Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuis
The induction of an effective immune response is critical for the success of mRNA-based therapeutics. Here, we developed a nanoadjuvant system compromised of Quil-A and DOTAP (dioleoyl 3 trimethylammonium propane), hence named QTAP, for the efficient delivery of mRNA vaccine constructs into cells. E...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-06-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1188754/full |
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| author | Bubacarr J.B. Touray Mostafa Hanafy Mostafa Hanafy Yashdeep Phanse Rachel Hildebrand Adel M. Talaat Adel M. Talaat Adel M. Talaat |
| author_facet | Bubacarr J.B. Touray Mostafa Hanafy Mostafa Hanafy Yashdeep Phanse Rachel Hildebrand Adel M. Talaat Adel M. Talaat Adel M. Talaat |
| author_sort | Bubacarr J.B. Touray |
| collection | DOAJ |
| description | The induction of an effective immune response is critical for the success of mRNA-based therapeutics. Here, we developed a nanoadjuvant system compromised of Quil-A and DOTAP (dioleoyl 3 trimethylammonium propane), hence named QTAP, for the efficient delivery of mRNA vaccine constructs into cells. Electron microscopy indicated that the complexation of mRNA with QTAP forms nanoparticles with an average size of 75 nm and which have ~90% encapsulation efficiency. The incorporation of pseudouridine-modified mRNA resulted in higher transfection efficiency and protein translation with low cytotoxicity than unmodified mRNA. When QTAP-mRNA or QTAP alone transfected macrophages, pro-inflammatory pathways (e.g., NLRP3, NF-kb, and MyD88) were upregulated, an indication of macrophage activation. In C57Bl/6 mice, QTAP nanovaccines encoding Ag85B and Hsp70 transcripts (QTAP-85B+H70) were able to elicit robust IgG antibody and IFN- ɣ, TNF-α, IL-2, and IL-17 cytokines responses. Following aerosol challenge with a clinical isolate of M. avium ss. hominissuis (M.ah), a significant reduction of mycobacterial counts was observed in lungs and spleens of only immunized animals at both 4- and 8-weeks post-challenge. As expected, reduced levels of M. ah were associated with diminished histological lesions and robust cell-mediated immunity. Interestingly, polyfunctional T-cells expressing IFN- ɣ, IL-2, and TNF- α were detected at 8 but not 4 weeks post-challenge. Overall, our analysis indicated that QTAP is a highly efficient transfection agent and could improve the immunogenicity of mRNA vaccines against pulmonary M. ah, an infection of significant public health importance, especially to the elderly and to those who are immune compromised. |
| first_indexed | 2024-03-13T06:44:07Z |
| format | Article |
| id | doaj.art-41541be1236241788b64f56e407ba16c |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-13T06:44:07Z |
| publishDate | 2023-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-41541be1236241788b64f56e407ba16c2023-06-08T11:29:06ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-06-011410.3389/fimmu.2023.11887541188754Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuisBubacarr J.B. Touray0Mostafa Hanafy1Mostafa Hanafy2Yashdeep Phanse3Rachel Hildebrand4Adel M. Talaat5Adel M. Talaat6Adel M. Talaat7Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI, United StatesDepartment of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI, United StatesDepartment of Microbiology and Immunology, Faculty of Veterinary Medicine, Cairo University, Giza, EgyptPan Genome Systems, Madison, WI, United StatesDepartment of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI, United StatesDepartment of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI, United StatesPan Genome Systems, Madison, WI, United StatesVireo Vaccines International, LLC, Madison, Wisconsin, United StatesThe induction of an effective immune response is critical for the success of mRNA-based therapeutics. Here, we developed a nanoadjuvant system compromised of Quil-A and DOTAP (dioleoyl 3 trimethylammonium propane), hence named QTAP, for the efficient delivery of mRNA vaccine constructs into cells. Electron microscopy indicated that the complexation of mRNA with QTAP forms nanoparticles with an average size of 75 nm and which have ~90% encapsulation efficiency. The incorporation of pseudouridine-modified mRNA resulted in higher transfection efficiency and protein translation with low cytotoxicity than unmodified mRNA. When QTAP-mRNA or QTAP alone transfected macrophages, pro-inflammatory pathways (e.g., NLRP3, NF-kb, and MyD88) were upregulated, an indication of macrophage activation. In C57Bl/6 mice, QTAP nanovaccines encoding Ag85B and Hsp70 transcripts (QTAP-85B+H70) were able to elicit robust IgG antibody and IFN- ɣ, TNF-α, IL-2, and IL-17 cytokines responses. Following aerosol challenge with a clinical isolate of M. avium ss. hominissuis (M.ah), a significant reduction of mycobacterial counts was observed in lungs and spleens of only immunized animals at both 4- and 8-weeks post-challenge. As expected, reduced levels of M. ah were associated with diminished histological lesions and robust cell-mediated immunity. Interestingly, polyfunctional T-cells expressing IFN- ɣ, IL-2, and TNF- α were detected at 8 but not 4 weeks post-challenge. Overall, our analysis indicated that QTAP is a highly efficient transfection agent and could improve the immunogenicity of mRNA vaccines against pulmonary M. ah, an infection of significant public health importance, especially to the elderly and to those who are immune compromised.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1188754/fullmRNA-vaccinesmycobacteriananoadjuvantsimmunologyNTM = nontuberculous mycobacteria |
| spellingShingle | Bubacarr J.B. Touray Mostafa Hanafy Mostafa Hanafy Yashdeep Phanse Rachel Hildebrand Adel M. Talaat Adel M. Talaat Adel M. Talaat Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuis Frontiers in Immunology mRNA-vaccines mycobacteria nanoadjuvants immunology NTM = nontuberculous mycobacteria |
| title | Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuis |
| title_full | Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuis |
| title_fullStr | Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuis |
| title_full_unstemmed | Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuis |
| title_short | Protective RNA nanovaccines against Mycobacterium avium subspecies hominissuis |
| title_sort | protective rna nanovaccines against mycobacterium avium subspecies hominissuis |
| topic | mRNA-vaccines mycobacteria nanoadjuvants immunology NTM = nontuberculous mycobacteria |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1188754/full |
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