猪乙型脑炎病毒在Vero细胞上的繁殖特性及其灭活疫苗的免疫原性研究

doi:10.16431/j.cnki.1671-7236.2018.12.031

Abstract

Abstract:

This study was aimed to investigate the reproductive characteristics of swine Japanese encephalitis virus (SA14-14-2 strains) in Vero cell and the immunogenicity of its inactivated vaccine.The swine Japanese encephalitis virus was cultured in Vero cell.The proliferation technology of swine Japanese encephalitis virus in Vero cell line was studied by the selection and optimization of the following seven culture conditions,including Vero cell incubation time,the amount of inoculated virus,adsorption time,adsorption temperature,pH of maintenance fluid,maintain culturing temperature and culture time.After one time of freezing and thawing,the virus titer was determined by means of viral plaque number determination.The inactivated vaccine of swine Japanese encephalitis virus was prepared by reproducing a batch of viruses under optimized conditions and inactivating it by means of inactivation of β-propiolactone and mixed with bidirectional adjuvant.Two immunizations (interval 14 d) were given to piglets with negative antibody to encephalitis,and the serum was collected before (0 d) and after 7,14,21,28,35 and 42 d for the first immunization,and serum neutralization antibodies were detected.We attacked piglets using the encephalitis P3 strain at the 28th after two immunizations,and the plasma was collected before (0 d) and after 1,2,3,5,7 and 9 d;14 d after the attack,piglets were dissected and brain tissues were collected,and detecting the swine Japanese encephalitis virus in plasma and brain tissue.The results showed that the cells were cultured in a cell culture flask,and the Vero cells were cultured for 48 h for virus inoculation,the inoculum size was 1 000 PFU/mL,the virus adsorption temperature was 37℃,the adsorption time was 90 min,and pH was 7.6 to 8.8 after adsorption.The maintenance solution was continued to be cultured at a temperature of 35℃.After 96 h of culture,the venom was harvested and frozen and thawed to obtain a higher titer.The serum antibody level of the inactivated vaccine prepared by immunization of piglets increased rapidly.No Japanese encephalitis virus was detected in plasma or brain tissue.The test piglets of immunized group could resist strong poisoning and obtain an effective immune protection.This results provided a basis for the production of swine encephalitis vaccine.

Key words: Japanese encephalitis virus (JEV); Vero cells; reproductive characteristics; virus titer; inactivated vaccine; immunogenicity

CLC Number:

S852.65

HAN Wei, ZHOU Jianmin, HAO Linyu, PAN Wen, PAN Jingxue. Study on the Reproductive Characteristics of Swine Japanese Encephalitis Virus in Vero Cells and the Immunogenicity of Its Inactivated Vaccine[J]. , 2018, 45(12): 3572-3578.

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Study on the Reproductive Characteristics of Swine Japanese Encephalitis Virus in Vero Cells and the Immunogenicity of Its Inactivated Vaccine

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