重组猪圆环病毒2型/白介素-2核酸疫苗的构建及免疫效果研究
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摘要
猪圆环病毒2型(Porcine circovirus type 2, PCV2)是引发猪断奶后多系统衰竭综合征的主要病原。PCV2主要由ORF1和ORF2构成,ORF2基因是PCV2的重要抗原基因,编码病毒的衣壳蛋白(Cap蛋白),在Cap蛋白上存在抗原表位,具有免疫原性,因此ORF2基因成为建立PCV2特异性血清学检测方法及核酸疫苗研究的目的基因。当前国内外尚没有PCV2疫苗可用,因此研制猪圆环病毒2型核酸疫苗对控制猪PCV2病的发生具有重要意义。
设计含双酶切位点的引物采用PCR方法扩增PCV2的ORF2全长基因并克隆入pGEM-T easy克隆载体;采用重叠延伸PCR(splicing by overlap extension-PCR,SOE-PCR)技术将PCV2的ORF2全长基因和猪白介素2成熟肽基因( PoIL-2 )构建成重组嵌合基因(PCV2-linker-PoIL-2)并克隆入pGEM-T easy克隆载体;筛选含PCV2-linker-PoIL-2、PCV2 ORF2的阳性克隆质粒经限制性内切酶双切后连接入经相同酶切的pcDNA3.1(+)真核表达载体中以构建重组表达质粒(rpcDNA3.1/ PCV2-linker-PoIL-2、rpcDNA3.1/ORF2)。筛选阳性重组表达质粒rpcDNA3.1/PCV2-linker-PoIL-2、rpcDNA3.1/ORF2瞬时转染COS-7细胞,分别采用PCV2抗原间接免疫荧光试验和PoIL-2蛋白ELISA试验方法检测COS-7细胞中表达的重组融合蛋白(rCap-linker-PoIL-2、rCap )免疫反应活性;提取rpcDNA3.1/PCV2-linker-PoIL-2质粒和rpcDNA3.1-OFR2质粒分别对Balb/c小鼠和猪进行三次免疫,采用PCV2抗体ELISA检测方法以评价其免疫效果。
结果表明:成功构建了单重组表达质粒pcDNA3.1/OFR2和PCV2-linker-PoIL-2嵌合基因及其共表达质粒rpcDNA3.1/PCV2-linker-PoIL-2 ; rpcDNA3.1/PCV2-linker-PoIL-2和rpcDNA3.1/OFR2质粒在COS-7细胞浆内进行了表达,表达的rCap-linker-PoIL-2蛋白可分别与抗PCV2和抗PoIL-2蛋白抗血清发生特异性免疫反应,表明rCap-linker-PoIL-2蛋白具有Cap蛋白和PoIL-2蛋白的双重免疫反应活性。rpcDNA3.1/PCV2-linker-PoIL-2质粒免疫小鼠后可诱导小鼠产生明显的抗PCV2抗体,第3次加强免疫后,免疫组抗体滴度最高是空白对照组的3.2倍,且显著高于rpcDNA3.1/OFR2质粒对照组; rpcDNA3.1/PCV2-linker-PoIL-2和rpcDNA3.1/ORF2质粒均可诱导猪体产生抗PCV2免疫应答,rpcDNA3.1/PCV2-linker-PoIL-2、rpcDNA3.1/ORF2质粒免疫组抗体滴度分别是空载体对照组的12.46倍、10.13倍,表明PCV2-linker-PoIL-2嵌合基因中的PoIL-2对rpcDNA3.1/PCV2-linker-PoIL-2质粒在小鼠体内的免疫起到了很好的免疫增强作用。
本研究成功构建了PCV2-linker-PoIL-2嵌合基因及其重组表达质粒pcDNA3.1/PCV2-linker-PoIL-2;重组表达质粒在COS-7细胞中表达了具有Cap蛋白和PoIL-2蛋白双重免疫反应活性的融合蛋白,免疫小鼠和猪后可诱导小鼠和猪产生高的抗PCV2抗体,且其诱导的抗体水平明显高于pcDNA3.1/OFR2重组质粒。本研究为PCV2的DNA疫苗研究奠定了基础。
Porcine circovirus type 2 (Porcine circovirus type 2, PCV2) are the main pathogens and trigger Postweaning multisystemic wasting syndrome.ORF1 and ORF2 are main open reading frames in PCV2,the ORF2 gene is an important antigen genes, the protein-coding (Cap protein), the existence of epitope in Cap epitope, with immunogenicity, so ORF2 gene is an objective gene which use establishing PCV2 specific serology and researching DNA vaccine of the PCV2. Current, no available PCV2 vaccine in domestic and foreign, the development of PCV2 DNA vaccine to control disease of PCV2 is of great significance.
The primers with double restriction site were designed and amplify ORF2 gene of PCV2.The ORF2 gene was cloned into pGEM-T easy cloning vector. The recombinant chimeric gene of PCV2-linker-PoIL-2 was constructed by PCV2 ORF2 gene linked porcine interleukin-2 (PoIL-2) mature peptide gene via a 15-amino acid glycine-rich linker [linker(,G4S)3] by SOE-PCR (splicing by overlap extension-PCR). The recombinant chimeric gene was cloned into pGEM-T Easy vector and subsequently into eukaryotic expression pcDNA3.1(+) vector. The positive recombinant expression plasmid rpcDNA3.1/PCV2-linker-PoIL-2 and r pcDNA3.1/ORF2 were transfected into COS-7 cells by lipofectamine. Indirect immunofluorescent assay (IFA) and porcine IL-2 ELISA assay was used to detect the bioactivities for PCV2 Cap protein and PoIL-2 protein of the expressed recombinant fusion protein (rCap-linker-PoIL-2 protein), respectively. The positive recombinant expression plasmid were extracted and inoculated Balb/c mice and pigs. The immune effects induced by rpcDNA3.1/PCV2-linker-PoIL-2 and rpcDNA3.1/OFR2 was evaluated by ELISA assay for detection PCV2 antibody in Balb/c mice and pigs.
The result showed the chimeric gene of PCV2-linker-PoIL-2 and its recombination expression plasmid were successfully constructed. The rCap-linker-PoIL-2 protein was expressed in the cytoplasm of COS-7 cells and displayed the specific immune responses for anti-PCV2 and anti-PoIL-2 antibodies, which indicated the rCap-linker-PoIL-2 protein possessing the duplex bio-activity of Cap protein and PoIL-2 protein. the mice have been immunited by rpcDNA3.1/PCV2-linker-PoIL-2 plasmid and have a clear anti-PCV2 antibodies,after the 3rd immunization,the immunity group antibody is the control group of 3.2 times, and significantly higher than the rpcDNA3.1/OFR2 plasmid control group; rpcDNA3.1/PCV2-linker-PoIL-2 and rpcDNA3.1/ORF2 plasmid can be induced in pigs production of anti-PCV2 immune response, rpcDNA3.1/PCV2-linker - PoIL-2, rpcDNA3.1/ORF2 plasmid immunohistochemistry antibody titers were empty vector control group of 12.46 times and 10.13 times that PoIL-2 in PCV2-linker-PoIL-2 chimeric gene played a very good immune-enhancing effect to rpcDNA3.1/ PCV2-linker-PoIL-2 plasmid immune in mice and pigs.
The successful construction of the PCV2-linker-PoIL-2 chimeric gene and its pcDNA3.1 eukaryotic expression plasmid rpcDNA3.1/PCV2-linker-PoIL-2; the recombinant expression plasmid have expressed biological activity fusion protein of Cap-and PoIL-2 in COS-7 cells .The recombinant plasmid can induce mice and pigs have a high anti-PCV2 antibody in mice and pigs and the induced antibody levels were significantly higher than the pcDNA3.1/OFR2 recombinant plasmid.This study for the PCV2 DNA vaccine research laid the foundation.
