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Human monoclonal ScFv that bind to different functional domains of M2 and inhibit H5N1 influenza virus replication

หน่วยงาน สถาบันวิจัยและให้คำปรึกษาแห่ง มหาวิทยาลัยธรรมศาสตร์

รายละเอียด

ชื่อเรื่อง : Human monoclonal ScFv that bind to different functional domains of M2 and inhibit H5N1 influenza virus replication
นักวิจัย : Tippawan Pissawong , Santi Maneewatch , Kanyarat Thueng-in , Potjanee Srimanote , Fonthip Dong-din-on , Jeeraphong Thanongsaksrikul , Thaweesak Songserm , Pongsri Tongtawe , Kunan Bangphoomi , Wanpen Chaicumpa
คำค้น : H5N1 , Human ScFv , Influenza virus , M2 protein , Mimotope , Molecular docking , Phage display , Quantitative real-time RT-PCR , Virus neutralization
หน่วยงาน : สถาบันวิจัยและให้คำปรึกษาแห่ง มหาวิทยาลัยธรรมศาสตร์
ผู้ร่วมงาน : -
ปีพิมพ์ : 2556
อ้างอิง : Virology journal. 10 (2013) Art. no. 148 , 1743-422X , http://dspace.library.tu.ac.th/handle/3517/6748
ที่มา : -
ความเชี่ยวชาญ : -
ความสัมพันธ์ : -
ขอบเขตของเนื้อหา : -
บทคัดย่อ/คำอธิบาย :

Background: Novel effective anti-influenza agent that tolerates influenza virus antigenic variation is needed. Highly conserved influenza virus M2 protein has multiple pivotal functions including ion channel activity for vRNP uncoating, anti-autophagy and virus assembly, morphogenesis and release. Thus, M2 is an attractive target of anti-influenza agents including small molecular drugs and specific antibodies. Methods. Fully human monoclonal single chain antibodies (HuScFv) specific to recombinant and native M2 proteins of A/H5N1 virus were produced from huscfv-phagemid transformed E. coli clones selected from a HuScFv phage display library using recombinant M2 of clade 1 A/H5N1 as panning antigen. The HuScFv were tested for their ability to inhibit replication of A/H5N1 of both homologous and heterologous clades. M2 domains bound by HuScFv of individual E. coli clones were identified by phage mimotope searching and computerized molecular docking. Results: HuScFv derived from four huscfv-phagemid transformed E. coli clones (no. 2, 19, 23 and 27) showed different amino acid sequences particularly at the CDRs. Cells infected with A/H5N1 influenza viruses (both adamantane sensitive and resistant) that had been exposed to the HuScFv had reduced virus release and intracellular virus. Phage peptide mimotope search and multiple alignments revealed that conformational epitopes of HuScFv2 located at the residues important for ion channel activity, anti-autophagy and M1 binding; epitopic residues of HuScFv19 located at the M2 amphipathic helix and cytoplasmic tail important for anti-autophagy, virus assembly, morphogenesis and release; epitope of HuScFv23 involved residues important for the M2 activities similar to HuScFv2 and also amphipathic helix residues for viral budding and release while HuScFv27 epitope spanned ectodomain, ion channel and anti-autophagy residues. Results of computerized homology modelling and molecular docking conformed to the epitope identification by phages. Conclusions: HuScFv that bound to highly conserved epitopes across influenza A subtypes and human pathogenic H5N1clades located on different functional domains of M2 were produced. The HuScFv reduced viral release and intracellular virus of infected cells. While the molecular mechanisms of the HuScFv await experimental validation, the small human antibody fragments have high potential for developing further as a safe, novel and mutation tolerable anti-influenza agent especially against drug resistant variants. © 2013 Pissawong et al.; licensee BioMed Central Ltd.

บรรณานุกรม :
Tippawan Pissawong , Santi Maneewatch , Kanyarat Thueng-in , Potjanee Srimanote , Fonthip Dong-din-on , Jeeraphong Thanongsaksrikul , Thaweesak Songserm , Pongsri Tongtawe , Kunan Bangphoomi , Wanpen Chaicumpa . (2556). Human monoclonal ScFv that bind to different functional domains of M2 and inhibit H5N1 influenza virus replication.
    กรุงเทพมหานคร : สถาบันวิจัยและให้คำปรึกษาแห่ง มหาวิทยาลัยธรรมศาสตร์ .
Tippawan Pissawong , Santi Maneewatch , Kanyarat Thueng-in , Potjanee Srimanote , Fonthip Dong-din-on , Jeeraphong Thanongsaksrikul , Thaweesak Songserm , Pongsri Tongtawe , Kunan Bangphoomi , Wanpen Chaicumpa . 2556. "Human monoclonal ScFv that bind to different functional domains of M2 and inhibit H5N1 influenza virus replication".
    กรุงเทพมหานคร : สถาบันวิจัยและให้คำปรึกษาแห่ง มหาวิทยาลัยธรรมศาสตร์ .
Tippawan Pissawong , Santi Maneewatch , Kanyarat Thueng-in , Potjanee Srimanote , Fonthip Dong-din-on , Jeeraphong Thanongsaksrikul , Thaweesak Songserm , Pongsri Tongtawe , Kunan Bangphoomi , Wanpen Chaicumpa . "Human monoclonal ScFv that bind to different functional domains of M2 and inhibit H5N1 influenza virus replication."
    กรุงเทพมหานคร : สถาบันวิจัยและให้คำปรึกษาแห่ง มหาวิทยาลัยธรรมศาสตร์ , 2556. Print.
Tippawan Pissawong , Santi Maneewatch , Kanyarat Thueng-in , Potjanee Srimanote , Fonthip Dong-din-on , Jeeraphong Thanongsaksrikul , Thaweesak Songserm , Pongsri Tongtawe , Kunan Bangphoomi , Wanpen Chaicumpa . Human monoclonal ScFv that bind to different functional domains of M2 and inhibit H5N1 influenza virus replication. กรุงเทพมหานคร : สถาบันวิจัยและให้คำปรึกษาแห่ง มหาวิทยาลัยธรรมศาสตร์ ; 2556.