Molecular biological characterization analysis of tick-borne encephalitis virus strain adapted to primary hamster kidney cells
10.13200/j.cnki.cjb.004568
- VernacularTitle:森林脑炎病毒原代地鼠肾细胞适应株分子生物学特性分析
- Author:
MIAO Hui
- Publication Type:Journal Article
- Keywords:
Tick-borne encephalitis virus(TBEV);
Primary hamster kidney(PHK) cells;
Sequence analysis;
Genetic stability;
Vaccine candidate strain
- From:
Chinese Journal of Biologicals
2025;38(10):1168-1174
- CountryChina
- Language:Chinese
-
Abstract:
Objective To analyze the molecular biological characteristics of tick-borne encephalitis virus(TBEV) strain adapted to primary hamster kidney(PHK) cells, and evaluate the genetic stability, so as to provide a basis for the screening of vaccine candidate strains.Methods The mouse brain-adapted TBEV original “Senzhang” strain was adapted on PHK cells for 10 generations(PT-1 to PT-10), and animals' method was used to determine the virus titers of each generation. The E protein gene of PT-1 to PT-10 passages and the whole genomes of PT-1, 3, 4, 7 and 10 passages were amplified and sequenced. Bioinformatics software was used for sequence alignment, homology analysis and E protein structure prediction.Results The virus titer of TBEV was stable at(8. 89 ± 0. 07) lgLD_(50)/mL after adaptation to PHK cells. The genome full length of TBEV strain adapted to PHK cells was 10 782 bp, encoding 3 414 amino acids. The length of E protein gene was 1 488 bp,encoding 496 amino acids. Compared to the original strain, the TBEV adapted to PHK cells exhibited four amino acid mutations at specific sites: E-203 from aspartic acid to alanine at PT-8, PT-9 and PT-10 generation, NS2A-163 from glutamine to lysine at PT-3 and PT-4 generations, NS4B-110 from isoleucine to phenylalanine at PT-3 and PT-4 generations, and NS4 B-40 from tryptophan to arginine at PT-4 generation. The amino acid sequence identity between the PHK cell-adapted TBEV strain and the original mouse brain TBEV stain was 99. 99%, and the simulated tertiary structure of E protein, the theoretical number and position of transmembrane domains as well as the secondary structure of the viral polyprotein were completely consistent.In addition, the virus showed the highest homology with the representative strain of the Far Eastern subtype.Conclusion “Senzhang” strain of TBEV demonstrates good genetic stability after adaptation to PHK cells with conserved key antigen structure,and is expected to serve as a TBEV vaccine candidate strain.
