The Generation of a H9N2 Avian Influenza Virus with HA and C3d-P29 Protein Fusions and Vaccine Development Applications

Background: Maternal-derived antibody (MDA) interferes with immune responses, leading to the failure of H9N2 avian influenza vaccinations in poultry. So far, none of the commercially available H9N2 avian influenza vaccines used in poultry have been able to overcome MDA interference. Methods: To deve...

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Main Authors: Xue Pan, Fan Zhou, Xiaona Shi, Qinfang Liu, Dawei Yan, Qiaoyang Teng, Chunxiu Yuan, Bangfeng Xu, Zhifei Zhang, Minghao Yan, Zejun Li
Format: Article
Language:English
Published: MDPI AG 2025-01-01
Series:Vaccines
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Online Access:https://www.mdpi.com/2076-393X/13/2/99
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author Xue Pan
Fan Zhou
Xiaona Shi
Qinfang Liu
Dawei Yan
Qiaoyang Teng
Chunxiu Yuan
Bangfeng Xu
Zhifei Zhang
Minghao Yan
Zejun Li
author_facet Xue Pan
Fan Zhou
Xiaona Shi
Qinfang Liu
Dawei Yan
Qiaoyang Teng
Chunxiu Yuan
Bangfeng Xu
Zhifei Zhang
Minghao Yan
Zejun Li
author_sort Xue Pan
collection DOAJ
description Background: Maternal-derived antibody (MDA) interferes with immune responses, leading to the failure of H9N2 avian influenza vaccinations in poultry. So far, none of the commercially available H9N2 avian influenza vaccines used in poultry have been able to overcome MDA interference. Methods: To develop a vaccine that can overcome MDA interference, one or multiple copies of the minimum-binding domain (P29) from the complement protein C3d were inserted in between the signal peptide and the head domain of the hemagglutinin (HA) protein on a H9N2 avian influenza virus (A/Chicken/Shanghai/H514/2017, named H514). Results: The HA proteins containing P29 stimulated stronger type I interferences than wild-type HA proteins in vitro. The modified viruses with the HA proteins containing one copy of P29 (rH514-P29.1) and two copies of P29.2 (rH514-P29.2) were successfully rescued using reverse genetics. The inactivated vaccines developed with rH514-P29.1 or rH514-P29.2 induced higher and faster humoral immune responses than the vaccine developed with rH514 in specific pathogen-free (SPF) chickens. To evaluate the vaccines’ efficacy in the presence of MDA and to ensure a uniform level of MDA, passively transferred antibody (PTA) was used as a model to mimic MDA in 1-day-old SPF chickens. Our results showed that the rH514-P29.2 inactivated vaccine induced significantly higher HI titers than the rH514 inactivated vaccine in the presence of PTA. More importantly, it reduced viral shedding after being challenged with H514 in the presence of PTA. Conclusions: Our results suggest that vaccine antigens fused with two copies of P29 can decrease the interference of MDA on immunity in chickens. Overall, our results provide a new strategy for overcoming MDA interference.
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spelling doaj-art-55a04bb368484bf4a933f7e6080a59432025-08-20T02:45:30ZengMDPI AGVaccines2076-393X2025-01-011329910.3390/vaccines13020099The Generation of a H9N2 Avian Influenza Virus with HA and C3d-P29 Protein Fusions and Vaccine Development ApplicationsXue Pan0Fan Zhou1Xiaona Shi2Qinfang Liu3Dawei Yan4Qiaoyang Teng5Chunxiu Yuan6Bangfeng Xu7Zhifei Zhang8Minghao Yan9Zejun Li10Shanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, ChinaBackground: Maternal-derived antibody (MDA) interferes with immune responses, leading to the failure of H9N2 avian influenza vaccinations in poultry. So far, none of the commercially available H9N2 avian influenza vaccines used in poultry have been able to overcome MDA interference. Methods: To develop a vaccine that can overcome MDA interference, one or multiple copies of the minimum-binding domain (P29) from the complement protein C3d were inserted in between the signal peptide and the head domain of the hemagglutinin (HA) protein on a H9N2 avian influenza virus (A/Chicken/Shanghai/H514/2017, named H514). Results: The HA proteins containing P29 stimulated stronger type I interferences than wild-type HA proteins in vitro. The modified viruses with the HA proteins containing one copy of P29 (rH514-P29.1) and two copies of P29.2 (rH514-P29.2) were successfully rescued using reverse genetics. The inactivated vaccines developed with rH514-P29.1 or rH514-P29.2 induced higher and faster humoral immune responses than the vaccine developed with rH514 in specific pathogen-free (SPF) chickens. To evaluate the vaccines’ efficacy in the presence of MDA and to ensure a uniform level of MDA, passively transferred antibody (PTA) was used as a model to mimic MDA in 1-day-old SPF chickens. Our results showed that the rH514-P29.2 inactivated vaccine induced significantly higher HI titers than the rH514 inactivated vaccine in the presence of PTA. More importantly, it reduced viral shedding after being challenged with H514 in the presence of PTA. Conclusions: Our results suggest that vaccine antigens fused with two copies of P29 can decrease the interference of MDA on immunity in chickens. Overall, our results provide a new strategy for overcoming MDA interference.https://www.mdpi.com/2076-393X/13/2/99maternal-derived antibodyvaccineH9N2 avian influenza virushemagglutininscomplement
spellingShingle Xue Pan
Fan Zhou
Xiaona Shi
Qinfang Liu
Dawei Yan
Qiaoyang Teng
Chunxiu Yuan
Bangfeng Xu
Zhifei Zhang
Minghao Yan
Zejun Li
The Generation of a H9N2 Avian Influenza Virus with HA and C3d-P29 Protein Fusions and Vaccine Development Applications
Vaccines
maternal-derived antibody
vaccine
H9N2 avian influenza virus
hemagglutinins
complement
title The Generation of a H9N2 Avian Influenza Virus with HA and C3d-P29 Protein Fusions and Vaccine Development Applications
title_full The Generation of a H9N2 Avian Influenza Virus with HA and C3d-P29 Protein Fusions and Vaccine Development Applications
title_fullStr The Generation of a H9N2 Avian Influenza Virus with HA and C3d-P29 Protein Fusions and Vaccine Development Applications
title_full_unstemmed The Generation of a H9N2 Avian Influenza Virus with HA and C3d-P29 Protein Fusions and Vaccine Development Applications
title_short The Generation of a H9N2 Avian Influenza Virus with HA and C3d-P29 Protein Fusions and Vaccine Development Applications
title_sort generation of a h9n2 avian influenza virus with ha and c3d p29 protein fusions and vaccine development applications
topic maternal-derived antibody
vaccine
H9N2 avian influenza virus
hemagglutinins
complement
url https://www.mdpi.com/2076-393X/13/2/99
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