接种疫苗防流感

<正>又到秋冬之交,我国很多地方开始进入流感高发期。流感是由不同类型的流感病毒引起的,历史上的流感大流行曾经造成数千万人的死亡。接种流感疫苗是控制与预防流感大规模流行的有效办法。     

H5亚型禽流感病毒HA基因昆虫/杆状病毒系统的表达及对BALB/c小鼠的免疫保护

经RT-PCR扩增了禽流感病毒A/Goose/Guangdong/1／96 H5N1亚型1．7kb HA基因的cDNA,将其克隆到pMD18-T中并测序。亚克隆到杆状病毒转移载体pMelBacA的蜜蜂蜂毒素分泌信号下游中,测序正确后与线性化的杆状病毒DNA(Bac-N-BlueTM DNA)共转染Sf9昆虫细胞。将重组杆状病毒感染HFive细胞,72h左右收获细胞,超声波裂解,SDS—PAGE结果表明HA基因在重组杆状病毒感染的HFive细胞中获得表达。蛋白胶薄层扫描分析显示:表达的HA蛋白占重组杆状病毒感染细胞总蛋白含量的17．1％。Western-blot 及血凝实验结果显示,表达的禽流感H5N1亚型病毒HA蛋白具有生物学活性。表达的H5 HA蛋白定量乳化后,皮下多点注射免疫SPF 级BALB/c雌性小鼠,免疫后产生了H5 HA特异抗体,并在三免前后达到并保持较高水平。用致死剂量的HPAIV H5N1攻击小鼠,免疫组小鼠提供了100％的保护力,而对照组小鼠先后发病且死亡:为研制禽流感H5N1亚型病毒亚单位疫苗,防制禽流感奠定了基础。     

体内注射植物血凝素提高蛇骨髓细胞分裂指数的研究

体内注射植物血凝素提高蛇骨髓细胞分裂指数的研究王钜＊李士怡（辽宁中医学院生物教研室，沈阳１１００３２）杨果杰（首都医科大学实验科学动物部，北京１０００５３）关键词植物性血凝素；蛇／分裂指数．１９６０年Ｎｏｗｅｌ发现植物血凝素（Ｐｈｙｔｏｈｅｍａｇｇｌ...     

禽流感病毒HSNl血凝素基因和神经氨酸酶基因在真核酵母菌中的表达毒

目的克隆、表达和鉴定禽流感病毒HSNI血凝素基因(hemaggludnin,HA)和神经氨酸酶基因(neuramidinase,NA) 序列,为制备抗体和基因T程疫苗打下基础。方法在成功克隆禽流感病毒H5NI全长HA、NA基因并测序的基础 上。将部分基因序列克隆到表达载体pMET A上,构建了重组表达质粒pMET A／HA(49一l 587 bp)、pMET A／NA (121—1 200 bp),电转化真核酵母菌pMADl6,甲醇诱导表达,利用Ni“亲和层析柱对重组蛋白进行纯化,并用 Western Blotting和ELISA方法检测其抗原性。结果重组蛋白在酵母菌中可以高效表达,SDS—PAGE显示蛋白表 达后形成了二聚体,蛋白纯度占总蛋白的95％以上,ELISA和Western Blotting实验证实,重组蛋白具有良好的抗原 性。结论本研究成功克隆和表达了禽流感病毒H5NI HA、NA基因序列,为禽流感病毒H5N1诊断试剂和疫苗的 开发等进一步的研究提供了依据。     

禽流感病毒H5N1血凝素基因和神经氨酸酶基因在真核酵母菌中的表达

目的克隆、表达和鉴定禽流感病毒H5N1血凝素基因（hemagglutinin,HA）和神经氨酸酶基因（neuramidinase,NA）序列,为制备抗体和基因工程疫苗打下基础。方法在成功克隆禽流感病毒H5N1全长HA、NA基因并测序的基础上,将部分基因序列克隆到表达载体pMET A上,构建了重组表达质粒pMET A/HA（49～1 587 bp）、pMET A/NA（121～1 200 bp）,电转化真核酵母菌pMAD16,甲醇诱导表达,利用Ni2＋亲和层析柱对重组蛋白进行纯化,并用Western Blotting和ELISA方法检测其抗原性。结果重组蛋白在酵母菌中可以高效表达,SDS-PAGE显示蛋白表达后形成了二聚体,蛋白纯度占总蛋白的95%以上,ELISA和Western Blotting实验证实,重组蛋白具有良好的抗原性。结论本研究成功克隆和表达了禽流感病毒H5N1 HA、NA基因序列,为禽流感病毒H5N1诊断试剂和疫苗的开发等进一步的研究提供了依据。     

H5N1亚型禽流感病毒血凝素裂解位点碱性氨基酸对应mRNA核苷酸的二级结构分析

以H5N1亚型禽流感病毒(avian influenza viruses, AIV)毒株血凝素(hemagglutinin, HA)蛋白裂解位点碱性氨基酸为研究对象, 研究其密码子的偏好性及对应mRNA序列折叠二级结构的特点. 将mRNA样本按照序列等步长递增的方法, 用RNAstructure 4.1程序预测这些样本的动态延伸折叠二级结构. 结果表明, 裂解位点的碱性氨基酸对富含腺嘌呤(A)的密码子有强烈偏好; 与碱性氨基酸对应的mRNA片段上的核苷酸主要位于折叠二级结构的单链环区, 极少数位于配对螺旋区. 此外, 本研究还从理论上提出了防治H5N1亚型AIV感染的对策, 考虑以位于环区的mRNA核苷酸, 运用RNAi的方法阻止H5N1亚型AIV的HA的表达.     

流感病毒传播元凶

美国疾病控制和预防中心的研究人员目前发现血凝素（红血球凝聚素）可能是人类流感病毒得以肆虐传播的元凶。     

Mutations in Hemagglutinin of a Novel Avian-Origin H7N9 Virus That Are Critical for Receptor Binding Specificity

A novel avian-origin H7N9 influenza virus was discovered in March in China and has caused a total of131 people infected including 39 deaths in China as of June 9, 2013. Adaptation of avian viruses to efficiently infect humans requires the viral hemagglutinin（HA） binding switches from avian to human type receptors with help of some mutations in HA. As such it is critical for pandemic assessment to discover these mutations as hallmarks of adaptation. To continue our previous study of this novel H7N9 virus, we identified two sets of mutations in HA. The first set of mutations are present in the current circulating strains of 2013 H7N9 in China, and the second set are potential mutations that were found when compared to the HAs of previous human H7 subtype. These two sets of mutations exhibited unique features. The first group of mutations, on average, enhanced the HA binding to human type receptors whereas reduced that to avian types. Further the reduction of avian binding was almost three times of the increase of the human binding. The second group increased the binding to both human and avian types.But the increase in human types was almost three times of that in the avian types. Though different in their way of changing the binding preference, these two sets of mutations both contained more mutations to decrease the avian binding and increase the human binding than those that did the opposite. Our research highlighted the pandemic potential of this novel virus by showing the important mutations that could potentially help it to adapt to human hosts. Our findings offered new insights into the current state of evolution of this virus, which might be helpful for the continued surveillance of the emergence of H7N9 strains having the ability of human-to-human transmission.     

韩国初步研制出禽流感疫苗毒株

韩国科学家最近研制出针对H5N1型禽流感病毒的疫苗毒株,并计划不久后用该疫苗毒株进行动物试验。