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Immunogenicity and protective efficacy of a tuberculosis DNA vaccine co-expressing pro-apoptotic caspase-3
Tatiana Gartner1, Marta Romano, Vanessa Suin
1Department of Molecular and Cellular Interactions, Vlaams Instituut voor Biotechnologie, Brussels, Belgium.
Abstract:
DNA vaccination is a potent means for inducing strong cell-mediated immune responses and protective immunity against viral, bacterial and parasite pathogens in rodents. In an attempt to increase cross-presentation through apoptosis, the DNA-encoding caspase-2 prodomain followed by wild-type or catalytically inactive mutated caspase-3 was inserted into a plasmid encoding the 32 kDa mycolyl transferase (Ag85A) from Mycobacterium tuberculosis. Transient transfection showed that the mutated caspase induced slow apoptosis, normal protein expression and NF-kappaB activation while wild-type caspase induced rapid apoptosis, lower protein expression and no NF-kappaB activation. Ag85A specific antibody production was increased by co-expressing the mutated and decreased by co-expressing the wild-type caspase. Vaccination with pro-apoptotic plasmids triggered more Ag85A specific IFN-gamma producing spleen cells, and more efficient IL-2 and IFN-gamma producing memory cells in spleen and lungs after M. tuberculosis challenge. Compared to DNA-encoding secreted Ag85A, vaccination with DNA co-expressing wild-type caspase increased protection after infection with M. tuberculosis, while vaccination with plasmid co-expressing mutated caspase was not protective, possibly due to the stimulation of IL-6, IL-10 and IL-17A production.
Insights
DNA vaccination using modified caspases enhances immune responses against Mycobacterium tuberculosis. Co-expressing wild-type caspase with Ag85A DNA improved protection, while mutated caspase did not confer protection.
Area of Science:
- Immunology
- Molecular Biology
- Vaccine Development
Background:
- DNA vaccination is effective for inducing cell-mediated immunity against various pathogens.
- Enhancing antigen cross-presentation via apoptosis is a strategy to boost vaccine efficacy.
- Mycobacterium tuberculosis (M. tuberculosis) infection necessitates improved vaccine strategies.
Purpose of the Study:
- To investigate the impact of co-expressing caspase-3 variants with Ag85A DNA on immune responses and protection against M. tuberculosis.
- To evaluate the role of apoptosis induction in DNA vaccine-mediated immunity.
Main Methods:
- Constructing plasmids encoding Ag85A with wild-type or mutated caspase-3.
- Transient transfection to assess protein expression, apoptosis, and NF-kappaB activation.
- Vaccination of rodents followed by M. tuberculosis challenge to evaluate immune responses (antibody, cytokine production) and protection.
Main Results:
- Mutated caspase-3 induced slow apoptosis, normal protein expression, and NF-kappaB activation, enhancing Ag85A antibody production.
- Wild-type caspase-3 induced rapid apoptosis, reduced protein expression, and decreased antibody production.
- Vaccination with pro-apoptotic plasmids increased Ag85A-specific IFN-gamma producing cells and memory cells.
- Co-expression of wild-type caspase-3 with Ag85A DNA improved protection against M. tuberculosis challenge.
- Mutated caspase-3 co-expression was not protective and associated with IL-6, IL-10, and IL-17A production.
Conclusions:
- Modulating apoptosis via caspase co-expression can influence DNA vaccine efficacy.
- Wild-type caspase-3 co-expression enhances protective immunity against M. tuberculosis, possibly through specific immune signaling pathways.
- Mutated caspase-3 co-expression does not confer protection and may skew the immune response towards less effective cytokines.
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