设计含双酶切位点的引物采用PCR方法扩增PCV2的ORF2全长基因并克隆入pGEM-T easy克隆载体;采用重叠延伸PCR(splicing by overlap extension-PCR,SOE-PCR)技术将PCV2的ORF2全长基因和猪白介素2成熟肽基因( PoIL-2 )构建成重组嵌合基因(PCV2-linker-PoIL-2)并克隆入pGEM-T easy克隆载体;筛选含PCV2-linker-PoIL-2、PCV2 ORF2的阳性克隆质粒经限制性内切酶双切后连接入经相同酶切的pcDNA3.1(+)真核表达载体中以构建重组表达质粒(rpcDNA3.1/ PCV2-linker-PoIL-2、rpcDNA3.1/ORF2)。筛选阳性重组表达质粒rpcDNA3.1/PCV2-linker-PoIL-2、rpcDNA3.1/ORF2瞬时转染COS-7细胞,分别采用PCV2抗原间接免疫荧光试验和PoIL-2蛋白ELISA试验方法检测COS-7细胞中表达的重组融合蛋白(rCap-linker-PoIL-2、rCap )免疫反应活性;提取rpcDNA3.1/PCV2-linker-PoIL-2质粒和rpcDNA3.1-OFR2质粒分别对Balb/c小鼠和猪进行三次免疫,采用PCV2抗体ELISA检测方法以评价其免疫效果。
结果表明:成功构建了单重组表达质粒pcDNA3.1/OFR2和PCV2-linker-PoIL-2嵌合基因及其共表达质粒rpcDNA3.1/PCV2-linker-PoIL-2 ; rpcDNA3.1/PCV2-linker-PoIL-2和rpcDNA3.1/OFR2质粒在COS-7细胞浆内进行了表达,表达的rCap-linker-PoIL-2蛋白可分别与抗PCV2和抗PoIL-2蛋白抗血清发生特异性免疫反应,表明rCap-linker-PoIL-2蛋白具有Cap蛋白和PoIL-2蛋白的双重免疫反应活性。rpcDNA3.1/PCV2-linker-PoIL-2质粒免疫小鼠后可诱导小鼠产生明显的抗PCV2抗体,第3次加强免疫后,免疫组抗体滴度最高是空白对照组的3.2倍,且显著高于rpcDNA3.1/OFR2质粒对照组; rpcDNA3.1/PCV2-linker-PoIL-2和rpcDNA3.1/ORF2质粒均可诱导猪体产生抗PCV2免疫应答,rpcDNA3.1/PCV2-linker-PoIL-2、rpcDNA3.1/ORF2质粒免疫组抗体滴度分别是空载体对照组的12.46倍、10.13倍,表明PCV2-linker-PoIL-2嵌合基因中的PoIL-2对rpcDNA3.1/PCV2-linker-PoIL-2质粒在小鼠体内的免疫起到了很好的免疫增强作用。
本研究成功构建了PCV2-linker-PoIL-2嵌合基因及其重组表达质粒pcDNA3.1/PCV2-linker-PoIL-2;重组表达质粒在COS-7细胞中表达了具有Cap蛋白和PoIL-2蛋白双重免疫反应活性的融合蛋白,免疫小鼠和猪后可诱导小鼠和猪产生高的抗PCV2抗体,且其诱导的抗体水平明显高于pcDNA3.1/OFR2重组质粒。本研究为PCV2的DNA疫苗研究奠定了基础。
Porcine circovirus type 2 (Porcine circovirus type 2, PCV2) are the main pathogens and trigger Postweaning multisystemic wasting syndrome.ORF1 and ORF2 are main open reading frames in PCV2,the ORF2 gene is an important antigen genes, the protein-coding (Cap protein), the existence of epitope in Cap epitope, with immunogenicity, so ORF2 gene is an objective gene which use establishing PCV2 specific serology and researching DNA vaccine of the PCV2. Current, no available PCV2 vaccine in domestic and foreign, the development of PCV2 DNA vaccine to control disease of PCV2 is of great significance.
The primers with double restriction site were designed and amplify ORF2 gene of PCV2.The ORF2 gene was cloned into pGEM-T easy cloning vector. The recombinant chimeric gene of PCV2-linker-PoIL-2 was constructed by PCV2 ORF2 gene linked porcine interleukin-2 (PoIL-2) mature peptide gene via a 15-amino acid glycine-rich linker [linker(,G4S)3] by SOE-PCR (splicing by overlap extension-PCR). The recombinant chimeric gene was cloned into pGEM-T Easy vector and subsequently into eukaryotic expression pcDNA3.1(+) vector. The positive recombinant expression plasmid rpcDNA3.1/PCV2-linker-PoIL-2 and r pcDNA3.1/ORF2 were transfected into COS-7 cells by lipofectamine. Indirect immunofluorescent assay (IFA) and porcine IL-2 ELISA assay was used to detect the bioactivities for PCV2 Cap protein and PoIL-2 protein of the expressed recombinant fusion protein (rCap-linker-PoIL-2 protein), respectively. The positive recombinant expression plasmid were extracted and inoculated Balb/c mice and pigs. The immune effects induced by rpcDNA3.1/PCV2-linker-PoIL-2 and rpcDNA3.1/OFR2 was evaluated by ELISA assay for detection PCV2 antibody in Balb/c mice and pigs.
The result showed the chimeric gene of PCV2-linker-PoIL-2 and its recombination expression plasmid were successfully constructed. The rCap-linker-PoIL-2 protein was expressed in the cytoplasm of COS-7 cells and displayed the specific immune responses for anti-PCV2 and anti-PoIL-2 antibodies, which indicated the rCap-linker-PoIL-2 protein possessing the duplex bio-activity of Cap protein and PoIL-2 protein. the mice have been immunited by rpcDNA3.1/PCV2-linker-PoIL-2 plasmid and have a clear anti-PCV2 antibodies,after the 3rd immunization,the immunity group antibody is the control group of 3.2 times, and significantly higher than the rpcDNA3.1/OFR2 plasmid control group; rpcDNA3.1/PCV2-linker-PoIL-2 and rpcDNA3.1/ORF2 plasmid can be induced in pigs production of anti-PCV2 immune response, rpcDNA3.1/PCV2-linker - PoIL-2, rpcDNA3.1/ORF2 plasmid immunohistochemistry antibody titers were empty vector control group of 12.46 times and 10.13 times that PoIL-2 in PCV2-linker-PoIL-2 chimeric gene played a very good immune-enhancing effect to rpcDNA3.1/ PCV2-linker-PoIL-2 plasmid immune in mice and pigs.
The successful construction of the PCV2-linker-PoIL-2 chimeric gene and its pcDNA3.1 eukaryotic expression plasmid rpcDNA3.1/PCV2-linker-PoIL-2; the recombinant expression plasmid have expressed biological activity fusion protein of Cap-and PoIL-2 in COS-7 cells .The recombinant plasmid can induce mice and pigs have a high anti-PCV2 antibody in mice and pigs and the induced antibody levels were significantly higher than the pcDNA3.1/OFR2 recombinant plasmid.This study for the PCV2 DNA vaccine research laid the foundation.
引文
[1]. Tischer I, Rasch R, Tochtermann G. Characterization of papovavirus-and picornavirus-like particles in permanent pig kidney cell lines[J]. Zbl Bakt Hyg, l Abt Orig A, 1974, 226: 153-167.
[2]. Tischer I, Gelderblom H, Vettermann W, Koch M A. A small porcine virus with circular single-stranded DNA[J]. Nature, 1982, 295: 64-66.
[3]. Islam MR,Johne R,Raue R,et al.,Sequence analysis of the full-length cloned DNA of a chicken anaemia virus(CAV)strain from Bangladesh:evidence for genetic grouping of CAV strains based on the deduced VP1 amino acid sequences[J].J Vet Med B Infect Dis Vet Public Health, 2002, 49(7):33-337.
[4]. Kiatipattanasakul-Banlunara W,Tantileartcharoen R,Katayama K, et al.,Psittacine beak and feather disease in three captive sulpHur-crested cockatoos(Cacatua galerita)inThailand[J].J Vet Med Sci, 2002, 64(6):527-529.
[5]. Niagro F D, Forsthoefel A N, Lawther R P, Kamalanathan L, Ritchie B W, Latimer K S, Lukert P D. Beak and ferther disease virus and porcine circovirus genomes: intermediates between thegeminiviruses and plant circoviruses[J]. Arch Irrol, 1998, 143:1723-1744.
[6]. Allan G M, McNeilly F, Cassidy J P, Reilly G A, Adair , Ellis W A, McNulty M S. Pathogenesis of porcinecircovirus: Experimental infections of colostrums deprived piglets and examination of pig foetal material[J]. Vet Microbiol, 1995, 44: 49-64.
[7]. Tischer I, Mields W, Wolff D, Vagt M, Griem W. Studies on epidemiology and pathogenicity of porcine circovirus[J]. Arch. Virol. 1986, 91: 271-276.
[8]. Clark E G. Post-weaning multisystemic wasting syndrome[J].Proc Am Assoc Swine Pract, 1997,28: 499-501.
[9]. Harding J C. Post-weaning multisystemic wasting syndrome (PMWS): preliminary epidemiology and clinical presentation. Swine Health Prod, 1998, 6: 249-254.
[10]. Allan G M, Meehan B, Todd D, Kennedy S, McNeilly F, Ellis J, Clark E C,Harding J, Espuna E,Botner A, Charreyre C. Novel porcine circoviruses from pigs with wasting disease syndromes[J].Vet Rec, 1998 a, 142: 467-468.
