Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens
Chicken coccidiosis caused by Eimeria spp. can occur on almost all poultry farms, causing huge economic losses to the industry. Genetically manipulated Eimeria parasites as a vaccine vector to deliver viral antigens have been reported. In our preliminary study, transgenic E. acervulina expressing a...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-04-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2024.1367912/full |
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| author | Qingbin Guo Qingbin Guo Ying Yu Jingxia Suo Xinming Tang Sixin Zhang Colin Crouch Beth Bruton Ian Tarpey Xianyong Liu Guanghui Zhao Xun Suo |
| author_facet | Qingbin Guo Qingbin Guo Ying Yu Jingxia Suo Xinming Tang Sixin Zhang Colin Crouch Beth Bruton Ian Tarpey Xianyong Liu Guanghui Zhao Xun Suo |
| author_sort | Qingbin Guo |
| collection | DOAJ |
| description | Chicken coccidiosis caused by Eimeria spp. can occur on almost all poultry farms, causing huge economic losses to the industry. Genetically manipulated Eimeria parasites as a vaccine vector to deliver viral antigens have been reported. In our preliminary study, transgenic E. acervulina expressing a VP2 gene (Ea-VP2) of the infectious bursal disease virus (IBDV) demonstrated partial protection against IBDV infection. To enhance immune responses, we aimed to increase the VP2 gene copy number in transgenic E. acervulina. In this study, we used a novel plasmid vector carrying a VP2 gene fused with three flag tags and a red fluorescent reporter gene (mCherry). The vector was introduced into Ea-VP2 sporozoites through nucleofection, leading to the generation of Ea-2VP2. Subsequent analysis revealed a notable escalation in the fluorescent rate, increasing from 0.11 to 95.1% following four consecutive passages facilitated by fluorescent-activated cell sorting. Verification via PCR, Western blot, and immunofluorescence confirmed the successful construction of the Ea-2VP2 population. Despite lower fecundity compared to wild-type E. acervulina, Ea-2VP2 maintained immunogenicity. Our research effectively created a transgenic E. acervulina strain transfected sequentially with two copies of the VP2 gene from IBDV. This modification resulted in an increased humoral immune response after primary immunization in chickens. Additionally, it demonstrated a degree of protection within the bursa against IBDV infection. Future studies will focus on further enhancing immune response levels. |
| first_indexed | 2024-04-24T11:26:40Z |
| format | Article |
| id | doaj.art-59c59319b78a4ba78c4e503fb48da786 |
| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-04-24T11:26:40Z |
| publishDate | 2024-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Veterinary Science |
| spelling | doaj.art-59c59319b78a4ba78c4e503fb48da7862024-04-10T13:40:28ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692024-04-011110.3389/fvets.2024.13679121367912Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickensQingbin Guo0Qingbin Guo1Ying Yu2Jingxia Suo3Xinming Tang4Sixin Zhang5Colin Crouch6Beth Bruton7Ian Tarpey8Xianyong Liu9Guanghui Zhao10Xun Suo11College of Veterinary Medicine, Northwest A&F University, Xianyang, ChinaNational Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing, ChinaNational Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing, ChinaNational Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing, ChinaKey Laboratory of Animal Biosafety Risk Prevention and Control (North) of MARA, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, ChinaNational Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing, ChinaMSD Animal Health, Milton Keynes, United KingdomMSD Animal Health, Milton Keynes, United KingdomMSD Animal Health, Milton Keynes, United KingdomNational Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing, ChinaCollege of Veterinary Medicine, Northwest A&F University, Xianyang, ChinaNational Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing, ChinaChicken coccidiosis caused by Eimeria spp. can occur on almost all poultry farms, causing huge economic losses to the industry. Genetically manipulated Eimeria parasites as a vaccine vector to deliver viral antigens have been reported. In our preliminary study, transgenic E. acervulina expressing a VP2 gene (Ea-VP2) of the infectious bursal disease virus (IBDV) demonstrated partial protection against IBDV infection. To enhance immune responses, we aimed to increase the VP2 gene copy number in transgenic E. acervulina. In this study, we used a novel plasmid vector carrying a VP2 gene fused with three flag tags and a red fluorescent reporter gene (mCherry). The vector was introduced into Ea-VP2 sporozoites through nucleofection, leading to the generation of Ea-2VP2. Subsequent analysis revealed a notable escalation in the fluorescent rate, increasing from 0.11 to 95.1% following four consecutive passages facilitated by fluorescent-activated cell sorting. Verification via PCR, Western blot, and immunofluorescence confirmed the successful construction of the Ea-2VP2 population. Despite lower fecundity compared to wild-type E. acervulina, Ea-2VP2 maintained immunogenicity. Our research effectively created a transgenic E. acervulina strain transfected sequentially with two copies of the VP2 gene from IBDV. This modification resulted in an increased humoral immune response after primary immunization in chickens. Additionally, it demonstrated a degree of protection within the bursa against IBDV infection. Future studies will focus on further enhancing immune response levels.https://www.frontiersin.org/articles/10.3389/fvets.2024.1367912/fullchickencoccidiadelivery vectorgenetic manipulationviral antigen |
| spellingShingle | Qingbin Guo Qingbin Guo Ying Yu Jingxia Suo Xinming Tang Sixin Zhang Colin Crouch Beth Bruton Ian Tarpey Xianyong Liu Guanghui Zhao Xun Suo Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens Frontiers in Veterinary Science chicken coccidia delivery vector genetic manipulation viral antigen |
| title | Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens |
| title_full | Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens |
| title_fullStr | Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens |
| title_full_unstemmed | Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens |
| title_short | Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens |
| title_sort | oral delivery of eimeria acervulina transfected sequentially with two copies of the vp2 gene induces immunity against infectious bursal disease virus in chickens |
| topic | chicken coccidia delivery vector genetic manipulation viral antigen |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2024.1367912/full |
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