鸭α干扰素原（真）核表达及其生物学活性研究
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摘要
本文对四川省内地方品种四川麻鸭的干扰素进行如下系列研究:对四川麻鸭Ⅰ型干扰素基因IFN-αORF进行克隆、鉴定和序列分析;构建IFN-αORF基因原核表达载体和真核表达载体;IFN-α原核表达及其生物学活性研究;利用雏鸭为模型,应用免疫组织化学方法研究pcDNA-SDIFN-α真核表达质粒在鸭体内的动态表达分布规律;同时对作为免疫佐剂的pcDNA-SDIFN-α真核表达质粒免疫调节活性进行研究,获得以下结果:
1.四川麻鸭IFN-αORF基因克隆、序列比对分析和蛋白结构预测
应用Oligo6.0软件设计了两对加入EcoRI和BamHI两个酶切位点的特异性引物对,采用PCR和nest-PCR的方法,从鸭肝脏组织基因组DNA扩增得到大小两个完全包含麻鸭α干扰素ORF基因的DNA片段。其中小片段更适合用于表达,并将其进行pGEM-T载体克隆与测序,对测序结果采用ClustalX软件进行序列分析以及与北京鸭、鸡、鹅等NCBI上序列进行了比较。结果显示四川麻鸭与北京鸭ORF基因序列十分相似,其核苷酸同源性达到99.13%,氨基酸同源性为98.96%。与鸡的核苷酸及氨基酸同源性分别为71.8%、49.97-50.47%,与鹅的核苷酸同源性为96%。同时用BioEdit、NetNGlyc、TreeView软件对该基因所推测的蛋白进行了分析,结果表明四川麻鸭有2个潜在糖基化位点,1个蛋白激酶C磷酸化位点(Protein kinase C phosphorylation)和1个酪蛋白激酶Ⅲ磷酸化位点(Casein kinaseⅢphosphorylation site),信号肽切割位点28-29位ANA和FS氨基酸之间,成熟的鸭α干扰素有5个疏水区,无跨膜区。
2.SDIFN-αORF基因原核表达载体pBV220-SDIFN-α的构建及表达产物的抗病毒活性
采用BamHI和EcoRI酶切,通过定向粘末段连接法,构建了原核表达载体pBV220-SDIFN-α质粒。并将质粒转化到原核表达菌株DH5α中进行表达,采用温度诱导的方式,对诱导后的菌液进行SDS-PAGE检测。结果显示工程菌进行了表达,产生的包涵体蛋白分子量为40KD左右。对纯化后的包涵体采用稀释复性的方法复性,采用VSV/DEF系统初步检测抗病毒活性,干扰素的抗病毒活性约为150U/ml,比活力为440U/mg。应用FQ-PCR检测方法,动态监测体内抑制DHBV和体外抑制DPV病毒核酸。结果发现,表达的干扰素具有强烈的病毒复制抑制作用,具有明显的生物学活性。
3.SDIFN-αORF基因真核表达载体pcDNA-SDIFN-α的构建及鉴定
利用合成在引物两端的EcoRI、BamHI酶切位点,将目的基因成功地从pGEM-T载体克隆重组体中切下,并通过粘端连接的方法,连接到pUC18定向位置上,然后用EcoRI和XbaI双酶切真核表达载体pcDNA3.1(+)和pUC18-SDIFN-α,实现了SDIFN-α定向克隆到pcDNA3.1(+)载体上。四川麻鸭IFN-α真核表达重组质粒pcDNA-SDIFN-α经BamHI、EcoRI和XbaI三种酶切验证,并测序证明四川麻鸭IFN-α的真核表达质粒pcDNA-SDIFN-α构建正确。
4.免疫组织化学检测真核表达质粒pcDNA-SDIFN-α在鸭体内表达方法的建立及动态表达规律的研究
以pcDNA-SDIFN-α真核质粒免疫鸭,分时间段采集鸭16种不同组织,建立了免疫组织化学检测方法,并应用该方法检测干扰素在体内的动态表达规律。pcDNA-SDIFN-α免疫鸭后,持续表达时间长达9周。能在肺脏细胞、心肌细胞、肝细胞、脾细胞、肠腺和肠绒毛细胞、肌肉细胞、皮肤内检测到表达产物,在14d-35d进行了高峰表达。胸腺、胰腺、法氏囊和哈德氏腺未检测到阳性黄色颗粒,肺脏、免疫部位最早出现阳性颗粒,脑组织细胞中阳性颗粒数量随时间变化较小。其中基因枪免疫鸭后,各组织中的含量和表达量呈现的总体规律为6μg组＞3μg组＞1μg组。肌肉注射免疫鸭后,各组织中的含量和表达量呈现的总体规律为200μg组＞100μg组＞50μg组,都具有一定的剂量相关性。基因枪轰击法较肌肉注射法IFN-α基因表达出现的时间早,3h就可以检测到表达产物。同时,相对于肌肉注射法免疫,基因枪轰击法所用的表达质粒用量更少。
5.真核表达质粒pcDNA-SDIFN-α在鸭体内免疫调节剂活性的初步研究
以pcDNA-SDIFN-α真核表达质粒为免疫调节剂,研究对DPV/DVH弱毒苗免疫活性调节作用。分不同时间段采血,采用MTT法测定鸭外周血T淋巴细胞转化效果、CD3+T淋巴细胞数量变化和特异性DPV中和抗体IgG水平。结果表明,一定剂量的pcDNA-SDIFN-α真核表达质粒作为免疫调节剂在一定时间内能促进和提高细胞免疫和体液免疫应答,发挥着有效的正向免疫调节作用。
In this paper,the study is on Sichuan Province local varieties of duck interferon. We conducted the following series of studies:Sichuan duck typeⅠinterferon IFN-αORF gene sequence was cloned、identified and analysis;IFN-αORF prokaryotic gene expression vector and eukaryotic expression vector were successful constructed; interferon-αhas been expressed by prokaryotic expression &.biological activity;Useing ducklings as model,by means of immunohistochemistry.We studied on pcDNA-SDIFN-αvector dynamic distribution of expression in ducks;At the same time as the immunoadjuvant,we test the pcDNA-SDIFN-αeukaryotic expression vector to carry on immunological regulation function in ducks.The results were as follows:
1.Sichuan duck IFN-αORF gene cloning,sequencing,analysis and protein structure prediction.
Application software Oligo6.0,designed two pairs of specific primers within BamHI and EcoRI restriction sites,by PCR and nest-PCR method,we get two size amplified from duck liver geome about completely duck interferon-αgene ORF DNA fragment,Small fragments of which is more suitable for expression.Using pGEM-T vector,we get clone about duck interferon-αgene.Using ClustalX sequence analysis software,as well as with beijing ducks,chickens,geese,and other sequences in the NCBI comparison,The results showed that Sichuan and Beijing duck ORF sequences are very similar,its nucleotide homology to 99.13%,amino acid homology of 98.96%.And chicken nucleotide and amino acid homology are 71.8%and 49.97-50.47%,the goose nucleotide homology of is 96%.At the same time using BioEdit,NetNGlyc,TreeView software speculate that the gene protein was analyzed.The results show that Sichuan duck has two potential glycosylation sites,a protein kinase C phosphorylation sites,and a casein kinaseⅡphosphorylation sites,the signal peptide cleavage 28-29(ANA and FS) between amino acids,and mature duck interferon-αhas five hydrophobic,no transmembrane region.
2.SDIFN-αORF prokaryotic gene expression vector pBV220-α-SDIFN Construction, expression and the anti-viral activity detection.
BamHI and EcoRt restriction,by the end of visco-directional link,prokaryotic expression vector pBV220-SDIFN-αplasmid was constructed.And the plasmid was transformed into prokaryotic expression strain Ecoli DH5α,using temperature-induced and SDS-PAGE.The results showed that product is 40 KD.By purified and refolding and VSV /DEF system,we detected the anti-viral activity of product.The results of antiviral activity are about 150 U / ml,vitality about 440 U / mg.Application FQ-PCR method,dynamic monitoring of the virus nucleic acid about DHBV in vivo and DPV in vitro.The results showed that the interferon has a strong inhibition of virus amplify.
3.SDIFN-αORF eukaryotic expression vector pcDNA-SDIFN-αconstruction and identification.
By means of primers with ends of the EcoRI &.BamHI restriction sites,we get gene from the pGEM-T vector,and link with the pUC 18 directional positions.And then use EcoRI &.Xbal double digestion of eukaryotic expression-vector pcDNA3.1(+) and pUC 18-SDIFN-α,we get pcDNA-SDIFN-αeukaryotic expression vector clone.By BamHI, EcoRI and XbaI three digestions and sequencing shows that it is correct about Sichuan duck IFN-αeukaryotic expression recombinant pcDNA-SDIFN-αplasmid.
4.Immunohistochemistry's Established and reasearch on eukaryotic expression plasmid pcDNA-SDIFN-αexpression laws in duck.
Using pcDNA-SDIFN-αeukaryotic vector immune duck,the 16 different duck organizations were collected at the different time.We established the immunohistochemistry method.And application the method,we tested the expression laws of interferon in vivo.pcDNA-SDIFN-αimmune duck,continued to express time up to nine weeks.In the lung cells,heart muscle cells,liver cells and spleen cells,intestinal glands and intestinal villi cells,muscle cells,skin cells successfully expressed,14 d-35d in a peak of expression.Thymus,pancreas,Bursa and Bemhard's glands were not detected yellow particles.The earliest site is in Lung &.immune tissue.Brain tissue's Positive reaction is almost no change with times.By Gene gun immunization method,the content of exprcssion shpws of Group 6μg＞3μg group＞1μg group.Intramuscular immunization ducks,the content of expression shows of 200μg group＞100μg group＞50μg group, the all of which have a dose related.Gene gun immunization method is earlier than intramuscular gene in expression;3 h can be detected expression products by Gene gun immunization method.At the same time,comparing to intramuscular injection method,the gene gun use less plasmids.
5.The study on Eukaryotic expression plasmid pcDNA-SDIFN-αas immune adjuvant.
pcDNA- SDIFN-αeukaryotic expression vector as for immune adjuvant,research on the DPV / DVH attenuated vaccine regulation of immune activity.At different times,by MTT assay determination of T lymphocytes,the number of CD3 + T lymphocytes and specific changes in the IgG antibody levels about DPV immuned duck.The results show that a certain dose of pcDNA-SDIFN-αeukaryotic expression vector as an immune adjuvants have positive role in regulating.
