鸡白细胞介素15基因的克隆及在大肠杆菌和昆虫细胞中的表达
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摘要
白细胞介素15(IL-15)是1994年才发现的一种能促进细胞生长和分化的细胞因子，主要由天然免疫细胞产生。它与白细胞介素2(IL-2)有相似的生物学活性，但其结构和组成与IL-2毫无相似之处。已有的研究表明，鸡细胞因子在功能活性方面与人类和哺乳动物的细胞因子相似，尤其是IL-2与干扰素γ(IFN-γ)均具有较强的免疫增强作用。鸡IL-15(ChIL-15)是一种新发现的且与IL-2活性相似的细胞因子，作为疫苗佐剂研究的重要候选细胞因子有巨大潜力。
参考已发表的ChIL-15基因序列，设计合成引物，应用RT-PCR技术从刀豆球蛋白A(ConA)活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA，将其与SmalⅠ酶切处理的pUC19载体连接，构建重组质粒pUC19ChIL-15；用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因，与pMD 18-T载体相连构建重组质粒pMDChIL-15；然后用KpnⅠ／PstⅠ双酶切下mChIL-15基因片段，定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~(TM)HTa中，构建重组质粒pP_RoChIL-15，对其测序确认读框正确后，转入大肠杆菌DH5α中诱导表达并进行纯化，用重组ChIL-15(rChIL-15)免疫豚鼠，制备多克隆抗体；再用HindⅢ／PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因，定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中，经酶切、PCR和序列测定鉴定后，与杆状病毒线形化DNA(Bac-N-Blue~(TM) DNA)共转染Sf9昆虫细胞，经三轮蚀斑纯化，构建重组杆状病毒rBac-ChIL-15，用该重组病毒感染处于对数生长期的Sf9细胞，按不同的时间收取样品，离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。
结果显示，本研究所用白来航鸡IL-15 cDNA 5’非编码区有两个框外ATG起始密码子，开放阅读框由564bp组成，编码187个氨基酸，其中N末端信号肽含有66个氨基酸残基，在第48、149和166位的天冬酰胺残基上有三个潜在的N-糖基化位点。mChIL-15含有4个高度保守的半胱氨酸残基，可形成两个链内二硫键。与已发表的ChIL-15基因序列相比，该序列编码区第365、397、407、457、474、556、557位的核苷酸发生了变化，分别由A、C、G、G、A、G、C依次变成了G、T、T、T、G、A、T，推导的氨基酸序列在第122、133、136、153和186位发生了改变，分别由原来的天冬氨酸、精氨酸、色氨酸、丙氨酸、丙氨酸依次变成了甘氨酸、色氨酸、亮氨酸、丝氨酸和异亮氨酸，其中第474位碱基的变化是同义突变，这表明ChIL-15可能存在多态性；rChIL-15融合蛋白在大肠杆菌中得到了成功表达，其分子量约为18 kDa，扫描分析含量在30％左右，用离子亲和树脂对rChIL-15进行了有效的纯化，并成功制备出了豚鼠抗rChIL-15多克隆抗体；重组杆状病毒rBac-ChIL-15感染Sf9细胞后24小时已表达rChIL-15蛋白，96小时表达量达到高峰，其分子量约为22 kDa，占细胞总蛋白的25％，在Western blot实验中，该重组蛋白可被豚鼠抗原核rChIL-15多克隆抗体识别。
Interleukin-15 (IL-15) is a multifunctional cytokine that plays a major role in promoting cell growth and differentiation. IL-15 is produced mainly by natural immunocyte. In mammals, IL-15 shares many in vitro functions with IL-2. however, both cytokines bind to receptors with uniqe a chians enabling IL-15 and IL-2 to mediate very different functions in vivo. Recent results have shown that chicken cytokines share similar biological activity with mammalian cytokines, especially, in immunological enhancement of IL-2 and IFN- . Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.
According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal ant
ibody. The mChIL-15 gene from recombinant plasmid pPRoChIL-15 was subcloned into transfer vector pMelBacB and a recombinant vector pMelChIL-15 was constructed. Co-transfection of Sf9 insect cells with pMelChIL-15 and baculovirus linear DNA was performed. After three times purification, a recombinant baculovirus designated as rBac-ChIL-15 was obtained. The infected cells were collected at different time post-infection and analyzed by SDS-PAGE and Western blot.
The results showed that the open reading frame of ChIL-15 cDNA encompassed 564 base pairs(bp) and encoded a protein of 187 amino acids with three potential N-linked glycosylation sites, four conserved cysteine residues, two out-of-frame ATG initiation codons in the 5' untranslated region, and a signal peptide consisting of 66 amino acids. When it was compared with the published sequence of ChIL-15 cDNA, 7 mutant sites were found, and 5 amino acids were changed in predicted amino acids, which indicated that ChIL-15 may be polymorphic. The rChIL-15 was expressed in E.coli and was approximately 18kDa in size, which accounted for 30% of total protein. The purified rChIL-15 and rChIL-15 polyclonal antibody was obtained. Expression of rChIL-15 in rBac-ChIL-15 infected Sf9 cells was detected by SDS-PAGE and Western blot, the expressed product had a molecular weight of approximately 22kDa and the expression level of the recombinant protein was up to 25% of total cell protein .