[11]. Kiupel M, Stevenson G W, Mittal S K, Clark E G, Haines D M. Circovirus-like viral associated disease in weaned pigs in lndiana[J]. Vet Pathol, 1998, 35: 303-307.
[12]. Nayar G P, Hamel A, Lin L. Detection and characterization of porcine circovirus associated with pastweaning multisystemic wasting syndrome in pigs[J]. Can Yet J, 1997,38: 385-386.
[13]. Daft B,Nordhausen R,Latimer K S,Niagro F D.Interstitial pneumonia and lymphadenophathy associated with circoviral infection in a six week-old pig[J]. Proc Am Assoc Vet Lab Diag,1996, 39: 32.
[14]. Segales J,Sitjar M,Domingo M,Ferro A,Latimer K S. Dee S,Del Pozo M,Noval R,Sacristan C,De las Heras A.First report of post-weaning multisysternic wasting syndrome in pigs in Spain[J].Vet Rec,1997, 141: 600-601.
[15]. Kennedy S, Allan G,McNeilly F, Adair B M, Hughes A, Spilane P. Porcine circovirus infection in Northern Ireland[J]. Vet Rec, 1998, 142: 495-496.
[16]. LeCann P, Albina E, Madec F, Cariolet R, Jestin A. Piglet wasting disease[J]. Vet Rec, 1997, 141:660.
[17]. Segales J, Piella J, Marco E, Mateu-de-Antonio E M, Espuna E, Domingo M. Porcine dermatitis and nephropathy syndrome in Spain[J]. Yet Rec, 1998, 142: 483-486.
[18]. Ellis J, Hassard L, Clark E, Harding J, Allan G, Willson P, Strokappe J, Martin K, McNeilly FMeehan B, Todd D, Haines D. Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome[J]. Can hetJ, 1998, 39: 44-51.
[19]. Nayar G P, Hamel A, Lin L. Detection and characterization of porcine circovirus associated with pastweaning multisystemic wasting syndrome in pigs[J]. Can Yet J, 1997,38: 385-386.
[20]. Allan G M, McNeilly F,Kennedy S, Daft B, Clarke E G, Ellis J A, Haines D M, Meehan B M, Adair B M. Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe[J]. J Yet Diagn Invest, 1998 b, 10: 3-10.
[21]. Allan G M, McNeilly F, Cassidy J P, Reilly G A, Adair B, Ellis W A, McNulty M S. Pathogenesis of porcinecircovirus: Experimental infections of colostrums deprived piglets and examination of pig foetal materia l [J]. Vet Microbiol, 1995, 44: 49-64.
[22]. Rosell C, Segales J, Plana-Duran J, Balasch M, Rodriguez-Arrioja G M, Kennedy S, Allan G M,McNeilly F, Latimer K S, Domingo M. Pathological, immunohistochemical, and in-situ hybridization studies of natural cases of postweaning multisystemic wasting syndrome (PMWS) in pigs[J]. J Comp Pathol, 1999, 120: 59-78.
[23]. Meehan B M, McNeilly F, Todd D, Kennedy S, Jewhurst V A, Ellis J, Hansard L E, Clark E G,Haines D M, Allan G M. Characterization of novel circovirus with wasting syndromes in pigs[J]. J Gen Yrol, 1998, 79: 2171-2179.
[24]. Allan G.M, Phenix K.V, Todd D, et al., Some biological and physico-chemical properties of porcine circovirus[J], J Vet Med.1994, 41:17-26.
[25]. Allan G M, Mackie D P, McNair J, et al. Production, preliminary characterisation and applications of monoclonal antibodies to porcine circovirus[J].Vet Immunol Immunopathol,1994, 43:357-371.
[26]. Tischer I,Peters D, Rasch R, et al.,Repl ication of poecine circovirus: induction byglucosamine and cell cycle dependence[J].Arch Virol,1987,96 (1-2):39-57.
[27]. Cheung AK, Bolin SR, Kinetics of porcine circovirus type 2 Replication[J]. Arch Virol. 2002,147(1):43-58.
[28]. Mankertz, A., F. Persson, et al.,Mapping and characterization of the origin of DNA replication of porcine circovirus[J] .ViroL,1997,71(3):2562-2566.
[29]. Mankertz J,H J Buhk, et al.,Transcription analysis of porcinecircovirus(PCV) [J].Virus Genes,1998,16(3):267-276.
[30]. Mankertz A; Persson F, Mankertz J, et al.Mapping and characterization of the origin of DNA replication of porcine circovinis[J].J Virol 1997, 32:2562-2566.
[31]. Meehan B M, Todd D, Creelan J L, et al.Investigation of the attenuation exhibited by a molecularly cloned chicken anemia virus isolate by utilizing a chimeric virus approach[J].J Virol,1997,71(11):8362-8367.
[32]. Hanley-Bowdoin L, Settlage S B, Orozco B M, et al.,Geminiviruses:models for plant DNA replication; transcription, and cell cycle regulation[J].Cirt Rev Biochem Mol Biol ,2000,35:105-140.
[33]. MankertzA, Hillenbrand B.Replieation of porcine circovirus type l requires two proteins encoded by the viral rep gene[J].Virology, 2001,279:429-438.
[34]. Steinfeldt T,Finsterbusch T,Mankertz A. Rep and Rep' protein of porcinecircovirus type 1 hind to the origin of replication in vitro[J].Virology,2001,291(1):152-160.
[35]. Liu Q, Tikoo S K, Babiuk L A. Nuclear localization of the ORF2 protein encoded by porcine circovirus type 2[J].Virology, 2001, 285(1): 91-98.
[36]. Annette M, Bettina M, Tobias S, Cornelia S, and Timm R .New Reporter Gene-Based Replication Assay Reveals Exchangeability of Replication hactc:rs of Porcine Circovirus Type1 and 2[J]. J Virol. 2003, 77(18): 9885-9893.
[37]. Mankertz A, Hillenbrand B.Replication of Porcine Circovirus Type 1 Requires Two Proteins Encoded by the Viral Rep Gene[J].Virology, 2001, 279(2): 429-438.
[38]. Steinfeldt T, Finsterbusch T, Mandertz A.Rep and Rep' protein of porcine circovirus type 1 bind to the origin of replication in vitro[J].Virology, 2001, 91(1): 152-160.
[39]. Andew K, Cheung.Detection of ramant nucleotide reversion at the origin of DNA replication of porcine circovirus type 1[J].Virology, 2005, 333: 22-30.
[40]. Andew K, Cheung.The essential and nonessential transcription units for viral protein synthesis and DNA replication of porcine circovirus type 2[J]. Virology, 2003,301: 452-459.
[41]. Truong C, Mahe D, Blanchard P, Madec F, 3estin A, and Albina E.Identification of an immunorelevant ORF2 epitope from porcine circovirus type 2 as a serological marker forexperimental and natural infection[J].Arch Virol, 2001, 146: 1197-1211.
[42]. Porntippa N, Lgor M,Steven R.Open reading frame 2 porcine circovirus type 2 encodes a major capsid protein[J].J Gen Virol.2000. 81: 2281-2287.
[43]. Hamel A L, Lin L L, Nayar G P. Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs[J]. J Virol, 1998, 72: 5262- 5267.
[44].曹胜波,孙敏轩,刘学芹等.嵌合猪圆环病毒PCV1-2的构建及其感染性初步鉴定[J].微生物学报.2006.46 (1) :158-161
[45].刘长明,陆月华,张超范等.猪圆环病毒2型重组Cap蛋白在昆虫杆状病毒中的表达[J].中国预防兽医学报.2005,27(6):479-486.
[46].王先炜,姜平,李玉峰等.猪圆环病毒Ⅱ型Cap蛋白重组腺病毒的构建与鉴定[J].中国病毒学.2006,21(1):29-33.
[47].宋云峰,肖少波,金梅林等.猪2型圆环病毒核酸疫苗免疫效应研究[J].畜牧兽医学报。2005,36(10):1049-1054.
[48].金宁一,秦晓冰,郑敏等.猪2型圆环病毒核酸疫苗的接种Balb/c小鼠实验免疫研究[J].中国生物工程杂志.2005,25(7):76-79.
[49]. Blanchard P , Mahe D , Cariolet R ,et al. Protection of swine against post - weaning multisystemic wasting syndrome(PMWS)by porcine circovirus type 2 (PCV2) proteins[J]. Vaccine .2003,21 (31):4565-4575.
[50]. Kamst rup S , Barfoed A M , Frimann T H ,et al. Immunisation against PCV2 structural protein by DNA vaccination of mice[J].Vaccine,2004,22 (11-12):1358-1361.
