Abstract
Human metapneumovirus (HMPV) has recently been identified as a significant cause of serious respiratory tract disease in humans. In particular, the emerging information on the contribution of HMPV to pediatric respiratory tract disease suggests that it will be important to develop a vaccine against this virus for use in conjunction with those being developed for human respiratory syncytial virus and the human parainfluenza viruses. A recently described reverse genetic system (S. Biacchesi, M. H. Skiadopoulos, K. C. Tran, B. R. Murphy, P. L. Collins, and U. J. Buchholz, Virology 321:247-259, 2004) was used to generate recombinant HMPVs (rHMPVs) that lack the G gene, the SH gene, or both. The DeltaSH, DeltaG, and DeltaSH/G deletion mutants were readily recovered and were found to replicate efficiently during multicycle growth in cell culture. Thus, the SH and G proteins are not essential for growth in cell culture. Apart from the absence of the deleted protein(s), the virions produced by the gene deletion mutants were similar by protein yield and gel electrophoresis protein profile to wild-type HMPV. When administered intranasally to hamsters, the DeltaG and DeltaSH/G mutants replicated in both the upper and lower respiratory tracts, showing that HMPV containing F as the sole viral surface protein is competent for replication in vivo. However, both viruses were at least 40-fold and 600-fold restricted in replication in the lower and upper respiratory tract, respectively, compared to wild-type rHMPV. They also induced high titers of HMPV-neutralizing serum antibodies and conferred complete protection against replication of wild-type HMPV challenge virus in the lungs. Surprisingly, G is dispensable for protection, and the DeltaG and DeltaSH/G viruses represent promising vaccine candidates. In contrast, DeltaSH replicated somewhat more efficiently in hamster lungs compared to wild-type rHMPV (20-fold increase on day 5 postinfection). This indicates that SH is completely dispensable in vivo and that its deletion does not confer an attenuating effect, at least in this rodent model.
MeSH Terms
Animals
Cricetinae
Gene Deletion
HN Protein/genetics,physiology
Mesocricetus
Metapneumovirus/immunology,physiology
RNA, Viral/biosynthesis
Recombination, Genetic
Viral Envelope Proteins
Viral Proteins/physiology
Viral Vaccines/immunology
Virus Replication
Chemicals
HN Protein
RNA, Viral
Viral Envelope Proteins
Viral Proteins
Viral Vaccines
attachment protein G
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Biacchesi Stéphane
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892-8007, USA.
Skiadopoulos Mario H
Yang Lijuan
Lamirande Elaine W
Tran Kim C
Murphy Brian R
Collins Peter L
Buchholz Ursula J
References (33)
33 references, click to expand
-
Effects of human metapneumovirus and respiratory syncytial virus antigen insertion in two 3' proximal genome positions of bovine/human parainfluenza virus type 3 on virus replication and immunogenicity.
J Virol. 2003 Oct;77(20):10819-28
PMID: 14512532
-
Viroporins.
FEBS Lett. 2003 Sep 18;552(1):28-34
PMID: 12972148
-
Modeling the structure of the respiratory syncytial virus small hydrophobic protein by silent-mutation analysis of global searching molecular dynamics.
Protein Sci. 2003 Dec;12(12):2668-74
PMID: 14627728
-
Clinical impact and diagnosis of human metapneumovirus infection.
Pediatr Infect Dis J. 2004 Jan;23(1 Suppl):S25-32
PMID: 14730267
-
Human metapneumovirus: a not so new virus.
Pediatr Infect Dis J. 2004 Jan;23(1):1-7; quiz 8-10
PMID: 14743038
-
Sequence polymorphism of the predicted human metapneumovirus G glycoprotein.
J Gen Virol. 2004 Mar;85(Pt 3):679-86
PMID: 14993653
-
Human metapneumovirus: a new player among respiratory viruses.
Clin Infect Dis. 2004 Apr 1;38(7):983-90
PMID: 15034830
-
Recovery of human metapneumovirus from cDNA: optimization of growth in vitro and expression of additional genes.
Virology. 2004 Apr 10;321(2):247-59
PMID: 15051385
-
Identification of small-animal and primate models for evaluation of vaccine candidates for human metapneumovirus (hMPV) and implications for hMPV vaccine design.
J Gen Virol. 2004 Jun;85(Pt 6):1655-63
PMID: 15166450
-
The two major human metapneumovirus genetic lineages are highly related antigenically, and the fusion (F) protein is a major contributor to this antigenic relatedness.
