为探讨中药复方多糖(compound Chinese herbal medicine polysaccharides,cCHMPS)对不同MHC B-LβⅡ基因型鸡免疫调节作用的影响,采用PCR-SSCP方法将200羽白羽肉鸡按不同MHC B-LβⅡ基因型分组,采集不同MHC B-LβⅡ基因型鸡的外周血淋...为探讨中药复方多糖(compound Chinese herbal medicine polysaccharides,cCHMPS)对不同MHC B-LβⅡ基因型鸡免疫调节作用的影响,采用PCR-SSCP方法将200羽白羽肉鸡按不同MHC B-LβⅡ基因型分组,采集不同MHC B-LβⅡ基因型鸡的外周血淋巴细胞,分别加入终剂量为100、75、50、0μg/mL的cCHMPS,共培养24h,采用实时荧光定量PCR方法检测cCHMPS对鸡淋巴细胞NF-κB、TNF-α、IL-6mRNA表达量的影响。结果显示:与对照组相比,cCHMPS能显著增强不同MHC B-LβⅡ基因型鸡淋巴细胞NF-κB、TNF-α、IL-6mRNA的表达量(P<0.05)。并且同一基因型鸡中,当cCHMPS剂量为50μg/mL时,AB、AA基因型鸡淋巴细胞NF-κB、TNF-α、IL-6mRNA表达量显著高于其他剂量组(P<0.05);AC基因型鸡淋巴细胞NF-κB、IL-6mRNA的表达量显著高于其他剂量组(P<0.05);AC基因型鸡淋巴细胞TNF-αmRNA的表达量在cCHMPS为100μg/mL时显著高于其他剂量组(P<0.05)。综上所述,cCHMPS能不同程度的促进各MHC B-LβⅡ基因型鸡NF-κB、TNF-α、IL-6mRNA的表达,且不同MHC B-LβⅡ基因型鸡淋巴细胞最适中药复方多糖免疫调节剂量不同。展开更多
[ Objective] To prepare the monoclonal antibody against chicken major histocompatibility complex class II molecules （MHC II）. [ Method ] The prokaryotic expression of the gene fragments of exons 2 -6 encoding alpha ...[ Objective] To prepare the monoclonal antibody against chicken major histocompatibility complex class II molecules （MHC II）. [ Method ] The prokaryotic expression of the gene fragments of exons 2 -6 encoding alpha chain of MHC II and exons 3 -6 encoding beta chain of MHC II were performed based on its protein sequences. After BALB/c mice were immunized with the purified fusion proteins, the mouse spleen cells were fused with mouse myeloma cells SP2/0. Then the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay （ELISA）. [ Result] One hybridoma cell strain secreting monoclonal antibody against alpha chain and two strains secreting monoclonal antibody against beta chain were obtained. These three hybridoma cell strains were named as MHC II alpha-4, MHC II betas-2 and MCH II betas-31, respectively. Their titers of ascites in indirect ELISA were 1 ： 256 000, 1 ： 256 000 and 1 ： 1 280 000, respectively. These antibodies could specifically recog- nize MHC II alpha chain or beta chain in western blotting. [ Conclusion] Three obtained hybridoma stains can stably produce the monoclonal antibody against chicken MHC class II molecules.展开更多
基金supported by the National Natural Science Foundation (30671537)
文摘[ Objective] To prepare the monoclonal antibody against chicken major histocompatibility complex class II molecules （MHC II）. [ Method ] The prokaryotic expression of the gene fragments of exons 2 -6 encoding alpha chain of MHC II and exons 3 -6 encoding beta chain of MHC II were performed based on its protein sequences. After BALB/c mice were immunized with the purified fusion proteins, the mouse spleen cells were fused with mouse myeloma cells SP2/0. Then the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay （ELISA）. [ Result] One hybridoma cell strain secreting monoclonal antibody against alpha chain and two strains secreting monoclonal antibody against beta chain were obtained. These three hybridoma cell strains were named as MHC II alpha-4, MHC II betas-2 and MCH II betas-31, respectively. Their titers of ascites in indirect ELISA were 1 ： 256 000, 1 ： 256 000 and 1 ： 1 280 000, respectively. These antibodies could specifically recog- nize MHC II alpha chain or beta chain in western blotting. [ Conclusion] Three obtained hybridoma stains can stably produce the monoclonal antibody against chicken MHC class II molecules.
