Alpha-galactosylceramide as mRNA vaccine adjuvant modulates cellular immunity against tuberculosis and contributes to antigen-specific lung-associated T cells with a TRM-associated phenotype following prime-pull strategy
6 affiliations
- 1Laboratory of Physical Pharmacy and General Biochemistry, Department of Pharmaceutics, Ghent Research Group on Nanomedicines, Ghent University
- 2Immunobiology and Therapy Unit, Inserm U1224, Institut Pasteur, Université Paris Cité
- 3Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University
- 4Department of Biomolecular Medicine, Ghent University
- 5Vlaams Instituut voor Biotechnologie (VIB) Center for Medical Biotechnology, VIB
- 6Translational Oncology Research Centre, Department of Biomedical Sciences, Vrije Universiteit Brussel (VUB)
Abstract
Tuberculosis (TB) remains the leading cause of death from a single infectious agent worldwide, highlighting the need for improved vaccines beyond the limited protection currently provided by the Bacillus Calmette-Guérin (BCG) vaccine. Here we investigated the immunological impact of incorporating alpha-Galactosylceramide (αGC) into mRNA lipid nanoparticles (mRNA-LNPs) as a vaccination strategy against Mycobacterium tuberculosis (Mtb), the pathogen causing TB. Using Mtb antigens ESAT-6 and Ag85B, we demonstrate that αGC-adjuvanted mRNA-LNPs induce controlled activation of invariant natural killer T (iNKT) cells, thereby reshaping vaccine-induced immunity. Specifically, αGC incorporation promoted the generation of CD44 ⁺ CD69 ⁺ CD62L ⁻ CD4 ⁺ T cells in the spleen and altered the CD4 ⁺ T -helper (T H ) profile through the addition of modest T H 2 and T H 17 responses alongside robust T H 1 immunity. Furthermore, antigen-specific CD8 ⁺ T-cell responses against ESAT-6 and Ag85B were observed exclusively in the genetically diverse CB6F1/J mouse model, highlighting the importance of host genetic background in evaluating mRNA vaccine immunogenicity after homologous vaccination strategies. Finally, we demonstrate that using a heterologous prime–pull strategy combining intramuscular mRNA vaccination with local antigen instillation increased the frequency of antigen-specific lung-associated CD4 ⁺ T cells displaying a tissue residency-associated phenotype, with additional contribution of αGC adjuvantation to this response. In summary, these findings provide novel insights into αGC adjuvant-mediated immune modulation of mRNA vaccines and support further testing of protective efficacy of this approach against TB.
Related
- The Pregnane X Receptor (PXR) contributes to phenotypic plasticity and modulates the sensitivity to targeted therapies in melanoma
- 1161 Novel IL1RAP expression gradient modulates the myeloid cell phenotype and T-cell infiltration patterns of the Ewing sarcoma immune microenvironment
- SPOCK2-mediated phenotype switch of vascular smooth muscle cells contributes to aortic aneurysm in Kawasaki disease
- Dual CD73/A2AR blockade modulates the neurotoxic astrocyte phenotype without disrupting core inflammatory signaling
- MFN2-associated astrocyte dysfunction contributes to neuronal hyperexcitability and depressive-like behaviors
BibTeX
@article{aernout2026alphagalactosylceramide,
title = {{Alpha-galactosylceramide as mRNA vaccine adjuvant modulates cellular immunity against tuberculosis and contributes to antigen-specific lung-associated T cells with a TRM-associated phenotype following prime-pull strategy}},
author = {Ilke Aernout and Thomas Ehouarne and Kevin Mwangi and Kia C. Ferrell and Margo De Velder and Janne Swinnen and Aaron Verplancke and Laura Van Moortel and Margaux De Meyer and Patrick Willems and Lorenzo Franceschini and Karine Breckpot and Stefaan C. De Smedt and Francis Impens and Caroline Demangel and Rein Verbeke and Ine Lentacker},
journal = {Frontiers in Immunology},
year = {2026},
month = oct,
doi = {10.3389/fimmu.2026.1929726},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1929726/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Alpha-galactosylceramide as mRNA vaccine adjuvant modulates cellular immunity against tuberculosis and contributes to antigen-specific lung-associated T cells with a TRM-associated phenotype following prime-pull strategy
AU - Ilke Aernout
AU - Thomas Ehouarne
AU - Kevin Mwangi
AU - Kia C. Ferrell
AU - Margo De Velder
AU - Janne Swinnen
AU - Aaron Verplancke
AU - Laura Van Moortel
AU - Margaux De Meyer
AU - Patrick Willems
AU - Lorenzo Franceschini
AU - Karine Breckpot
AU - Stefaan C. De Smedt
AU - Francis Impens
AU - Caroline Demangel
AU - Rein Verbeke
AU - Ine Lentacker
PY - 2026
DA - 2026/10/05
JO - Frontiers in Immunology
DO - 10.3389/fimmu.2026.1929726
UR - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1929726/full
AB - Tuberculosis (TB) remains the leading cause of death from a single infectious agent worldwide, highlighting the need for improved vaccines beyond the limited protection currently provided by the Bacillus Calmette-Guérin (BCG) vaccine. Here we investigated the immunological impact of incorporating alpha-Galactosylceramide (αGC) into mRNA lipid nanoparticles (mRNA-LNPs) as a vaccination strategy against Mycobacterium tuberculosis (Mtb), the pathogen causing TB. Using Mtb antigens ESAT-6 and Ag85B, we demonstrate that αGC-adjuvanted mRNA-LNPs induce controlled activation of invariant natural killer T (iNKT) cells, thereby reshaping vaccine-induced immunity. Specifically, αGC incorporation promoted the generation of CD44 ⁺ CD69 ⁺ CD62L ⁻ CD4 ⁺ T cells in the spleen and altered the CD4 ⁺ T -helper (T H ) profile through the addition of modest T H 2 and T H 17 responses alongside robust T H 1 immunity. Furthermore, antigen-specific CD8 ⁺ T-cell responses against ESAT-6 and Ag85B were observed exclusively in the genetically diverse CB6F1/J mouse model, highlighting the importance of host genetic background in evaluating mRNA vaccine immunogenicity after homologous vaccination strategies. Finally, we demonstrate that using a heterologous prime–pull strategy combining intramuscular mRNA vaccination with local antigen instillation increased the frequency of antigen-specific lung-associated CD4 ⁺ T cells displaying a tissue residency-associated phenotype, with additional contribution of αGC adjuvantation to this response. In summary, these findings provide novel insights into αGC adjuvant-mediated immune modulation of mRNA vaccines and support further testing of protective efficacy of this approach against TB.
SN - 1664-3224
ER -