1.四川麻鸭IFN-αORF基因克隆、序列比对分析和蛋白结构预测
应用Oligo6.0软件设计了两对加入EcoRI和BamHI两个酶切位点的特异性引物对,采用PCR和nest-PCR的方法,从鸭肝脏组织基因组DNA扩增得到大小两个完全包含麻鸭α干扰素ORF基因的DNA片段。其中小片段更适合用于表达,并将其进行pGEM-T载体克隆与测序,对测序结果采用ClustalX软件进行序列分析以及与北京鸭、鸡、鹅等NCBI上序列进行了比较。结果显示四川麻鸭与北京鸭ORF基因序列十分相似,其核苷酸同源性达到99.13%,氨基酸同源性为98.96%。与鸡的核苷酸及氨基酸同源性分别为71.8%、49.97-50.47%,与鹅的核苷酸同源性为96%。同时用BioEdit、NetNGlyc、TreeView软件对该基因所推测的蛋白进行了分析,结果表明四川麻鸭有2个潜在糖基化位点,1个蛋白激酶C磷酸化位点(Protein kinase C phosphorylation)和1个酪蛋白激酶Ⅲ磷酸化位点(Casein kinaseⅢphosphorylation site),信号肽切割位点28-29位ANA和FS氨基酸之间,成熟的鸭α干扰素有5个疏水区,无跨膜区。
2.SDIFN-αORF基因原核表达载体pBV220-SDIFN-α的构建及表达产物的抗病毒活性
采用BamHI和EcoRI酶切,通过定向粘末段连接法,构建了原核表达载体pBV220-SDIFN-α质粒。并将质粒转化到原核表达菌株DH5α中进行表达,采用温度诱导的方式,对诱导后的菌液进行SDS-PAGE检测。结果显示工程菌进行了表达,产生的包涵体蛋白分子量为40KD左右。对纯化后的包涵体采用稀释复性的方法复性,采用VSV/DEF系统初步检测抗病毒活性,干扰素的抗病毒活性约为150U/ml,比活力为440U/mg。应用FQ-PCR检测方法,动态监测体内抑制DHBV和体外抑制DPV病毒核酸。结果发现,表达的干扰素具有强烈的病毒复制抑制作用,具有明显的生物学活性。
3.SDIFN-αORF基因真核表达载体pcDNA-SDIFN-α的构建及鉴定
利用合成在引物两端的EcoRI、BamHI酶切位点,将目的基因成功地从pGEM-T载体克隆重组体中切下,并通过粘端连接的方法,连接到pUC18定向位置上,然后用EcoRI和XbaI双酶切真核表达载体pcDNA3.1(+)和pUC18-SDIFN-α,实现了SDIFN-α定向克隆到pcDNA3.1(+)载体上。四川麻鸭IFN-α真核表达重组质粒pcDNA-SDIFN-α经BamHI、EcoRI和XbaI三种酶切验证,并测序证明四川麻鸭IFN-α的真核表达质粒pcDNA-SDIFN-α构建正确。
4.免疫组织化学检测真核表达质粒pcDNA-SDIFN-α在鸭体内表达方法的建立及动态表达规律的研究
以pcDNA-SDIFN-α真核质粒免疫鸭,分时间段采集鸭16种不同组织,建立了免疫组织化学检测方法,并应用该方法检测干扰素在体内的动态表达规律。pcDNA-SDIFN-α免疫鸭后,持续表达时间长达9周。能在肺脏细胞、心肌细胞、肝细胞、脾细胞、肠腺和肠绒毛细胞、肌肉细胞、皮肤内检测到表达产物,在14d-35d进行了高峰表达。胸腺、胰腺、法氏囊和哈德氏腺未检测到阳性黄色颗粒,肺脏、免疫部位最早出现阳性颗粒,脑组织细胞中阳性颗粒数量随时间变化较小。其中基因枪免疫鸭后,各组织中的含量和表达量呈现的总体规律为6μg组＞3μg组＞1μg组。肌肉注射免疫鸭后,各组织中的含量和表达量呈现的总体规律为200μg组＞100μg组＞50μg组,都具有一定的剂量相关性。基因枪轰击法较肌肉注射法IFN-α基因表达出现的时间早,3h就可以检测到表达产物。同时,相对于肌肉注射法免疫,基因枪轰击法所用的表达质粒用量更少。
5.真核表达质粒pcDNA-SDIFN-α在鸭体内免疫调节剂活性的初步研究
以pcDNA-SDIFN-α真核表达质粒为免疫调节剂,研究对DPV/DVH弱毒苗免疫活性调节作用。分不同时间段采血,采用MTT法测定鸭外周血T淋巴细胞转化效果、CD3+T淋巴细胞数量变化和特异性DPV中和抗体IgG水平。结果表明,一定剂量的pcDNA-SDIFN-α真核表达质粒作为免疫调节剂在一定时间内能促进和提高细胞免疫和体液免疫应答,发挥着有效的正向免疫调节作用。
In this paper,the study is on Sichuan Province local varieties of duck interferon. We conducted the following series of studies:Sichuan duck typeⅠinterferon IFN-αORF gene sequence was cloned、identified and analysis;IFN-αORF prokaryotic gene expression vector and eukaryotic expression vector were successful constructed; interferon-αhas been expressed by prokaryotic expression &.biological activity;Useing ducklings as model,by means of immunohistochemistry.We studied on pcDNA-SDIFN-αvector dynamic distribution of expression in ducks;At the same time as the immunoadjuvant,we test the pcDNA-SDIFN-αeukaryotic expression vector to carry on immunological regulation function in ducks.The results were as follows:
1.Sichuan duck IFN-αORF gene cloning,sequencing,analysis and protein structure prediction.
Application software Oligo6.0,designed two pairs of specific primers within BamHI and EcoRI restriction sites,by PCR and nest-PCR method,we get two size amplified from duck liver geome about completely duck interferon-αgene ORF DNA fragment,Small fragments of which is more suitable for expression.Using pGEM-T vector,we get clone about duck interferon-αgene.Using ClustalX sequence analysis software,as well as with beijing ducks,chickens,geese,and other sequences in the NCBI comparison,The results showed that Sichuan and Beijing duck ORF sequences are very similar,its nucleotide homology to 99.13%,amino acid homology of 98.96%.And chicken nucleotide and amino acid homology are 71.8%and 49.97-50.47%,the goose nucleotide homology of is 96%.At the same time using BioEdit,NetNGlyc,TreeView software speculate that the gene protein was analyzed.The results show that Sichuan duck has two potential glycosylation sites,a protein kinase C phosphorylation sites,and a casein kinaseⅡphosphorylation sites,the signal peptide cleavage 28-29(ANA and FS) between amino acids,and mature duck interferon-αhas five hydrophobic,no transmembrane region.
2.SDIFN-αORF prokaryotic gene expression vector pBV220-α-SDIFN Construction, expression and the anti-viral activity detection.
BamHI and EcoRt restriction,by the end of visco-directional link,prokaryotic expression vector pBV220-SDIFN-αplasmid was constructed.And the plasmid was transformed into prokaryotic expression strain Ecoli DH5α,using temperature-induced and SDS-PAGE.The results showed that product is 40 KD.By purified and refolding and VSV /DEF system,we detected the anti-viral activity of product.The results of antiviral activity are about 150 U / ml,vitality about 440 U / mg.Application FQ-PCR method,dynamic monitoring of the virus nucleic acid about DHBV in vivo and DPV in vitro.The results showed that the interferon has a strong inhibition of virus amplify.
3.SDIFN-αORF eukaryotic expression vector pcDNA-SDIFN-αconstruction and identification.
By means of primers with ends of the EcoRI &.BamHI restriction sites,we get gene from the pGEM-T vector,and link with the pUC 18 directional positions.And then use EcoRI &.Xbal double digestion of eukaryotic expression-vector pcDNA3.1(+) and pUC 18-SDIFN-α,we get pcDNA-SDIFN-αeukaryotic expression vector clone.By BamHI, EcoRI and XbaI three digestions and sequencing shows that it is correct about Sichuan duck IFN-αeukaryotic expression recombinant pcDNA-SDIFN-αplasmid.
4.Immunohistochemistry's Established and reasearch on eukaryotic expression plasmid pcDNA-SDIFN-αexpression laws in duck.
Using pcDNA-SDIFN-αeukaryotic vector immune duck,the 16 different duck organizations were collected at the different time.We established the immunohistochemistry method.And application the method,we tested the expression laws of interferon in vivo.pcDNA-SDIFN-αimmune duck,continued to express time up to nine weeks.In the lung cells,heart muscle cells,liver cells and spleen cells,intestinal glands and intestinal villi cells,muscle cells,skin cells successfully expressed,14 d-35d in a peak of expression.Thymus,pancreas,Bursa and Bemhard's glands were not detected yellow particles.The earliest site is in Lung &.immune tissue.Brain tissue's Positive reaction is almost no change with times.By Gene gun immunization method,the content of exprcssion shpws of Group 6μg＞3μg group＞1μg group.Intramuscular immunization ducks,the content of expression shows of 200μg group＞100μg group＞50μg group, the all of which have a dose related.Gene gun immunization method is earlier than intramuscular gene in expression;3 h can be detected expression products by Gene gun immunization method.At the same time,comparing to intramuscular injection method,the gene gun use less plasmids.
5.The study on Eukaryotic expression plasmid pcDNA-SDIFN-αas immune adjuvant.
pcDNA- SDIFN-αeukaryotic expression vector as for immune adjuvant,research on the DPV / DVH attenuated vaccine regulation of immune activity.At different times,by MTT assay determination of T lymphocytes,the number of CD3 + T lymphocytes and specific changes in the IgG antibody levels about DPV immuned duck.The results show that a certain dose of pcDNA-SDIFN-αeukaryotic expression vector as an immune adjuvants have positive role in regulating.
引文
[1]Weissmann C,and Weber H.the interferon genes,Prog Nucleic Acide Res.Mol Biol,1986,33:251-300.
[2]Issacs A,Lindanmann J.Virus interference,I.the interferon[J].Proc R Soc Ser B.1957,147:258-267.
[3]刘胜旺、卢景良、张万旗.鸡干扰素及其相关蛋白分子生物学研究进展。预防兽医学进展,2000,2(1):17-19。
[4]孙志宏,陈正礼,范光丽。干扰素研究进展。延安大学学报(自然科学版),2000,19(1):76-79。
[5]Bazer F W.Mediators of maternal recognition of pregnancy in mammals[J].Proc Soc Exp Bio Med,1992,199:373
[6]Steinmann G G,Rosenkaimer F,Leitz G.Clinical experience with interferon-α and interferon-γ[J].Int Rev Exp Pathol,1993,34B:193
[7]Stanton G J,Weigent D A,Fleischmann W R.et al.Interferon Review[J],Investigative Radiology,1987,22:259-273.
[8]金伯泉,细胞和分子免疫学(第二版),北京,科学出版社,2000。
[9]陈坤华。细胞因子对下丘脑—垂体—肾上腺皮质轴之间调节作用[J].生理科学进展,1993,34(2):113-117
[10]吴玉厚、吴冰洁、周国利等.干扰素研究进展,生物学教学,2007,32(7):2-4。
[11]邢利和,王福生。干扰素产生细胞的研究进展,国外医学免疫学分册[J].2002,25(2):76-78.
[12]卢建.细胞因子受体组成、结构功能及信号传导机制[J].生理科学进展,1995,26(4):305-311.
[13]杨元忠,试述干扰素的主要作用.畜牧兽医杂志,1997,16(3):34-35。
[14]程坚、王立俊、杨波。a干扰素治疗肿瘤机制的研究进展。农业生物技术学报,2000,8(3):236-238。
[15]Sekellick MJ,Ferrandino AF,Hopkins DA,et al.Chicken interferon gene:cloning,expression and analysis[J].J Interferon research,1994,14:71-79
[16]H W Murray,A.Szuro-Sudol,D wellner.et al,Role of ryptophan degradation in respiratory burst-independent antimicrobial activity of gamma interferon-stimulated human macrophages.Infect.Immun.1989,57:845-849.
[17]马良,干扰素与寄生虫感染.重庆医科大学学报.1995,30(3):241-244
[18]M A Orellana,Y Suzuki,F Araujo,et al.Role of beta interferon in resistance to Toxoplasma gondii infection.Infect.Immun.1991,59:3287-3290.
[19]郭定宗,祝海宝。干扰素研究进展及其在兽医临床上的应用[J],养殖与饲料,2006,2:5-8。
[20]杜念兴.兽医免疫学[M].第2版,北京:中国农业出版社.1997,74-76.