[51]. Harding J C. Post-weaning multisystemic wasting syndrome (PMWS): preliminary epidemiology and clinical presentation[J]. Swine Health Prod, 1998, 6: 249-254.
[52]. Cottrell T, Friendship R M, Dewey C E, 3osephson G, Allan G M, McNeilly F, Walker I. Epidemiology of postweaning multisystemic wasting syndrome in Ontario [J]. Proc Am Assoc Swine Pract, 1999, 389-390.
[53]. Ellis J, Hassard L, Clark E, Harding J, Allan G, Willson P, Strokappe J, Martin K, McNeilly F,Meehan B, Todd D, Haines D. Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome[J]. Can VetJ, 1998, 39: 44-51.
[54]. Edwards S, Sands J J. Evidence of circovirus infection in Britishpigs[J]. Vet Rec, 1994, 134: 680-681.
[55]. Magar R, Muller P, Larochelle R. Retrospective serological survey of antibodies to porcine circovirus type 1 and type 2[J]. Can J Yet Res, 2000 a, 64 (3): 184-186.
[56].朗洪武,张广川,吴发权.断奶猪多系统衰弱综合征血清抗体检测[J].中国兽医科技,2000, 30(3): 3-5
[57]. Larochelle R, Antaya M, Moran M. Typing of porcine circovirus in clinical specimense by multiplex PCR[J]. J Trrol Meth, 1999 a, 80: 69-75.
[58]. AIlan G M, Kennedy S, McNeilly F, Foster J C, Ellis J A, S. TCrakowka J, Meehan B M, Adair B M. Experimental reproduction of severe wasting disease bco-infection of pigs with porcine circovirus and porcine parvovirus[J]. J Comp Pathol, 1999, 121: 1-11.
[59]. West K H, Bystrom J M, Wojnarowicz C. Mycoardits and abortion associated with intrauterine infection of sows with porcine circovirus 2[J]. J Vet Diagn Invest, 1999, 11: 530-532.
[60]. Allan G M, McNeilly F, McNair I. Absence of evidence for porcine circovirus type 2 in cattle and humans, and lack of seroconversion or lesions in experimentally infected sheep[J]. Arch Yzrol,2000 d, 145 (4): 853-857.
[61]. Jake W .Where circovirus is suspected[J]. Pig Inter, 2000, 4: 19-22.
[62]. LeCann P, Albina E, Madec F, Cariolet R, Jestin A. Piglet wasting disease[J]. Vet Rec, 1997, 141:660.
[63]. Allan G M, McNeilly F, Ellis J, Botner A., McCullough K, Nauwynck H.,Kennedy S.PMWS: experimental model and co-infections[J]. Vet Microbiol, 2004, 96 (2): 165-168.
[64]. Mori M, Sato K Akachi S. Retrospective study of porcine circovirus 2 infection in Japan: seven cases in 1989[J]. Vet Pathol, 2000, 37 (6): 667-669.
[65]. Sato K, Shibahara T, Ishikawa Y. Evidence of porcine circovirus infection in pigs with wasting disease syndrome from 1985 to 1999 in Hokkaido, Japan[J]. J Vet Med Sci, 2000, 62 (6): 627-33.
[66]. Kenndey S,Moffett D,McNeilly F,etal.Reproduction of lesions of post weaning multi systemic wasting syndrome by infection of conventional pigs with porcine circovirus type 2 alone or in combination with porcine parvovirus[J].J Comp Pathol, 2000, 122:9-24.
[67]. Allan G M, John A. Porcine circovirus:a review[J].J Vet Diagn nvest, 2000,12:3-14.
[68]. Onuki A, ABEK, Togashi K, et al. Detection of porcine circovirus from lesions of pig with wasting Disease in Japan[J].J Vet Med Sci,1999,61(10):1119-1123.
[69].吴德铭,曹永长,毕英佐等.猪圆环病毒研究进展[J].中国预防兽医学报,2004, 26(1) :76-77
[70].崔尚金,李曦,符芳等.猪断奶后多系统衰竭综合征诊断方法及标准的建立[J].中国兽医科技,2003,33(11):68-72.
[71].高骏,胡建华,孙凤萍等.猪圆环病毒研究进展.上海交通大学学报(农业科学版),2005,(23)1:107-110.
[72]. Kim J, Han DU, Choi C, et al., Simultaneous detection and differentiation between porcinecircovirus and porcine parvovirus in boar semen by multiplex seminested polymerase chain reaction[J], J Vet Med Sci. 2003, 65(6):741-744.
[73]. Larochelle R, Antaya M, Morin M, et al.,Typing of porcine clinical specimens by multiplex PCR[J], J Virol Methods,1999, 80(1):69-75.
[74]. Ouardani M, Wilson L, Jette R, et aL, Mul如lex PCR far detection and of porcine circoviruses[J].J Clin Microbiol,1999,37(12):3917-24.
[75]. Yang JS, Song DS, Kim SY, et al.,Detection of porcine circovirus type 2 in feces of pigs with or yvithout enteric disease by polymerise chain reaction[J], J Vet Diagn invest ,2003,15(4):369-73.
[76]. Choi C. Chae C.Coloealization of porcine Reproductive and respiratory syndrome virus and porcine circovirus 2 in porcine dermatitis and nephrology syndrome by double-labeling technique[J], Vet Pathol. 2001, 38(4):436-441.
[77]. Sirinarumitr T, Sorden SD, Morozov I, et al., Double in situ hybridization for simultaneous detection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus (PCV) [J], J Vet Diagn Invest,2001,13(1):68-71.
[78]. Kim J, Chae C. Simultaneous detection of porcine circovirus 2 and parvovirus in naturally and experimentally coinfected pigs by double in situ hybridization[J], J Vet Diagn Invest ,2002, 14(3):236-240.
[79]. Porntippa Nawagitgul, Perry A. Harms, Igor Morozov, et al.,Modified indirect porcine circovirus (PCV) type 2-based and recombinant capsid protein (ORF2)-based enzyme-linked immunosorbent assays for detection of antibodies to PCV[J],Clin Diagn Lab Immure,2002,9(1):57-68.
[80].王涛,王子轼,彭德旺.猪圆环病毒感染与断奶仔猪多系统衰竭综合征[J].中国兽医学报,2002, 22 (3): 314-316
[81].陈曦,刘焕奇,魏萍.一种新的病毒病一断奶仔猪多系统衰竭综合征.黑龙江畜牧兽医.2002,9: 49-50.
[82].踞春梅.猪2型圆环病毒ELISA诊断方法及猪2型圆环病毒-伪狂犬病病毒二价基因工程疫苗研究(博士论文).2005,39-40.
[83]. Broom J, Heys S D, Whiting P H, Park K q Strachan A, Rothnie I, Franks C R, Eremin O Interleukin 2 therapy in cancer: identification of responders. Br J Cancer, 1992, 66:1185-1187
[84]. Morgan D A,Ruscctti,F W, Gallo R C. Selective in vitro growth of T lymphocytes from normal human bone marrow[J]s. Science, 1976, 193:1007-1008.
[85]. Robb R I. Tell growth factor receptors,quantitation, specificity and biological relevance[J]. J ExP Med, 1981, 154: 1455～1457.
[86].王瑜,王健等.白细胞介素-2受体结构及功能研究进展[J].中国现代医学杂志,2006,16(2):24 1-245.
[87]. Hughes-Fulford M, Sugano E, Schopper T, et al., Early immune response and regulation of IL-2 receptor subunits[J].Cellular Signalling, 2005, 17: 1111-1124.
[88]. Leonard W J. A monoclonal antibody that appears to recognize the receptor for human T cell growth factor: partial characterization of the receptor[J] . Nature, 1982, 300: 287-296.
[89]. Smith K A. Production and characterization of monoclonal antibodies to human intereukin2: strategy and tactics[J]. J Tmmunol. 1983. 131: 1808-1815.
[90]. CantrellD A,Smith K A .Transient expression of intereukin2 receptor[J].J Exp Med. 1983, 18:1895-1911.
[91]. Cox G W Mathieson B J,Giardina S L et al.,Characterization of IL-2 receptor expression and function on murine macrophages[J].J Immunol. 1990 145(6):1719-1726.
[92]. TaniguchiT,MinamiY.The IL-2/IL-2 receptor system:acurrent overview[J].Cell. 1993, 73 (1):5-8.
[93]. Ellery J M, Nicholls P J. Aiternate signaling pathways from the interleukin-2 receptor[J]. Cytokine & Growth Factor Reviews, 2002, 13:27-40.
[94]. Good M F, Pombo D, Lunde M N, Maloy W L, Halenbeck R, Koths K, Miller L H, Berzofsky J A.Recombinant human IL-2 overcomes genetic nonresponsivenes to malaria sporozoite peptides.Correlation of effect with biologic activity of IL-2[J]. J Immunol, 1988, 141:972-977.
