Development of a Ferritin-Based Nano-Allergen and Its Immunological Effects in vitro and in vivo

Dongmei Zhou,1,* Yaning Ren,1,* Ying Zhou,2 Yuanfen Liao,1 Qi Cheng,1 Jinni Chen,3 Cunyin Yuan,1 Dan Zeng,4 Yubao Cui1 1Clinical Research Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Wuxi, 214023, People’s Republic of China; 2Department of Pedi...

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Main Authors: Zhou D, Ren Y, Zhou Y, Liao Y, Cheng Q, Chen J, Yuan C, Zeng D, Cui Y
Format: Article
Language:English
Published: Dove Medical Press 2025-07-01
Series:International Journal of Nanomedicine
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Online Access:https://www.dovepress.com/development-of-a-ferritin-based-nano-allergen-and-its-immunological-ef-peer-reviewed-fulltext-article-IJN
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author Zhou D
Ren Y
Zhou Y
Liao Y
Cheng Q
Chen J
Yuan C
Zeng D
Cui Y
author_facet Zhou D
Ren Y
Zhou Y
Liao Y
Cheng Q
Chen J
Yuan C
Zeng D
Cui Y
author_sort Zhou D
collection DOAJ
description Dongmei Zhou,1,* Yaning Ren,1,* Ying Zhou,2 Yuanfen Liao,1 Qi Cheng,1 Jinni Chen,3 Cunyin Yuan,1 Dan Zeng,4 Yubao Cui1 1Clinical Research Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Wuxi, 214023, People’s Republic of China; 2Department of Pediatrics Laboratory, The Affiliated Children´s Hospital of Jiangnan University, Wuxi, 214023, People’s Republic of China; 3Department of Respiratory, Hainan Women and Children’s Medical Center, Affiliated Pediatrics Clinical College of Hainan Medical University, Haikou, 570100, People’s Republic of China; 4Department of Allergy, Chongqing General Hospital, Chongqing University, Chongqing, 401147, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yubao Cui; Dan Zeng, Email ybcui1975@hotmail.com; zengdan@cqu.edu.cnIntroduction: We aimed to develop recombinant allergens on the nanoscale by using self-assembly property of ferritin and to investigate their immune response and protective effect.Methods: The cDNA encoding Ftn-Blo t 2 was synthesized and cloned into pET-21a (+) vector, then expressed in E. coli BL21 (DE3). Recombinant proteins (rBlo t 2, rFtn, and rFtn-Blo t 2) were purified and their immunoreactivity confirmed for immunoreactivity by IgE-ELISA and Western blot. In vitro, the immunogenicity of rFtn-Blo t 2 was assessed using BEAS-2B human bronchial epithelial cells by measuring epithelial cytokine responses. In vivo, a murine airway inflammation model was established via sensitization and challenge with crude B. tropicalis extract, followed by treatment with rFtn-Blo t 2 to investigate its effects on airway inflammation, Th1/Th2 cytokine profiles, and allergen-specific antibody production.Results: The average diameter of rFtn and rFtn-Blo t 2 particles are (11.24 ± 1.31) nm and (16.00 ± 1.59) nm. The IgE- binding rates of rFtn-Blo t 2 and rBlo t 2 were 62.5% (15/24) and 58.3% (12/24), respectively. The expressions of IL-25, IL-33, and TSLP increased by 3.29, 3.43, and 2.22 folds after the addition of rFtn-Blo t 2 in co-culture with BEAS-2B. Mice challenged with rBlo t 2 had lower levels of cytokines IL-13 and IL-4 but higher levels of IFN-γ and TGF-β in alveolar lavage fluid compared to controls (p 
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series International Journal of Nanomedicine
