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乙肝表面抗原-CD40L胞外段融合蛋白的设计和生物活性预测 总被引:1,自引:1,他引:0
目的:探讨乙肝表面抗原-CD40L胞外段融合蛋白设计的合理性。方法:应用Gene Constructionkit2.5、DNAStar软件和WWW.expasy.org网站提供的分析方案分析重组体的开放读框以及融合蛋白的柔性、亲水性、抗原性、表位等性质,并作了二级结构模拟分析。结果:重组体CMV启动子下游有完整的目的基因ORF,融合蛋白二级结构水平未出现新的抗原性及表位,亲水性无改变,Linker部位抗原性低,呈中性且柔性高,不影响两端的蛋白质二级结构及融合蛋白空间构象。结论:重组体设计合理,融合蛋白很大可能保留了乙肝表面抗原和CD40L胞外段的生物学活性,为进一步研究提供了理论依据。 相似文献
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目的:构建携铜绿假单胞菌外毒素PE38基因及甲胎蛋白(AFP)启动子的重组载体,并研究其在体外对AFP阳性肝细胞癌(hepatocellularcarcinoma,HCC)的靶向杀伤作用。方法:构建重组免疫毒素表达质粒pAFP-PE38,通过转染细胞观察其作用,RT-PCR法测定PE38 mRNA表达,CCK-8法检测细胞毒性。结果:各组细胞转染质粒pAFP-PE38后仅AFP阳性的HepG2细胞表达PE38 mRNA,且形态发生改变,生长显著受抑(P〈0.01),48 h和72 h抑制率分别为27.2%、58.3%,而AFP阴性的PC-3和HeLa细胞未受明显影响。结论:PE38基因真核表达载体可实现HCC基因治疗的靶向性和高效性,有望成为肝细胞癌靶向基因治疗的有力工具。 相似文献
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Objective To investigate the effect and the potential mechanism of splenic artery coarctation on the expression of iNOS and Th1/Th2 cytokines in spleen of cirrhotic rats with portal hypertension (PHT). Methods Cirrhotic rats were randomized into 3 groups (n= 10):sham operation group (SOG), splenic artery coarctation group (SAC) and splenic artery ligation group (SAL). Ten normal rats treated with sham operation were employed to serve as normal control group (NCG). Immunohistochemial staining was used to observe iNOS. RT-PCR was used to detect IFN-γ and IL-4mRNA. The Pearson's correlation analysis was used to investigate the relationship between iNOS and IFN-γ or IL-4. Results The expression of iNOS was increased significantly in spleen of cirrhotic rats as compared with NCG(P<0. 01). It was decreased after SAC and SAL compared with SOG (P<0. 01). The expression of IFN-γmRNA and IFN-γ/IL-4 of SOG were decreased but IL-4mRNA increased significantly than that of NCG(P<0.01). IFN-γmRNA was increased after SAC compared with SOG (P<0.05). IL-4mRNA was decreased and IFN-γ/IL-4 increased after SAC and SAL compared with SOG (P<0. 05). The expression of iNOS was negatively correlated with the expression of IFN-γmRNA(r=-0.672, P< 0.01 ) and positively correlated with the expression of IL-4 mRNA (r=0.634,P<0. 01). Conclusion The expression of iNOS is decreased and IFN-γ/IL-4 increased after SAC in spleen of cirrhotic rats with PHT and it may improve Th1/Th2 polarization by reducing the expression of iNOS. 相似文献
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目的:检测HBVS-ecdCD40L表达对乙肝转基因小鼠树突状细胞(DC)成熟分化和功能的影响及其可能机制。方法:乙肝转基因小鼠分别肌肉注射质粒pcDNA3.1-S—ecdCD40L、pcDNA3.1-S、pcDNA3.1或PBS,每只100.g/25g,2、4周后各加强免疫一次。首次免疫6周后,取血,分离DC,检测细胞表型及功能,并用荧光定量PCR法检测血清HBVDNA含量。结果:注射pcDNA3.1-S-ecdCD40L小鼠Dc中CD80、CD86、MHC CⅡ及Toll样受体4(TLR4)的表达,白介素-12(IL-12)的分泌及刺激淋巴细胞的能力均高于其余组小鼠,而平均血清HBVDNA含量明显低于其余组小鼠。结论:pcDNA3.1-S-ecdCD40L重组表达载体介导的HBVS-ecdCD40L表达能有效地促进转基因小鼠的Dc成熟分化和功能提高,激活Dc中的TLR4可能是其机制之一。 相似文献
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