[21]孙圣兰,江青艳,傅伟龙.干扰素的研究进展[J].动物医学进展,2002,23(6):39-41.
[22]马飞.干扰素的研究进展[J].中国畜牧兽医,2005,32,12:36-37.
[23]Towata T,Hayashi N,Katayama K,et al.Effects of cytokines on HLA class Ⅰ antigen expression on Huh6 and HB611 cells[J].Gastroenterol,1993,28(2):242-248.
[24]Luft T,Pang K C,Thomas E,et al.Type Ⅰ IFNs enhance the terminal differentiation of dendritic cells[J].J Immunol,1998,161:1947-1953.
[25]Radvanyi L G,Banerjee A,Weir M,et al.Low levels of interferon-α induce CD86(B7.2)expression and accelerates dendritic cell maturation from human peripheral blood mononuclear cells[J].Stand J Immunol,1999,50(5):499-509.
[26]Santini S M,Lapenta C,Logozzi M,et al.Type 1 interferon as a powerful adjuvant for monocytederived dendritic cell development and activity in vitro and in Hu-PBL-scid mice[J].J Exp Med,2000,191(10):1777-1788.
[27]柴玉波、孙海峰、李春源。干扰素治疗慢性乙型肝炎的疗效分析,生物工程学报。1997,13(4):406-408
[28]陈晓初,谢佩蓉,郑蒲.IFN—γ和IL—6或IL—1联合应用对人肝癌细胞系CLINH合成的作用[J],免疫学杂志,1999,15(4):239-241。
[29]刘高生,卢升云,干扰素的免疫作用机制。中国兽医科技,2004,34(10):72-74
[30]朱长庚,李正莉,细胞因子与免疫—神经—内分泌调节网络[J],解剖学报,1996,27(4):339-344
[31]Chao C C,Hu S,Sheng W S,et al.Lytokine stimulated astrocytes damage human neurous via a nitri-oxide mechanism[J],Glia,1996,16:276-284
[32]Kiefer R,Keutzberg G W.Distribution of cytokines in nerous system[J].Nenro sci.1990,35(3):725-732.
[33]Gray P W,Peung D W,Pennica D,et al.Expression of human immune interferon cDNA in E.coli and monkey cells[J].Nature,1982,295:503-506.
[34]Goeddel D V,YeIverton E,Uurich A,et al.Human leukocyte interferon produced by E.coli is biologically active[J].nature,1980,287,2:4411-4415.
[35]Gray PW,GoeddeL D V,Cloning and expression of murine immune interferon cDNA[J].Proc Natl Acad Sci USA.1983.80:5842-5846.
[36]Dijkmans R,Vandenbroeck K,Beuken E,et al.Sequence of the porcine interferon-gamma gene[J].Nucleic Acids Research,1990,18,42-59.
[37]侯云德,分子病毒学[M],北京:学苑出版社,1990,598-642.
[38]Lillehoj HS,Kaspers B.Jenkins MC,et al.Avian interferon and interleukin-2.A review by comparison with mammalian homologues[J].Poultry Science Review,1992,4,67-85.
[39]Lowenthal JW,Digby MR,York JJ.Production of interferon-γ by chicken T cells[J].Journal of interferon and Cylokine Research.1995,15:933-938
[40]Sick C,Schuhz U,Staeheli P,A family of genes coding for two serologicaIly distince chicken interferon[J],The JournaI of Biological Chemistry.1996.271.I3:7636-7639
[41]Schultz U,Ridderle C,Sekeliick MJ.Recombinant chicken interferon from E.coli and transfected COS 7 cells is biologically active[J].European Journal biochemistry,1995,229,73-76
[42]Suresh M,Karaca K,Foster D.Molecular and functional characterization of turkey interferon[J],J Virol,1995,69,12:8159-8163
[43]夏春,汪明,朱凌云等。惠阳胡须鸡干扰素基因分子克隆与序列丹析[J],畜牧兽医学报,2000,31(6):563-566
[44]夏春,汪明,杨琪等。丝羽乌骨鸡干扰索基因分子克隆与字列丹析[J]。中国免疫学杂志,2000,5:22-24
[45]曾永长,吕英姿.鸡α干扰素基因的克隆和鉴定[J].中国预防兽医学报,2001,23(4):259-261.
[46]汪明、吴志光,夏春。肉鸡IFN-alpha基因的克隆、序列分析以及在大肠杆菌中的表达,农业生物技术学报,2000,8(4)377-381。
[47]韦琴,彭贵青,金梅林,等.鸡α千扰素基因的克隆、原核表达及抗病毒效果研究[J].生物工程学报,2006.22(5):737-743.
[48]Schultz U,Kock J,Schlicht HJ,et al.Recombinant auck interferon:A new reagent for studying the mode of interferon action against hepatitis B virus[J].Virology,1995,212:641-649
[49]Schultz U,Chisari FV.Recombinant duck interferon gamma inhibits duck hepatitis.B virus replication in primary hepatocytest[J],J Virol,1999,73(4):3162-3168
[50]夏春,万建青,吴志光等,北京鸭Ⅰ型干扰素基因分子克隆与序列分析,畜牧兽医学报,2000,31(6):567-570
[51]阮小飞,林常有,杨天耀,等.北京鸭干扰素-α基因的分子克隆与表达[J].中国兽医杂,2004,40(12):11-13.
[52]王春霞,王林川.番鸭IFN-α基因的克隆与序列分析[J].华南农业大学学报,2002,23(4):68-70.
[53]王春霞.王林川.鄢志强等,藩鸭IFN-α基因在毕赤酵母中的分泌表达,中国兽医科技,33(7):44-48
[54]叶伟成,张存,王一成,等.绍兴鸭α-干扰素基因克隆、表达及活性测定[J].浙江农业学报,2005,17(3):115-119.
[55]Hung yueh yeh,Barbara J,Winslow et al,In vitro effects of recombinant chicken interferon-r on immune cells[J].Journal of interferon and cytokine res.1999,19:687-691.
[56]Digby MR.Lowenthal JW.cloning and expression of the chicken interferon-γ gene[J].Journal of Interferon and Cytokine Research.1995.15:939-949.
[57]Song KD,Lillehoj HS,Choi KD,et al.Expression and functional characterization of recombinant chicken Interferon-gamma[J].Veterinary Immunology and Immunopathology,1997,58:321-333.
[58]Kaisr P,Wain HM.Rothwell L.Structure of the chicken Interferon-γ gene and comparison to mammalian homo]ogues[J].Gene,1998,207:25-23.
[59]Kaiser P.Sonnemans D.Smith LM.Avian IFN-γ Sequence Analysis Suggests Probable Cross Species Reactivity among Galliforms[J].Journal of Interferon and Cytokine,1998,18:711-719
[60]刘胜旺,陈洪岩,孔宪刚等,鸡γ-干扰素基因的分子克隆与序列测定,中国兽医学报,2000,20(3):228-230。
[61]吕英姿,毕英佐,曹永长,石歧杂鸡γ-干扰素基因的克隆与序列分析,华南农业大学学报(自然科学版),2002,23(4):61-63。
[62]吴志光,夏春,汪明。北京鸭Ⅱ型干扰索基因分子克隆与序列分析,中国兽医科技,2001,31(3):7-9。
[63]夏春,刘津,杨琪等,猪干扰素β基因的分子克隆与测序,中国兽医杂志,2000,26:6-7。
[64]郭瀛军,吴丹,陈蕊雯等,猪γ干扰素基因的克隆、表达及其纯化,生物工程学报,2001,17(2):184-186。
[65]赵英杰、徐宏军、李尚波等,长白猪γ干扰素基因的原核表达与分析,中国兽医科技,2006,36(1):46-51。
[66]曹瑞兵、包晶晶,周海霞等,猪β干扰素的原核表达及其对猪流行性腹泻病毒的抑制作用研究,中国病毒学报,2004,19(4):364-368。
[67]刘万钧,张道永,马思齐等.猪白细胞干扰素防治猪流行性腹泻试验.中国兽医杂志.1999,25(2):13-14。
[68]夏春、汪明,拉布拉多犬和德国牧羊犬IFN-α基因的分子克隆和测序,农业生物技术学报,1999,7(3):245-247。
[69]李学仁,王震海,葛菲菲等,奶牛γ干扰素基因的高效表达及活性测定,中国病毒学,2004,19(4)356-359。
[70]史喜菊、夏春、汪明,水牛α-干扰素基因的克隆与表达及其抗病毒活性,农业生物技术学报,2004,12(3):288-293。
[71]李玉萍,景志忠,蒲训等,牦牛β干扰素基因的克隆与表达,中国兽医科学,2006,36(1):40-45。
[72]Dimier IH.Quere P.Naciri M.et al.Inhibition of Eimeriatenel la development in vitro mediated by chicken macrophages and fibroblasts treated with chicken cell supernatants with IFN γactivity[J].Avian Diseases.1998.42:239-247
[73]Lowenthal JW,Onel TE,Broadway M,et al.Coadministration of IFN γ enhance antibody responses in chickens[J],Journal of interferon and Cytokine Research,1998.28:617-622.
[74]Plachy J.Weining KC,Kremmer MJ,et al.Proctive effects of type Ⅰ and type Ⅱ interferons toward Rous Sarcoma Virus induced tumors in chicken[J],Virology,1999,256,85-91.
[75]Marcus PI,Heide VD,Sekellick MJ.Interferon action on Avian viruses I,Oral administration of chicken interferon a Ameliorates Newcastle Disease[J],Journal of interferon and Cytokine Research,1999,19:881-885
[76]侯俊玲、郑亚东、景志忠等,DNA疫苗免疫佐剂的研究进展,中国兽医科技,2005,35(2):153-156。
[77]王克坚,殷震,白细胞介素类细胞因子的免疫佐剂作用,预防兽医学进展.2002,2(1):30-33。
[78]Cox J C,Coulter A R.Adjuvants a classification and review of their modes of action[J].Vaccine,1997,15(3):248-256
[79]张悦,戚晓红,冯振卿,免疫佐剂的研究紧展,中国血吸虫病防治杂志.2004,16(3)236-238。
[80]吴乃虎,基因工程原理.科学出版社,北京,2001。
[81]A Chaudhuri,S Orme,S Eilam et al,CD40-mediated signals inhibit the binding of TNF receptor-associated factor 2 to the CD40 cytoplasmic,domain[J],J Immunol,1997,159:4244-4251.
[82]Ihata A,Watabe S,Saski S,et al.Immunomodulatory effect of a plasmid expressing CD40ligand,on DNA vaccination against human immunodeficiency virus type-1[J].Immunology,1999,98(3):436-442
[83]Dierks SE,Bartlett WC,Edmeades RL,et al.The oligomeric nature of the murine FC epsilon RⅡ/CD23.implications for function[J],J Immunol,1993,150:2372-2382.
[84]Mendoza RB,Cantwell MJ,Kpps TJ.Immunostimulatory effects of a plasmid expressing CD40ligand(CD154) on gene immunization[J],J Immunol,1997,59(12):5777-5781.
[85]Kreing AM,Yiak,Klinman DM.CpG motif in bacterial DNA trigger direct B-cell activation[J].Nature,1995,374:546-549.
[86]Chace JH,Hooker NA,Coedery JS.Bacterial DNA induced NK cell IFN-γ production is dependent on macrophage secretion IL-12[J].Clin immunol immunopath,1997,84:185-193。
[87]Kojima Y,Xin K Q,Ooki T,et al.Adjuvant effect of multi-CpG motifs on an HIV-1 DNA vaccine[J],Vaccine,2002,20(23-24):2857-2865
[88]Wang X,Jiang P,Deen S,et al.Efficacy of DNA vaccines enhanced by coadministration,with CpG oligodeoxynucleotide[J],Avian Disc,2003,47(4):1305-1312.