参考已发表的ChIL-15基因序列，设计合成引物，应用RT-PCR技术从刀豆球蛋白A(ConA)活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA，将其与SmalⅠ酶切处理的pUC19载体连接，构建重组质粒pUC19ChIL-15；用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因，与pMD 18-T载体相连构建重组质粒pMDChIL-15；然后用KpnⅠ／PstⅠ双酶切下mChIL-15基因片段，定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~(TM)HTa中，构建重组质粒pP_RoChIL-15，对其测序确认读框正确后，转入大肠杆菌DH5α中诱导表达并进行纯化，用重组ChIL-15(rChIL-15)免疫豚鼠，制备多克隆抗体；再用HindⅢ／PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因，定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中，经酶切、PCR和序列测定鉴定后，与杆状病毒线形化DNA(Bac-N-Blue~(TM) DNA)共转染Sf9昆虫细胞，经三轮蚀斑纯化，构建重组杆状病毒rBac-ChIL-15，用该重组病毒感染处于对数生长期的Sf9细胞，按不同的时间收取样品，离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。
结果显示，本研究所用白来航鸡IL-15 cDNA 5’非编码区有两个框外ATG起始密码子，开放阅读框由564bp组成，编码187个氨基酸，其中N末端信号肽含有66个氨基酸残基，在第48、149和166位的天冬酰胺残基上有三个潜在的N-糖基化位点。mChIL-15含有4个高度保守的半胱氨酸残基，可形成两个链内二硫键。与已发表的ChIL-15基因序列相比，该序列编码区第365、397、407、457、474、556、557位的核苷酸发生了变化，分别由A、C、G、G、A、G、C依次变成了G、T、T、T、G、A、T，推导的氨基酸序列在第122、133、136、153和186位发生了改变，分别由原来的天冬氨酸、精氨酸、色氨酸、丙氨酸、丙氨酸依次变成了甘氨酸、色氨酸、亮氨酸、丝氨酸和异亮氨酸，其中第474位碱基的变化是同义突变，这表明ChIL-15可能存在多态性；rChIL-15融合蛋白在大肠杆菌中得到了成功表达，其分子量约为18 kDa，扫描分析含量在30％左右，用离子亲和树脂对rChIL-15进行了有效的纯化，并成功制备出了豚鼠抗rChIL-15多克隆抗体；重组杆状病毒rBac-ChIL-15感染Sf9细胞后24小时已表达rChIL-15蛋白，96小时表达量达到高峰，其分子量约为22 kDa，占细胞总蛋白的25％，在Western blot实验中，该重组蛋白可被豚鼠抗原核rChIL-15多克隆抗体识别。
Interleukin-15 (IL-15) is a multifunctional cytokine that plays a major role in promoting cell growth and differentiation. IL-15 is produced mainly by natural immunocyte. In mammals, IL-15 shares many in vitro functions with IL-2. however, both cytokines bind to receptors with uniqe a chians enabling IL-15 and IL-2 to mediate very different functions in vivo. Recent results have shown that chicken cytokines share similar biological activity with mammalian cytokines, especially, in immunological enhancement of IL-2 and IFN- . Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.
According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal ant
ibody. The mChIL-15 gene from recombinant plasmid pPRoChIL-15 was subcloned into transfer vector pMelBacB and a recombinant vector pMelChIL-15 was constructed. Co-transfection of Sf9 insect cells with pMelChIL-15 and baculovirus linear DNA was performed. After three times purification, a recombinant baculovirus designated as rBac-ChIL-15 was obtained. The infected cells were collected at different time post-infection and analyzed by SDS-PAGE and Western blot.
The results showed that the open reading frame of ChIL-15 cDNA encompassed 564 base pairs(bp) and encoded a protein of 187 amino acids with three potential N-linked glycosylation sites, four conserved cysteine residues, two out-of-frame ATG initiation codons in the 5' untranslated region, and a signal peptide consisting of 66 amino acids. When it was compared with the published sequence of ChIL-15 cDNA, 7 mutant sites were found, and 5 amino acids were changed in predicted amino acids, which indicated that ChIL-15 may be polymorphic. The rChIL-15 was expressed in E.coli and was approximately 18kDa in size, which accounted for 30% of total protein. The purified rChIL-15 and rChIL-15 polyclonal antibody was obtained. Expression of rChIL-15 in rBac-ChIL-15 infected Sf9 cells was detected by SDS-PAGE and Western blot, the expressed product had a molecular weight of approximately 22kDa and the expression level of the recombinant protein was up to 25% of total cell protein .
引文
[1] Isaacs A,Lindanmann J.Virus interference.I: the interferon.Proc R Soc Ser B, 1957, 147:258-267.
[2] 王世若，王兴龙，韩文瑜．现代动物免疫学．吉林：吉林科学技术出版社，2001，73-96．
[3] 金伯泉．细胞与分子免疫学．北京：世界图书出版公司，1995，95-175．
[4] 马大龙．新一轮的白细胞介素克隆化高潮——兼论计算机克隆的重要贡献．中国免疫学杂志，2001．17(3)：113-114．
[5] 刘胜旺．鸡细胞因子研究进展．中国预防兽医学报，2000，22(6)：474-476．
[6] Byrnes S, Eaton R, Kogut M.In vitro interleukin-1 and tumor necrosis factor-alpha production by macrophages from chickens infected with either Eimeria maxima or Eimeria tenella. Int J Parasitol, 1993,23(5):639-645.
[7] Cieszynski JA, Qureshi MA, Taylor RL.Calcium dependency of interleukin-1 secretion by a chicken macrophage cell line. Poult Sci, 1999,78(1):70-74.
[8] Dil N, Qureshi MA.Interleukin-1beta does not contribute to genetic strain-based differences in iNOS expression and activity in chicken macrophages. Dev Comp Immunol, 2003,27(2): 137-146.
[9] Weining K C,Sick C,Kaspers B,et al.A chickenhomolog of mammalian interleukin-1 beta:cDNA cloning and purification of active recombinant protein.Eur J Biochem, 1998,258: 994-1000.
[10] Guide S,Heguy A,Melli M.Gene, 1993,111:239-243.
[11] Heguy A,Baldari CT, Macchia C, et al.J Biol Chem,1992,267:2605-2609.
[12] Taniguchi T, matsui H,fujita T, et al.Structure and expression of a cloned cDNA for human interleukin-2. nature, 1983,302:305-310.
[13] Kaplan MH,Smith DI,Sundick RS.Identification of a G protein coupled receptor induced in activated T cells.J Immunol,1993,151:628-636.
[14] Myers TJ,Lillehoj HS,Fetterer RH.Partial purification and charaterization of chicken interleukin-2. Vet Immunol Immunopathol, 1992,34:97-114.
[15] Sundick RS, Gill-Dixon C.A cloned chicken lymphokine homologous to both mammalian IL-2 and IL-15. J Immunol, 1997.158:720-725.
[16] Kaiser P, Mariani EPromoter sequenee,exon:intron structure,and synteny of genetic location show that a chicken cytokine with T-cell proliferative activity is IL-2 and not IL-15. Immunogenetics, 1999,49:26-35.
[17] Stepaniak JA,Shuster JE, Wanping Hu,et al.Production and in vitro characterization of recombinant chicken interleukin-2.J Interferon Cytokine Res, 1999,19:515-526.
[18] Hala K, Schauenstein K,Neu N,et al.A monoclonal antibody reacting with a membrane determinant expressed on activated chicken T lymphocytes.Eur J Immunol,1986,16:1331-1336.
[19] Lowenthal JW, Oneil TE,David A,et al.Cytokine therapy: a natural alternative for disease control. Vet Immunol Immunolpathol, 1999,72:183-188.
[20] Lynagh GR, Bailey M, Kaiser P.Interleukin-6 is produced during both murine and avian Eimeria infections. Vet Immunol Immunopathol,2000,76(1-2):89-102.
[21] Kaiser P, Underwood G, Davison F.Differential cytokine responses following Marek's disease virusinfection of chickens differing in resistance to Marek's disease. J Virol,2003,77(1): 762-768.
