An RNA aptamer that specifically binds to the glycosylated hemagglutinin of avian influenza virus and suppresses viral infection in cells.
The influenza virus surface glycoprotein hemagglutinin (HA) is responsible for viral attachment to sialic acid-containing host cell receptors and it facilitates the initial stage of viral infection. In the present study, we isolated an RNA aptamer specific to the glycosylated receptor-binding domain of the HA protein (gHA1) after 12 cycles of the systematic evolution of ligands by exponential enrichment procedure (SELEX), and we then investigated if the selected aptamer suppresses viral infection in host cells. Nitrocellulose filter binding and enzyme-linked immunosorbent assay (ELISA) experiments revealed that 1 RNA aptamer, HA12-16, bound specifically to the gHA1 protein. Cell viability assay showed that the HA12-16 RNA aptamer suppressed viral infection in host cells by enhancing cell viability. Immunofluorescence microscopic analysis further demonstrated that the HA12-16 RNA aptamer suppresses viral attachment to host cells by neutralizing the receptor-binding site of influenza virus HA. These results indicate that the isolated RNA aptamer can be developed as an antiviral reagent against influenza through appropriate therapeutic formulation.
Department of Bioscience and Biotechnology, Konkuk University, Seoul, Republic of Korea.
Hyun-Mi
HM
Kwon
Department of Bioscience and Biotechnology, Konkuk University, Seoul, Republic of Korea.
Kwang Hyun
KH
Lee
Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, Republic of Korea.
Byung Woo
BW
Han
Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, Republic of Korea.
Mi Ra
MR
Han
Department of Pediatrics, Korea Cancer Center Hospital, Seoul, Republic of Korea.
Dong Ho
DH
Kim
Department of Bioscience and Biotechnology, Konkuk University, Seoul, Republic of Korea.
Dong-Eun
DE
Kim
D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism* / Q000627:therapeutic use; D052157:Aptamers, Nucleotide / Q000378:metabolism* / Q000627:therapeutic use; D001483:Base Sequence; D004285:Dogs; D006031:Glycosylation; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000378:metabolism*; D053122:Influenza A Virus, H3N2 Subtype / Q000502:physiology; D053125:Influenza A Virus, H5N2 Subtype / Q000378:metabolism*; D061985:Madin Darby Canine Kidney Cells; D008969:Molecular Sequence Data; D009976:Orthomyxoviridae Infections / Q000188:drug therapy*; D011485:Protein Binding; D012313:RNA / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D052156:SELEX Aptamer Technique; D061987:Sf9 Cells; D018411:Spodoptera
D000998:Antiviral Agents; D052157:Aptamers, Nucleotide; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins; C109963:hemagglutinin, avian influenza A virus; D012313:RNA