[89]Lipford GB,Bamer M,Heeg K.Oligonucleotides protomer B and cytotoxic T cell responses to protein antigen:a new class of vaccine adjuvant[J].Eur J Immunol,1997,27:2340-2344.
[90]Heather LD,Ridini W,Krieg AM.CpG DNA is a porent enhancer and specific immunity in mice immunized with recombinant hepatitis B surface antigen[J].J Immunol,1998,160(2):870-876
[91]Moldoveanu Z,Love-Homan L,Wen QH,et al.CpG DNA,a novel immune enhancer for systemic and mucosal immunization with influenza virus,vaccine,1998,16(11-12):1216-1224.
[92]朱建中,朱鸿飞,鲁春,李富光。免疫佐剂CpG DNA的研究进展。生命的化学,2002,22(2):155-157。
[93]Okuda K,Xin KQ,Haruki A,et al.Transplacental genetic immunization after intravenous delivery of plasmid DNA to pregnant mice[J],J Immunol,2001,167(9):5478-5484.
[94]陈吉刚。郑肖娟,齐静,禽类细胞因子——新型免疫佐剂和免疫治疗剂,饲料工业,2005,26(1):35-41。
[95]Heath AW.细胞因子:新型免疫佐剂。国外医学,预防、诊断,治疗用生物制品分册,1993,16(3):110-113
[96]Karaca K,Sharma J M,Winslow B J,et al.Recombinant fowlpox virus coexpressing chicken type Ⅰ IFN and Newcastle disease virus HN and F genes:influence of IFN on protrctive efficacy and humoral responses of chickens following in ovo or post-hatch administration of recombinant virus[J].Vaccine,1998,16(16):1496-1503.
[97]Sekellick M J,Lowenthal J W,Oneil T E,et al.Chicken interferon types Ⅰ and Ⅱ enhance synergistically the antiviral state and nitric oxide secretion[J],J Interferon Cytokine Res,1998,18(6):407-414.
[98]Schijins VE,Weining KC,Nuijten P,et al.Immunoadjuvant activities of E.coli-and plasmid -expressed recombinant chicken IFN-alpha/beta,IFN-gamma and IL-1beta in 1-day and 3-week old chickens[J].Vaccine,2000,18(20):2147-2154
[99]邹丽容、陈则。DNA疫苗免疫佐剂的研究进展,激光生物学报,2002,11(5):395-400。
[100]张晋举,钱平。表达干扰素作为佐剂的重组鸡痘病毒胚胎免疫火鸡防制新城疫的研究。预防兽医学进展,2000,2(2):39-41。
[101]程坚、刘秀梵、彭大新等,表达鸡Ⅱ型干扰素基因的重组鸡痘病毒的构建,农业生物技术学报,2002,10(2):152-155。
[102]Warren T L,Weiner G J.Uses of granulocyte macophage colony-stimulating factor in vaccine development[J].Curr Opinhenratol,2000,7(3):168-173
[103]Lowenthal JW,Lambrecht B,van den Berg T,at al.Avain cytokines-the natural approach to therapeutics[J].Dev comp immunol,2000,24(2-3):355-365.
[104]Lowenthal JW,York JJ,Oneil TE.et al.In vivo effects of chicken interferon-gamma during infection with Eimeria[J].J interferon cytokine res,1997,17(9):551-558
[105]Guo H,Chen G,Lu F,et al.Immunity induced by DNA vaccine of plasmid encoding the rhoptry protein Ⅰ gene combined with the genetic adjuvant of pclFN-γ against toxoplasma gondii in mice[J],Chin med J(EngL),2001,114(3):317-320.
[106]彭大新,表达马立克氏病毒糖蛋白gB基因及共表达糖蛋白B和鸡γ干扰素重组鸡痘病毒的构建及其免疫保护作用,扬州大学博士论文,2001。
[107]于涟,李建荣,黄耀伟等。鸡白细胞介素2增强传染性法氏囊病病毒多聚蛋白DNA疫苗免疫原性的研究[J],生物工程学报,2001,17(6):653-657。
[108]Bu J,Song Y,Rompatio G,et al.Codelievery of IL-2 or liposomes augment the responses of mice to a DNA vaccine for pseudorabies virus IE180[J].Comp Immunol Microbio Intfect Dis,2003,26(3):175-187
[109]杨汉春、姚火春、王君伟等。动物免疫学(第二版),北京,中国农业大学出版社,2003,8。
[110]Reed SG,Pihid L,et al.IL-1 as adjuvant:role of T cell in the augmentation of specific antibody production by recombinant human IL-1 alpha[J].Immunol,1989,142:3129-3133.
[111]Kim J L,Maguire H C,Nottingham L K,et al.Coadministration of IL-12 or IL-10 expression oassettes drives immune responses towards a Th1 phenotype[J].Interferon Cytokin Res,1998,18(7):537-547
[112]Van Snick J.Interleukin-6:an overview[J].Annu Res Immunol,1990.8:253-278
[113]Schoenhaut D S,Chua A O,Wo litzky A G,et al.Cloning and expression of murine IL-12[J].J Immunnol,1992,148:3433-34440
[114]杜德伟,周永兴,冯志华,等.IL-12的真核表达载体促进小鼠对DNA疫苗的免疫应答[J].中华微生物学核免疫学杂志,2000,20(6):543-544
[115]刘卓宝,人类战胜疾病的新武器——DNA疫苗[M],上海科学技术出版社,2001。
[116]卢铀,田聆,魏于全等,DNA疫苗抗肿瘤研究进展[J],中华医学遗传学杂志,2000,17(4):288-290。
[117]Sedegrah M,Weiss W,Saci J B,et al.Improving protective immunity induced by DNA-based immunization:priming with antigen and GM-CSF-encoding plasmid DNA and boosting with antigen-expressing recombinant poxvirus[J].J Immunol,2000,164(11):5905-5912.
[118]Geissler M,Gesien A,Wands JR,Inhibitory effects of chronic ethanol consumption on cellular immune responses to hepatitis C virus core protein are reversed by genetic immunizations augmented with cytokine-expressing plasmids.J Immunol,1997,159(10):5107-5113
[119]Operschall E,Schuh T,Heinzerling L,Pavlovic J,et al.Enhanced protection against virual infection by co-administration of plasmid DNA coding for ritual antigen and cytokines in mice.J Clin Virol,1999,13(1-2):17-27
[120]Lu Y,Xin KQ,Hamajima K,Tsuji T,et al.Macrophage inflammatory protein-lalpha(MIP-lalpha)expression plasmid enhances DNA vaccine-induced immune response against HIV-1.Clin Exp Immunol,1999,115(2):335-341
[121]晏菊芳。DNA疫苗的分子佐剂研究进展。国外医学免疫学分册,2000,24(4):197-201
[122]尹荣兰、李昌、张乃生。DNA疫苗免疫佐剂研究进展,中国兽医杂志,2008,44(4):77-79
[123]杜德伟,周永兴,吴学勤,冯志华等,IL-12的真核表达载体对乙肝基因疫苗的免疫佐剂效应。中国现代医学杂志,2000,10(7):1-3。
[124]赫然,DNA疫苗的佐剂研究综述,中山大学研究生学刊(自然科学、医学版),2003,24(2):9-15。
[125]Weiss w R,Ishii K J,Hedstrom R C,et al.A plasmid encoding murine granulocyte-macrophage colony stimulating factor increases protection conferred by a malaria DNA vaccine[J].J Immunol,1998,161(5):2325-2332.
[126]Adel,patient K,Creminon C,et al.Genetic immunization with bovine beta-lactoglobulin cDNA induces a preventive and persistent inhibition of specific anti-BLG IgE response in mice[J].Int Arch Allergy Immunol,2001,26(1):59-67
[127]Chen Z,Sahasho Y,Matsuo K,et al.Comparison of the ability of viral protein-expressing plasmid DNAs to protect against influenza[J].Vaccine,1998,16(16):1544-1549.
[128]Chen Z,Natsuo K,Asanuma H,et al.Enhanced protection against A lethal influenza virus challenge by immunization with both hemagglutinin and neuraminidase-expressing DNAs[J],Vaccine,1999,17:653-659
[129]Sick C,Schuhz U、Staeheli P,A family of genes coding for two serologicaIly distince chicken interferon[J],The JournaI of Biological Chemistry.1996.271.13:7636-7639
[130]萨姆布鲁克,E.F.弗里奇,T曼尼阿蒂斯.分子克隆实验指南[M].(第二版)北京:科学出版社。
[131]陈斌,程安春,汪铭书,等.麻鸭α干扰素基因克隆与序列分析[J].中国兽医学报,2006,26(4):401-404.
[132]吴琦,枯草芽孢杆菌植酸酶phyC基因的克隆及其表达研究。四川大学博士学位论文,2004。
[133]Stewart W E.Interferon nomenclature[J].Nature,1980,286:110
[134]李伍举、吴加金.pBV220载体中外源基因表达水平定量分析。病毒学报,1997,13(2):125-133。
[135]龚永强,鸭α-干扰素基因的原核表达及抗病毒活性研究,雅安,四川农业大学硕士论文,2007。
[136]杨梅,重组鸭a-干扰素抗鸭乙型肝炎病毒实验感染研究。四川农业大学硕士论文,2007。
[137]郭宇飞.鸭病毒性肝炎病毒CH强毒株部分生物学特性的研究及荧光实时定量PCR检测方法的建立和应用[学位论文].四川雅安.四川农业大学.2005
[138]Gold L.Posttranscriptional regulatory mechanisms in Escherichia coli[J].Ann Rev Biophs Chem,1988,57(2):199-223.
[139]Tessier LH,Sondermeyer P,Faure T,et al.The influence of mRNA primary and secondary structure on human IFN-gamma gene expression in E.coli[J].Nucl Acids Res,1984,12(20):7663-7675.
[140]DE Smitmh,Van Duin J.Control of translation by mRNA secondary structure in Escherichia coli.A quantitative analysis of literature data[J].J Mol Biol,1994,244(2):144-150.
[141]任启生.王嘉玺,邹民吉。PBV-IL-6重组质粒的构建及其在E.coli细胞中的超高表达.中国免疫学杂志,1992,8:133-135。
[142]张智清、姚立红、候云德。含PrPl启动子的原核表达载体的组建及其应用。病毒学报,1990,6(2):111-116
[143]李武平、宋瑜丽、张成海等。重组人干扰素—β(rhIFN-β)中试工艺研究。中国生物制品学杂志,2004,17(1):33-35。
[144]张成海、李武平、衣作安等。人干扰素ψ(huIFN-ψ)的高效表达、纯化及生物学活性。病毒学报,2004.20(2):124-127。
[145]Stephen R.H.,Piotr Bobrowicz,Beata Bobrowicz,et al..Production of complex human glycoproteins in yeast[J],science,2003,301:1244-1246.
[146]Byung-Kwon Choi,Piotr Bobrowicz,Robert C.Davidson,et al..Use of combinatorial genetic libraries to humanize N-linked glycosylation in the yeast Pichia pastoris[J].Proceedings of Natinal Academy of sciences of the USA,2003,100(9):5022-5027.
[147]Billiau A.Interferon-gamma:biology and role in pathogenesis[J].Adv Immunol,1996,62:61-130.
[148]周海钧.干扰素国际标准品的标定及存在问题[J].中国生物制品学杂志,1996,9(2):97-99.