[22] Rath NC, Huff WE, Bayyari GR, et al. Identification of transforming growth factor-beta and interleukin-6 in chicken ascites fluid. Avian Dis, 1995,39(2):382-389.
[23] Schneider K, Klass R,Kaspers B,et al.chicken interleukin-6 cDNA structure and biological properties.Eur J Immunol,2001,268:4200-4206.
[24] Samad F, Bergtrom G, Eissa H, et al.Stimulation of chick hepatocyte fibronectin production by fibroblast-conditioned medium is due to interleukin 6. Biochim Biophys Acta, 1993, 1181(3):207-213.
[25] Barker KA, Hampe A, Stoeckle MY, et al.Transformation-associated cytokine 9E3/CEF4 ischemotactic
for chicken peripheral blood mononuclear cells. J Virol, 1993,67(6):3528-3533.
[26] Kaiser P, Hughes S,Bumstead N.The chicken 9E3/CEF4 CXC chemokine is the avian orthologue of IL-8 and maps to chick chromosome 4 syntenic with genes flanking the mammalian chemokine cluster. Immunogenetics, 1999,49:673-684.
[27] Sick C, Schneider K, Staeheli P, et al.Novel chicken CXC and CC chemokines. Cytokine, 2000,12(3): 181-186.
[28] Parcells MS, Lin SF, Dienglewicz RL, et al.Marek's disease virus (MDV) encodes an interleukin-8 homolog (vIL-8): characterization of the vIL-8 protein and a vIL-8 deletion mutant MDV. J Virol, 2001, 75(11):5159-5173.
[29] Okamura H, Tsutsui H, Komatsu T, et al. Cloning of a new cytokine that induces IFN-γ production by T cells. Nature,1995,378:88-91.
[30] Ushio S, Namba M, Okura T, et al. Cloning of the cDNA for human IFN-gamma-inducing factor, expression in Escherichia coli, and studies on the biologic activities of the protein.J Immunol,1996, 156:4274-4279.
[31] Schneider K, Puehler F, Baeubrle D, et al. cDNA cloning of biologically active chicken interleukin-18. J Interferon Cytokine Res,2000, 20: 879-883.
[32] Pete Kaiser. Turkey and chicken interleukin-18 (IL-18) share high sequence identity, but have different polyadenylation sites in their 3'UTR.Dev Comp Immunol, 2002,26:681-687.
[33] Gu Y, Kuida K, Tsutsui H, et al. Activation of interferon-gamma-inducing factor mediated by interleukin-1 converting enzyme. Science, 1997, 275:206-209.
[34] Bazan JF, Timans JC, Kastelein RA. A newly defined interleukin-1?. Nature, 1996,379:591.
[35] 林学颜，张玲．现代细胞与分子免疫学．北京：科学出版社，1999，233-261．
[36] Sekellick MJ,Ferrandino AF, Hopkins DA,et al.Chicken interferon gene:cloning.expression,and analysis.J interferon Reseach, 1994,14:71-79.
[37] Sick C,Schultz U,Staeheli P.A family of genes coding for two serologically distince chicken interferon.J Biol Chem, 1996,271(13):7636-7639.
[38] Schultz U, Chisari F V. Recombinant duck interferon:A new reagent for studying the mode of interferon action against hepatitis B virus.Virology, 1995,212:641-649.
[39] Suresh M,Karaca K,Fostor D.molecular and functional characterization of turkey interferon.J Virol, 1995,69(12):8159-8163.
[40] Schultz U,Ridderle C,Sekellick MJ.Recombinant duck interferon from E.coli and transfected COS cells is biologically active.Eur J Biochemistry, 1995,229:73-76.
[41] Digby MR, Lowenthal JW.Cloning and expression of the chicken interferon-γ gene. J Interferon Cytokine Res, 1995,15:939-949.
[42] Kaiser P, wain HM,Rothwell L.Structure of the chicken interferon-γ gene and comparison to mammalian homologues.Gene, 1998,207:23-25.
[43] Schultz U,Chisari FV.Recombinant duck interferon gamma inhibits duck hepatitis B virus replication in primary hepatocytes.J Virol, 1999,73(4):3162-3168.
[44] Kaiser P, Sonnemans D,Smith LM.Avian IFN-γ sequence analysis suggests probable cross-species reactivity among galliforms.J Interferon and Cytokine, 1998,18:711-719.
[45] Leutz A,Damm K, Sterneck E,et al.Molecular cloning of the chicken myelomonocytic growth factor(cMGF) reveals relationship to interleukin-6 and granulocyte colony stimulating factor.EMBO J,1989,8:175-181.
[46] Van't Hof RJ, von Lindern M, Nijweide PJ, et al. Stem cell factor stimulates chicken osteoclast activity in vitro. FASEB J, 1997,11(4):287-293.
[47] Woldman I, Mellitzer G, Kieslinger M,et al.STAT5 involvement in the differentiation response of primary chicken myeloid progenitor cells to chicken myelomonocytic growth factor. Immunol, 1997, 159(2):877-886.
[48] Sterneck E, Blattner C, Graf T, et al.Structure of the chicken myelomonocytic growth factor gene and specific activation of its promoter in avian myelomonocytic cells by protein kinases. Mol Cell
Biol, 1992,12(4):1728-1735.
[49] Djeraba A, Musset E, Lowenthal JW, et al.Protective effect of avian myelomonocytic growth factor in infection with Marek's disease virus. J Virol, 2002,76(3): 1062-1070.
[50] Burt DW, Paton IR.Evolutionary origins of the transforming growth factor-beta gene family. DNA Cell Biol, 1992,11(7):497-510.
[51] Jakowlew SB, Dillard PJ, Sporn MB, et al.Complementary deoxyribonucleic acid cloning of an mRNA encoding transforming growth factor-beta 2 from chicken embryo chondrocytes. Growth Factors, 1990,2(2-3):123-133.
[52] Burt DW, Paton IR.Molecular cloning and primary structure of the chicken transforming growth factor-beta2 gene. DNA Cell Biol, 1991, 10(10):723-734.
[53] Burt DW, Dey BR, Paton IR, et al.The chicken transforming growth factor-beta 3 gene: genomic structure, transcriptional analysis, and chromosomal location. DNA Cell Biol, 1995, 14(2): 111-123.
[54] Byrnes S, Eaton R, Kogut M.In vitro interleukin-1 and tumor necrosis factor-alpha production by macrophages from chickens infected with either Eimeria maxima or Eimeria tenella. Int J Parasitol, 1993, 23(5):639-645.