[95]. Hyun B H, Song J Y, Lee J B, Cha S H, Park J H, Lee J O, An D J, Kang Y W. Cloning and expression of porcine interleukin-2 gene in Escherichia coli.RDA J Vet Sci, 1997, 39:42-50
[96]. Heys S D, Franks C R, Eremin O. lnterleukin 2 therapy: current role in surgical oncological practice[J]. Br J Surg, 1993, 80:155-162.
[97]. Fehniger T A, Cooper M A, Caligiuri M A. Interleukin-2 and interleukin-15: immunotherapy for cancer[J]. Cytokine&Growth Factor Reviews, 2002, 13:169-183.
[98]. Figlin R A. Renal cell carcinoma: current status and future plans[J]. Cancer J Sci Am, 2000, 6:45-51.
[99]. Broom J, Heys S D, Whiting P H, Park K G, Strachan A, Rothnie I, Franks C R, Eremin O Interleukin 2 therapy in cancer: identification of responders[J]. Br J Cancer, 1992, 66:1185-1187.
[100]. Rosenberg S A. Progress in human tumour immunology and immunotherapy[J]. Nature, 2001,4(11):380-384.
[101]. Prentice H G,Macdonald I D, Hamon M D. The role of immunotherapy in the treatment ofacute myeloblastic leukemia: from allogeneic bone marrow transplantation to the application of interleukin 2[J].Cancer Treat Research, 1993, 64:121-134.
[102]. Hazama M A,Mayumi Aono N,Asakawa S,et al.Vaccine,1993, 629-636.
[103]. Reddy DN, Reddy PG.Immuopotentiation of bovine respiratory disease virus vaccines by interleukinl b and interleukin2[J]. Veterinary Immuology and Immuopathology,1993,37:25.
[104]. Hughs HRA, Campos M. Immuopotentiaton of bovine herpevirus subunit vaccinationation by interleukin2[J]. Immuology,1991.74:461.
[105].李宏梅,李祥瑞.白细胞介素-2的研究进展及应用[J].山东畜牧兽医:38-39
[106]. Nunberg J H, Doyle M V, York S et al, Interleukine 2 acts as adjuvant to increase the potency of inactivated rabies virus vaccine[J], Proc Natl Acad SCI. 1989,86(6):4240-4243.
[107]. Campos M, Rossi CR. Characterization and activation requirement of bovine lymphocytes acquiring cytotoxicativity after interleukin2 treatment[J]. Veterinary Irnmuology and Immuopathology,1992;32:205.
[108]. Brown WC, Grab BJ. Biological and biochemical characterization of bovine interleukin2 studies with cloned bovine T cells[J].The Journal of Immuology,1985,35(5):3184.
[109]. Arunachalam B,Subba Raomv. Bovine interleukin2 biochemical and biological charactrization[J].Veterinary Immuology and Immupathology,1989;23:377.
[110]. Chow Y H, Huang W L, Chi W et al, Improvement of hepatitis B virus DNA vaccine by plasmids coexpressing hepatitis B surface antigen and Interleukine 2[J], J of Virology, 1997, 1:169-178.
[111]. Geissler M, Gesien A, Wands J R. Inhibitory effects of chronic ethanolconsuption on cellular immune response to hepatitis C virus core protein are reversed by genetic immunizations augmented with cytokine expression plasmid[J].J Immunol,1997, 159: 5107-5113.
[112]. Ramshaw IA, Woodsworth M.The in vitro culture of spleen cells from mice expressing humotal immunity leads to the development of T cells that mediate delayed-type hypersensitivity[J]. Cell Immunol.1981,15,57(2):468-476.
[113]. Flexner C, Hugin A, Moss B.Prevention of vaccin in virus infection in immunodeficient mice by vector-directed IL-2 expression[J]. Nature, 1987,330:259-261.
[114].姜永厚,陈奖励,宋秀龙等。鸡新城疫病毒F基因和鸡IL-2重组疫苗的构建[J]中国预防兽医学报,2001, (2): 81-83.
[115]. Hazama M, Mayumi-Aono A, Asakawa N, Kuroda S, Hinuma S, Fujisawa Y. Adjuvant-independent enhanced immune responses to recombinant herpes simplex virus type 1 glycoprotein D by fusion with biologically active interleukin-2[J].Vaccine, 1993,11(6):629-636.
[116] Tischer,Geldblon H, Vettermann W, et al. A very small porcine virus with cireular single-stranded DNA[J].Nature, 1982,(295):64-66
[117] Tischer I ,Rasch R ,Tochcermana G. Characterizationof papovavirus and picornavirus like particle in Permanent pig kidney cell lines [J ] . ZblBakt Hyg A ,I Abt Ovig A ,1974 ,326 :153-167.
[118] Post-weaning multisystemic Waning multisystemic syndrome [J ] . Proc Am Assoc Swine Pract ,1997 ,28 :499-501.
[119]黄平等.预防输血传播的感染[J ] .海峡预防医学杂志,2000 ,6(5) :19-21.
[120] Blanchard P,Mahe D,Cariolet R, et al. Protection of swine against post-weaning multisystemic wasting syndrome(PMWS) by porcine circovirus type (PCV2) proteins[J]. Vaccine,2003,21:4565-4575.
[121] Chow YH , Chiang BL , Lee YL , et al.Development of Th1 and Th2 populations and the nature of immune responses to hepatitis B virus DNA vaccines can be modulated by codelivery of various cytokine genes [J] .Immunol , 1998 , 160 : 1320–1329.
[122] Kim JJ,Ayyavoo V,Bagarazzi ML, et al.In vivo engineering of a cellular immune response by coadministration of IL-2 expression vector with a DNA immunogen. [J].Immunol ,1997, 158:816-826.
[123]. Kamstrup S,Barfoed A M,Frimann TH, et al.,Immunisation against PCV2 structural protein by DNA vaccination of mice[J].Vaccine,2004,22:1358-1361.
[124]. Blanchard P,Mahe D,Carioet R,et al.Protection of swine against post-weaning multisystemic wasting syndrome(PMWS) by porcine circovirus type 2 (PCV2) proteins[J]. Vaccine,2003,21:4565-4575.
[125]. Kim JJ,Yang JS,Manson KH, et al.,.Modulation of antigen-specific cellular immune renponses to DNA vaccination in rhesus macaques through the use of IL-2,IFN-γor IL-4 gene adjuvants[J].Vaccine,2001,19:2496-2505.
[126]. McNeilly F, Allan G M, Foster J C, Adair B M, McNulty M S and Pollock J. Effect of porcine circovirus infection on porcine alveolar macrophage function[J]. Veterinary Immunology and Immunopathology, 1996, 49(4): 295~306.
[127]. Segales J, Domingo M, Chianini F, Majo N, Dominguez J, Darwich L and Mateu E. Immunosuppression in postweaning multisystemic wasting syndrome affected pigs[J]. Veterinary Microbiology, 2004, 98(2): 151~158.
[128]. Premenko-Lanier M, Rota P A, Rhodes G., Verhoeven D, Barouch D H, Lerche N W, Letvin N L, Bellini W J and McChesney M B. DNA vaccination of infants in the presence ofmaternal antibody: a measles model in the primate[J]. Virology, 2003, 307(1): 67~75.
[129].王镜岩,朱圣庚,徐长法.生物化学[M](第三版).北京高教出版社, 2002.
[130].萨姆布鲁克J,弗里奇E F,曼尼阿蒂斯T.金冬雁,黎孟枫,等译.分子克隆指南[M].北京:科学出版社,1997
[2]. Tischer I, Gelderblom H, Vettermann W, Koch M A. A small porcine virus with circular single-stranded DNA[J]. Nature, 1982, 295: 64-66.
[3]. Islam MR,Johne R,Raue R,et al.,Sequence analysis of the full-length cloned DNA of a chicken anaemia virus(CAV)strain from Bangladesh:evidence for genetic grouping of CAV strains based on the deduced VP1 amino acid sequences[J].J Vet Med B Infect Dis Vet Public Health, 2002, 49(7):33-337.
[4]. Kiatipattanasakul-Banlunara W,Tantileartcharoen R,Katayama K, et al.,Psittacine beak and feather disease in three captive sulpHur-crested cockatoos(Cacatua galerita)inThailand[J].J Vet Med Sci, 2002, 64(6):527-529.
[5]. Niagro F D, Forsthoefel A N, Lawther R P, Kamalanathan L, Ritchie B W, Latimer K S, Lukert P D. Beak and ferther disease virus and porcine circovirus genomes: intermediates between thegeminiviruses and plant circoviruses[J]. Arch Irrol, 1998, 143:1723-1744.
