Clin Exp Vaccine Res.  2020 Jan;9(1):56-63. 10.7774/cevr.2020.9.1.56.

Comparison of antigenic mutation during egg and cell passage cultivation of H3N2 influenza virus

• Park YW
• Kim YH
• Jung HU
• Jeong OS
• Hong EJ
• Kim H
• Lee JI

• KMID: 2470838
• DOI: http://doi.org/10.7774/cevr.2020.9.1.56

Abstract

PURPOSE
When influenza viruses are cultured in eggs, amino acid mutations of the hemagglutinin may occur through egg adaptation. On the other hand, when influenza viruses are cultured in animal cells, no antigenic mutation occurs unlike in eggs. Therefore, we examined whether the antigenic mutations actually occurred after passage of H3N2 (A/Texas/50/2012) virus up to 15 times in eggs and MDCK-Sky3851 cells.
MATERIALS AND METHODS
Prototype A/Texas/50/2012 (H3N2) influenza virus which was isolated from clinical patient, not passaged in egg, was obtained and propagated using the specific pathogen free egg and the MDCK-Sky3851 cell line up to 15 passage, and the changes in the antigen sequence of the influenza viruses were confirmed by gene sequencing and protein structure analysis.
RESULTS
In term of the hemagglutination titer of influenza virus, the reactivity to chicken and guinea pig red blood cell showed different results between egg propagated and cell propagated viruses. In the sequence analysis results for hemagglutinin and neuraminidase, no antigenic mutation was observed throughout all passages when cultured in MDCK-Sky3851 cells. On the other hand, mutations occurred in three amino acid sequences (H156R, G186S, S219F) in hemagglutinin up to 15 passages when cultured in eggs.
CONCLUSION
H3N2 influenza virus cultured in eggs could lead mutations in amino acid sequence of hemagglutinin, distinct from the corresponding virus cultured in cells for which no antigenic mutation was observed. These findings suggest that cell culture is a more stable and effective way of production with lower risk of antigenic mutations for the manufacture of influenza vaccines.

Keyword

Egg adaptation; Influenza hemagglutinin; Antigenic mutation; Cell culture influenza vaccine

MeSH Terms

Amino Acid Sequence
Animals
Cell Culture Techniques
Cell Line
Chickens
Eggs
Erythrocytes
Guinea Pigs
Hand
Hemagglutination
Hemagglutinins
Humans
Influenza Vaccines
Influenza, Human*
Neuraminidase
Orthomyxoviridae*
Ovum*
Sequence Analysis
Specific Pathogen-Free Organisms
Hemagglutinins
Influenza Vaccines
Neuraminidase

Figure

• 

  Fig. 1 Structure of hemagglutinin and mutation points of amino acid during egg cultivation of H3N2 influenza virus (A/Texas/50/2012). (A) All three mutated amino acid were located near the receptor binding site of hemagglutinin. (B) Detail structures were compared for mutated amino acid during egg cultivation. Red color shows the original hemagglutinin structure of parent virus and blue color shows the mutated structure by egg adaptation. Surface view and backbone structure showing corresponding amino acid residue were presented for each mutation.

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