Oral Delivery of <i>Lactococcus lactis</i> Expressing Full-Length S Protein via Alginate–Chitosan Capsules Induces Immune Protection Against PEDV Infection in Mice

<b>Background/Objectives</b>: Porcine epidemic diarrhea (PED) is a highly contagious enteric infectious disease that causes severe morbidity and mortality in piglets, posing significant economic losses to the swine industry worldwide. Oral vaccines based on <i>Lactococcus lactis<...

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Main Authors: Miaoyan Yang, Denglong Xie, Wei Ji, Shu Jeffrey Zhu, Yongqi Zhou
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Vaccines
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Online Access:https://www.mdpi.com/2076-393X/13/4/421
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author Miaoyan Yang
Denglong Xie
Wei Ji
Shu Jeffrey Zhu
Yongqi Zhou
author_facet Miaoyan Yang
Denglong Xie
Wei Ji
Shu Jeffrey Zhu
Yongqi Zhou
author_sort Miaoyan Yang
collection DOAJ
description <b>Background/Objectives</b>: Porcine epidemic diarrhea (PED) is a highly contagious enteric infectious disease that causes severe morbidity and mortality in piglets, posing significant economic losses to the swine industry worldwide. Oral vaccines based on <i>Lactococcus lactis</i> offer a promising approach due to their safety and genetic manipulability. This study aims to develop and evaluate an oral <i>L. lactis</i>-based vaccine expressing the full-length PEDV S protein. <b>Methods</b>: A recombinant <i>L. lactis</i> strain expressing the PEDV S protein was constructed and encapsulated in alginate–chitosan microcapsules. Vaccine stability was tested in simulated digestive fluids, and mice were orally immunized. Immune responses were evaluated by measuring specific antibodies, cytokines, and lymphocyte proliferation. <b>Results</b>: The recombinant <i>L. lactis</i> NZ3900/pNZ8149-S strain successfully expressed the full-length PEDV S protein and maintained stable plasmid inheritance. Oral immunization in mice induced detectable PEDV-specific immune responses. Both encapsulated and non-encapsulated vaccines stimulated the production of IgG and sIgA antibodies, as well as cytokines associated with Th1 and Th2 responses. Notably, encapsulation with alginate–chitosan significantly enhanced bacterial survival in digestive conditions and further amplified immune responses, including higher antibody titers, elevated levels of IFN-γ, IL-4, and IL-10, and greater lymphocyte proliferation, indicating improved immune memory. <b>Conclusions</b>: The oral <i>L. lactis</i> NZ3900/pNZ8149-S vaccine expressing the PEDV S protein effectively induced systemic and mucosal immunity in mice. Encapsulation with alginate–chitosan further enhanced its immunogenicity and stability in gastrointestinal conditions. These results suggest that both the engineered <i>L. lactis</i> strain and the encapsulation strategy contribute to the development of a promising oral vaccine platform for controlling PEDV in swine populations.
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spelling doaj-art-8d57258b5f594ca0b0ada8e0e2de2fc62025-08-20T02:18:10ZengMDPI AGVaccines2076-393X2025-04-0113442110.3390/vaccines13040421Oral Delivery of <i>Lactococcus lactis</i> Expressing Full-Length S Protein via Alginate–Chitosan Capsules Induces Immune Protection Against PEDV Infection in MiceMiaoyan Yang0Denglong Xie1Wei Ji2Shu Jeffrey Zhu3Yongqi Zhou4Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaZhejiang Hisun Animal Healthcare Products Co., Ltd., Hangzhou 311400, ChinaZhejiang Hisun Animal Healthcare Products Co., Ltd., Hangzhou 311400, ChinaDepartment of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaZhejiang Hisun Animal Healthcare Products Co., Ltd., Hangzhou 311400, China<b>Background/Objectives</b>: Porcine epidemic diarrhea (PED) is a highly contagious enteric infectious disease that causes severe morbidity and mortality in piglets, posing significant economic losses to the swine industry worldwide. Oral vaccines based on <i>Lactococcus lactis</i> offer a promising approach due to their safety and genetic manipulability. This study aims to develop and evaluate an oral <i>L. lactis</i>-based vaccine expressing the full-length PEDV S protein. <b>Methods</b>: A recombinant <i>L. lactis</i> strain expressing the PEDV S protein was constructed and encapsulated in alginate–chitosan microcapsules. Vaccine stability was tested in simulated digestive fluids, and mice were orally immunized. Immune responses were evaluated by measuring specific antibodies, cytokines, and lymphocyte proliferation. <b>Results</b>: The recombinant <i>L. lactis</i> NZ3900/pNZ8149-S strain successfully expressed the full-length PEDV S protein and maintained stable plasmid inheritance. Oral immunization in mice induced detectable PEDV-specific immune responses. Both encapsulated and non-encapsulated vaccines stimulated the production of IgG and sIgA antibodies, as well as cytokines associated with Th1 and Th2 responses. Notably, encapsulation with alginate–chitosan significantly enhanced bacterial survival in digestive conditions and further amplified immune responses, including higher antibody titers, elevated levels of IFN-γ, IL-4, and IL-10, and greater lymphocyte proliferation, indicating improved immune memory. <b>Conclusions</b>: The oral <i>L. lactis</i> NZ3900/pNZ8149-S vaccine expressing the PEDV S protein effectively induced systemic and mucosal immunity in mice. Encapsulation with alginate–chitosan further enhanced its immunogenicity and stability in gastrointestinal conditions. These results suggest that both the engineered <i>L. lactis</i> strain and the encapsulation strategy contribute to the development of a promising oral vaccine platform for controlling PEDV in swine populations.https://www.mdpi.com/2076-393X/13/4/421adjuvantimmune response<i>Lactococcus lactis</i>oral vaccinePEDV
spellingShingle Miaoyan Yang
Denglong Xie
Wei Ji
Shu Jeffrey Zhu
Yongqi Zhou
Oral Delivery of <i>Lactococcus lactis</i> Expressing Full-Length S Protein via Alginate–Chitosan Capsules Induces Immune Protection Against PEDV Infection in Mice
Vaccines
adjuvant
immune response
<i>Lactococcus lactis</i>
oral vaccine
PEDV
title Oral Delivery of <i>Lactococcus lactis</i> Expressing Full-Length S Protein via Alginate–Chitosan Capsules Induces Immune Protection Against PEDV Infection in Mice
title_full Oral Delivery of <i>Lactococcus lactis</i> Expressing Full-Length S Protein via Alginate–Chitosan Capsules Induces Immune Protection Against PEDV Infection in Mice
title_fullStr Oral Delivery of <i>Lactococcus lactis</i> Expressing Full-Length S Protein via Alginate–Chitosan Capsules Induces Immune Protection Against PEDV Infection in Mice
title_full_unstemmed Oral Delivery of <i>Lactococcus lactis</i> Expressing Full-Length S Protein via Alginate–Chitosan Capsules Induces Immune Protection Against PEDV Infection in Mice
title_short Oral Delivery of <i>Lactococcus lactis</i> Expressing Full-Length S Protein via Alginate–Chitosan Capsules Induces Immune Protection Against PEDV Infection in Mice
title_sort oral delivery of i lactococcus lactis i expressing full length s protein via alginate chitosan capsules induces immune protection against pedv infection in mice
topic adjuvant
immune response
<i>Lactococcus lactis</i>
oral vaccine
PEDV
url https://www.mdpi.com/2076-393X/13/4/421
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