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Lactobacilli as a Vector for Delivery of Nanobodies against Norovirus Infection

Passive administration of neutralizing antibodies (Abs) is an attractive strategy for the control of gastrointestinal infections. However, an unanswered practical concern is the need to assure the stability of sufficient amounts of orally administered neutralizing Abs against intestinal pathogens (e...

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Autores principales: Yuki, Yoshikazu, Zuo, Fanglei, Kurokawa, Shiho, Uchida, Yohei, Sato, Shintaro, Sakon, Naomi, Hammarström, Lennart, Kiyono, Hiroshi, Marcotte, Harold
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9863548/
https://www.ncbi.nlm.nih.gov/pubmed/36678692
http://dx.doi.org/10.3390/pharmaceutics15010063
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author Yuki, Yoshikazu
Zuo, Fanglei
Kurokawa, Shiho
Uchida, Yohei
Sato, Shintaro
Sakon, Naomi
Hammarström, Lennart
Kiyono, Hiroshi
Marcotte, Harold
author_facet Yuki, Yoshikazu
Zuo, Fanglei
Kurokawa, Shiho
Uchida, Yohei
Sato, Shintaro
Sakon, Naomi
Hammarström, Lennart
Kiyono, Hiroshi
Marcotte, Harold
author_sort Yuki, Yoshikazu
collection PubMed
description Passive administration of neutralizing antibodies (Abs) is an attractive strategy for the control of gastrointestinal infections. However, an unanswered practical concern is the need to assure the stability of sufficient amounts of orally administered neutralizing Abs against intestinal pathogens (e.g., norovirus) in the harsh environment of the gastrointestinal tract. To this end, we expressed a single-domain Ab (VHH, nanobody) against norovirus on the cell surface of Lactobacillus, a natural and beneficial commensal component of the gut microbiome. First, we used intestinal epithelial cells generated from human induced pluripotent stem cells to confirm that VHH 1E4 showed neutralizing activity against GII.17 norovirus. We then expressed VHH 1E4 as a cell-wall–anchored form in Lactobacillus paracasei BL23. Flow cytometry confirmed the expression of VHH 1E4 on the surface of lactobacilli, and L. paracasei that expressed VHH 1E4 inhibited the replication of GII.17 norovirus in vitro. We then orally administered VHH 1E4-expressing L. paracasei BL23 to germ-free BALB/c mice and confirmed the presence of lactobacilli with neutralizing activity in the intestine for at least 10 days after administration. Thus, cell-wall-anchored VHH-displaying lactobacilli are attractive oral nanobody deliver vectors for passive immunization against norovirus infection.
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spelling pubmed-98635482023-01-22 Lactobacilli as a Vector for Delivery of Nanobodies against Norovirus Infection Yuki, Yoshikazu Zuo, Fanglei Kurokawa, Shiho Uchida, Yohei Sato, Shintaro Sakon, Naomi Hammarström, Lennart Kiyono, Hiroshi Marcotte, Harold Pharmaceutics Article Passive administration of neutralizing antibodies (Abs) is an attractive strategy for the control of gastrointestinal infections. However, an unanswered practical concern is the need to assure the stability of sufficient amounts of orally administered neutralizing Abs against intestinal pathogens (e.g., norovirus) in the harsh environment of the gastrointestinal tract. To this end, we expressed a single-domain Ab (VHH, nanobody) against norovirus on the cell surface of Lactobacillus, a natural and beneficial commensal component of the gut microbiome. First, we used intestinal epithelial cells generated from human induced pluripotent stem cells to confirm that VHH 1E4 showed neutralizing activity against GII.17 norovirus. We then expressed VHH 1E4 as a cell-wall–anchored form in Lactobacillus paracasei BL23. Flow cytometry confirmed the expression of VHH 1E4 on the surface of lactobacilli, and L. paracasei that expressed VHH 1E4 inhibited the replication of GII.17 norovirus in vitro. We then orally administered VHH 1E4-expressing L. paracasei BL23 to germ-free BALB/c mice and confirmed the presence of lactobacilli with neutralizing activity in the intestine for at least 10 days after administration. Thus, cell-wall-anchored VHH-displaying lactobacilli are attractive oral nanobody deliver vectors for passive immunization against norovirus infection. MDPI 2022-12-25 /pmc/articles/PMC9863548/ /pubmed/36678692 http://dx.doi.org/10.3390/pharmaceutics15010063 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yuki, Yoshikazu
Zuo, Fanglei
Kurokawa, Shiho
Uchida, Yohei
Sato, Shintaro
Sakon, Naomi
Hammarström, Lennart
Kiyono, Hiroshi
Marcotte, Harold
Lactobacilli as a Vector for Delivery of Nanobodies against Norovirus Infection
title Lactobacilli as a Vector for Delivery of Nanobodies against Norovirus Infection
title_full Lactobacilli as a Vector for Delivery of Nanobodies against Norovirus Infection
title_fullStr Lactobacilli as a Vector for Delivery of Nanobodies against Norovirus Infection
title_full_unstemmed Lactobacilli as a Vector for Delivery of Nanobodies against Norovirus Infection
title_short Lactobacilli as a Vector for Delivery of Nanobodies against Norovirus Infection
title_sort lactobacilli as a vector for delivery of nanobodies against norovirus infection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9863548/
https://www.ncbi.nlm.nih.gov/pubmed/36678692
http://dx.doi.org/10.3390/pharmaceutics15010063
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