[149]Latina MA.Gamma-interferon effects on human fibroblasts from Tenon's capsule[J].Invest ophthalmol Vis Scl,1991,32(10):2806-2815.
[150]Luft T,Pang K C,Thomas E,et al.Type Ⅰ IFNs enhance the terminal differentiation of dendritic cells[J].J Immunol,1998.161:1947-1953.
[151]Radvanyi L G,Banerjee A,Weir M,et al.Low levels of interferon-α induce CD86(B7.2)expression and accelerates dendritic cell maturation from human peripheral blood mononuclear cells[J].Scand.J.Immunol,1999,50(5):499-509.
[152]Santini S M,Lapenta C,Logozzi M,et al.Type Ⅰ interferon as a powerful adjuvant for monocyte-derived dendritic cell development and activity in vitro and in Hu-PBL-scid mice[J].J Exp Med,2000,191(10):1777-1788.
[153]Tamir A,Jordan W J,Ritter M,et al.Interferon-alpha 2a is sufficient for promoting dendritic cell immunogenicity[J].Clin Exp Immunol,2005,142(3):471-80.
[154]Tompkins W A.Immunomodulation and Therapeutic Effects of the Oral Use of Interferon-alpha:Mechanism of Action[J].Journal of Interferon & Cytokine Research,1999,19(8):817-828
[155]朱玉贤、李毅。现代分子生物学。高等教育出版社,北京,2000,4
[156]陈广南,梁希若,吴淑华.含巨细胞病毒早期启动子的EPO逆转录病毒表达载体的构建[J].广州医学院学报.1997,25(2):1-4
[157]杨红宇,曾耀英,曹艳英,等.穿梭质粒pGFP-1与CMV启动子重组体的构建[J].暨南大学学报.2000,21(2):12-17
[158]Cheng L,Ziegelhoffer PR,Yang NS,et al.In vivo promoter activity and transgene expression in mammlian somatic tissuse evaluated by using particle bombardment[J].Proc Natl Acid Sci,1993,90:4455-4459.
[159]Xiang ZQ,Spitalnik SL,Cheng J,et al.Immune responses to nuceic acid vaccines to rabies virus[J].Virology,1995,209(2):569-579
[160]蔡文琴,王伯坛.实用免疫细胞化学与核酸分子杂交技术[Mj.成都:四川科学技术出版社.1994.1-5
[161]Funaro A,Horenstein A L,Ghisolfi G,et al.Identification of a 220-kDa membrane Tumor-Associated antigen by Human Anti-UK 114 monoclonal antibodies selected from the Immunoglobulin Repertoire of a cancer patient[J].Experimental cell research,1999247(2):44-46
[162]Morag C M,Breeman S,Simon P,et al.Immunohistochemical localization of Cytochrome P450CYP1B1 in Breast Cancer with monoclonal antibodies specific for CYP1B1[J].Journal of Histochemistry and Cytochemistry,1999,47(11):1457-1464
[163]沈关心、周汝麟,现代免疫学实验技术,武汉,湖北科学技术出版社,1998。
[164]孙树汉、戴建新、孙平武等,核酸疫苗,上海,第二军医大学出版社,1999。
[165]金振晓,刘维永,胡军,等.人甘露糖结合凝集素在小鼠肝细胞内表达[J].中国免疫学杂志,2002,18:828-831.
[166]韩双艳,李杰生,郭勇,等.DNA疫苗经基因枪介导的免疫研究[J].华南理工大学学报(自然科学版)2004,3,32(3):33-36.
[167]黎敏,程安春,汪铭书等.小鹅瘟病毒VP3基因疫苗在免疫鸭体内动态分布及表达规律的研究[硕士学位论文].四川雅安:四川农业大学,2007.6.
[168]刘小东,程安春,汪铭书等.小鹅瘟病毒VP3基因疫苗在小鼠体内动态分布与表达的研究[硕士学位论文].四川雅安:四川农业大学,2007.6。
[169]Michella M,Mckinney,Andrew Parkinson.A simple,non-chromatographic procedure to purify immunoglobulins from serum and ascites fluid.Journal of immunological methods,1987,96:271-278
[170]希尼尼根,猪繁殖与呼吸综合征病毒ORF5核酸疫苗的研究,四川雅安,四川农业大学博士学业论文,2004。
[171]曹澍泽,郭玉璞,董国雄,等.兽医微生物学及免疫检测技术[M].北京农业大学出版社,1992.5.
[172]周春宇.间接免疫组化和间接免疫荧光检测石蜡切片中鹅细小病毒方法的建立及其在鸭体内病毒分布规律的研究[硕士学位论文].四川雅安:四川农业大学,2005.6.
[173]汪洋,含有IBV基因的大肠杆菌发酵和质粒DNA抽提技术的研究,雅安,四川农业大学硕士论文,2000。
[174]陈创夫,余兴龙,马正海,等.猪瘟基因疫苗在小鼠肌肉中的动态分布表达[J].畜牧兽医学报.2002,33(3):280-284.
[175]龚焱宏,刘秀文,朱宝珍,等.乙型肝炎疫苗在BALB/c小鼠体内的生物分布情况[J].军事医学院院刊.2003,27(2):103-106.
[176]W Walther,T Minow et al.Uptake,Biodistribution,and Time Course of Naked Plasmid DNA Trafficking After Intratumoral In Vivo Jet Injection[J].human gene therpy,2006,17(6):611-624.
[177]G Acsadi,G Dickson,DR Love,et al.Direct gene transfer and expression into rat heart[J].New Biol.1991,3(1):71-81.
[178]Jukka H,Michael S,Francine C,et al.An improved plasmid DNA Expression vector for direct injection into skeletal muscle[J].Hum Gene Ther,1996(7):1205-1217.
[179]段坤,鸭α-干扰素核酸疫苗在鸭体内的动态分布及表达产物抗鸭瘟强毒效果初探,四川雅安,四川农业大学硕士学位论文,2007。
[180]陈红、孙红立、李震等,细胞因子的应用研究进展,上海畜牧兽医通讯,2003,5,6-8
[181]Weining K,Schuhz U,Munster U,et al.Biological properties of recombinant chicken interferon-gamma[J].Eur J Immunol,1996,26:2440-2447.
[182]Spector D L,Goldman R D,Leinwand L A.Cell:A Laboratory Manual[M].Third Edition.New York:Cold Spring Harbor Laboratory Press,1998.44-50
[183]Edward M B,Wilkinson R G,Lee B A,et al.Identification of duck T lymphocytes using an anti-human T cell(CD3) antiserum[J].Veterinary Immunology and Immunopathology,1996,51:353-363.
[184]程志萍,程安春,汪铭书,陈斌等.鸭IFN-α真核表达质粒基因枪免疫对鸭瘟弱毒疫苗免疫鸭细胞免疫调节作用初探,畜牧兽医学报,2007,38(10):1066-1071
[185]Roitt I,Brostoff J,Male D.Immunology[M].sixth edition.Harcourt Asia Pte Ltd,2001,113-125.
[186]GUO H C,LIU Z X,SUN S Q,et al.The Effect of Bovine IFN-α on the Immune Response in Guinea Pigs.Vaccinated with DNA Vaccine of Foot-and-Mouth Disease Virus[J].Acta Biochimica et Biophysica Sinica,2004,36(10):701-706.
[187]Krieg A M,Yi A K,Sehorr J,et al.rhe role CpG dinucleotides in DNA vaccines[J].Trends Microbiol,1998,(6):23-27.
[188]Jones M,Cordell JL,Beyers AD,et al.Detection of T and B cells in many animal species using cross reactive anti-peptide antibodies[J].Immunol,1993,150:5429-5435.
[189]Gazzinelli RT,Hakim FT,Hieny S,et al.Synergistic mle of CD4 and CD8 T lymphocytes in IFN-gamma production and protective immunity induced by an attenuated Toxoplasma gondii vaccine[J].Immunol,1991,146(1):286-292.
[190]C.Janeway.Immunobiology(5th Ed)[M].New York:Garland Publishing,2004,81-90
[191]陈诗书,汤雪明.医学细胞与分子生物学[M].上海:上海医科大学出版社,1995.2-4.
[192]Edward M B,Ray G W,Belinda A L,et al.Identification of duck T lymphocytes using an anti-human T cell(CD3) antiserum[J].Veterinary Immunology and Immunopathology,1996,7(51):353-363.
[193]Tang D C,Devit M,Johnston SA,et al.Genetic immunization in a single method for eliciting an immune response[J].Nature,1992,365:152-154.
[194]Fynan E F,Webster R G,Fuller D H,et al.DNA vaccines;protective immunizations by parenteral,mucoral and gene-gun inoculations[J].Proc Natl Acad Sci USA,1993,90:11478-11482.
[195]Chattergoon M A,Robinson T M,Boyer J D,et al.Specific Immune Induction Following DNA-Based Immunization Through In Vivo Transfectionand Activation of Macrophages/Antigen-Presenting Cells[J].J Immunology,1998,160(12):5707-5718.
[196]王军阳、沈桂香、胜利等,DNA核酸疫苗诱导小鼠免疫应答的作用,西安医科大学学报,2002,23(3):249-251
[197]Enrico P,Laura B,Simona P,et al.Type Ⅰ IFN as a Natural Adjuvant for a Protective Immune Response:Lessons from the Influenza Vaccine Model[J].J Immunology,2002,169:375-383.
[198]金宁一,王洪军,王继群,等.HIV-1gag/IFN-2b表达产物的生物活性研究[J].生物技术,2004,14(1):14-16.
[199]Wells D J,Goldspink G.Age and sex influence expression of plasmid DNA directly injected into mouse skeletal muscle[J].FEBS Lett.1992.306(2-3):203-205.
[200]徐新女,基因疫苗的研究概况,中华综合临床医学杂志,2004,7(6):152-153
[201]张惠星,张维,罗玉芳,等.鸭外周血淋巴细胞转化试验的研究[J].上海免疫学杂志,1990,10(5):262-264.
[202]Cauchy L,Malkinson M.The mitogenic response to concanavalin A of Muscovy duck peripheral blood lymphoeytes in the early posthatching period[J].Avian Pathol,1981,10(1):77-81.
[2]Issacs A,Lindanmann J.Virus interference,I.the interferon[J].Proc R Soc Ser B.1957,147:258-267.
[3]刘胜旺、卢景良、张万旗.鸡干扰素及其相关蛋白分子生物学研究进展。预防兽医学进展,2000,2(1):17-19。
[4]孙志宏,陈正礼,范光丽。干扰素研究进展。延安大学学报(自然科学版),2000,19(1):76-79。
[5]Bazer F W.Mediators of maternal recognition of pregnancy in mammals[J].Proc Soc Exp Bio Med,1992,199:373
[6]Steinmann G G,Rosenkaimer F,Leitz G.Clinical experience with interferon-α and interferon-γ[J].Int Rev Exp Pathol,1993,34B:193
[7]Stanton G J,Weigent D A,Fleischmann W R.et al.Interferon Review[J],Investigative Radiology,1987,22:259-273.
[8]金伯泉,细胞和分子免疫学(第二版),北京,科学出版社,2000。
[9]陈坤华。细胞因子对下丘脑—垂体—肾上腺皮质轴之间调节作用[J].生理科学进展,1993,34(2):113-117
[10]吴玉厚、吴冰洁、周国利等.干扰素研究进展,生物学教学,2007,32(7):2-4。
[11]邢利和,王福生。干扰素产生细胞的研究进展,国外医学免疫学分册[J].2002,25(2):76-78.