[55] Wride MA, Sanders EJ.Expression of tumor necrosis factor-alpha (TNF alpha)-cross-reactive proteins during early chick embryo development. Dev Dyn, 1993,198(3):225-239.
[56] Zhang S, Lillehoj HS, Ruff MD.In vivo role of tumor necrosis-like factor in Eimeria tenella infection. Avian Dis, 1995, 39(4):859-866.
[57] Rautenschlein S, Subramanian A, Sharma JM.Bioactivities of a tumour necrosis-like factor released by chicken macrophages. Dev Comp Immunol, 1999, 23(7-8):629-640.
[58] Zhou JH,Ohtaki M,Sakurai M.Sequence of a cDNA encoding chicken stem cell factor.gene,1993, 127:269-270.
[59] Zhou JH, Ohtaki M, Inumaru S, et al.Purification and activity of recombinant chicken stem cell factor produced by using a baculovirus vector. J Vet Med Sci, 1996,58(2):173-175.
[60] Bartunek P, Pichlikova L, Stengl G, et al.Avian stem cell factor (SCF): production and characterization of the recombinant His-tagged SCF of chicken and its neutralizing antibody. Cytokine, 1996, J8(1):14-20.
[61] Sasaki E, Etches RJ.Expression of protein tyrosine kinases and stem cell factor in chicken blastodermal cells. Poult Sci, 2001, 80(2): 161-171.
[62] petitte JN,Kulik MJ.cloning and characterization of cDNAs encoding two forms of avian stem cell factor.Biochem Biophys Acta, 1996,1307:149-151.
[63] D'Costa S, Kulik MJ, Petitte JN.Expression and purification of biologically active recombinant quail stem cell factor in E. coli. Cell Biol Int, 2000,24(5):311-317.
[64] Grabstein KH,Eisenman J,Shanebeck K,et al.Cloning of a T cell growth factor that interacts with the β chain of the interleukin-2 receptor.Science, 1994, 264: 965-968.
[65] Burton JD,Bamford RN,Peter C,et al.A lymphokine,provisionally designated IL-T leukemia line,stimulates T-cell proliferation and induction of lymphokine-activated killer cells. Proc Natl Acad Sci USA,1994,91:4935-4939.
[66] Bamford RN,Grant AJ,Burton JD,et al.The interleukin(IL)2 receptor beta chain is shared by IL-2 and a cytokine provisionally designated IL-T, that stimulates T-cell proliferation and the induction of lymphokine-activated killer cells.Proc Natl Acad Sci USA, 1994,91:4940-4944.
[67] Krause H, Jandrig B, Wernicke C,et al.Genomic structure and chromosomal localization of the human interleukin 15 gene (IL-15). Cytokine,1996, 8(9): 667-674.
[68] Choi KD,Lillehoj HS,Song KD,et al.Molecular and functional characterization of chicken IL-15. Dep Comp Immunol, 1999,23(2): 165-177.
[69] Lillehoj HS,Min W, Choi KD,et al.Molecular, cellular, and functional characterization of chicken cytokines homologous to mammalian IL-15 and IL-2. Vet Immunol Immunopathol,2001, 82(1-2): 229-244.
[70] Meazza RS,Verdiani R,biassoni M,et al.Identification of a novel interleukin-15 (IL-15) transcript
isoform generated by alternative splicing in human cell lung cancer cell lines.Oncogene, 1996,12:2187.
[71] Onu A,Pohl T, Krause H,et al.Regulation of the IL-15 secretion Via the leader peptide of two IL-15 isoform.J Immunol,1997,158:255-262.
[72] Nishimura H,Washizu J,Nakamura N,et al.Translational efficiency is up-regulated by alternative exon in murine IL-15 mRNA.J Immunol, 1998,160:936-942.
[73] Kimura K,Nishimura H,Hirose K,et al.Immunogene therapy of murine fibrosarcoma using Il-15 gene with high translation efficiency.Eur.J Immunol, 1999,29:1532-1542.
[74] Giri JG, Kumaki S,Ahdieh M,et al.Utilization of the β and γ chains of the IL-2 receptor by the novel cytokine IL-15.EMBO J, 1994,13:2822-2830.
[75] Bamford RN,Battiata JD,Burton H,et al.Intrleukin(IL)15/IL-T production by the adult T-cell leukemia cell line HuT-102 is associated with a human T-cell lymphotrophic virus type I R region/IL-15 fusion message that lacks many upstream AUGs that normally attenuate IL-15 mRNA translation. Proc Natl Acad Sci USA,1996,93:2897.
[76] Bamford RN,Defilippis AP, Azimin N,et al.The 5' untranslated region,signal peptide,and the coding sequence of the carboxyl terminus of IL-15 participate in its multifaceted translational control.J Immunol, 1998,160.4418-4426.
[77] Meazza R,Gaggero,Neglia F, et al.Expression of two interleukin-15 mRNA isoforms in human tumors does not correlate with secretion: role of different signal peptides. Eur J Immunol, 1997,27:1049-1050.
[78] Jong JL,Farner NL,Widmer MB,et al.Interaction of IL-15 with the shared IL-2 receptor β and γ subunit.J Immunol,1996,156:1339.
[79] Wei XQ,Orchardson M,Gracie JA,et al.The sushi domain of soluble IL-15 receptor α is essential for binding IL-15 and inhibiting inflammatory and allogenic rseponses in Vitro and Vivo. J Immunol, 2001,167: 277-282.
[80] Li G X,Lillehoj HS,Min W.Production and characterization of monoclonal antibodies reactive with the chicken interleukin-15 receptor alpha chian.Vet Immunol Immunopathol, 2001,82(1-2):215-227.
[81] Tirunagaru VG, Sofer L,Cui J,et al.The multifaceted regulation of interleukin-15 expression and the role of this cytokine in NK cell differentiation and host response to intracellular pathogens.Annu Rev Immunol, 1999,17:19-49.
[82] 苏琴．白细胞介素-15研究进展．国外医学分子生物学分册，2000，22(2)：101-104．
[83] Waldmann TA,Dubois S,Tagaya Y, et al.Contrasting roles of IL-2 and IL-15 in the life and death of lymphocytes: implications for immunotherapy.Immunity,2001,14(2): 105-110.
[84] Chu C L,Chen S S,Wu T S,et al.Differential Effects of IL-2 and IL-15 on the Death and survival of Activated TCRr δ + intestinal intraepithelial lymphocyte. J Immunol, 1999,1896-1903.
[85] Leclercq G, Debacker V, Smedt MD,et al.Differential effects of interleukin-15 and interleukin-2 on differentiation of bipotential T/natural killer progenitor cells.J Exp Med, 1996,184:325.
