1. Recombinant Bacillus subtilis spores for the delivery of Mycobacterium tuberculosis Ag85B-CFP10 secretory antigens.
- Author
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Das K, Thomas T, Garnica O, and Dhandayuthapani S
- Subjects
- Acyltransferases biosynthesis, Acyltransferases genetics, Acyltransferases immunology, Administration, Intranasal, Animals, Antibodies, Bacterial blood, Antigens, Bacterial biosynthesis, Antigens, Bacterial genetics, Antigens, Bacterial immunology, Bacillus subtilis genetics, Bacillus subtilis immunology, Bacterial Proteins biosynthesis, Bacterial Proteins genetics, Bacterial Proteins immunology, Cells, Cultured, Female, Immunity, Cellular, Immunity, Humoral, Immunization, Immunoglobulin G blood, Interferon-gamma immunology, Interferon-gamma metabolism, Macrophages immunology, Macrophages metabolism, Mice, Inbred C57BL, Recombinant Fusion Proteins administration & dosage, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins immunology, Spleen immunology, Spleen metabolism, Tuberculosis Vaccines biosynthesis, Tuberculosis Vaccines genetics, Tuberculosis Vaccines immunology, Vaccines, Synthetic administration & dosage, Vaccines, Synthetic biosynthesis, Vaccines, Synthetic immunology, Acyltransferases administration & dosage, Antigens, Bacterial administration & dosage, Bacillus subtilis metabolism, Bacterial Proteins administration & dosage, Genetic Vectors, Spores, Bacterial, Tuberculosis Vaccines administration & dosage
- Abstract
Tuberculosis continues to be a great cause of morbidity and mortality in different parts of the world. Unfortunately, the current BCG vaccine being administered is not fully protective against tuberculosis; therefore, there is a great need for alternate vaccines. With an aim to develop such vaccines, we have analyzed the utility of Bacillus subtilis spores for the expression of two major immunodominant antigens of Mycobacterium tuberculosis, Ag85B and CFP10. We created three recombinant B. subtilis strains to express a truncated fusion of Ag85B
191-325 and CFP101-70 antigens (T85BCFP), either on the spore coat (MTAG1 strain) or in the cytosol of B. subtilis (MTAG 2 and MTAG 3 strains). Examination of spores isolated from these strains revealed successful expression of T85BCFP antigens on the spore coat of MTAG1 as well as in the cytosol of vegetatively grown cells of MTAG2 and MTAG3, indicating that spores can indeed express M. tuberculosis antigens. In vitro antigen presentation assays with spore-infected mouse bone marrow derived macrophages (BMDM) showed that all three recombinant spores could deliver these antigens to antigen presenting cells (APCs). Mice immunized with recombinant spores displayed significantly higher levels of Ag85B specific IFN-γ producing cells in the spleen than in mice immunized with wild-type (non-recombinant) spores. In addition, these mice showed relatively higher levels of Ag85B specific IgG antibodies in the serum in comparison to mice immunized with non-recombinant spores, thus providing additional evidence that recombinant spores can deliver these antigens in vivo. These results suggest that B. subtilis spores are ideal vehicles for antigen delivery and have great potential in the development of primary and booster vaccines against tuberculosis., (Copyright © 2016 Elsevier Ltd. All rights reserved.)- Published
- 2016
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