[12]卢建.细胞因子受体组成、结构功能及信号传导机制[J].生理科学进展,1995,26(4):305-311.
[13]杨元忠,试述干扰素的主要作用.畜牧兽医杂志,1997,16(3):34-35。
[14]程坚、王立俊、杨波。a干扰素治疗肿瘤机制的研究进展。农业生物技术学报,2000,8(3):236-238。
[15]Sekellick MJ,Ferrandino AF,Hopkins DA,et al.Chicken interferon gene:cloning,expression and analysis[J].J Interferon research,1994,14:71-79
[16]H W Murray,A.Szuro-Sudol,D wellner.et al,Role of ryptophan degradation in respiratory burst-independent antimicrobial activity of gamma interferon-stimulated human macrophages.Infect.Immun.1989,57:845-849.
[17]马良,干扰素与寄生虫感染.重庆医科大学学报.1995,30(3):241-244
[18]M A Orellana,Y Suzuki,F Araujo,et al.Role of beta interferon in resistance to Toxoplasma gondii infection.Infect.Immun.1991,59:3287-3290.
[19]郭定宗,祝海宝。干扰素研究进展及其在兽医临床上的应用[J],养殖与饲料,2006,2:5-8。
[20]杜念兴.兽医免疫学[M].第2版,北京:中国农业出版社.1997,74-76.
[21]孙圣兰,江青艳,傅伟龙.干扰素的研究进展[J].动物医学进展,2002,23(6):39-41.
[22]马飞.干扰素的研究进展[J].中国畜牧兽医,2005,32,12:36-37.
[23]Towata T,Hayashi N,Katayama K,et al.Effects of cytokines on HLA class Ⅰ antigen expression on Huh6 and HB611 cells[J].Gastroenterol,1993,28(2):242-248.
[24]Luft T,Pang K C,Thomas E,et al.Type Ⅰ IFNs enhance the terminal differentiation of dendritic cells[J].J Immunol,1998,161:1947-1953.
[25]Radvanyi L G,Banerjee A,Weir M,et al.Low levels of interferon-α induce CD86(B7.2)expression and accelerates dendritic cell maturation from human peripheral blood mononuclear cells[J].Stand J Immunol,1999,50(5):499-509.
[26]Santini S M,Lapenta C,Logozzi M,et al.Type 1 interferon as a powerful adjuvant for monocytederived dendritic cell development and activity in vitro and in Hu-PBL-scid mice[J].J Exp Med,2000,191(10):1777-1788.
[27]柴玉波、孙海峰、李春源。干扰素治疗慢性乙型肝炎的疗效分析,生物工程学报。1997,13(4):406-408
[28]陈晓初,谢佩蓉,郑蒲.IFN—γ和IL—6或IL—1联合应用对人肝癌细胞系CLINH合成的作用[J],免疫学杂志,1999,15(4):239-241。
[29]刘高生,卢升云,干扰素的免疫作用机制。中国兽医科技,2004,34(10):72-74
[30]朱长庚,李正莉,细胞因子与免疫—神经—内分泌调节网络[J],解剖学报,1996,27(4):339-344
[31]Chao C C,Hu S,Sheng W S,et al.Lytokine stimulated astrocytes damage human neurous via a nitri-oxide mechanism[J],Glia,1996,16:276-284
[32]Kiefer R,Keutzberg G W.Distribution of cytokines in nerous system[J].Nenro sci.1990,35(3):725-732.
[33]Gray P W,Peung D W,Pennica D,et al.Expression of human immune interferon cDNA in E.coli and monkey cells[J].Nature,1982,295:503-506.
[34]Goeddel D V,YeIverton E,Uurich A,et al.Human leukocyte interferon produced by E.coli is biologically active[J].nature,1980,287,2:4411-4415.
[35]Gray PW,GoeddeL D V,Cloning and expression of murine immune interferon cDNA[J].Proc Natl Acad Sci USA.1983.80:5842-5846.
[36]Dijkmans R,Vandenbroeck K,Beuken E,et al.Sequence of the porcine interferon-gamma gene[J].Nucleic Acids Research,1990,18,42-59.
[37]侯云德,分子病毒学[M],北京:学苑出版社,1990,598-642.
[38]Lillehoj HS,Kaspers B.Jenkins MC,et al.Avian interferon and interleukin-2.A review by comparison with mammalian homologues[J].Poultry Science Review,1992,4,67-85.
[39]Lowenthal JW,Digby MR,York JJ.Production of interferon-γ by chicken T cells[J].Journal of interferon and Cylokine Research.1995,15:933-938
[40]Sick C,Schuhz U,Staeheli P,A family of genes coding for two serologicaIly distince chicken interferon[J],The JournaI of Biological Chemistry.1996.271.I3:7636-7639
[41]Schultz U,Ridderle C,Sekeliick MJ.Recombinant chicken interferon from E.coli and transfected COS 7 cells is biologically active[J].European Journal biochemistry,1995,229,73-76
[42]Suresh M,Karaca K,Foster D.Molecular and functional characterization of turkey interferon[J],J Virol,1995,69,12:8159-8163
[43]夏春,汪明,朱凌云等。惠阳胡须鸡干扰素基因分子克隆与序列丹析[J],畜牧兽医学报,2000,31(6):563-566
[44]夏春,汪明,杨琪等。丝羽乌骨鸡干扰索基因分子克隆与字列丹析[J]。中国免疫学杂志,2000,5:22-24
[45]曾永长,吕英姿.鸡α干扰素基因的克隆和鉴定[J].中国预防兽医学报,2001,23(4):259-261.
[46]汪明、吴志光,夏春。肉鸡IFN-alpha基因的克隆、序列分析以及在大肠杆菌中的表达,农业生物技术学报,2000,8(4)377-381。
[47]韦琴,彭贵青,金梅林,等.鸡α千扰素基因的克隆、原核表达及抗病毒效果研究[J].生物工程学报,2006.22(5):737-743.
[48]Schultz U,Kock J,Schlicht HJ,et al.Recombinant auck interferon:A new reagent for studying the mode of interferon action against hepatitis B virus[J].Virology,1995,212:641-649
[49]Schultz U,Chisari FV.Recombinant duck interferon gamma inhibits duck hepatitis.B virus replication in primary hepatocytest[J],J Virol,1999,73(4):3162-3168
[50]夏春,万建青,吴志光等,北京鸭Ⅰ型干扰素基因分子克隆与序列分析,畜牧兽医学报,2000,31(6):567-570
[51]阮小飞,林常有,杨天耀,等.北京鸭干扰素-α基因的分子克隆与表达[J].中国兽医杂,2004,40(12):11-13.
[52]王春霞,王林川.番鸭IFN-α基因的克隆与序列分析[J].华南农业大学学报,2002,23(4):68-70.
[53]王春霞.王林川.鄢志强等,藩鸭IFN-α基因在毕赤酵母中的分泌表达,中国兽医科技,33(7):44-48
[54]叶伟成,张存,王一成,等.绍兴鸭α-干扰素基因克隆、表达及活性测定[J].浙江农业学报,2005,17(3):115-119.
[55]Hung yueh yeh,Barbara J,Winslow et al,In vitro effects of recombinant chicken interferon-r on immune cells[J].Journal of interferon and cytokine res.1999,19:687-691.
[56]Digby MR.Lowenthal JW.cloning and expression of the chicken interferon-γ gene[J].Journal of Interferon and Cytokine Research.1995.15:939-949.
[57]Song KD,Lillehoj HS,Choi KD,et al.Expression and functional characterization of recombinant chicken Interferon-gamma[J].Veterinary Immunology and Immunopathology,1997,58:321-333.
[58]Kaisr P,Wain HM.Rothwell L.Structure of the chicken Interferon-γ gene and comparison to mammalian homo]ogues[J].Gene,1998,207:25-23.
[59]Kaiser P.Sonnemans D.Smith LM.Avian IFN-γ Sequence Analysis Suggests Probable Cross Species Reactivity among Galliforms[J].Journal of Interferon and Cytokine,1998,18:711-719
[60]刘胜旺,陈洪岩,孔宪刚等,鸡γ-干扰素基因的分子克隆与序列测定,中国兽医学报,2000,20(3):228-230。
[61]吕英姿,毕英佐,曹永长,石歧杂鸡γ-干扰素基因的克隆与序列分析,华南农业大学学报(自然科学版),2002,23(4):61-63。
[62]吴志光,夏春,汪明。北京鸭Ⅱ型干扰索基因分子克隆与序列分析,中国兽医科技,2001,31(3):7-9。
[63]夏春,刘津,杨琪等,猪干扰素β基因的分子克隆与测序,中国兽医杂志,2000,26:6-7。
[64]郭瀛军,吴丹,陈蕊雯等,猪γ干扰素基因的克隆、表达及其纯化,生物工程学报,2001,17(2):184-186。
[65]赵英杰、徐宏军、李尚波等,长白猪γ干扰素基因的原核表达与分析,中国兽医科技,2006,36(1):46-51。
[66]曹瑞兵、包晶晶,周海霞等,猪β干扰素的原核表达及其对猪流行性腹泻病毒的抑制作用研究,中国病毒学报,2004,19(4):364-368。
[67]刘万钧,张道永,马思齐等.猪白细胞干扰素防治猪流行性腹泻试验.中国兽医杂志.1999,25(2):13-14。
[68]夏春、汪明,拉布拉多犬和德国牧羊犬IFN-α基因的分子克隆和测序,农业生物技术学报,1999,7(3):245-247。
[69]李学仁,王震海,葛菲菲等,奶牛γ干扰素基因的高效表达及活性测定,中国病毒学,2004,19(4)356-359。
[70]史喜菊、夏春、汪明,水牛α-干扰素基因的克隆与表达及其抗病毒活性,农业生物技术学报,2004,12(3):288-293。
[71]李玉萍,景志忠,蒲训等,牦牛β干扰素基因的克隆与表达,中国兽医科学,2006,36(1):40-45。
[72]Dimier IH.Quere P.Naciri M.et al.Inhibition of Eimeriatenel la development in vitro mediated by chicken macrophages and fibroblasts treated with chicken cell supernatants with IFN γactivity[J].Avian Diseases.1998.42:239-247
[73]Lowenthal JW,Onel TE,Broadway M,et al.Coadministration of IFN γ enhance antibody responses in chickens[J],Journal of interferon and Cytokine Research,1998.28:617-622.
[74]Plachy J.Weining KC,Kremmer MJ,et al.Proctive effects of type Ⅰ and type Ⅱ interferons toward Rous Sarcoma Virus induced tumors in chicken[J],Virology,1999,256,85-91.
[75]Marcus PI,Heide VD,Sekellick MJ.Interferon action on Avian viruses I,Oral administration of chicken interferon a Ameliorates Newcastle Disease[J],Journal of interferon and Cytokine Research,1999,19:881-885
[76]侯俊玲、郑亚东、景志忠等,DNA疫苗免疫佐剂的研究进展,中国兽医科技,2005,35(2):153-156。
[77]王克坚,殷震,白细胞介素类细胞因子的免疫佐剂作用,预防兽医学进展.2002,2(1):30-33。
[78]Cox J C,Coulter A R.Adjuvants a classification and review of their modes of action[J].Vaccine,1997,15(3):248-256
[79]张悦,戚晓红,冯振卿,免疫佐剂的研究紧展,中国血吸虫病防治杂志.2004,16(3)236-238。
[80]吴乃虎,基因工程原理.科学出版社,北京,2001。
[81]A Chaudhuri,S Orme,S Eilam et al,CD40-mediated signals inhibit the binding of TNF receptor-associated factor 2 to the CD40 cytoplasmic,domain[J],J Immunol,1997,159:4244-4251.