[86] Schluns KS,Williams K,Ma A,et al.Requirement for IL-15 in the generation of primary antigen-specific CD8 T cells.J Immunol,2002,168:4827-4831.
[87] Armitage R T, Macduff B M,Eisenman J,et al.IL-15 has stimulatory activity for induction of B cell proliferation and differentiation.J Immunol,1995,154:483.
[88] Liu C C,Perussia B,Young JD.The emerging role of IL-15 in NK-cell development.Immunolgy today,2000,21(3): 113-115.
[89] Stepaniak JA.Shuster JE,Hu WP, et al.Production and in vitro characterization of recombinant chicken interleukin-2. J Interferon Cytokine Res, 1999,19:515-526.
[90] Min W, Lillehoj HS,Burnside J, et al.Adjuvant effects of IL-1β,IL-2,IL-8,IL-15,IFN-α, IFN-γ,TGF-β4 and lymphotactin on DNA vaccination against Eimeria acervulina, vaccine, 2002,20:267-274.
[91] Seder RA.High-dose IL-2 and IL-15 enhance the in vitro priming of nave CD4+ T cells for IFN-γbut have differential effects on priming for IL-4.J Immunol. 1996,156:2413.
[92] Chehimi J,Marshall JD,salvucci O,et al.IL-15 enhances immune functions during HIV infection.J Immunol, 1997,5979-5987.
[93] Meazza R,Basso S,Gaggero A,et al.Interleukin-15 induces survival and proliferation of the
growth-dependent acute myeloid leukemia M-07e through the IL-2 receptor beta/gamma.Int J cancer, 1998,78(2):189-195.
[94] Ajjan RA,Watson PF, Weetman AP, et al.Detection of IL-12,IL-13,and IL-15 Messager ribonucleic acid in the thyroid of patients with autoimmune thyroid disease.J Clin Endocrinol Metab,1997,82:666-669.
[95] Kaiser P, Rothwell L,Dusan V, et al.A role for IL-15 in driving the onset of spontaneous autoimmune thyroiditis.J Immunol,2002,168:4216-4220.
[96] Min W, Lillehoj HS.Development and characterization of monoclonal antibodies to chicken interleukin-15. Vet Immunol Immunopathol,2002,88:49-56.
[97] Williams PE. The European ban of the prophylactic use of antibiotics as growth promoters in animal nutrition:political and economic aspects.In;Proceedings of the Australian Poultry Science Symposium, 2001,83-93.
[98] World Health Organisation.report of a Meeting to Review the Use of Fluoroquinolones in Animal Agriculture and its Implications for Human Health.WHO,Geneva.Switzerland.
[99] Schijns VE,Weining KC,Nuijten P.Immunoadjuvant activities of E.coli and plasmid-expressed recombinant chicken IFN-α/β,IFN-γand 1L-1β in 1 day and 3-week-old chickens. Vaccine,2000,18: 2147-2154.
[100] Lowenthal JW, Oneil TE,Broadway M,et al.Coadministration of chicken interferon-gamma enhances antibody responses in chickens.J interferon Cytokine Res, 1998,18:617-622.
[101] Lowenthal JW, Lambrecht B,Van denBerg TP, et al.Avian cytokine-the natural approach to therapeutics.Dev Comp Immunol,2000,24:355-365.
[102] Lillehoj HS,Choi KD.Recombinant chicken interferon-gamma-mediated inhibition of Eimeria tenella development in vitro and reduction of oocyst production and body weight loss following Eimeria acervulina challenge infection. Avian Dis, 1998,42:307-314.
[103] Marcus PI,Van der HL,Sekellick MJ.interferon action on avian virus.I.Oral administration of chicken interferon-alpha ameliorates Newcastle disease.J Interferon Cytokine Res, 1999, 19:881-885.
[104] Karaca K,Sharma JM,Winstow BJ,et al.Recombinant fowlpox viruses coexpressing chicken type Ⅰ IFN and Newcastle disease virus HN and F genes:influence of IFN on protective efficacy and humoral responses of chickens following in ovo or post-hatch administration of recombinant viruses.Vaccine, 1998,16:1496-1503.
[105] 姜永厚，刘忠贵，陈奖励，等．鸡IL-2在新城疫病毒F基因疫苗中的作用．中国兽医学报，2002，22(1)：22-24．
[106] 于链，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疫苗免疫原性的研究．生物工程学报，2001，17(6)：652-657．
[107] 萨姆布鲁克 J，弗里奇 E F，曼尼阿蒂斯 T，等．分子克隆实验指南，第二版．金冬雁，黎孟枫，侯云德译．北京：科学出版社，1992，19-22．
[108] 李迎秋，王循章，龙絮新．乙型肝炎病毒preC／C基因在杆状病毒载体系统中的表达．病毒学报，1997，13(4)：325-331．
[109] 尹长城，刘晶，黄华梁．昆虫表达系统及其在基因工程抗体中的应用．生物工程进展，2001，21(3)：66-70．
[2] 王世若，王兴龙，韩文瑜．现代动物免疫学．吉林：吉林科学技术出版社，2001，73-96．
[3] 金伯泉．细胞与分子免疫学．北京：世界图书出版公司，1995，95-175．
[4] 马大龙．新一轮的白细胞介素克隆化高潮——兼论计算机克隆的重要贡献．中国免疫学杂志，2001．17(3)：113-114．
[5] 刘胜旺．鸡细胞因子研究进展．中国预防兽医学报，2000，22(6)：474-476．
[6] Byrnes S, Eaton R, Kogut M.In vitro interleukin-1 and tumor necrosis factor-alpha production by macrophages from chickens infected with either Eimeria maxima or Eimeria tenella. Int J Parasitol, 1993,23(5):639-645.
[7] Cieszynski JA, Qureshi MA, Taylor RL.Calcium dependency of interleukin-1 secretion by a chicken macrophage cell line. Poult Sci, 1999,78(1):70-74.
[8] Dil N, Qureshi MA.Interleukin-1beta does not contribute to genetic strain-based differences in iNOS expression and activity in chicken macrophages. Dev Comp Immunol, 2003,27(2): 137-146.
[9] Weining K C,Sick C,Kaspers B,et al.A chickenhomolog of mammalian interleukin-1 beta:cDNA cloning and purification of active recombinant protein.Eur J Biochem, 1998,258: 994-1000.
[10] Guide S,Heguy A,Melli M.Gene, 1993,111:239-243.
[11] Heguy A,Baldari CT, Macchia C, et al.J Biol Chem,1992,267:2605-2609.
[12] Taniguchi T, matsui H,fujita T, et al.Structure and expression of a cloned cDNA for human interleukin-2. nature, 1983,302:305-310.