[6]. Allan G M, McNeilly F, Cassidy J P, Reilly G A, Adair , Ellis W A, McNulty M S. Pathogenesis of porcinecircovirus: Experimental infections of colostrums deprived piglets and examination of pig foetal material[J]. Vet Microbiol, 1995, 44: 49-64.
[7]. Tischer I, Mields W, Wolff D, Vagt M, Griem W. Studies on epidemiology and pathogenicity of porcine circovirus[J]. Arch. Virol. 1986, 91: 271-276.
[8]. Clark E G. Post-weaning multisystemic wasting syndrome[J].Proc Am Assoc Swine Pract, 1997,28: 499-501.
[9]. Harding J C. Post-weaning multisystemic wasting syndrome (PMWS): preliminary epidemiology and clinical presentation. Swine Health Prod, 1998, 6: 249-254.
[10]. Allan G M, Meehan B, Todd D, Kennedy S, McNeilly F, Ellis J, Clark E C,Harding J, Espuna E,Botner A, Charreyre C. Novel porcine circoviruses from pigs with wasting disease syndromes[J].Vet Rec, 1998 a, 142: 467-468.
[11]. Kiupel M, Stevenson G W, Mittal S K, Clark E G, Haines D M. Circovirus-like viral associated disease in weaned pigs in lndiana[J]. Vet Pathol, 1998, 35: 303-307.
[12]. Nayar G P, Hamel A, Lin L. Detection and characterization of porcine circovirus associated with pastweaning multisystemic wasting syndrome in pigs[J]. Can Yet J, 1997,38: 385-386.
[13]. Daft B,Nordhausen R,Latimer K S,Niagro F D.Interstitial pneumonia and lymphadenophathy associated with circoviral infection in a six week-old pig[J]. Proc Am Assoc Vet Lab Diag,1996, 39: 32.
[14]. Segales J,Sitjar M,Domingo M,Ferro A,Latimer K S. Dee S,Del Pozo M,Noval R,Sacristan C,De las Heras A.First report of post-weaning multisysternic wasting syndrome in pigs in Spain[J].Vet Rec,1997, 141: 600-601.
[15]. Kennedy S, Allan G,McNeilly F, Adair B M, Hughes A, Spilane P. Porcine circovirus infection in Northern Ireland[J]. Vet Rec, 1998, 142: 495-496.
[16]. LeCann P, Albina E, Madec F, Cariolet R, Jestin A. Piglet wasting disease[J]. Vet Rec, 1997, 141:660.
[17]. Segales J, Piella J, Marco E, Mateu-de-Antonio E M, Espuna E, Domingo M. Porcine dermatitis and nephropathy syndrome in Spain[J]. Yet Rec, 1998, 142: 483-486.
[18]. Ellis J, Hassard L, Clark E, Harding J, Allan G, Willson P, Strokappe J, Martin K, McNeilly FMeehan B, Todd D, Haines D. Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome[J]. Can hetJ, 1998, 39: 44-51.
[19]. Nayar G P, Hamel A, Lin L. Detection and characterization of porcine circovirus associated with pastweaning multisystemic wasting syndrome in pigs[J]. Can Yet J, 1997,38: 385-386.
[20]. Allan G M, McNeilly F,Kennedy S, Daft B, Clarke E G, Ellis J A, Haines D M, Meehan B M, Adair B M. Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe[J]. J Yet Diagn Invest, 1998 b, 10: 3-10.
[21]. Allan G M, McNeilly F, Cassidy J P, Reilly G A, Adair B, Ellis W A, McNulty M S. Pathogenesis of porcinecircovirus: Experimental infections of colostrums deprived piglets and examination of pig foetal materia l [J]. Vet Microbiol, 1995, 44: 49-64.
[22]. Rosell C, Segales J, Plana-Duran J, Balasch M, Rodriguez-Arrioja G M, Kennedy S, Allan G M,McNeilly F, Latimer K S, Domingo M. Pathological, immunohistochemical, and in-situ hybridization studies of natural cases of postweaning multisystemic wasting syndrome (PMWS) in pigs[J]. J Comp Pathol, 1999, 120: 59-78.
[23]. Meehan B M, McNeilly F, Todd D, Kennedy S, Jewhurst V A, Ellis J, Hansard L E, Clark E G,Haines D M, Allan G M. Characterization of novel circovirus with wasting syndromes in pigs[J]. J Gen Yrol, 1998, 79: 2171-2179.
[24]. Allan G.M, Phenix K.V, Todd D, et al., Some biological and physico-chemical properties of porcine circovirus[J], J Vet Med.1994, 41:17-26.
[25]. Allan G M, Mackie D P, McNair J, et al. Production, preliminary characterisation and applications of monoclonal antibodies to porcine circovirus[J].Vet Immunol Immunopathol,1994, 43:357-371.
[26]. Tischer I,Peters D, Rasch R, et al.,Repl ication of poecine circovirus: induction byglucosamine and cell cycle dependence[J].Arch Virol,1987,96 (1-2):39-57.
[27]. Cheung AK, Bolin SR, Kinetics of porcine circovirus type 2 Replication[J]. Arch Virol. 2002,147(1):43-58.
[28]. Mankertz, A., F. Persson, et al.,Mapping and characterization of the origin of DNA replication of porcine circovirus[J] .ViroL,1997,71(3):2562-2566.
[29]. Mankertz J,H J Buhk, et al.,Transcription analysis of porcinecircovirus(PCV) [J].Virus Genes,1998,16(3):267-276.
[30]. Mankertz A; Persson F, Mankertz J, et al.Mapping and characterization of the origin of DNA replication of porcine circovinis[J].J Virol 1997, 32:2562-2566.
[31]. Meehan B M, Todd D, Creelan J L, et al.Investigation of the attenuation exhibited by a molecularly cloned chicken anemia virus isolate by utilizing a chimeric virus approach[J].J Virol,1997,71(11):8362-8367.
[32]. Hanley-Bowdoin L, Settlage S B, Orozco B M, et al.,Geminiviruses:models for plant DNA replication; transcription, and cell cycle regulation[J].Cirt Rev Biochem Mol Biol ,2000,35:105-140.
[33]. MankertzA, Hillenbrand B.Replieation of porcine circovirus type l requires two proteins encoded by the viral rep gene[J].Virology, 2001,279:429-438.
[34]. Steinfeldt T,Finsterbusch T,Mankertz A. Rep and Rep' protein of porcinecircovirus type 1 hind to the origin of replication in vitro[J].Virology,2001,291(1):152-160.
[35]. Liu Q, Tikoo S K, Babiuk L A. Nuclear localization of the ORF2 protein encoded by porcine circovirus type 2[J].Virology, 2001, 285(1): 91-98.
[36]. Annette M, Bettina M, Tobias S, Cornelia S, and Timm R .New Reporter Gene-Based Replication Assay Reveals Exchangeability of Replication hactc:rs of Porcine Circovirus Type1 and 2[J]. J Virol. 2003, 77(18): 9885-9893.
[37]. Mankertz A, Hillenbrand B.Replication of Porcine Circovirus Type 1 Requires Two Proteins Encoded by the Viral Rep Gene[J].Virology, 2001, 279(2): 429-438.
[38]. Steinfeldt T, Finsterbusch T, Mandertz A.Rep and Rep' protein of porcine circovirus type 1 bind to the origin of replication in vitro[J].Virology, 2001, 91(1): 152-160.
[39]. Andew K, Cheung.Detection of ramant nucleotide reversion at the origin of DNA replication of porcine circovirus type 1[J].Virology, 2005, 333: 22-30.
[40]. Andew K, Cheung.The essential and nonessential transcription units for viral protein synthesis and DNA replication of porcine circovirus type 2[J]. Virology, 2003,301: 452-459.
[41]. Truong C, Mahe D, Blanchard P, Madec F, 3estin A, and Albina E.Identification of an immunorelevant ORF2 epitope from porcine circovirus type 2 as a serological marker forexperimental and natural infection[J].Arch Virol, 2001, 146: 1197-1211.
[42]. Porntippa N, Lgor M,Steven R.Open reading frame 2 porcine circovirus type 2 encodes a major capsid protein[J].J Gen Virol.2000. 81: 2281-2287.
[43]. Hamel A L, Lin L L, Nayar G P. Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs[J]. J Virol, 1998, 72: 5262- 5267.
[44].曹胜波,孙敏轩,刘学芹等.嵌合猪圆环病毒PCV1-2的构建及其感染性初步鉴定[J].微生物学报.2006.46 (1) :158-161
[45].刘长明,陆月华,张超范等.猪圆环病毒2型重组Cap蛋白在昆虫杆状病毒中的表达[J].中国预防兽医学报.2005,27(6):479-486.
