A recombinant BCG with surface-displayed antigen induces humoral and cellular immune responses

Abstract Bacillus Calmette-Guérin (BCG) is an attenuated vaccine widely used for tuberculosis prevention. While BCG has long been perceived as an intracellular candidate vector for delivering antigens against infectious diseases and cancers, challenges persist in inducing durable immune responses, p...

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Main Authors: Jin-Yu Zhang, Zhi-Dong Hu, Li-Xiao Xing, Zhen-Yan Chen, Jin-Chuan Xu, Qiao-Yu Wu, Juan Wu, Guo-Ping Zhao, Xiao-Yong Fan, Liang-Dong Lyu
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-00553-x
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author Jin-Yu Zhang
Zhi-Dong Hu
Li-Xiao Xing
Zhen-Yan Chen
Jin-Chuan Xu
Qiao-Yu Wu
Juan Wu
Guo-Ping Zhao
Xiao-Yong Fan
Liang-Dong Lyu
author_facet Jin-Yu Zhang
Zhi-Dong Hu
Li-Xiao Xing
Zhen-Yan Chen
Jin-Chuan Xu
Qiao-Yu Wu
Juan Wu
Guo-Ping Zhao
Xiao-Yong Fan
Liang-Dong Lyu
author_sort Jin-Yu Zhang
collection DOAJ
description Abstract Bacillus Calmette-Guérin (BCG) is an attenuated vaccine widely used for tuberculosis prevention. While BCG has long been perceived as an intracellular candidate vector for delivering antigens against infectious diseases and cancers, challenges persist in inducing durable immune responses, particularly high-titer neutralizing antibodies (Nabs). Here we show that displaying antigens in the surface of BCG is a promising strategy to induce long-lasting Nabs production and T-cell responses. We constructed a recombinant BCG expressing the SARS-CoV-2 receptor-binding domain (RBD) antigen on its cell wall, termed CW-rBCG::RBD, which achieved an antigen yield approaching 850 nanograms per 107 colony-forming unit. Compared with both the parental BCG and the RBD protein subunit vaccine (RBDAS01), intravenous administration of CW-rBCG::RBD followed by a booster dose significantly enhanced Nab production and increased the frequencies of RBD-specific central memory T cells (Tcm) and T follicular helper (Tfh) cells in the spleen. In mice primed with a single dose of CW-rBCG::RBD and boosted with RBDAS01, we also observed elevated Nab titers and detectable levels of RBD-specific IgG2a antibodies at 8 weeks post-priming, responses that were not observed in the BCG-primed or RBDAS01-only groups. Furthermore, subcutaneous co-administration of CW-rBCG::RBD and RBDAS01 sustained Nab production for up to 31 weeks and maintained higher Tfh and Tcm cell frequencies compared to both BCG co-administration with RBDAS01 and RBDAS01 alone. These findings highlight an effective strategy for optimizing BCG-based vaccination and immunotherapy platforms. Subject terms: recombinant BCG; immune response; vaccines; cell wall; SARS-CoV-2 RBD.
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spelling doaj-art-bea35c36fbd8408ba49a798f87de19fe2025-08-20T01:51:38ZengNature PortfolioScientific Reports2045-23222025-05-0115111510.1038/s41598-025-00553-xA recombinant BCG with surface-displayed antigen induces humoral and cellular immune responsesJin-Yu Zhang0Zhi-Dong Hu1Li-Xiao Xing2Zhen-Yan Chen3Jin-Chuan Xu4Qiao-Yu Wu5Juan Wu6Guo-Ping Zhao7Xiao-Yong Fan8Liang-Dong Lyu9Key Laboratory of Medical Molecular Virology of the Ministry of Education/National Health Commission, School of Basic Medical Sciences and Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan UniversityShanghai Public Health Clinical Center, Fudan UniversityKey Laboratory of Medical Molecular Virology of the Ministry of Education/National Health Commission, School of Basic Medical Sciences and Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan UniversityShanghai Public Health Clinical Center, Fudan UniversityShanghai Public Health Clinical Center, Fudan UniversityShanghai Public Health Clinical Center, Fudan UniversityShanghai Public Health Clinical Center, Fudan UniversityKey Laboratory of Medical Molecular Virology of the Ministry of Education/National Health Commission, School of Basic Medical Sciences and Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan UniversityShanghai Public Health Clinical Center, Fudan UniversityKey Laboratory of Medical Molecular Virology of the Ministry of Education/National Health Commission, School of Basic Medical Sciences and Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan UniversityAbstract Bacillus Calmette-Guérin (BCG) is an attenuated vaccine widely used for tuberculosis prevention. While BCG has long been perceived as an intracellular candidate vector for delivering antigens against infectious diseases and cancers, challenges persist in inducing durable immune responses, particularly high-titer neutralizing antibodies (Nabs). Here we show that displaying antigens in the surface of BCG is a promising strategy to induce long-lasting Nabs production and T-cell responses. We constructed a recombinant BCG expressing the SARS-CoV-2 receptor-binding domain (RBD) antigen on its cell wall, termed CW-rBCG::RBD, which achieved an antigen yield approaching 850 nanograms per 107 colony-forming unit. Compared with both the parental BCG and the RBD protein subunit vaccine (RBDAS01), intravenous administration of CW-rBCG::RBD followed by a booster dose significantly enhanced Nab production and increased the frequencies of RBD-specific central memory T cells (Tcm) and T follicular helper (Tfh) cells in the spleen. In mice primed with a single dose of CW-rBCG::RBD and boosted with RBDAS01, we also observed elevated Nab titers and detectable levels of RBD-specific IgG2a antibodies at 8 weeks post-priming, responses that were not observed in the BCG-primed or RBDAS01-only groups. Furthermore, subcutaneous co-administration of CW-rBCG::RBD and RBDAS01 sustained Nab production for up to 31 weeks and maintained higher Tfh and Tcm cell frequencies compared to both BCG co-administration with RBDAS01 and RBDAS01 alone. These findings highlight an effective strategy for optimizing BCG-based vaccination and immunotherapy platforms. Subject terms: recombinant BCG; immune response; vaccines; cell wall; SARS-CoV-2 RBD.https://doi.org/10.1038/s41598-025-00553-x
spellingShingle Jin-Yu Zhang
Zhi-Dong Hu
Li-Xiao Xing
Zhen-Yan Chen
Jin-Chuan Xu
Qiao-Yu Wu
Juan Wu
Guo-Ping Zhao
Xiao-Yong Fan
Liang-Dong Lyu
A recombinant BCG with surface-displayed antigen induces humoral and cellular immune responses
Scientific Reports
title A recombinant BCG with surface-displayed antigen induces humoral and cellular immune responses
title_full A recombinant BCG with surface-displayed antigen induces humoral and cellular immune responses
title_fullStr A recombinant BCG with surface-displayed antigen induces humoral and cellular immune responses
title_full_unstemmed A recombinant BCG with surface-displayed antigen induces humoral and cellular immune responses
title_short A recombinant BCG with surface-displayed antigen induces humoral and cellular immune responses
title_sort recombinant bcg with surface displayed antigen induces humoral and cellular immune responses
url https://doi.org/10.1038/s41598-025-00553-x
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