[82]Ihata A,Watabe S,Saski S,et al.Immunomodulatory effect of a plasmid expressing CD40ligand,on DNA vaccination against human immunodeficiency virus type-1[J].Immunology,1999,98(3):436-442
[83]Dierks SE,Bartlett WC,Edmeades RL,et al.The oligomeric nature of the murine FC epsilon RⅡ/CD23.implications for function[J],J Immunol,1993,150:2372-2382.
[84]Mendoza RB,Cantwell MJ,Kpps TJ.Immunostimulatory effects of a plasmid expressing CD40ligand(CD154) on gene immunization[J],J Immunol,1997,59(12):5777-5781.
[85]Kreing AM,Yiak,Klinman DM.CpG motif in bacterial DNA trigger direct B-cell activation[J].Nature,1995,374:546-549.
[86]Chace JH,Hooker NA,Coedery JS.Bacterial DNA induced NK cell IFN-γ production is dependent on macrophage secretion IL-12[J].Clin immunol immunopath,1997,84:185-193。
[87]Kojima Y,Xin K Q,Ooki T,et al.Adjuvant effect of multi-CpG motifs on an HIV-1 DNA vaccine[J],Vaccine,2002,20(23-24):2857-2865
[88]Wang X,Jiang P,Deen S,et al.Efficacy of DNA vaccines enhanced by coadministration,with CpG oligodeoxynucleotide[J],Avian Disc,2003,47(4):1305-1312.
[89]Lipford GB,Bamer M,Heeg K.Oligonucleotides protomer B and cytotoxic T cell responses to protein antigen:a new class of vaccine adjuvant[J].Eur J Immunol,1997,27:2340-2344.
[90]Heather LD,Ridini W,Krieg AM.CpG DNA is a porent enhancer and specific immunity in mice immunized with recombinant hepatitis B surface antigen[J].J Immunol,1998,160(2):870-876
[91]Moldoveanu Z,Love-Homan L,Wen QH,et al.CpG DNA,a novel immune enhancer for systemic and mucosal immunization with influenza virus,vaccine,1998,16(11-12):1216-1224.
[92]朱建中,朱鸿飞,鲁春,李富光。免疫佐剂CpG DNA的研究进展。生命的化学,2002,22(2):155-157。
[93]Okuda K,Xin KQ,Haruki A,et al.Transplacental genetic immunization after intravenous delivery of plasmid DNA to pregnant mice[J],J Immunol,2001,167(9):5478-5484.
[94]陈吉刚。郑肖娟,齐静,禽类细胞因子——新型免疫佐剂和免疫治疗剂,饲料工业,2005,26(1):35-41。
[95]Heath AW.细胞因子:新型免疫佐剂。国外医学,预防、诊断,治疗用生物制品分册,1993,16(3):110-113
[96]Karaca K,Sharma J M,Winslow B J,et al.Recombinant fowlpox virus coexpressing chicken type Ⅰ IFN and Newcastle disease virus HN and F genes:influence of IFN on protrctive efficacy and humoral responses of chickens following in ovo or post-hatch administration of recombinant virus[J].Vaccine,1998,16(16):1496-1503.
[97]Sekellick M J,Lowenthal J W,Oneil T E,et al.Chicken interferon types Ⅰ and Ⅱ enhance synergistically the antiviral state and nitric oxide secretion[J],J Interferon Cytokine Res,1998,18(6):407-414.
[98]Schijins VE,Weining KC,Nuijten P,et al.Immunoadjuvant activities of E.coli-and plasmid -expressed recombinant chicken IFN-alpha/beta,IFN-gamma and IL-1beta in 1-day and 3-week old chickens[J].Vaccine,2000,18(20):2147-2154
[99]邹丽容、陈则。DNA疫苗免疫佐剂的研究进展,激光生物学报,2002,11(5):395-400。
[100]张晋举,钱平。表达干扰素作为佐剂的重组鸡痘病毒胚胎免疫火鸡防制新城疫的研究。预防兽医学进展,2000,2(2):39-41。
[101]程坚、刘秀梵、彭大新等,表达鸡Ⅱ型干扰素基因的重组鸡痘病毒的构建,农业生物技术学报,2002,10(2):152-155。
[102]Warren T L,Weiner G J.Uses of granulocyte macophage colony-stimulating factor in vaccine development[J].Curr Opinhenratol,2000,7(3):168-173
[103]Lowenthal JW,Lambrecht B,van den Berg T,at al.Avain cytokines-the natural approach to therapeutics[J].Dev comp immunol,2000,24(2-3):355-365.
[104]Lowenthal JW,York JJ,Oneil TE.et al.In vivo effects of chicken interferon-gamma during infection with Eimeria[J].J interferon cytokine res,1997,17(9):551-558
[105]Guo H,Chen G,Lu F,et al.Immunity induced by DNA vaccine of plasmid encoding the rhoptry protein Ⅰ gene combined with the genetic adjuvant of pclFN-γ against toxoplasma gondii in mice[J],Chin med J(EngL),2001,114(3):317-320.
[106]彭大新,表达马立克氏病毒糖蛋白gB基因及共表达糖蛋白B和鸡γ干扰素重组鸡痘病毒的构建及其免疫保护作用,扬州大学博士论文,2001。
[107]于涟,李建荣,黄耀伟等。鸡白细胞介素2增强传染性法氏囊病病毒多聚蛋白DNA疫苗免疫原性的研究[J],生物工程学报,2001,17(6):653-657。
[108]Bu J,Song Y,Rompatio G,et al.Codelievery of IL-2 or liposomes augment the responses of mice to a DNA vaccine for pseudorabies virus IE180[J].Comp Immunol Microbio Intfect Dis,2003,26(3):175-187
[109]杨汉春、姚火春、王君伟等。动物免疫学(第二版),北京,中国农业大学出版社,2003,8。
[110]Reed SG,Pihid L,et al.IL-1 as adjuvant:role of T cell in the augmentation of specific antibody production by recombinant human IL-1 alpha[J].Immunol,1989,142:3129-3133.
[111]Kim J L,Maguire H C,Nottingham L K,et al.Coadministration of IL-12 or IL-10 expression oassettes drives immune responses towards a Th1 phenotype[J].Interferon Cytokin Res,1998,18(7):537-547
[112]Van Snick J.Interleukin-6:an overview[J].Annu Res Immunol,1990.8:253-278
[113]Schoenhaut D S,Chua A O,Wo litzky A G,et al.Cloning and expression of murine IL-12[J].J Immunnol,1992,148:3433-34440
[114]杜德伟,周永兴,冯志华,等.IL-12的真核表达载体促进小鼠对DNA疫苗的免疫应答[J].中华微生物学核免疫学杂志,2000,20(6):543-544
[115]刘卓宝,人类战胜疾病的新武器——DNA疫苗[M],上海科学技术出版社,2001。
[116]卢铀,田聆,魏于全等,DNA疫苗抗肿瘤研究进展[J],中华医学遗传学杂志,2000,17(4):288-290。
[117]Sedegrah M,Weiss W,Saci J B,et al.Improving protective immunity induced by DNA-based immunization:priming with antigen and GM-CSF-encoding plasmid DNA and boosting with antigen-expressing recombinant poxvirus[J].J Immunol,2000,164(11):5905-5912.
[118]Geissler M,Gesien A,Wands JR,Inhibitory effects of chronic ethanol consumption on cellular immune responses to hepatitis C virus core protein are reversed by genetic immunizations augmented with cytokine-expressing plasmids.J Immunol,1997,159(10):5107-5113
[119]Operschall E,Schuh T,Heinzerling L,Pavlovic J,et al.Enhanced protection against virual infection by co-administration of plasmid DNA coding for ritual antigen and cytokines in mice.J Clin Virol,1999,13(1-2):17-27
[120]Lu Y,Xin KQ,Hamajima K,Tsuji T,et al.Macrophage inflammatory protein-lalpha(MIP-lalpha)expression plasmid enhances DNA vaccine-induced immune response against HIV-1.Clin Exp Immunol,1999,115(2):335-341
[121]晏菊芳。DNA疫苗的分子佐剂研究进展。国外医学免疫学分册,2000,24(4):197-201
[122]尹荣兰、李昌、张乃生。DNA疫苗免疫佐剂研究进展,中国兽医杂志,2008,44(4):77-79
[123]杜德伟,周永兴,吴学勤,冯志华等,IL-12的真核表达载体对乙肝基因疫苗的免疫佐剂效应。中国现代医学杂志,2000,10(7):1-3。
[124]赫然,DNA疫苗的佐剂研究综述,中山大学研究生学刊(自然科学、医学版),2003,24(2):9-15。
[125]Weiss w R,Ishii K J,Hedstrom R C,et al.A plasmid encoding murine granulocyte-macrophage colony stimulating factor increases protection conferred by a malaria DNA vaccine[J].J Immunol,1998,161(5):2325-2332.
[126]Adel,patient K,Creminon C,et al.Genetic immunization with bovine beta-lactoglobulin cDNA induces a preventive and persistent inhibition of specific anti-BLG IgE response in mice[J].Int Arch Allergy Immunol,2001,26(1):59-67
[127]Chen Z,Sahasho Y,Matsuo K,et al.Comparison of the ability of viral protein-expressing plasmid DNAs to protect against influenza[J].Vaccine,1998,16(16):1544-1549.
[128]Chen Z,Natsuo K,Asanuma H,et al.Enhanced protection against A lethal influenza virus challenge by immunization with both hemagglutinin and neuraminidase-expressing DNAs[J],Vaccine,1999,17:653-659
[129]Sick C,Schuhz U、Staeheli P,A family of genes coding for two serologicaIly distince chicken interferon[J],The JournaI of Biological Chemistry.1996.271.13:7636-7639
[130]萨姆布鲁克,E.F.弗里奇,T曼尼阿蒂斯.分子克隆实验指南[M].(第二版)北京:科学出版社。
[131]陈斌,程安春,汪铭书,等.麻鸭α干扰素基因克隆与序列分析[J].中国兽医学报,2006,26(4):401-404.
[132]吴琦,枯草芽孢杆菌植酸酶phyC基因的克隆及其表达研究。四川大学博士学位论文,2004。
[133]Stewart W E.Interferon nomenclature[J].Nature,1980,286:110
[134]李伍举、吴加金.pBV220载体中外源基因表达水平定量分析。病毒学报,1997,13(2):125-133。
[135]龚永强,鸭α-干扰素基因的原核表达及抗病毒活性研究,雅安,四川农业大学硕士论文,2007。
[136]杨梅,重组鸭a-干扰素抗鸭乙型肝炎病毒实验感染研究。四川农业大学硕士论文,2007。
[137]郭宇飞.鸭病毒性肝炎病毒CH强毒株部分生物学特性的研究及荧光实时定量PCR检测方法的建立和应用[学位论文].四川雅安.四川农业大学.2005
[138]Gold L.Posttranscriptional regulatory mechanisms in Escherichia coli[J].Ann Rev Biophs Chem,1988,57(2):199-223.
[139]Tessier LH,Sondermeyer P,Faure T,et al.The influence of mRNA primary and secondary structure on human IFN-gamma gene expression in E.coli[J].Nucl Acids Res,1984,12(20):7663-7675.
[140]DE Smitmh,Van Duin J.Control of translation by mRNA secondary structure in Escherichia coli.A quantitative analysis of literature data[J].J Mol Biol,1994,244(2):144-150.
