PMID- 25056882 OWN - NLM STAT- MEDLINE DCOM- 20141111 LR - 20211021 IS - 1098-5514 (Electronic) IS - 0022-538X (Print) IS - 0022-538X (Linking) VI - 88 IP - 19 DP - 2014 Oct TI - Rational design of human metapneumovirus live attenuated vaccine candidates by inhibiting viral mRNA cap methyltransferase. PG - 11411-29 LID - 10.1128/JVI.00876-14 [doi] AB - The paramyxoviruses human respiratory syncytial virus (hRSV), human metapneumovirus (hMPV), and human parainfluenza virus type 3 (hPIV3) are responsible for the majority of pediatric respiratory diseases and inflict significant economic loss, health care costs, and emotional burdens. Despite major efforts, there are no vaccines available for these viruses. The conserved region VI (CR VI) of the large (L) polymerase proteins of paramyxoviruses catalyzes methyltransferase (MTase) activities that typically methylate viral mRNAs at positions guanine N-7 (G-N-7) and ribose 2'-O. In this study, we generated a panel of recombinant hMPVs carrying mutations in the S-adenosylmethionine (SAM) binding site in CR VI of L protein. These recombinant viruses were specifically defective in ribose 2'-O methylation but not G-N-7 methylation and were genetically stable and highly attenuated in cell culture and viral replication in the upper and lower respiratory tracts of cotton rats. Importantly, vaccination of cotton rats with these recombinant hMPVs (rhMPVs) with defective MTases triggered a high level of neutralizing antibody, and the rats were completely protected from challenge with wild-type rhMPV. Collectively, our results indicate that (i) amino acid residues in the SAM binding site in the hMPV L protein are essential for 2'-O methylation and (ii) inhibition of mRNA cap MTase can serve as a novel target to rationally design live attenuated vaccines for hMPV and perhaps other paramyxoviruses, such as hRSV and hPIV3. IMPORTANCE: Human paramyxoviruses, including hRSV, hMPV, and hPIV3, cause the majority of acute upper and lower respiratory tract infections in humans, particularly in infants, children, the elderly, and immunocompromised individuals. Currently, there is no licensed vaccine available. A formalin-inactivated vaccine is not suitable for these viruses because it causes enhanced lung damage upon reinfection with the same virus. A live attenuated vaccine is the most promising vaccine strategy for human paramyxoviruses. However, it remains a challenge to identify an attenuated virus strain that has an optimal balance between attenuation and immunogenicity. Using reverse genetics, we generated a panel of recombinant hMPVs that were specifically defective in ribose 2'-O methyltransferase (MTase) but not G-N-7 MTase. These MTase-defective hMPVs were genetically stable and sufficiently attenuated but retained high immunogenicity. This work highlights a critical role of 2'-O MTase in paramyxovirus replication and pathogenesis and a new avenue for the development of safe and efficacious live attenuated vaccines for hMPV and other human paramyxoviruses. CI - Copyright (c) 2014, American Society for Microbiology. All Rights Reserved. FAU - Zhang, Yu AU - Zhang Y AD - Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA Program in Food Science and Technology, The Ohio State University, Columbus, Ohio, USA. FAU - Wei, Yongwei AU - Wei Y AD - Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA. FAU - Zhang, Xiaodong AU - Zhang X AD - Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA. FAU - Cai, Hui AU - Cai H AD - Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA. FAU - Niewiesk, Stefan AU - Niewiesk S AD - Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA. FAU - Li, Jianrong AU - Li J AD - Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA li.926@osu.edu. LA - eng GR - R01 AI090060/AI/NIAID NIH HHS/United States GR - R56 AI090060/AI/NIAID NIH HHS/United States GR - R01AI090060/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20140723 PL - United States TA - J Virol JT - Journal of virology JID - 0113724 RN - 0 (Antibodies, Neutralizing) RN - 0 (Antibodies, Viral) RN - 0 (Recombinant Proteins) RN - 0 (Vaccines, Attenuated) RN - 0 (Viral Proteins) RN - 0 (Viral Vaccines) RN - 7LP2MPO46S (S-Adenosylmethionine) RN - EC 2.1.1.- (Methyltransferases) SB - IM MH - Animals MH - Antibodies, Neutralizing/biosynthesis MH - Antibodies, Viral/*biosynthesis MH - Binding Sites MH - Female MH - Gene Expression MH - Humans MH - Immunity, Active MH - Metapneumovirus/enzymology/genetics/*immunology MH - Methyltransferases/chemistry/genetics/*immunology MH - Paramyxoviridae Infections/immunology/*prevention & control MH - Protein Binding MH - Recombinant Proteins/chemistry/genetics/immunology MH - S-Adenosylmethionine/chemistry/metabolism MH - Sigmodontinae MH - Vaccination MH - Vaccines, Attenuated MH - Viral Proteins/chemistry/genetics/*immunology MH - Viral Vaccines/administration & dosage/*immunology PMC - PMC4178811 EDAT- 2014/07/25 06:00 MHDA- 2014/11/12 06:00 PMCR- 2015/04/01 CRDT- 2014/07/25 06:00 PHST- 2014/07/25 06:00 [entrez] PHST- 2014/07/25 06:00 [pubmed] PHST- 2014/11/12 06:00 [medline] PHST- 2015/04/01 00:00 [pmc-release] AID - JVI.00876-14 [pii] AID - 00876-14 [pii] AID - 10.1128/JVI.00876-14 [doi] PST - ppublish SO - J Virol. 2014 Oct;88(19):11411-29. doi: 10.1128/JVI.00876-14. Epub 2014 Jul 23.