Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea.
Group A Rotavirus (RVA) is the leading cause of severe diarrhea in children. The aims of the present study were to determine the neutralizing activity of VP6-specific llama-derived single domain nanoantibodies (VHH nanoAbs) against different RVA strains in vitro and to evaluate the ability of G6P[1]...
| Main Authors: | , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
|
| Series: | PLoS Pathogens |
| Online Access: | http://europepmc.org/articles/PMC3642062?pdf=render |
| _version_ | 1818981442867167232 |
|---|---|
| author | Celina G Vega Marina Bok Anastasia N Vlasova Kuldeep S Chattha Silvia Gómez-Sebastián Carmen Nuñez Carmen Alvarado Rodrigo Lasa José M Escribano Lorena L Garaicoechea Fernando Fernandez Karin Bok Andrés Wigdorovitz Linda J Saif Viviana Parreño |
| author_facet | Celina G Vega Marina Bok Anastasia N Vlasova Kuldeep S Chattha Silvia Gómez-Sebastián Carmen Nuñez Carmen Alvarado Rodrigo Lasa José M Escribano Lorena L Garaicoechea Fernando Fernandez Karin Bok Andrés Wigdorovitz Linda J Saif Viviana Parreño |
| author_sort | Celina G Vega |
| collection | DOAJ |
| description | Group A Rotavirus (RVA) is the leading cause of severe diarrhea in children. The aims of the present study were to determine the neutralizing activity of VP6-specific llama-derived single domain nanoantibodies (VHH nanoAbs) against different RVA strains in vitro and to evaluate the ability of G6P[1] VP6-specific llama-derived single domain nanoantibodies (VHH) to protect against human rotavirus in gnotobiotic (Gn) piglets experimentally inoculated with virulent Wa G1P[8] rotavirus. Supplementation of the daily milk diet with 3B2 VHH clone produced using a baculovirus vector expression system (final ELISA antibody -Ab- titer of 4096; virus neutralization -VN- titer of 256) for 9 days conferred full protection against rotavirus associated diarrhea and significantly reduced virus shedding. The administration of comparable levels of porcine IgG Abs only protected 4 out of 6 of the animals from human RVA diarrhea but significantly reduced virus shedding. In contrast, G6P[1]-VP6 rotavirus-specific IgY Abs purified from eggs of hyperimmunized hens failed to protect piglets against human RVA-induced diarrhea or virus shedding when administering similar quantities of Abs. The oral administration of VHH nanoAb neither interfered with the host's isotype profiles of the Ab secreting cell responses to rotavirus, nor induced detectable host Ab responses to the treatment in serum or intestinal contents. This study shows that the oral administration of rotavirus VP6-VHH nanoAb is a broadly reactive and effective treatment against rotavirus-induced diarrhea in neonatal pigs. Our findings highlight the potential value of a broad neutralizing VP6-specific VHH nanoAb as a treatment that can complement or be used as an alternative to the current strain-specific RVA vaccines. Nanobodies could also be scaled-up to develop pediatric medication or functional food like infant milk formulas that might help treat RVA diarrhea. |
| first_indexed | 2024-12-20T17:31:24Z |
| format | Article |
| id | doaj.art-4d85d24c4607410ba310e98165981f63 |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-20T17:31:24Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-4d85d24c4607410ba310e98165981f632022-12-21T19:31:22ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742013-01-0195e100333410.1371/journal.ppat.1003334Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea.Celina G VegaMarina BokAnastasia N VlasovaKuldeep S ChatthaSilvia Gómez-SebastiánCarmen NuñezCarmen AlvaradoRodrigo LasaJosé M EscribanoLorena L GaraicoecheaFernando FernandezKarin BokAndrés WigdorovitzLinda J SaifViviana ParreñoGroup A Rotavirus (RVA) is the leading cause of severe diarrhea in children. The aims of the present study were to determine the neutralizing activity of VP6-specific llama-derived single domain nanoantibodies (VHH nanoAbs) against different RVA strains in vitro and to evaluate the ability of G6P[1] VP6-specific llama-derived single domain nanoantibodies (VHH) to protect against human rotavirus in gnotobiotic (Gn) piglets experimentally inoculated with virulent Wa G1P[8] rotavirus. Supplementation of the daily milk diet with 3B2 VHH clone produced using a baculovirus vector expression system (final ELISA antibody -Ab- titer of 4096; virus neutralization -VN- titer of 256) for 9 days conferred full protection against rotavirus associated diarrhea and significantly reduced virus shedding. The administration of comparable levels of porcine IgG Abs only protected 4 out of 6 of the animals from human RVA diarrhea but significantly reduced virus shedding. In contrast, G6P[1]-VP6 rotavirus-specific IgY Abs purified from eggs of hyperimmunized hens failed to protect piglets against human RVA-induced diarrhea or virus shedding when administering similar quantities of Abs. The oral administration of VHH nanoAb neither interfered with the host's isotype profiles of the Ab secreting cell responses to rotavirus, nor induced detectable host Ab responses to the treatment in serum or intestinal contents. This study shows that the oral administration of rotavirus VP6-VHH nanoAb is a broadly reactive and effective treatment against rotavirus-induced diarrhea in neonatal pigs. Our findings highlight the potential value of a broad neutralizing VP6-specific VHH nanoAb as a treatment that can complement or be used as an alternative to the current strain-specific RVA vaccines. Nanobodies could also be scaled-up to develop pediatric medication or functional food like infant milk formulas that might help treat RVA diarrhea.http://europepmc.org/articles/PMC3642062?pdf=render |
| spellingShingle | Celina G Vega Marina Bok Anastasia N Vlasova Kuldeep S Chattha Silvia Gómez-Sebastián Carmen Nuñez Carmen Alvarado Rodrigo Lasa José M Escribano Lorena L Garaicoechea Fernando Fernandez Karin Bok Andrés Wigdorovitz Linda J Saif Viviana Parreño Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea. PLoS Pathogens |
| title | Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea. |
| title_full | Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea. |
| title_fullStr | Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea. |
| title_full_unstemmed | Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea. |
| title_short | Recombinant monovalent llama-derived antibody fragments (VHH) to rotavirus VP6 protect neonatal gnotobiotic piglets against human rotavirus-induced diarrhea. |
| title_sort | recombinant monovalent llama derived antibody fragments vhh to rotavirus vp6 protect neonatal gnotobiotic piglets against human rotavirus induced diarrhea |
| url | http://europepmc.org/articles/PMC3642062?pdf=render |
| work_keys_str_mv | AT celinagvega recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT marinabok recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT anastasianvlasova recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT kuldeepschattha recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT silviagomezsebastian recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT carmennunez recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT carmenalvarado recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT rodrigolasa recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT josemescribano recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT lorenalgaraicoechea recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT fernandofernandez recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT karinbok recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT andreswigdorovitz recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT lindajsaif recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea AT vivianaparreno recombinantmonovalentllamaderivedantibodyfragmentsvhhtorotavirusvp6protectneonatalgnotobioticpigletsagainsthumanrotavirusinduceddiarrhea |