Evaluation of Immunogenicity of <i>Mycobacterium tuberculosis ag85ab</i> DNA Vaccine Delivered by Pulmonary Administration

<b>Background:</b> Tuberculosis (TB) is a respiratory infectious disease, and the current TB vaccine has low local lung protection. We aim to optimize immune pathways to improve the immunogenicity of vaccines. <b>Methods:</b> In the immunogenicity study, 50 BALB/c mice were r...

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Main Authors: Haimei Zhao, Zhen Zhang, Yong Xue, Nan Wang, Yinping Liu, Xihui Ma, Lan Wang, Xiaoou Wang, Danyang Zhang, Junxian Zhang, Xueqiong Wu, Yan Liang
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Language:English
Published: MDPI AG 2025-04-01
Series:Vaccines
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Online Access:https://www.mdpi.com/2076-393X/13/5/442
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author Haimei Zhao
Zhen Zhang
Yong Xue
Nan Wang
Yinping Liu
Xihui Ma
Lan Wang
Xiaoou Wang
Danyang Zhang
Junxian Zhang
Xueqiong Wu
Yan Liang
author_facet Haimei Zhao
Zhen Zhang
Yong Xue
Nan Wang
Yinping Liu
Xihui Ma
Lan Wang
Xiaoou Wang
Danyang Zhang
Junxian Zhang
Xueqiong Wu
Yan Liang
author_sort Haimei Zhao
collection DOAJ
description <b>Background:</b> Tuberculosis (TB) is a respiratory infectious disease, and the current TB vaccine has low local lung protection. We aim to optimize immune pathways to improve the immunogenicity of vaccines. <b>Methods:</b> In the immunogenicity study, 50 BALB/c mice were randomly divided into the following: (1) phosphate buffered saline (PBS)+intramuscular injection combined with electroporation (EP) group (100 μL), (2) pVAX1+EP group (50 μg/100 μL), (3) <i>ag85ab</i>+EP group (50 μg/100 μL), (4) pVAX1+pulmonary delivery (PD) group (50 μg/50 μL), and (5) <i>ag85ab</i>+PD group (50 μg/50 μL). Immunization was given once every 2 weeks for a total of three times. The number of IFN-γ-secreting lung and spleen lymphocytes was determined by enzyme-linked immunospot assay (ELISPOT). The levels of Th1, Th2, and Th17 cytokines in the culture supernatants of lung and spleen lymphocytes were detected with the Luminex method. The proportion of FoxP3 regulatory T cells in splenocytes was determined by flow cytometry. The levels of IgG-, IgG1-, and IgG2a-specific antibodies in plasma and IgA antibody in bronchoalveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assay (ELISA). <b>Results:</b> The PD and EP routes of <i>Mycobacterium tuberculosis</i> (<i>M. tb</i>) <i>ag85ab</i> DNA vaccine can effectively induce the responses of IFN-γ-secreting lung and spleen lymphocytes, and induce dominant Th1 and Th17 cell immune responses. The PD route can induce earlier, greater numbers and stronger responses of pulmonary effector T cells, with higher levels of the specific antibody IgA detected in BALF. High levels of the specific antibodies IgG, IgG1, and IgG2α were detected in the plasma of mice immunized by the EP route. <b>Conclusions:</b> The PD route of DNA vaccines can more effectively stimulate the body to produce strong cellular and mucosal immunity than the EP route, especially local cellular immunity in the lungs, which can provide early protection for the lungs. It can significantly improve the immunogenicity of the <i>ag85ab</i> DNA vaccine, suggesting a feasible and effective approach to DNA immunization.