[13] Kaplan MH,Smith DI,Sundick RS.Identification of a G protein coupled receptor induced in activated T cells.J Immunol,1993,151:628-636.
[14] Myers TJ,Lillehoj HS,Fetterer RH.Partial purification and charaterization of chicken interleukin-2. Vet Immunol Immunopathol, 1992,34:97-114.
[15] Sundick RS, Gill-Dixon C.A cloned chicken lymphokine homologous to both mammalian IL-2 and IL-15. J Immunol, 1997.158:720-725.
[16] Kaiser P, Mariani EPromoter sequenee,exon:intron structure,and synteny of genetic location show that a chicken cytokine with T-cell proliferative activity is IL-2 and not IL-15. Immunogenetics, 1999,49:26-35.
[17] Stepaniak JA,Shuster JE, Wanping Hu,et al.Production and in vitro characterization of recombinant chicken interleukin-2.J Interferon Cytokine Res, 1999,19:515-526.
[18] Hala K, Schauenstein K,Neu N,et al.A monoclonal antibody reacting with a membrane determinant expressed on activated chicken T lymphocytes.Eur J Immunol,1986,16:1331-1336.
[19] Lowenthal JW, Oneil TE,David A,et al.Cytokine therapy: a natural alternative for disease control. Vet Immunol Immunolpathol, 1999,72:183-188.
[20] Lynagh GR, Bailey M, Kaiser P.Interleukin-6 is produced during both murine and avian Eimeria infections. Vet Immunol Immunopathol,2000,76(1-2):89-102.
[21] Kaiser P, Underwood G, Davison F.Differential cytokine responses following Marek's disease virusinfection of chickens differing in resistance to Marek's disease. J Virol,2003,77(1): 762-768.
[22] Rath NC, Huff WE, Bayyari GR, et al. Identification of transforming growth factor-beta and interleukin-6 in chicken ascites fluid. Avian Dis, 1995,39(2):382-389.
[23] Schneider K, Klass R,Kaspers B,et al.chicken interleukin-6 cDNA structure and biological properties.Eur J Immunol,2001,268:4200-4206.
[24] Samad F, Bergtrom G, Eissa H, et al.Stimulation of chick hepatocyte fibronectin production by fibroblast-conditioned medium is due to interleukin 6. Biochim Biophys Acta, 1993, 1181(3):207-213.
[25] Barker KA, Hampe A, Stoeckle MY, et al.Transformation-associated cytokine 9E3/CEF4 ischemotactic
for chicken peripheral blood mononuclear cells. J Virol, 1993,67(6):3528-3533.
[26] Kaiser P, Hughes S,Bumstead N.The chicken 9E3/CEF4 CXC chemokine is the avian orthologue of IL-8 and maps to chick chromosome 4 syntenic with genes flanking the mammalian chemokine cluster. Immunogenetics, 1999,49:673-684.
[27] Sick C, Schneider K, Staeheli P, et al.Novel chicken CXC and CC chemokines. Cytokine, 2000,12(3): 181-186.
[28] Parcells MS, Lin SF, Dienglewicz RL, et al.Marek's disease virus (MDV) encodes an interleukin-8 homolog (vIL-8): characterization of the vIL-8 protein and a vIL-8 deletion mutant MDV. J Virol, 2001, 75(11):5159-5173.
[29] Okamura H, Tsutsui H, Komatsu T, et al. Cloning of a new cytokine that induces IFN-γ production by T cells. Nature,1995,378:88-91.
[30] Ushio S, Namba M, Okura T, et al. Cloning of the cDNA for human IFN-gamma-inducing factor, expression in Escherichia coli, and studies on the biologic activities of the protein.J Immunol,1996, 156:4274-4279.
[31] Schneider K, Puehler F, Baeubrle D, et al. cDNA cloning of biologically active chicken interleukin-18. J Interferon Cytokine Res,2000, 20: 879-883.
[32] Pete Kaiser. Turkey and chicken interleukin-18 (IL-18) share high sequence identity, but have different polyadenylation sites in their 3'UTR.Dev Comp Immunol, 2002,26:681-687.
[33] Gu Y, Kuida K, Tsutsui H, et al. Activation of interferon-gamma-inducing factor mediated by interleukin-1 converting enzyme. Science, 1997, 275:206-209.
[34] Bazan JF, Timans JC, Kastelein RA. A newly defined interleukin-1?. Nature, 1996,379:591.
[35] 林学颜，张玲．现代细胞与分子免疫学．北京：科学出版社，1999，233-261．
[36] Sekellick MJ,Ferrandino AF, Hopkins DA,et al.Chicken interferon gene:cloning.expression,and analysis.J interferon Reseach, 1994,14:71-79.
[37] Sick C,Schultz U,Staeheli P.A family of genes coding for two serologically distince chicken interferon.J Biol Chem, 1996,271(13):7636-7639.
[38] Schultz U, Chisari F V. Recombinant duck interferon:A new reagent for studying the mode of interferon action against hepatitis B virus.Virology, 1995,212:641-649.
[39] Suresh M,Karaca K,Fostor D.molecular and functional characterization of turkey interferon.J Virol, 1995,69(12):8159-8163.
[40] Schultz U,Ridderle C,Sekellick MJ.Recombinant duck interferon from E.coli and transfected COS cells is biologically active.Eur J Biochemistry, 1995,229:73-76.
[41] Digby MR, Lowenthal JW.Cloning and expression of the chicken interferon-γ gene. J Interferon Cytokine Res, 1995,15:939-949.
[42] Kaiser P, wain HM,Rothwell L.Structure of the chicken interferon-γ gene and comparison to mammalian homologues.Gene, 1998,207:23-25.
[43] Schultz U,Chisari FV.Recombinant duck interferon gamma inhibits duck hepatitis B virus replication in primary hepatocytes.J Virol, 1999,73(4):3162-3168.
[44] Kaiser P, Sonnemans D,Smith LM.Avian IFN-γ sequence analysis suggests probable cross-species reactivity among galliforms.J Interferon and Cytokine, 1998,18:711-719.
[45] Leutz A,Damm K, Sterneck E,et al.Molecular cloning of the chicken myelomonocytic growth factor(cMGF) reveals relationship to interleukin-6 and granulocyte colony stimulating factor.EMBO J,1989,8:175-181.
[46] Van't Hof RJ, von Lindern M, Nijweide PJ, et al. Stem cell factor stimulates chicken osteoclast activity in vitro. FASEB J, 1997,11(4):287-293.