J Virol. 2004 Jul;78(13):6927-37
PMID: 15194769
-
Antigenic and genetic variability of human metapneumoviruses.
Emerg Infect Dis. 2004 Apr;10(4):658-66
PMID: 15200856
-
The G glycoprotein of human respiratory syncytial viruses of subgroups A and B: extensive sequence divergence between antigenically related proteins.
Proc Natl Acad Sci U S A. 1987 Aug;84(16):5625-9
PMID: 2441388
-
The 1A protein of respiratory syncytial virus is an integral membrane protein present as multiple, structurally distinct species.
J Virol. 1989 May;63(5):2019-29
PMID: 2649692
-
Genetic diversity of the attachment protein of subgroup B respiratory syncytial viruses.
J Virol. 1991 Oct;65(10):5425-34
PMID: 1895391
-
Membrane orientation and oligomerization of the small hydrophobic protein of human respiratory syncytial virus.
J Gen Virol. 1993 Jul;74 ( Pt 7):1445-50
PMID: 8336126
-
Membrane permeability changes induced in Escherichia coli by the SH protein of human respiratory syncytial virus.
Virology. 1997 Sep 1;235(2):342-51
PMID: 9281514
-
Recombinant respiratory syncytial virus from which the entire SH gene has been deleted grows efficiently in cell culture and exhibits site-specific attenuation in the respiratory tract of the mouse.
J Virol. 1997 Dec;71(12):8973-82
PMID: 9371553
-
Respiratory syncytial virus (RSV) SH and G proteins are not essential for viral replication in vitro: clinical evaluation and molecular characterization of a cold-passaged, attenuated RSV subgroup B mutant.
Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13961-6
PMID: 9391135
-
Identification of the respiratory syncytial virus proteins required for formation and passage of helper-dependent infectious particles.
J Virol. 1998 Jul;72(7):5707-16
PMID: 9621029
-
Generation of bovine respiratory syncytial virus (BRSV) from cDNA: BRSV NS2 is not essential for virus replication in tissue culture, and the human RSV leader region acts as a functional BRSV genome promoter.
J Virol. 1999 Jan;73(1):251-9
PMID: 9847328
-
Recombinant respiratory syncytial virus bearing a deletion of either the NS2 or SH gene is attenuated in chimpanzees.
J Virol. 1999 Apr;73(4):3438-42
PMID: 10074199
-
Recombinant bovine respiratory syncytial virus with deletions of the G or SH genes: G and F proteins bind heparin.
J Gen Virol. 2001 Mar;82(Pt 3):631-40
PMID: 11172105
-
A newly discovered human pneumovirus isolated from young children with respiratory tract disease.
Nat Med. 2001 Jun;7(6):719-24
PMID: 11385510
-
Functional analysis of recombinant respiratory syncytial virus deletion mutants lacking the small hydrophobic and/or attachment glycoprotein gene.
J Virol. 2001 Aug;75(15):6825-34
PMID: 11435561
-
Cleavage of the human respiratory syncytial virus fusion protein at two distinct sites is required for activation of membrane fusion.
Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):9859-64
PMID: 11493675
-
Proteolytic activation of respiratory syncytial virus fusion protein. Cleavage at two furin consensus sequences.
J Biol Chem. 2001 Aug 24;276(34):31642-50
PMID: 11418598
-
Contribution of the respiratory syncytial virus G glycoprotein and its secreted and membrane-bound forms to virus replication in vitro and in vivo.
Virology. 2001 Oct 25;289(2):283-96
PMID: 11689051
-
Human respiratory syncytial virus surface glycoproteins F, G and SH form an oligomeric complex.
Arch Virol. 2001 Dec;146(12):2369-83
PMID: 11811686
-
The central conserved cystine noose of the attachment G protein of human respiratory syncytial virus is not required for efficient viral infection in vitro or in vivo.
J Virol. 2002 Jun;76(12):6164-71
PMID: 12021350
-
Characterization of human metapneumoviruses isolated from patients in North America.
J Infect Dis. 2002 Jun 1;185(11):1660-3
PMID: 12023774
-
Analysis of the genomic sequence of a human metapneumovirus.
Virology. 2002 Mar 30;295(1):119-32
PMID: 12033771
-
Mucosal immunization with live recombinant bovine respiratory syncytial virus (BRSV) and recombinant BRSV lacking the envelope glycoprotein G protects against challenge with wild-type BRSV.
J Virol. 2002 Dec;76(23):12355-9
PMID: 12414977
-
Genetic diversity between human metapneumovirus subgroups.
Virology. 2003 Oct 10;315(1):1-9
PMID: 14592754