TY - JOUR
T1 - A mutation in the receptor binding site enhances infectivity of 2009 H1N1 influenza hemagglutinin pseudotypes without changing antigenicity
AU - Wang, Wei
AU - Castelán-Vega, Juan A.
AU - Jiménez-Alberto, Alicia
AU - Vassell, Russell
AU - Ye, Zhiping
AU - Weiss, Carol D.
N1 - Funding Information:
We thank the following persons for generously supplying key reagents for these studies: Galina Vodieko and Christine Anderson (FDA, Bethesda, MD) for sheep reference antisera; Dr. Kanta Subbarao (NIH, Bethesda, MD) for ferret A/California/04/2009 antiserum; and Dr. Gary Nabel (NIH, Bethesda, MD) for CMV/R 8κB and HIV-luciferase expression vectors. We are also grateful to Drs. Vladimir Lugostev and Judy Beeler (FDA, Bethesda, MD) for critical review of the manuscript and Dr. Juan Luis Arciniega (FDA, Bethesda, MD) for providing laboratory space for some of this research. This work was supported by institutional research funds from the FDA for WW, JC-V, AJ-A, RV, ZY, and CDW and from COTEPABE (Instituto Politécnico Nacional, México) and CONACYT for JC-V and AJ-A.
PY - 2010/11/25
Y1 - 2010/11/25
N2 - The 2009 H1N1 pandemic highlights the need to better understand influenza A infectivity and antigenicity. Relative to other recent seasonal H1N1 influenza strains, the 2009 H1N1 virus grew less efficiently in eggs, which hindered efforts to rapidly supply vaccine. Using lentiviral pseudotypes bearing influenza hemagglutinin (HA-pseudotypes) we evaluated a glutamine to arginine mutation at position 223 (Q223R) and glycosylation at residue 276 in HA for their effects on infectivity and neutralization. Q223R emerged during propagation in eggs and lies in the receptor binding site. We found that the Q223R mutation greatly enhanced infectivity of HA-pseudotypes in human cells, which was further augmented by inclusion of the viral neuraminidase (NA) and M2 proteins. Loss of glycosylation at residue 276 did not alter infectivity. None of these modifications affected neutralization. These findings provide information for increasing 2009 H1N1HA-pseudotype titers without altering antigenicity and offer insights into receptor use.
AB - The 2009 H1N1 pandemic highlights the need to better understand influenza A infectivity and antigenicity. Relative to other recent seasonal H1N1 influenza strains, the 2009 H1N1 virus grew less efficiently in eggs, which hindered efforts to rapidly supply vaccine. Using lentiviral pseudotypes bearing influenza hemagglutinin (HA-pseudotypes) we evaluated a glutamine to arginine mutation at position 223 (Q223R) and glycosylation at residue 276 in HA for their effects on infectivity and neutralization. Q223R emerged during propagation in eggs and lies in the receptor binding site. We found that the Q223R mutation greatly enhanced infectivity of HA-pseudotypes in human cells, which was further augmented by inclusion of the viral neuraminidase (NA) and M2 proteins. Loss of glycosylation at residue 276 did not alter infectivity. None of these modifications affected neutralization. These findings provide information for increasing 2009 H1N1HA-pseudotype titers without altering antigenicity and offer insights into receptor use.
KW - Hemagglutinin
KW - Hemagglutinin pseudotypes
KW - Influenza A 2009 H1N1 virus
KW - Influenza A antigenicity
KW - Influenza infectivity
KW - Pseudotype neutralization
UR - http://www.scopus.com/inward/record.url?scp=77957688042&partnerID=8YFLogxK
U2 - 10.1016/j.virol.2010.08.027
DO - 10.1016/j.virol.2010.08.027
M3 - Artículo
C2 - 20869738
SN - 0042-6822
VL - 407
SP - 374
EP - 380
JO - Virology
JF - Virology
IS - 2
ER -