[47] Woldman I, Mellitzer G, Kieslinger M,et al.STAT5 involvement in the differentiation response of primary chicken myeloid progenitor cells to chicken myelomonocytic growth factor. Immunol, 1997, 159(2):877-886.
[48] Sterneck E, Blattner C, Graf T, et al.Structure of the chicken myelomonocytic growth factor gene and specific activation of its promoter in avian myelomonocytic cells by protein kinases. Mol Cell
Biol, 1992,12(4):1728-1735.
[49] Djeraba A, Musset E, Lowenthal JW, et al.Protective effect of avian myelomonocytic growth factor in infection with Marek's disease virus. J Virol, 2002,76(3): 1062-1070.
[50] Burt DW, Paton IR.Evolutionary origins of the transforming growth factor-beta gene family. DNA Cell Biol, 1992,11(7):497-510.
[51] Jakowlew SB, Dillard PJ, Sporn MB, et al.Complementary deoxyribonucleic acid cloning of an mRNA encoding transforming growth factor-beta 2 from chicken embryo chondrocytes. Growth Factors, 1990,2(2-3):123-133.
[52] Burt DW, Paton IR.Molecular cloning and primary structure of the chicken transforming growth factor-beta2 gene. DNA Cell Biol, 1991, 10(10):723-734.
[53] Burt DW, Dey BR, Paton IR, et al.The chicken transforming growth factor-beta 3 gene: genomic structure, transcriptional analysis, and chromosomal location. DNA Cell Biol, 1995, 14(2): 111-123.
[54] Byrnes S, Eaton R, Kogut M.In vitro interleukin-1 and tumor necrosis factor-alpha production by macrophages from chickens infected with either Eimeria maxima or Eimeria tenella. Int J Parasitol, 1993, 23(5):639-645.
[55] Wride MA, Sanders EJ.Expression of tumor necrosis factor-alpha (TNF alpha)-cross-reactive proteins during early chick embryo development. Dev Dyn, 1993,198(3):225-239.
[56] Zhang S, Lillehoj HS, Ruff MD.In vivo role of tumor necrosis-like factor in Eimeria tenella infection. Avian Dis, 1995, 39(4):859-866.
[57] Rautenschlein S, Subramanian A, Sharma JM.Bioactivities of a tumour necrosis-like factor released by chicken macrophages. Dev Comp Immunol, 1999, 23(7-8):629-640.
[58] Zhou JH,Ohtaki M,Sakurai M.Sequence of a cDNA encoding chicken stem cell factor.gene,1993, 127:269-270.
[59] Zhou JH, Ohtaki M, Inumaru S, et al.Purification and activity of recombinant chicken stem cell factor produced by using a baculovirus vector. J Vet Med Sci, 1996,58(2):173-175.
[60] Bartunek P, Pichlikova L, Stengl G, et al.Avian stem cell factor (SCF): production and characterization of the recombinant His-tagged SCF of chicken and its neutralizing antibody. Cytokine, 1996, J8(1):14-20.
[61] Sasaki E, Etches RJ.Expression of protein tyrosine kinases and stem cell factor in chicken blastodermal cells. Poult Sci, 2001, 80(2): 161-171.
[62] petitte JN,Kulik MJ.cloning and characterization of cDNAs encoding two forms of avian stem cell factor.Biochem Biophys Acta, 1996,1307:149-151.
[63] D'Costa S, Kulik MJ, Petitte JN.Expression and purification of biologically active recombinant quail stem cell factor in E. coli. Cell Biol Int, 2000,24(5):311-317.
[64] Grabstein KH,Eisenman J,Shanebeck K,et al.Cloning of a T cell growth factor that interacts with the β chain of the interleukin-2 receptor.Science, 1994, 264: 965-968.
[65] Burton JD,Bamford RN,Peter C,et al.A lymphokine,provisionally designated IL-T leukemia line,stimulates T-cell proliferation and induction of lymphokine-activated killer cells. Proc Natl Acad Sci USA,1994,91:4935-4939.
[66] Bamford RN,Grant AJ,Burton JD,et al.The interleukin(IL)2 receptor beta chain is shared by IL-2 and a cytokine provisionally designated IL-T, that stimulates T-cell proliferation and the induction of lymphokine-activated killer cells.Proc Natl Acad Sci USA, 1994,91:4940-4944.
[67] Krause H, Jandrig B, Wernicke C,et al.Genomic structure and chromosomal localization of the human interleukin 15 gene (IL-15). Cytokine,1996, 8(9): 667-674.
[68] Choi KD,Lillehoj HS,Song KD,et al.Molecular and functional characterization of chicken IL-15. Dep Comp Immunol, 1999,23(2): 165-177.
[69] Lillehoj HS,Min W, Choi KD,et al.Molecular, cellular, and functional characterization of chicken cytokines homologous to mammalian IL-15 and IL-2. Vet Immunol Immunopathol,2001, 82(1-2): 229-244.
[70] Meazza RS,Verdiani R,biassoni M,et al.Identification of a novel interleukin-15 (IL-15) transcript
isoform generated by alternative splicing in human cell lung cancer cell lines.Oncogene, 1996,12:2187.
[71] Onu A,Pohl T, Krause H,et al.Regulation of the IL-15 secretion Via the leader peptide of two IL-15 isoform.J Immunol,1997,158:255-262.
[72] Nishimura H,Washizu J,Nakamura N,et al.Translational efficiency is up-regulated by alternative exon in murine IL-15 mRNA.J Immunol, 1998,160:936-942.
[73] Kimura K,Nishimura H,Hirose K,et al.Immunogene therapy of murine fibrosarcoma using Il-15 gene with high translation efficiency.Eur.J Immunol, 1999,29:1532-1542.
[74] Giri JG, Kumaki S,Ahdieh M,et al.Utilization of the β and γ chains of the IL-2 receptor by the novel cytokine IL-15.EMBO J, 1994,13:2822-2830.
[75] Bamford RN,Battiata JD,Burton H,et al.Intrleukin(IL)15/IL-T production by the adult T-cell leukemia cell line HuT-102 is associated with a human T-cell lymphotrophic virus type I R region/IL-15 fusion message that lacks many upstream AUGs that normally attenuate IL-15 mRNA translation. Proc Natl Acad Sci USA,1996,93:2897.
[76] Bamford RN,Defilippis AP, Azimin N,et al.The 5' untranslated region,signal peptide,and the coding sequence of the carboxyl terminus of IL-15 participate in its multifaceted translational control.J Immunol, 1998,160.4418-4426.
[77] Meazza R,Gaggero,Neglia F, et al.Expression of two interleukin-15 mRNA isoforms in human tumors does not correlate with secretion: role of different signal peptides. Eur J Immunol, 1997,27:1049-1050.
