vaccinia virus vaccines. Study of three vaccinia virus vaccinalstrains and several virus lines derived from them[J]. Vaccine,1995,13(5):487-493.
[233] CARROLL MW, MOSS B. Host range and cytopathogenicity of the highlyattenuated MVA strain of vaccinia virus: propagation and generation of recombinantviruses in a nonhuman mammalian cell line[J]. Virology,1997,238(2):198-211.
[234] YU W, FANG Q, ZHU W, et al. One time intranasal vaccination with amodified vaccinia Tiantan strain MVTT(ZCI) protects animals against pathogenicviral challenge[J]. Vaccine,2010,28(9):2088-2096.
[235] TURNER PC, MOYER RW. Orthopoxvirus fusion inhibitor glycoproteinSPI-3(open reading frame K2L) contains motifs characteristic of serine proteinaseinhibitors that are not required for control of cell fusion[J]. J Virol,1995,69(10):5978-5987.
[236] LUDVIKOVA V, KUTINOVA L, SIMONOVA V, OTAVOVA M.Evaluation of various virulence tests with low virulence vaccinia virus in mice[J].Biologicals,1994,22(2):187-190.
[237] LI G, CHEN N, FENG Z, et al. Genomic sequence and analysis of avaccinia virus isolate from a patient with a smallpox vaccine-related complication[J].Virology journal,2006,3:88.
[238]董树林.天坛株痘苗毒种创始人齐长庆[J].微生物学及免疫学进展,2009,37(3):1-3.
[239]金奇,陈南海,陈淑霞,黄静,冯泽华等.痘苗病毒天坛株全基因组结构特点的分析[J].中国科学(C辑),1997,27(6):563-567.
[240] MARIES VAN DEN BROEK MFB, GABRIELE KO¨HLER, MARIJKEBARNER. IL-4and IL-10Antagonize IL-12-Mediated Protection Against AcuteVaccinia Virus Infection with a Limited Role of IFN-g and Nitric Oxide Synthetase2[J]. The Journal of Immunology,2000,164:371–378.
[241] BORN TL, MORRISON LA, ESTEBAN DJ, et al. A poxvirus protein thatbinds to and inactivates IL-18, and inhibits NK cell response[J]. J Immunol,2000,164(6):3246-3254.
[242] ANDERSON KP, LIE YS, LOW MA, FENNIE EH. Effects of tumornecrosis factor-alpha treatment on mortality in murine cytomegalovirus-infectedmice[J]. Antiviral research,1993,21(4):343-355.
[243] ZINKERNAGEL RM. Immunology taught by viruses[J]. Science (NewYork, NY,1996,271(5246):173-178.
[244] MALONEY G, SCHRODER M, BOWIE AG. Vaccinia virus protein A52Ractivates p38mitogen-activated protein kinase and potentiateslipopolysaccharide-induced interleukin-10[J]. The Journal of biological chemistry,2005,280(35):30838-30844.
[245] KURILLA MG, SWAMINATHAN S, WELSH RM, KIEFF E,BRUTKIEWICZ RR. Effects of virally expressed interleukin-10on vaccinia virusinfection in mice[J]. Journal of virology,1993,67(12):7623-7628.
[246] VAN DEN BROEK M, BACHMANN MF, KOHLER G, et al. IL-4andIL-10antagonize IL-12-mediated protection against acute vaccinia virus infectionwith a limited role of IFN-gamma and nitric oxide synthetase2[J]. J Immunol,2000,164(1):371-378.
[247] IAN A. RAMSHAW MEA, SUSAN M. PHILLIPS, DAVID B. BOYL.Recovery of immunodeficient mice from a vaccinia virus/IL-2recombinantinfection[J]. Nature,1987,329:545-546.
[248] KIDOKORO M, TASHIRO M, SHIDA H. Genetically stable and fullyeffective smallpox vaccine strain constructed from highly attenuated vacciniaLC16m8[J]. Proceedings of the National Academy of Sciences of the United States ofAmerica,2005,102(11):4152-4157.
[249] PHELPS AL, GATES AJ, HILLIER M, EASTAUGH L, ULAETO DO.Comparative efficacy of modified vaccinia Ankara (MVA) as a potential replacementsmallpox vaccine[J]. Vaccine,2007,25(1):34-42.
[250] BERHANU A, WILSON RL, KIRKWOOD-WATTS DL, et al. Vaccinationof BALB/c mice with Escherichia coli-expressed vaccinia virus proteins A27L, B5R,and D8L protects mice from lethal vaccinia virus challenge[J]. Journal of virology,2008,82(7):3517-3529.