title: Generation of a safe and effective live viral vaccine by virus self-attenuation using species-specific artificial microRNA.
abstract: Vaccination with live attenuated vaccines (LAVs) is an effective way for prevention of infectious disease. While several methods are employed to create them, efficacy and safety are still a challenge. In this study, we evaluated the feasibility of creating a self-attenuated RNA virus expressing a functional species-specific artificial microRNA. Using influenza virus as a model, we produced an attenuated virus carrying a mammalian-specific miR-93 expression cassette that expresses a viral nucleoprotein (NP)-specific artificial microRNA from an insertion site within the non-structural (NS) gene segment. The resulting engineered live-attenuated influenza virus, PR8-amiR-93NP, produced mature and functional artificial microRNA against NP in mammalian cells, but not in avian cells. Furthermore, PR8-amiR-93NP was attenuated by 10(4) fold in mice compared with its wild-type counterpart. Importantly, intranasal immunization with PR8-amiR-93NP conferred cross-protective immunity against heterologous influenza virus strains. In short, this method provides a safe and effective platform for creation of live attenuated RNA viral vaccines.
authors: Center of Excellence for Infectious Diseases, Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, 5001 El Paso Drive, El Paso, TX 79905, USA.
authors: Junwei
authors: J
authors: Li
authors: Center of Excellence for Infectious Diseases, Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, 5001 El Paso Drive, El Paso, TX 79905, USA.
authors: Maria T
authors: MT
authors: Arévalo
authors: Center of Excellence for Infectious Diseases, Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, 5001 El Paso Drive, El Paso, TX 79905, USA.
authors: Diana
authors: D
authors: Diaz-Arévalo
authors: Center of Excellence for Infectious Diseases, Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, 5001 El Paso Drive, El Paso, TX 79905, USA.
authors: Yanping
authors: Y
authors: Chen
authors: Center of Excellence for Infectious Diseases, Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, 5001 El Paso Drive, El Paso, TX 79905, USA.
authors: Jang-Gi
authors: JG
authors: Choi
authors: Center of Excellence for Infectious Diseases, Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, 5001 El Paso Drive, El Paso, TX 79905, USA. Electronic address: mt.zeng@ttuhsc.edu.
authors: Mingtao
authors: M
authors: Zeng
mesh_terms: D000281:Administration, Intranasal; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015415:Biomarkers / Q000097:blood; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D004285:Dogs; D005240:Feasibility Studies; D057809:HEK293 Cells; D006801:Humans; D056724:Immunity, Humoral / Q000187:drug effects; D007114:Immunization; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000276:immunology; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000276:immunology; D009976:Orthomyxoviridae Infections / Q000097:blood / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D012328:RNA Viruses / Q000235:genetics* / Q000276:immunology; D012367:RNA, Viral / Q000235:genetics* / Q000276:immunology; D013997:Time Factors; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D015415:Biomarkers; D007252:Influenza Vaccines; D035683:MicroRNAs; D012367:RNA, Viral; D014613:Vaccines, Attenuated; D017361:Viral Nonstructural Proteins
keywords: Artificial microRNA; Cross-protection; Influenza virus; Live attenuated vaccine; Safety