流感病毒（H5N1）Vero细胞适应株制备

发布时间：2018-03-31 18:38

本文选题：Vero细胞　切入点：重组流感病毒　出处：《中国协和医科大学》2009年硕士论文

【摘要】： H5N1禽流感在全球各地爆发以来,到2007年底,WHO已经证实感染禽流感致死的人数超过200人,致死率高于50%。WHO的专家预测禽流感病毒在人体的逐步适应很可能引起一场全球性的流感大爆发,推荐各国开发生产H5N1人用流感疫苗。目前H5N1人用流感疫苗主要是用鸡胚为培养基质开发生产,H5N1禽流感爆发时难于在短期内提供足够的鸡胚生产疫苗,另外,鸡胚中可能带有的外源性因子也影响疫苗的质量。Vero细胞是WHO推荐用于生产疫苗的细胞,能够利用生物反应器进行大规模培养。与鸡胚来源的疫苗比较,在Vero细胞上得到的流感疫苗可以产生较强的体液免疫反应和细胞免疫反应。如果研究出一种用Vero细胞培养的H5N1禽流感疫苗毒株,将能较好的解决现在禽流感疫苗生产存在的一些问题,对防治禽流感有重大意义。该实验选用H5N1禽流感疫苗株与本实验室筛选的H3N2 Vero适应株同时接种鸡胚,24小时后取尿囊液,加入H3N2 Vero细胞适应株的抗血清中和后在Vero细胞上传代,获得H5N1 Vero细胞高产适应株,利用空斑实验纯化毒株。连续传代20代后,应用血凝抑制实验与基因测序鉴定该毒株,并进一步测定该H5N1 Vero细胞适应株的致病性、免疫原性等生物学特性。获得的H5N1 Vero细胞适应株可用于H5N1禽流感人用疫苗的开发。
[Abstract]:Since the outbreak of H5N1 bird flu around the world, by the end of 2007, the WHO had confirmed that more than 200 people had died from bird flu. Experts with a fatality rate higher than 50%.WHO predict that the gradual adaptation of the bird flu virus in humans is likely to lead to a global influenza outbreak. It is recommended that various countries develop and produce H5N1 human influenza vaccine. At present, H5N1 human influenza vaccine is mainly developed with chicken embryos as the culture substrate to produce enough chicken embryos to produce vaccines for the outbreak of H5N1 avian influenza in a short period of time. In addition, it is difficult to provide sufficient chicken embryos to produce vaccines in the short term. The exogenous factors in chicken embryo may also affect the quality of vaccine. Vero cells are recommended by WHO for vaccine production and can be cultured on a large scale using bioreactor. The influenza vaccine obtained on Vero cells can produce strong humoral and cellular immune responses. If a strain of H5N1 avian influenza vaccine cultured with Vero cells is developed, It is of great significance to control avian influenza. In this experiment, H5N1 avian influenza vaccine strain and H3N2 Vero adaptive strain selected by our laboratory were inoculated with chicken embryo for 24 hours to get allantoic fluid. The high yield adaptive strain of H5N1 Vero cells was obtained by adding the antiserum neutralization of H3N2 Vero cell acclimation strain to Vero cells. The strain was purified by plaque test. After 20 consecutive passages, the strain was identified by hemagglutination inhibition test and gene sequencing. The pathogenicity, immunogenicity and other biological characteristics of the H5N1 Vero cell adaptive strain were further determined. The obtained H5N1 Vero cell adaptive strain could be used in the development of human vaccine for H5N1 avian influenza.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392

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