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Intranasal Administration of Integrase Defective Lentiviral Vectors Expressing mAbs Protects from H5 Influenza Virus Challenge In Vivo

BACKGROUND: Despite medical advances, we are often unable to rapidly protect non-immune populations from infectious agents. Passive immunotherapy is a fast method of protection, but large-scale administration of monoclonal antibodies (mAbs) in unpractical. The delivery of mAbs using a viral vector c...

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Autores principales: Michelini, Zuleika, Fontana, Judith, Yang, Jianjun, Negri, Donatella, Cara, Andrea, Salvatore, Mirella
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7107124/
http://dx.doi.org/10.1093/ofid/ofx163.1355
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author Michelini, Zuleika
Fontana, Judith
Yang, Jianjun
Negri, Donatella
Cara, Andrea
Salvatore, Mirella
author_facet Michelini, Zuleika
Fontana, Judith
Yang, Jianjun
Negri, Donatella
Cara, Andrea
Salvatore, Mirella
author_sort Michelini, Zuleika
collection PubMed
description BACKGROUND: Despite medical advances, we are often unable to rapidly protect non-immune populations from infectious agents. Passive immunotherapy is a fast method of protection, but large-scale administration of monoclonal antibodies (mAbs) in unpractical. The delivery of mAbs using a viral vector can be an attractive alternative to direct mAbs injection. Integrase-defective lentiviral vectors (IDLV) have several advantages including the absence of pre-existing anti-vector immunity and the safety features of non-integration and non-replication. IDLV are maintained in non-dividing cells, and can express steady levels of functional proteins in vivo. We engineered IDLV to express mAbs against the influenza A virus (IAV) hemagglutinin, and tested their ability to protect from IAV in vivo. METHODS: IDLV were produced by co-transfection of transfer, packaging, and envelope plasmids in 293T cells and purification on sucrose gradients. IDLV were normalized using a colorimetric reverse transcriptase assay. Plasmid expressing mAb VN04-2 was provided by B. Hanson. mAb in the supernatant of transduced cells were detected by western blot and quantified by the Easy-Titer Human IgG Assay Kit. For in vivo studies, groups of 6–8 weeks old mice received IDLV either by intranasal (in) or intramuscular (im) route. mAb production was detected by western blot and ELISA. Mice were challenged using the recombinant IAV VNH5N1-PR8/CDC-RG derived from IAV A/Vietnam/1203/2004. RESULTS: We engineered IDLV producing the humanized mAb VN04-2 (IDLV-VN4-2), which is broadly neutralizing against H5 IAV. We found that after transduction of 293T cell with different dosages IDLV-VN4-2, the production of mAb was time and dose dependent. mAb were also functional, and bind specifically H5 HA but not other IAV proteins. We also measured VN04-2 production in the serum of mice 3, 6, 9, 14, 21 and 30 days after in or im administration of IDLV-VN4-2. We found that levels of mAb were sustained. In separate experiments 5/5 mice receiving IDLV-VN4-2 by the in route and 2/5 mice receiving it by the im route were protected from lethal IAV challenge. CONCLUSION: Our data suggest that IDLV may represent an attractive candidate for vector-mediated immunization against infectious disease. DISCLOSURES: All authors: No reported disclosures.
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spelling pubmed-71071242020-04-02 Intranasal Administration of Integrase Defective Lentiviral Vectors Expressing mAbs Protects from H5 Influenza Virus Challenge In Vivo Michelini, Zuleika Fontana, Judith Yang, Jianjun Negri, Donatella Cara, Andrea Salvatore, Mirella Open Forum Infect Dis Abstracts BACKGROUND: Despite medical advances, we are often unable to rapidly protect non-immune populations from infectious agents. Passive immunotherapy is a fast method of protection, but large-scale administration of monoclonal antibodies (mAbs) in unpractical. The delivery of mAbs using a viral vector can be an attractive alternative to direct mAbs injection. Integrase-defective lentiviral vectors (IDLV) have several advantages including the absence of pre-existing anti-vector immunity and the safety features of non-integration and non-replication. IDLV are maintained in non-dividing cells, and can express steady levels of functional proteins in vivo. We engineered IDLV to express mAbs against the influenza A virus (IAV) hemagglutinin, and tested their ability to protect from IAV in vivo. METHODS: IDLV were produced by co-transfection of transfer, packaging, and envelope plasmids in 293T cells and purification on sucrose gradients. IDLV were normalized using a colorimetric reverse transcriptase assay. Plasmid expressing mAb VN04-2 was provided by B. Hanson. mAb in the supernatant of transduced cells were detected by western blot and quantified by the Easy-Titer Human IgG Assay Kit. For in vivo studies, groups of 6–8 weeks old mice received IDLV either by intranasal (in) or intramuscular (im) route. mAb production was detected by western blot and ELISA. Mice were challenged using the recombinant IAV VNH5N1-PR8/CDC-RG derived from IAV A/Vietnam/1203/2004. RESULTS: We engineered IDLV producing the humanized mAb VN04-2 (IDLV-VN4-2), which is broadly neutralizing against H5 IAV. We found that after transduction of 293T cell with different dosages IDLV-VN4-2, the production of mAb was time and dose dependent. mAb were also functional, and bind specifically H5 HA but not other IAV proteins. We also measured VN04-2 production in the serum of mice 3, 6, 9, 14, 21 and 30 days after in or im administration of IDLV-VN4-2. We found that levels of mAb were sustained. In separate experiments 5/5 mice receiving IDLV-VN4-2 by the in route and 2/5 mice receiving it by the im route were protected from lethal IAV challenge. CONCLUSION: Our data suggest that IDLV may represent an attractive candidate for vector-mediated immunization against infectious disease. DISCLOSURES: All authors: No reported disclosures. Oxford University Press 2017-10-04 /pmc/articles/PMC7107124/ http://dx.doi.org/10.1093/ofid/ofx163.1355 Text en © The Author 2017. Published by Oxford University Press on behalf of Infectious Diseases Society of America. http://creativecommons.org/licenses/by-nc-nd/4.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Abstracts
Michelini, Zuleika
Fontana, Judith
Yang, Jianjun
Negri, Donatella
Cara, Andrea
Salvatore, Mirella
Intranasal Administration of Integrase Defective Lentiviral Vectors Expressing mAbs Protects from H5 Influenza Virus Challenge In Vivo
title Intranasal Administration of Integrase Defective Lentiviral Vectors Expressing mAbs Protects from H5 Influenza Virus Challenge In Vivo
title_full Intranasal Administration of Integrase Defective Lentiviral Vectors Expressing mAbs Protects from H5 Influenza Virus Challenge In Vivo
title_fullStr Intranasal Administration of Integrase Defective Lentiviral Vectors Expressing mAbs Protects from H5 Influenza Virus Challenge In Vivo
title_full_unstemmed Intranasal Administration of Integrase Defective Lentiviral Vectors Expressing mAbs Protects from H5 Influenza Virus Challenge In Vivo
title_short Intranasal Administration of Integrase Defective Lentiviral Vectors Expressing mAbs Protects from H5 Influenza Virus Challenge In Vivo
title_sort intranasal administration of integrase defective lentiviral vectors expressing mabs protects from h5 influenza virus challenge in vivo
topic Abstracts
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7107124/
http://dx.doi.org/10.1093/ofid/ofx163.1355
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