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spelling doaj-art-3fcd3fef37174ebc9b50acd0d1f2248e2025-08-20T01:56:42ZengMDPI AGVaccines2076-393X2025-04-0113544210.3390/vaccines13050442Evaluation of Immunogenicity of <i>Mycobacterium tuberculosis ag85ab</i> DNA Vaccine Delivered by Pulmonary AdministrationHaimei Zhao0Zhen Zhang1Yong Xue2Nan Wang3Yinping Liu4Xihui Ma5Lan Wang6Xiaoou Wang7Danyang Zhang8Junxian Zhang9Xueqiong Wu10Yan Liang11Beijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaGraduate School, Hebei North University, Zhangjiakou 075000, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, ChinaBeijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, 100091 Beijing, China<b>Background:</b> Tuberculosis (TB) is a respiratory infectious disease, and the current TB vaccine has low local lung protection. We aim to optimize immune pathways to improve the immunogenicity of vaccines. <b>Methods:</b> In the immunogenicity study, 50 BALB/c mice were randomly divided into the following: (1) phosphate buffered saline (PBS)+intramuscular injection combined with electroporation (EP) group (100 μL), (2) pVAX1+EP group (50 μg/100 μL), (3) <i>ag85ab</i>+EP group (50 μg/100 μL), (4) pVAX1+pulmonary delivery (PD) group (50 μg/50 μL), and (5) <i>ag85ab</i>+PD group (50 μg/50 μL). Immunization was given once every 2 weeks for a total of three times. The number of IFN-γ-secreting lung and spleen lymphocytes was determined by enzyme-linked immunospot assay (ELISPOT). The levels of Th1, Th2, and Th17 cytokines in the culture supernatants of lung and spleen lymphocytes were detected with the Luminex method. The proportion of FoxP3 regulatory T cells in splenocytes was determined by flow cytometry. The levels of IgG-, IgG1-, and IgG2a-specific antibodies in plasma and IgA antibody in bronchoalveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assay (ELISA). <b>Results:</b> The PD and EP routes of <i>Mycobacterium tuberculosis</i> (<i>M. tb</i>) <i>ag85ab</i> DNA vaccine can effectively induce the responses of IFN-γ-secreting lung and spleen lymphocytes, and induce dominant Th1 and Th17 cell immune responses. The PD route can induce earlier, greater numbers and stronger responses of pulmonary effector T cells, with higher levels of the specific antibody IgA detected in BALF. High levels of the specific antibodies IgG, IgG1, and IgG2α were detected in the plasma of mice immunized by the EP route. <b>Conclusions:</b> The PD route of DNA vaccines can more effectively stimulate the body to produce strong cellular and mucosal immunity than the EP route, especially local cellular immunity in the lungs, which can provide early protection for the lungs. It can significantly improve the immunogenicity of the <i>ag85ab</i> DNA vaccine, suggesting a feasible and effective approach to DNA immunization.https://www.mdpi.com/2076-393X/13/5/442<i>Mycobacterium tuberculosis</i>DNA vaccinepulmonary deliveryelectroporationimmunogenicity
spellingShingle Haimei Zhao
Zhen Zhang
Yong Xue
Nan Wang
Yinping Liu
Xihui Ma
Lan Wang
Xiaoou Wang
Danyang Zhang
Junxian Zhang
Xueqiong Wu
Yan Liang
Evaluation of Immunogenicity of <i>Mycobacterium tuberculosis ag85ab</i> DNA Vaccine Delivered by Pulmonary Administration
Vaccines
<i>Mycobacterium tuberculosis</i>
DNA vaccine
pulmonary delivery
electroporation
immunogenicity
title Evaluation of Immunogenicity of <i>Mycobacterium tuberculosis ag85ab</i> DNA Vaccine Delivered by Pulmonary Administration
title_full Evaluation of Immunogenicity of <i>Mycobacterium tuberculosis ag85ab</i> DNA Vaccine Delivered by Pulmonary Administration
title_fullStr Evaluation of Immunogenicity of <i>Mycobacterium tuberculosis ag85ab</i> DNA Vaccine Delivered by Pulmonary Administration
title_full_unstemmed Evaluation of Immunogenicity of <i>Mycobacterium tuberculosis ag85ab</i> DNA Vaccine Delivered by Pulmonary Administration
title_short Evaluation of Immunogenicity of <i>Mycobacterium tuberculosis ag85ab</i> DNA Vaccine Delivered by Pulmonary Administration
title_sort evaluation of immunogenicity of i mycobacterium tuberculosis ag85ab i dna vaccine delivered by pulmonary administration
topic <i>Mycobacterium tuberculosis</i>
DNA vaccine
pulmonary delivery
electroporation
immunogenicity
url https://www.mdpi.com/2076-393X/13/5/442
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