[46].王先炜,姜平,李玉峰等.猪圆环病毒Ⅱ型Cap蛋白重组腺病毒的构建与鉴定[J].中国病毒学.2006,21(1):29-33.
[47].宋云峰,肖少波,金梅林等.猪2型圆环病毒核酸疫苗免疫效应研究[J].畜牧兽医学报。2005,36(10):1049-1054.
[48].金宁一,秦晓冰,郑敏等.猪2型圆环病毒核酸疫苗的接种Balb/c小鼠实验免疫研究[J].中国生物工程杂志.2005,25(7):76-79.
[49]. Blanchard P , Mahe D , Cariolet R ,et al. Protection of swine against post - weaning multisystemic wasting syndrome(PMWS)by porcine circovirus type 2 (PCV2) proteins[J]. Vaccine .2003,21 (31):4565-4575.
[50]. Kamst rup S , Barfoed A M , Frimann T H ,et al. Immunisation against PCV2 structural protein by DNA vaccination of mice[J].Vaccine,2004,22 (11-12):1358-1361.
[51]. Harding J C. Post-weaning multisystemic wasting syndrome (PMWS): preliminary epidemiology and clinical presentation[J]. Swine Health Prod, 1998, 6: 249-254.
[52]. Cottrell T, Friendship R M, Dewey C E, 3osephson G, Allan G M, McNeilly F, Walker I. Epidemiology of postweaning multisystemic wasting syndrome in Ontario [J]. Proc Am Assoc Swine Pract, 1999, 389-390.
[53]. Ellis J, Hassard L, Clark E, Harding J, Allan G, Willson P, Strokappe J, Martin K, McNeilly F,Meehan B, Todd D, Haines D. Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome[J]. Can VetJ, 1998, 39: 44-51.
[54]. Edwards S, Sands J J. Evidence of circovirus infection in Britishpigs[J]. Vet Rec, 1994, 134: 680-681.
[55]. Magar R, Muller P, Larochelle R. Retrospective serological survey of antibodies to porcine circovirus type 1 and type 2[J]. Can J Yet Res, 2000 a, 64 (3): 184-186.
[56].朗洪武,张广川,吴发权.断奶猪多系统衰弱综合征血清抗体检测[J].中国兽医科技,2000, 30(3): 3-5
[57]. Larochelle R, Antaya M, Moran M. Typing of porcine circovirus in clinical specimense by multiplex PCR[J]. J Trrol Meth, 1999 a, 80: 69-75.
[58]. AIlan G M, Kennedy S, McNeilly F, Foster J C, Ellis J A, S. TCrakowka J, Meehan B M, Adair B M. Experimental reproduction of severe wasting disease bco-infection of pigs with porcine circovirus and porcine parvovirus[J]. J Comp Pathol, 1999, 121: 1-11.
[59]. West K H, Bystrom J M, Wojnarowicz C. Mycoardits and abortion associated with intrauterine infection of sows with porcine circovirus 2[J]. J Vet Diagn Invest, 1999, 11: 530-532.
[60]. Allan G M, McNeilly F, McNair I. Absence of evidence for porcine circovirus type 2 in cattle and humans, and lack of seroconversion or lesions in experimentally infected sheep[J]. Arch Yzrol,2000 d, 145 (4): 853-857.
[61]. Jake W .Where circovirus is suspected[J]. Pig Inter, 2000, 4: 19-22.
[62]. LeCann P, Albina E, Madec F, Cariolet R, Jestin A. Piglet wasting disease[J]. Vet Rec, 1997, 141:660.
[63]. Allan G M, McNeilly F, Ellis J, Botner A., McCullough K, Nauwynck H.,Kennedy S.PMWS: experimental model and co-infections[J]. Vet Microbiol, 2004, 96 (2): 165-168.
[64]. Mori M, Sato K Akachi S. Retrospective study of porcine circovirus 2 infection in Japan: seven cases in 1989[J]. Vet Pathol, 2000, 37 (6): 667-669.
[65]. Sato K, Shibahara T, Ishikawa Y. Evidence of porcine circovirus infection in pigs with wasting disease syndrome from 1985 to 1999 in Hokkaido, Japan[J]. J Vet Med Sci, 2000, 62 (6): 627-33.
[66]. Kenndey S,Moffett D,McNeilly F,etal.Reproduction of lesions of post weaning multi systemic wasting syndrome by infection of conventional pigs with porcine circovirus type 2 alone or in combination with porcine parvovirus[J].J Comp Pathol, 2000, 122:9-24.
[67]. Allan G M, John A. Porcine circovirus:a review[J].J Vet Diagn nvest, 2000,12:3-14.
[68]. Onuki A, ABEK, Togashi K, et al. Detection of porcine circovirus from lesions of pig with wasting Disease in Japan[J].J Vet Med Sci,1999,61(10):1119-1123.
[69].吴德铭,曹永长,毕英佐等.猪圆环病毒研究进展[J].中国预防兽医学报,2004, 26(1) :76-77
[70].崔尚金,李曦,符芳等.猪断奶后多系统衰竭综合征诊断方法及标准的建立[J].中国兽医科技,2003,33(11):68-72.
[71].高骏,胡建华,孙凤萍等.猪圆环病毒研究进展.上海交通大学学报(农业科学版),2005,(23)1:107-110.
[72]. Kim J, Han DU, Choi C, et al., Simultaneous detection and differentiation between porcinecircovirus and porcine parvovirus in boar semen by multiplex seminested polymerase chain reaction[J], J Vet Med Sci. 2003, 65(6):741-744.
[73]. Larochelle R, Antaya M, Morin M, et al.,Typing of porcine clinical specimens by multiplex PCR[J], J Virol Methods,1999, 80(1):69-75.
[74]. Ouardani M, Wilson L, Jette R, et aL, Mul如lex PCR far detection and of porcine circoviruses[J].J Clin Microbiol,1999,37(12):3917-24.
[75]. Yang JS, Song DS, Kim SY, et al.,Detection of porcine circovirus type 2 in feces of pigs with or yvithout enteric disease by polymerise chain reaction[J], J Vet Diagn invest ,2003,15(4):369-73.
[76]. Choi C. Chae C.Coloealization of porcine Reproductive and respiratory syndrome virus and porcine circovirus 2 in porcine dermatitis and nephrology syndrome by double-labeling technique[J], Vet Pathol. 2001, 38(4):436-441.
[77]. Sirinarumitr T, Sorden SD, Morozov I, et al., Double in situ hybridization for simultaneous detection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus (PCV) [J], J Vet Diagn Invest,2001,13(1):68-71.
[78]. Kim J, Chae C. Simultaneous detection of porcine circovirus 2 and parvovirus in naturally and experimentally coinfected pigs by double in situ hybridization[J], J Vet Diagn Invest ,2002, 14(3):236-240.
[79]. Porntippa Nawagitgul, Perry A. Harms, Igor Morozov, et al.,Modified indirect porcine circovirus (PCV) type 2-based and recombinant capsid protein (ORF2)-based enzyme-linked immunosorbent assays for detection of antibodies to PCV[J],Clin Diagn Lab Immure,2002,9(1):57-68.
[80].王涛,王子轼,彭德旺.猪圆环病毒感染与断奶仔猪多系统衰竭综合征[J].中国兽医学报,2002, 22 (3): 314-316
[81].陈曦,刘焕奇,魏萍.一种新的病毒病一断奶仔猪多系统衰竭综合征.黑龙江畜牧兽医.2002,9: 49-50.
[82].踞春梅.猪2型圆环病毒ELISA诊断方法及猪2型圆环病毒-伪狂犬病病毒二价基因工程疫苗研究(博士论文).2005,39-40.
[83]. Broom J, Heys S D, Whiting P H, Park K q Strachan A, Rothnie I, Franks C R, Eremin O Interleukin 2 therapy in cancer: identification of responders. Br J Cancer, 1992, 66:1185-1187
[84]. Morgan D A,Ruscctti,F W, Gallo R C. Selective in vitro growth of T lymphocytes from normal human bone marrow[J]s. Science, 1976, 193:1007-1008.
[85]. Robb R I. Tell growth factor receptors,quantitation, specificity and biological relevance[J]. J ExP Med, 1981, 154: 1455～1457.
[86].王瑜,王健等.白细胞介素-2受体结构及功能研究进展[J].中国现代医学杂志,2006,16(2):24 1-245.
[87]. Hughes-Fulford M, Sugano E, Schopper T, et al., Early immune response and regulation of IL-2 receptor subunits[J].Cellular Signalling, 2005, 17: 1111-1124.
[88]. Leonard W J. A monoclonal antibody that appears to recognize the receptor for human T cell growth factor: partial characterization of the receptor[J] . Nature, 1982, 300: 287-296.