[141]任启生.王嘉玺,邹民吉。PBV-IL-6重组质粒的构建及其在E.coli细胞中的超高表达.中国免疫学杂志,1992,8:133-135。
[142]张智清、姚立红、候云德。含PrPl启动子的原核表达载体的组建及其应用。病毒学报,1990,6(2):111-116
[143]李武平、宋瑜丽、张成海等。重组人干扰素—β(rhIFN-β)中试工艺研究。中国生物制品学杂志,2004,17(1):33-35。
[144]张成海、李武平、衣作安等。人干扰素ψ(huIFN-ψ)的高效表达、纯化及生物学活性。病毒学报,2004.20(2):124-127。
[145]Stephen R.H.,Piotr Bobrowicz,Beata Bobrowicz,et al..Production of complex human glycoproteins in yeast[J],science,2003,301:1244-1246.
[146]Byung-Kwon Choi,Piotr Bobrowicz,Robert C.Davidson,et al..Use of combinatorial genetic libraries to humanize N-linked glycosylation in the yeast Pichia pastoris[J].Proceedings of Natinal Academy of sciences of the USA,2003,100(9):5022-5027.
[147]Billiau A.Interferon-gamma:biology and role in pathogenesis[J].Adv Immunol,1996,62:61-130.
[148]周海钧.干扰素国际标准品的标定及存在问题[J].中国生物制品学杂志,1996,9(2):97-99.
[149]Latina MA.Gamma-interferon effects on human fibroblasts from Tenon's capsule[J].Invest ophthalmol Vis Scl,1991,32(10):2806-2815.
[150]Luft T,Pang K C,Thomas E,et al.Type Ⅰ IFNs enhance the terminal differentiation of dendritic cells[J].J Immunol,1998.161:1947-1953.
[151]Radvanyi L G,Banerjee A,Weir M,et al.Low levels of interferon-α induce CD86(B7.2)expression and accelerates dendritic cell maturation from human peripheral blood mononuclear cells[J].Scand.J.Immunol,1999,50(5):499-509.
[152]Santini S M,Lapenta C,Logozzi M,et al.Type Ⅰ interferon as a powerful adjuvant for monocyte-derived dendritic cell development and activity in vitro and in Hu-PBL-scid mice[J].J Exp Med,2000,191(10):1777-1788.
[153]Tamir A,Jordan W J,Ritter M,et al.Interferon-alpha 2a is sufficient for promoting dendritic cell immunogenicity[J].Clin Exp Immunol,2005,142(3):471-80.
[154]Tompkins W A.Immunomodulation and Therapeutic Effects of the Oral Use of Interferon-alpha:Mechanism of Action[J].Journal of Interferon & Cytokine Research,1999,19(8):817-828
[155]朱玉贤、李毅。现代分子生物学。高等教育出版社,北京,2000,4
[156]陈广南,梁希若,吴淑华.含巨细胞病毒早期启动子的EPO逆转录病毒表达载体的构建[J].广州医学院学报.1997,25(2):1-4
[157]杨红宇,曾耀英,曹艳英,等.穿梭质粒pGFP-1与CMV启动子重组体的构建[J].暨南大学学报.2000,21(2):12-17
[158]Cheng L,Ziegelhoffer PR,Yang NS,et al.In vivo promoter activity and transgene expression in mammlian somatic tissuse evaluated by using particle bombardment[J].Proc Natl Acid Sci,1993,90:4455-4459.
[159]Xiang ZQ,Spitalnik SL,Cheng J,et al.Immune responses to nuceic acid vaccines to rabies virus[J].Virology,1995,209(2):569-579
[160]蔡文琴,王伯坛.实用免疫细胞化学与核酸分子杂交技术[Mj.成都:四川科学技术出版社.1994.1-5
[161]Funaro A,Horenstein A L,Ghisolfi G,et al.Identification of a 220-kDa membrane Tumor-Associated antigen by Human Anti-UK 114 monoclonal antibodies selected from the Immunoglobulin Repertoire of a cancer patient[J].Experimental cell research,1999247(2):44-46
[162]Morag C M,Breeman S,Simon P,et al.Immunohistochemical localization of Cytochrome P450CYP1B1 in Breast Cancer with monoclonal antibodies specific for CYP1B1[J].Journal of Histochemistry and Cytochemistry,1999,47(11):1457-1464
[163]沈关心、周汝麟,现代免疫学实验技术,武汉,湖北科学技术出版社,1998。
[164]孙树汉、戴建新、孙平武等,核酸疫苗,上海,第二军医大学出版社,1999。
[165]金振晓,刘维永,胡军,等.人甘露糖结合凝集素在小鼠肝细胞内表达[J].中国免疫学杂志,2002,18:828-831.
[166]韩双艳,李杰生,郭勇,等.DNA疫苗经基因枪介导的免疫研究[J].华南理工大学学报(自然科学版)2004,3,32(3):33-36.
[167]黎敏,程安春,汪铭书等.小鹅瘟病毒VP3基因疫苗在免疫鸭体内动态分布及表达规律的研究[硕士学位论文].四川雅安:四川农业大学,2007.6.
[168]刘小东,程安春,汪铭书等.小鹅瘟病毒VP3基因疫苗在小鼠体内动态分布与表达的研究[硕士学位论文].四川雅安:四川农业大学,2007.6。
[169]Michella M,Mckinney,Andrew Parkinson.A simple,non-chromatographic procedure to purify immunoglobulins from serum and ascites fluid.Journal of immunological methods,1987,96:271-278
[170]希尼尼根,猪繁殖与呼吸综合征病毒ORF5核酸疫苗的研究,四川雅安,四川农业大学博士学业论文,2004。
[171]曹澍泽,郭玉璞,董国雄,等.兽医微生物学及免疫检测技术[M].北京农业大学出版社,1992.5.
[172]周春宇.间接免疫组化和间接免疫荧光检测石蜡切片中鹅细小病毒方法的建立及其在鸭体内病毒分布规律的研究[硕士学位论文].四川雅安:四川农业大学,2005.6.
[173]汪洋,含有IBV基因的大肠杆菌发酵和质粒DNA抽提技术的研究,雅安,四川农业大学硕士论文,2000。
[174]陈创夫,余兴龙,马正海,等.猪瘟基因疫苗在小鼠肌肉中的动态分布表达[J].畜牧兽医学报.2002,33(3):280-284.
[175]龚焱宏,刘秀文,朱宝珍,等.乙型肝炎疫苗在BALB/c小鼠体内的生物分布情况[J].军事医学院院刊.2003,27(2):103-106.
[176]W Walther,T Minow et al.Uptake,Biodistribution,and Time Course of Naked Plasmid DNA Trafficking After Intratumoral In Vivo Jet Injection[J].human gene therpy,2006,17(6):611-624.
[177]G Acsadi,G Dickson,DR Love,et al.Direct gene transfer and expression into rat heart[J].New Biol.1991,3(1):71-81.
[178]Jukka H,Michael S,Francine C,et al.An improved plasmid DNA Expression vector for direct injection into skeletal muscle[J].Hum Gene Ther,1996(7):1205-1217.
[179]段坤,鸭α-干扰素核酸疫苗在鸭体内的动态分布及表达产物抗鸭瘟强毒效果初探,四川雅安,四川农业大学硕士学位论文,2007。
[180]陈红、孙红立、李震等,细胞因子的应用研究进展,上海畜牧兽医通讯,2003,5,6-8
[181]Weining K,Schuhz U,Munster U,et al.Biological properties of recombinant chicken interferon-gamma[J].Eur J Immunol,1996,26:2440-2447.
[182]Spector D L,Goldman R D,Leinwand L A.Cell:A Laboratory Manual[M].Third Edition.New York:Cold Spring Harbor Laboratory Press,1998.44-50
[183]Edward M B,Wilkinson R G,Lee B A,et al.Identification of duck T lymphocytes using an anti-human T cell(CD3) antiserum[J].Veterinary Immunology and Immunopathology,1996,51:353-363.
[184]程志萍,程安春,汪铭书,陈斌等.鸭IFN-α真核表达质粒基因枪免疫对鸭瘟弱毒疫苗免疫鸭细胞免疫调节作用初探,畜牧兽医学报,2007,38(10):1066-1071
[185]Roitt I,Brostoff J,Male D.Immunology[M].sixth edition.Harcourt Asia Pte Ltd,2001,113-125.
[186]GUO H C,LIU Z X,SUN S Q,et al.The Effect of Bovine IFN-α on the Immune Response in Guinea Pigs.Vaccinated with DNA Vaccine of Foot-and-Mouth Disease Virus[J].Acta Biochimica et Biophysica Sinica,2004,36(10):701-706.
[187]Krieg A M,Yi A K,Sehorr J,et al.rhe role CpG dinucleotides in DNA vaccines[J].Trends Microbiol,1998,(6):23-27.
[188]Jones M,Cordell JL,Beyers AD,et al.Detection of T and B cells in many animal species using cross reactive anti-peptide antibodies[J].Immunol,1993,150:5429-5435.
[189]Gazzinelli RT,Hakim FT,Hieny S,et al.Synergistic mle of CD4 and CD8 T lymphocytes in IFN-gamma production and protective immunity induced by an attenuated Toxoplasma gondii vaccine[J].Immunol,1991,146(1):286-292.
[190]C.Janeway.Immunobiology(5th Ed)[M].New York:Garland Publishing,2004,81-90
[191]陈诗书,汤雪明.医学细胞与分子生物学[M].上海:上海医科大学出版社,1995.2-4.
[192]Edward M B,Ray G W,Belinda A L,et al.Identification of duck T lymphocytes using an anti-human T cell(CD3) antiserum[J].Veterinary Immunology and Immunopathology,1996,7(51):353-363.
[193]Tang D C,Devit M,Johnston SA,et al.Genetic immunization in a single method for eliciting an immune response[J].Nature,1992,365:152-154.
[194]Fynan E F,Webster R G,Fuller D H,et al.DNA vaccines;protective immunizations by parenteral,mucoral and gene-gun inoculations[J].Proc Natl Acad Sci USA,1993,90:11478-11482.
[195]Chattergoon M A,Robinson T M,Boyer J D,et al.Specific Immune Induction Following DNA-Based Immunization Through In Vivo Transfectionand Activation of Macrophages/Antigen-Presenting Cells[J].J Immunology,1998,160(12):5707-5718.
[196]王军阳、沈桂香、胜利等,DNA核酸疫苗诱导小鼠免疫应答的作用,西安医科大学学报,2002,23(3):249-251
[197]Enrico P,Laura B,Simona P,et al.Type Ⅰ IFN as a Natural Adjuvant for a Protective Immune Response:Lessons from the Influenza Vaccine Model[J].J Immunology,2002,169:375-383.
[198]金宁一,王洪军,王继群,等.HIV-1gag/IFN-2b表达产物的生物活性研究[J].生物技术,2004,14(1):14-16.
[199]Wells D J,Goldspink G.Age and sex influence expression of plasmid DNA directly injected into mouse skeletal muscle[J].FEBS Lett.1992.306(2-3):203-205.
[200]徐新女,基因疫苗的研究概况,中华综合临床医学杂志,2004,7(6):152-153
[201]张惠星,张维,罗玉芳,等.鸭外周血淋巴细胞转化试验的研究[J].上海免疫学杂志,1990,10(5):262-264.
[202]Cauchy L,Malkinson M.The mitogenic response to concanavalin A of Muscovy duck peripheral blood lymphoeytes in the early posthatching period[J].Avian Pathol,1981,10(1):77-81.