Giuseppe
Emeritus
[Source: Antimicrobial Agents and Chemotherapy, full page: (LINK). Abstract, edited.]
Targeted siRNA-immunoliposomes as a promising therapeutic agent against highly pathogenic avian influenza A (H5N1) virus infection
Kannika Khantasup a,b, Phikulthong Kopermsub b, Kridsada Chaichoun c and Tararaj Dharakul a,b*
Author Affiliations: <SUP>a</SUP>Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand <SUP>b</SUP>National Nanotechnology Center, National Science and Technology Development Agency, 111 Thailand Science Park, Pathum Thani 12120, Thailand <SUP>c</SUP>Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand
ABSTRACT
This study describes a proof of concept study on the use of siRNA-immunoliposomes as a therapeutic agent against H5N1 influenza virus infection. SiRNA specific for influenza virus nucleoprotein (NP) mRNA was employed as the key antiviral agent to inhibit viral replication in this study. A humanized single-chain Fv antibody (huscFv) against the hemagglutinin (HA) of H5N1 highly pathogenic avian influenza (HPAI) was used as the targeting molecule to HA of H5N1 virus, which is abundantly expressed on the surface of infected cells. The huscFv was decorated onto cationic PEGylated DC-Chol/DOPE liposomes to generate immunoliposomes for siRNA delivery. The immunoliposomes were shown to specifically bind HA-expressing Sf9 cells and demonstrated enhanced siRNA transfection efficiency. The siRNA transfection efficiency was significantly reduced after pre-incubation of the HA target cells with an excess amount of free huscFv. These results therefore demonstrated that the enhanced siRNA delivery of immunoliposomes was mediated via targeting by huscFv. Furthermore, the siRNA silencing effect was more pronounced when the immunoliposomes were administered 6-12 h post H5N1 infection in MDCK cells, compared with the non-targeted liposomes. This proof of concept study may contribute to the future design and development of a siRNA delivery system for combating viral infectious diseases in humans.
FOOTNOTES
*Corresponding author. Address: Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand, Tel.: +662419700x6682; fax: +6624181626, E-mail address:sitdr@mahidol.ac.th
Copyright ? 2014, American Society for Microbiology. All Rights Reserved.
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Targeted siRNA-immunoliposomes as a promising therapeutic agent against highly pathogenic avian influenza A (H5N1) virus infection
Kannika Khantasup a,b, Phikulthong Kopermsub b, Kridsada Chaichoun c and Tararaj Dharakul a,b*
Author Affiliations: <SUP>a</SUP>Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand <SUP>b</SUP>National Nanotechnology Center, National Science and Technology Development Agency, 111 Thailand Science Park, Pathum Thani 12120, Thailand <SUP>c</SUP>Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand
ABSTRACT
This study describes a proof of concept study on the use of siRNA-immunoliposomes as a therapeutic agent against H5N1 influenza virus infection. SiRNA specific for influenza virus nucleoprotein (NP) mRNA was employed as the key antiviral agent to inhibit viral replication in this study. A humanized single-chain Fv antibody (huscFv) against the hemagglutinin (HA) of H5N1 highly pathogenic avian influenza (HPAI) was used as the targeting molecule to HA of H5N1 virus, which is abundantly expressed on the surface of infected cells. The huscFv was decorated onto cationic PEGylated DC-Chol/DOPE liposomes to generate immunoliposomes for siRNA delivery. The immunoliposomes were shown to specifically bind HA-expressing Sf9 cells and demonstrated enhanced siRNA transfection efficiency. The siRNA transfection efficiency was significantly reduced after pre-incubation of the HA target cells with an excess amount of free huscFv. These results therefore demonstrated that the enhanced siRNA delivery of immunoliposomes was mediated via targeting by huscFv. Furthermore, the siRNA silencing effect was more pronounced when the immunoliposomes were administered 6-12 h post H5N1 infection in MDCK cells, compared with the non-targeted liposomes. This proof of concept study may contribute to the future design and development of a siRNA delivery system for combating viral infectious diseases in humans.
FOOTNOTES
*Corresponding author. Address: Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand, Tel.: +662419700x6682; fax: +6624181626, E-mail address:sitdr@mahidol.ac.th
Copyright ? 2014, American Society for Microbiology. All Rights Reserved.
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