DNA Aptamers against the Receptor Binding Region of Hemagglutinin Prevent Avian Influenza Viral Infection

• Published in: Molecules and cells 2011, 32(6), , pp.527-533
• Main Authors: Choi, S.K., Hallym University, Chuncheon, Republic of Korea, Lee, C.J., Seoul National University, Seoul, Republic of Korea, Lee, K.S., Hallym University, Chuncheon, Republic of Korea, Choe, S.Y., Chungbuk National University, Cheongju, Republic of Korea, Mo, I.P., Chungbuk National University, Cheongju, Republic of Korea, Seong, R.H., Seoul National University, Seoul, Republic of Korea, Hong, S.M., Sejong University, Seoul, Republic of Korea, Jeon, S.H., Hallym University, Chuncheon, Republic of Korea
• Format: Article
• Language: English
• Subjects: AGGLUTININE; AGGLUTININS; AGLUTININAS; Amino Acid Sequence; Animals; anti-viral effect; Antiviral Agents - chemistry; Antiviral Agents - pharmacology; Aptamers, Nucleotide - chemistry; Aptamers, Nucleotide - pharmacology; Base Sequence; Biochemistry; Biomedical and Life Sciences; Biomedicine; Biotechnology; Cell Biology; Cell Line; Cell Survival; Cloning, Molecular; Consensus Sequence; DNA aptamer; Dogs; H9N2 avian influenza virus; Hemagglutinins - chemistry; Hemagglutinins - immunology; Immune Sera - chemistry; Influenza A Virus, H9N2 Subtype; Life Sciences; Mice; Molecular Sequence Data; Orthomyxoviridae Infections - prevention & control; Peptide Fragments - chemistry; Peptide Fragments - immunology; Protein Binding; Protein Interaction Domains and Motifs; SELEX Aptamer Technique; Sequence Analysis, DNA; 생물학
• ISSN: 1016-8478; 0219-1032
• DOI: 10.1007/s10059-011-0156-x

The entrance of influenza virus into host cells is facilitated by the attachment of the globular region of viral hemagglutinin to the sialic acid receptors on host cell surfaces. In this study, we have cloned the cDNA fragment encoding the entire globular region (residues 101_57) of hemagglutinin of the H9N2 type avian influenza virus (A/ck/Korea/ms96/96). The protein segment (denoted as the H9 peptide), which was expressed and purified in E. coli, was used for the immunization of BALB/c mice to obtain the anti-H9 antiserum. To identify specific DNA aptamers with high affinity to H9 peptide, we conducted the SELEX method; 19 aptamers were newly isolated. A random mixture of these aptamers showed an increased level of binding affinity to the H9 peptide. The sequence alignment analysis of these aptamers revealed that 6 aptamers have highly conserved consensus sequences. Among these, aptamer C7 showed the highest similarity to the consensus sequences. Therefore, based on the C7 aptamer, we synthesized a new modified aptamer designated as C7-35M. This new aptamer showed strong binding capability to the viral particles. Furthermore, it could prevent MDCK cells from viral infection by strong binding to the viral particles. These results suggest that our aptamers can recognize the hemagglutinin protein of avian influenza virus and inhibit the binding of the virus to target receptors required for the penetration of host cells.