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Consensus and variations in cell line specificity among human metapneumovirus strains

Naganori Nao¹, Ko Sato², Junya Yamagishi³

¹Department of Virology 3, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan.

|April 24, 2019

Abstract:

Human metapneumovirus (HMPV) has been a notable etiological agent of acute respiratory infection in humans, but it was not discovered until 2001, because HMPV replicates only in a limited number of cell lines and the cytopathic effect (CPE) is often mild. To promote the study of HMPV, several groups have generated green fluorescent protein (GFP)-expressing recombinant HMPV strains (HMPVGFP). However, the growing evidence has complicated the understanding of cell line specificity of HMPV, because it seems to vary notably among HMPV strains. In addition, unique A2b clade HMPV strains with a 180-nucleotide duplication in the G gene (HMPV A2b180nt-dup strains) have recently been detected. In this study, we re-evaluated and compared the cell line specificity of clinical isolates of HMPV strains, including the novel HMPV A2b180nt-dup strains, and six recombinant HMPVGFP strains, including the newly generated recombinant HMPV A2b180nt-dup strain, MG0256-EGFP. Our data demonstrate that VeroE6 and LLC-MK2 cells generally showed the highest infectivity with any clinical isolates and recombinant HMPVGFP strains. Other human-derived cell lines (BEAS-2B, A549, HEK293, MNT-1, and HeLa cells) showed certain levels of infectivity with HMPV, but these were significantly lower than those of VeroE6 and LLC-MK2 cells. Also, the infectivity in these suboptimal cell lines varied greatly among HMPV strains. The variations were not directly related to HMPV genotypes, cell lines used for isolation and propagation, specific genome mutations, or nucleotide duplications in the G gene. Thus, these variations in suboptimal cell lines are likely intrinsic to particular HMPV strains.

Insights

Human metapneumovirus (HMPV) infects cells differently depending on the strain. VeroE6 and LLC-MK2 cells show the highest HMPV infectivity, while other cell lines exhibit variable results not linked to specific viral traits.

Area of Science:

Virology
Cell Biology
Respiratory Infections

Background:

Human metapneumovirus (HMPV) is a significant cause of acute respiratory illness.
HMPV discovery was delayed due to limited cell line replication and mild cytopathic effects (CPE).
Recombinant HMPV strains expressing green fluorescent protein (GFP) aid HMPV research, but cell line specificity varies among strains.

Purpose of the Study:

To re-evaluate and compare the cell line specificity of various HMPV clinical isolates and recombinant strains.
To investigate the infectivity of novel HMPV A2b strains with a 180-nucleotide duplication in the G gene.
To understand the factors influencing HMPV replication efficiency across different cell lines.

Main Methods:

Infectivity assays using clinical HMPV isolates and recombinant HMPV-GFP strains.
Comparison of HMPV replication in multiple human-derived cell lines (BEAS-2B, A549, HEK293, MNT-1, HeLa) and standard cell lines (VeroE6, LLC-MK2).
Analysis of infectivity variations in relation to HMPV genotypes, isolation/propagation cell lines, genome mutations, and G gene duplications.

Main Results:

VeroE6 and LLC-MK2 cells demonstrated the highest infectivity for all tested HMPV strains.
Human-derived cell lines showed significantly lower and highly variable HMPV infectivity.
Infectivity variations in suboptimal cell lines were strain-specific and not directly correlated with known genetic factors like G gene duplications or genotypes.

Conclusions:

VeroE6 and LLC-MK2 cells are optimal for HMPV propagation and study.
The intrinsic properties of specific HMPV strains significantly influence their infectivity in non-optimal cell lines.
Further research is needed to elucidate the mechanisms behind strain-specific variations in HMPV cell tropism.

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