[78] Jong JL,Farner NL,Widmer MB,et al.Interaction of IL-15 with the shared IL-2 receptor β and γ subunit.J Immunol,1996,156:1339.
[79] Wei XQ,Orchardson M,Gracie JA,et al.The sushi domain of soluble IL-15 receptor α is essential for binding IL-15 and inhibiting inflammatory and allogenic rseponses in Vitro and Vivo. J Immunol, 2001,167: 277-282.
[80] Li G X,Lillehoj HS,Min W.Production and characterization of monoclonal antibodies reactive with the chicken interleukin-15 receptor alpha chian.Vet Immunol Immunopathol, 2001,82(1-2):215-227.
[81] Tirunagaru VG, Sofer L,Cui J,et al.The multifaceted regulation of interleukin-15 expression and the role of this cytokine in NK cell differentiation and host response to intracellular pathogens.Annu Rev Immunol, 1999,17:19-49.
[82] 苏琴．白细胞介素-15研究进展．国外医学分子生物学分册，2000，22(2)：101-104．
[83] Waldmann TA,Dubois S,Tagaya Y, et al.Contrasting roles of IL-2 and IL-15 in the life and death of lymphocytes: implications for immunotherapy.Immunity,2001,14(2): 105-110.
[84] Chu C L,Chen S S,Wu T S,et al.Differential Effects of IL-2 and IL-15 on the Death and survival of Activated TCRr δ + intestinal intraepithelial lymphocyte. J Immunol, 1999,1896-1903.
[85] Leclercq G, Debacker V, Smedt MD,et al.Differential effects of interleukin-15 and interleukin-2 on differentiation of bipotential T/natural killer progenitor cells.J Exp Med, 1996,184:325.
[86] Schluns KS,Williams K,Ma A,et al.Requirement for IL-15 in the generation of primary antigen-specific CD8 T cells.J Immunol,2002,168:4827-4831.
[87] Armitage R T, Macduff B M,Eisenman J,et al.IL-15 has stimulatory activity for induction of B cell proliferation and differentiation.J Immunol,1995,154:483.
[88] Liu C C,Perussia B,Young JD.The emerging role of IL-15 in NK-cell development.Immunolgy today,2000,21(3): 113-115.
[89] Stepaniak JA.Shuster JE,Hu WP, et al.Production and in vitro characterization of recombinant chicken interleukin-2. J Interferon Cytokine Res, 1999,19:515-526.
[90] Min W, Lillehoj HS,Burnside J, et al.Adjuvant effects of IL-1β,IL-2,IL-8,IL-15,IFN-α, IFN-γ,TGF-β4 and lymphotactin on DNA vaccination against Eimeria acervulina, vaccine, 2002,20:267-274.
[91] Seder RA.High-dose IL-2 and IL-15 enhance the in vitro priming of nave CD4+ T cells for IFN-γbut have differential effects on priming for IL-4.J Immunol. 1996,156:2413.
[92] Chehimi J,Marshall JD,salvucci O,et al.IL-15 enhances immune functions during HIV infection.J Immunol, 1997,5979-5987.
[93] Meazza R,Basso S,Gaggero A,et al.Interleukin-15 induces survival and proliferation of the
growth-dependent acute myeloid leukemia M-07e through the IL-2 receptor beta/gamma.Int J cancer, 1998,78(2):189-195.
[94] Ajjan RA,Watson PF, Weetman AP, et al.Detection of IL-12,IL-13,and IL-15 Messager ribonucleic acid in the thyroid of patients with autoimmune thyroid disease.J Clin Endocrinol Metab,1997,82:666-669.
[95] Kaiser P, Rothwell L,Dusan V, et al.A role for IL-15 in driving the onset of spontaneous autoimmune thyroiditis.J Immunol,2002,168:4216-4220.
[96] Min W, Lillehoj HS.Development and characterization of monoclonal antibodies to chicken interleukin-15. Vet Immunol Immunopathol,2002,88:49-56.
[97] Williams PE. The European ban of the prophylactic use of antibiotics as growth promoters in animal nutrition:political and economic aspects.In;Proceedings of the Australian Poultry Science Symposium, 2001,83-93.
[98] World Health Organisation.report of a Meeting to Review the Use of Fluoroquinolones in Animal Agriculture and its Implications for Human Health.WHO,Geneva.Switzerland.
[99] Schijns VE,Weining KC,Nuijten P.Immunoadjuvant activities of E.coli and plasmid-expressed recombinant chicken IFN-α/β,IFN-γand 1L-1β in 1 day and 3-week-old chickens. Vaccine,2000,18: 2147-2154.
[100] Lowenthal JW, Oneil TE,Broadway M,et al.Coadministration of chicken interferon-gamma enhances antibody responses in chickens.J interferon Cytokine Res, 1998,18:617-622.
[101] Lowenthal JW, Lambrecht B,Van denBerg TP, et al.Avian cytokine-the natural approach to therapeutics.Dev Comp Immunol,2000,24:355-365.
[102] Lillehoj HS,Choi KD.Recombinant chicken interferon-gamma-mediated inhibition of Eimeria tenella development in vitro and reduction of oocyst production and body weight loss following Eimeria acervulina challenge infection. Avian Dis, 1998,42:307-314.
[103] Marcus PI,Van der HL,Sekellick MJ.interferon action on avian virus.I.Oral administration of chicken interferon-alpha ameliorates Newcastle disease.J Interferon Cytokine Res, 1999, 19:881-885.
[104] Karaca K,Sharma JM,Winstow BJ,et al.Recombinant fowlpox viruses coexpressing chicken type Ⅰ IFN and Newcastle disease virus HN and F genes:influence of IFN on protective efficacy and humoral responses of chickens following in ovo or post-hatch administration of recombinant viruses.Vaccine, 1998,16:1496-1503.
[105] 姜永厚，刘忠贵，陈奖励，等．鸡IL-2在新城疫病毒F基因疫苗中的作用．中国兽医学报，2002，22(1)：22-24．
[106] 于链，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疫苗免疫原性的研究．生物工程学报，2001，17(6)：652-657．
[107] 萨姆布鲁克 J，弗里奇 E F，曼尼阿蒂斯 T，等．分子克隆实验指南，第二版．金冬雁，黎孟枫，侯云德译．北京：科学出版社，1992，19-22．
[108] 李迎秋，王循章，龙絮新．乙型肝炎病毒preC／C基因在杆状病毒载体系统中的表达．病毒学报，1997，13(4)：325-331．
[109] 尹长城，刘晶，黄华梁．昆虫表达系统及其在基因工程抗体中的应用．生物工程进展，2001，21(3)：66-70．