[89]. Smith K A. Production and characterization of monoclonal antibodies to human intereukin2: strategy and tactics[J]. J Tmmunol. 1983. 131: 1808-1815.
[90]. CantrellD A,Smith K A .Transient expression of intereukin2 receptor[J].J Exp Med. 1983, 18:1895-1911.
[91]. Cox G W Mathieson B J,Giardina S L et al.,Characterization of IL-2 receptor expression and function on murine macrophages[J].J Immunol. 1990 145(6):1719-1726.
[92]. TaniguchiT,MinamiY.The IL-2/IL-2 receptor system:acurrent overview[J].Cell. 1993, 73 (1):5-8.
[93]. Ellery J M, Nicholls P J. Aiternate signaling pathways from the interleukin-2 receptor[J]. Cytokine & Growth Factor Reviews, 2002, 13:27-40.
[94]. Good M F, Pombo D, Lunde M N, Maloy W L, Halenbeck R, Koths K, Miller L H, Berzofsky J A.Recombinant human IL-2 overcomes genetic nonresponsivenes to malaria sporozoite peptides.Correlation of effect with biologic activity of IL-2[J]. J Immunol, 1988, 141:972-977.
[95]. Hyun B H, Song J Y, Lee J B, Cha S H, Park J H, Lee J O, An D J, Kang Y W. Cloning and expression of porcine interleukin-2 gene in Escherichia coli.RDA J Vet Sci, 1997, 39:42-50
[96]. Heys S D, Franks C R, Eremin O. lnterleukin 2 therapy: current role in surgical oncological practice[J]. Br J Surg, 1993, 80:155-162.
[97]. Fehniger T A, Cooper M A, Caligiuri M A. Interleukin-2 and interleukin-15: immunotherapy for cancer[J]. Cytokine&Growth Factor Reviews, 2002, 13:169-183.
[98]. Figlin R A. Renal cell carcinoma: current status and future plans[J]. Cancer J Sci Am, 2000, 6:45-51.
[99]. Broom J, Heys S D, Whiting P H, Park K G, Strachan A, Rothnie I, Franks C R, Eremin O Interleukin 2 therapy in cancer: identification of responders[J]. Br J Cancer, 1992, 66:1185-1187.
[100]. Rosenberg S A. Progress in human tumour immunology and immunotherapy[J]. Nature, 2001,4(11):380-384.
[101]. Prentice H G,Macdonald I D, Hamon M D. The role of immunotherapy in the treatment ofacute myeloblastic leukemia: from allogeneic bone marrow transplantation to the application of interleukin 2[J].Cancer Treat Research, 1993, 64:121-134.
[102]. Hazama M A,Mayumi Aono N,Asakawa S,et al.Vaccine,1993, 629-636.
[103]. Reddy DN, Reddy PG.Immuopotentiation of bovine respiratory disease virus vaccines by interleukinl b and interleukin2[J]. Veterinary Immuology and Immuopathology,1993,37:25.
[104]. Hughs HRA, Campos M. Immuopotentiaton of bovine herpevirus subunit vaccinationation by interleukin2[J]. Immuology,1991.74:461.
[105].李宏梅,李祥瑞.白细胞介素-2的研究进展及应用[J].山东畜牧兽医:38-39
[106]. Nunberg J H, Doyle M V, York S et al, Interleukine 2 acts as adjuvant to increase the potency of inactivated rabies virus vaccine[J], Proc Natl Acad SCI. 1989,86(6):4240-4243.
[107]. Campos M, Rossi CR. Characterization and activation requirement of bovine lymphocytes acquiring cytotoxicativity after interleukin2 treatment[J]. Veterinary Irnmuology and Immuopathology,1992;32:205.
[108]. Brown WC, Grab BJ. Biological and biochemical characterization of bovine interleukin2 studies with cloned bovine T cells[J].The Journal of Immuology,1985,35(5):3184.
[109]. Arunachalam B,Subba Raomv. Bovine interleukin2 biochemical and biological charactrization[J].Veterinary Immuology and Immupathology,1989;23:377.
[110]. Chow Y H, Huang W L, Chi W et al, Improvement of hepatitis B virus DNA vaccine by plasmids coexpressing hepatitis B surface antigen and Interleukine 2[J], J of Virology, 1997, 1:169-178.
[111]. Geissler M, Gesien A, Wands J R. Inhibitory effects of chronic ethanolconsuption on cellular immune response to hepatitis C virus core protein are reversed by genetic immunizations augmented with cytokine expression plasmid[J].J Immunol,1997, 159: 5107-5113.
[112]. Ramshaw IA, Woodsworth M.The in vitro culture of spleen cells from mice expressing humotal immunity leads to the development of T cells that mediate delayed-type hypersensitivity[J]. Cell Immunol.1981,15,57(2):468-476.
[113]. Flexner C, Hugin A, Moss B.Prevention of vaccin in virus infection in immunodeficient mice by vector-directed IL-2 expression[J]. Nature, 1987,330:259-261.
[114].姜永厚,陈奖励,宋秀龙等。鸡新城疫病毒F基因和鸡IL-2重组疫苗的构建[J]中国预防兽医学报,2001, (2): 81-83.
[115]. Hazama M, Mayumi-Aono A, Asakawa N, Kuroda S, Hinuma S, Fujisawa Y. Adjuvant-independent enhanced immune responses to recombinant herpes simplex virus type 1 glycoprotein D by fusion with biologically active interleukin-2[J].Vaccine, 1993,11(6):629-636.
[116] Tischer,Geldblon H, Vettermann W, et al. A very small porcine virus with cireular single-stranded DNA[J].Nature, 1982,(295):64-66
[117] Tischer I ,Rasch R ,Tochcermana G. Characterizationof papovavirus and picornavirus like particle in Permanent pig kidney cell lines [J ] . ZblBakt Hyg A ,I Abt Ovig A ,1974 ,326 :153-167.
[118] Post-weaning multisystemic Waning multisystemic syndrome [J ] . Proc Am Assoc Swine Pract ,1997 ,28 :499-501.
[119]黄平等.预防输血传播的感染[J ] .海峡预防医学杂志,2000 ,6(5) :19-21.
[120] Blanchard P,Mahe D,Cariolet R, et al. Protection of swine against post-weaning multisystemic wasting syndrome(PMWS) by porcine circovirus type (PCV2) proteins[J]. Vaccine,2003,21:4565-4575.
[121] Chow YH , Chiang BL , Lee YL , et al.Development of Th1 and Th2 populations and the nature of immune responses to hepatitis B virus DNA vaccines can be modulated by codelivery of various cytokine genes [J] .Immunol , 1998 , 160 : 1320–1329.
[122] Kim JJ,Ayyavoo V,Bagarazzi ML, et al.In vivo engineering of a cellular immune response by coadministration of IL-2 expression vector with a DNA immunogen. [J].Immunol ,1997, 158:816-826.
[123]. Kamstrup S,Barfoed A M,Frimann TH, et al.,Immunisation against PCV2 structural protein by DNA vaccination of mice[J].Vaccine,2004,22:1358-1361.
[124]. Blanchard P,Mahe D,Carioet R,et al.Protection of swine against post-weaning multisystemic wasting syndrome(PMWS) by porcine circovirus type 2 (PCV2) proteins[J]. Vaccine,2003,21:4565-4575.
[125]. Kim JJ,Yang JS,Manson KH, et al.,.Modulation of antigen-specific cellular immune renponses to DNA vaccination in rhesus macaques through the use of IL-2,IFN-γor IL-4 gene adjuvants[J].Vaccine,2001,19:2496-2505.
[126]. McNeilly F, Allan G M, Foster J C, Adair B M, McNulty M S and Pollock J. Effect of porcine circovirus infection on porcine alveolar macrophage function[J]. Veterinary Immunology and Immunopathology, 1996, 49(4): 295~306.
[127]. Segales J, Domingo M, Chianini F, Majo N, Dominguez J, Darwich L and Mateu E. Immunosuppression in postweaning multisystemic wasting syndrome affected pigs[J]. Veterinary Microbiology, 2004, 98(2): 151~158.
[128]. Premenko-Lanier M, Rota P A, Rhodes G., Verhoeven D, Barouch D H, Lerche N W, Letvin N L, Bellini W J and McChesney M B. DNA vaccination of infants in the presence ofmaternal antibody: a measles model in the primate[J]. Virology, 2003, 307(1): 67~75.
[129].王镜岩,朱圣庚,徐长法.生物化学[M](第三版).北京高教出版社, 2002.
[130].萨姆布鲁克J,弗里奇E F,曼尼阿蒂斯T.金冬雁,黎孟枫,等译.分子克隆指南[M].北京:科学出版社,1997
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