spelling doaj-art-77b06f65c3aa49af9ce9da70cc94fa4a2025-08-20T03:16:15ZengDove Medical PressInternational Journal of Nanomedicine1178-20132025-07-01Volume 20Issue 195059516105295Development of a Ferritin-Based Nano-Allergen and Its Immunological Effects in vitro and in vivoZhou D0Ren YZhou Y1Liao Y2Cheng Q3Chen J4Yuan C5Zeng D6Cui Y7Clinical Research CenterDepartment of Pediatrics LaboratoryClinical Research CenterClinical Research CenterDepartment of RespirologyClinical Research CenterDepartment of AllergyClinical Research CenterDongmei Zhou,1,* Yaning Ren,1,* Ying Zhou,2 Yuanfen Liao,1 Qi Cheng,1 Jinni Chen,3 Cunyin Yuan,1 Dan Zeng,4 Yubao Cui1 1Clinical Research Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Wuxi, 214023, People’s Republic of China; 2Department of Pediatrics Laboratory, The Affiliated Children´s Hospital of Jiangnan University, Wuxi, 214023, People’s Republic of China; 3Department of Respiratory, Hainan Women and Children’s Medical Center, Affiliated Pediatrics Clinical College of Hainan Medical University, Haikou, 570100, People’s Republic of China; 4Department of Allergy, Chongqing General Hospital, Chongqing University, Chongqing, 401147, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yubao Cui; Dan Zeng, Email ybcui1975@hotmail.com; zengdan@cqu.edu.cnIntroduction: We aimed to develop recombinant allergens on the nanoscale by using self-assembly property of ferritin and to investigate their immune response and protective effect.Methods: The cDNA encoding Ftn-Blo t 2 was synthesized and cloned into pET-21a (+) vector, then expressed in E. coli BL21 (DE3). Recombinant proteins (rBlo t 2, rFtn, and rFtn-Blo t 2) were purified and their immunoreactivity confirmed for immunoreactivity by IgE-ELISA and Western blot. In vitro, the immunogenicity of rFtn-Blo t 2 was assessed using BEAS-2B human bronchial epithelial cells by measuring epithelial cytokine responses. In vivo, a murine airway inflammation model was established via sensitization and challenge with crude B. tropicalis extract, followed by treatment with rFtn-Blo t 2 to investigate its effects on airway inflammation, Th1/Th2 cytokine profiles, and allergen-specific antibody production.Results: The average diameter of rFtn and rFtn-Blo t 2 particles are (11.24 ± 1.31) nm and (16.00 ± 1.59) nm. The IgE- binding rates of rFtn-Blo t 2 and rBlo t 2 were 62.5% (15/24) and 58.3% (12/24), respectively. The expressions of IL-25, IL-33, and TSLP increased by 3.29, 3.43, and 2.22 folds after the addition of rFtn-Blo t 2 in co-culture with BEAS-2B. Mice challenged with rBlo t 2 had lower levels of cytokines IL-13 and IL-4 but higher levels of IFN-γ and TGF-β in alveolar lavage fluid compared to controls (p https://www.dovepress.com/development-of-a-ferritin-based-nano-allergen-and-its-immunological-ef-peer-reviewed-fulltext-article-IJNBlomia tropicalisnanometerferritinAllergic diseaseBlo t 2
spellingShingle Zhou D
Ren Y
Zhou Y
Liao Y
Cheng Q
Chen J
Yuan C
Zeng D
Cui Y
Development of a Ferritin-Based Nano-Allergen and Its Immunological Effects in vitro and in vivo
International Journal of Nanomedicine
Blomia tropicalis
nanometer
ferritin
Allergic disease
Blo t 2
title Development of a Ferritin-Based Nano-Allergen and Its Immunological Effects in vitro and in vivo
title_full Development of a Ferritin-Based Nano-Allergen and Its Immunological Effects in vitro and in vivo
title_fullStr Development of a Ferritin-Based Nano-Allergen and Its Immunological Effects in vitro and in vivo
title_full_unstemmed Development of a Ferritin-Based Nano-Allergen and Its Immunological Effects in vitro and in vivo
title_short Development of a Ferritin-Based Nano-Allergen and Its Immunological Effects in vitro and in vivo
title_sort development of a ferritin based nano allergen and its immunological effects in vitro and in vivo
topic Blomia tropicalis
nanometer
ferritin
Allergic disease
Blo t 2
url https://www.dovepress.com/development-of-a-ferritin-based-nano-allergen-and-its-immunological-ef-peer-reviewed-fulltext-article-IJN
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