ORIGINAL RESEARCH article
Front. Immunol.
Sec. Vaccines and Molecular Therapeutics
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1623405
AS04 drives superior cross-protective antibody response by increased NOTCH signaling of Dendritic Cells and proliferation of memory B cells
Provisionally accepted
Valentino D'Onofrio1*
Ana-Carolina Santana2Marthe Pauwels2
Gwenn Waerlop1Anthony Willems1
Fien De Boever3Martin Müller4
Peter Sehr5
Tim Waterboer4Isabel Leroux-Roels1
Ashish A Sharma2
Rafick Pierre Sekaly2
Geert Leroux-Roels1- 1Ghent University, Ghent, Belgium
- 2Emory University, Atlanta, United States
- 3Ghent University Hospital, Gent, Belgium
- 4DKFZ, Heidelberg, Germany
- 5EMBL, Heidelberg, Germany
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Introduction. The Gardasil-4® vaccine targets HPV types 6, 11, 16 and 18 and is formulated with amorphous alum. Cervarix® targets HPV types 16 and 18 using AS04 (Al(OH)3 + TLR4 agonist MPL) to enhance immune response. Cervarix elicits higher cross-protection against other high-risk HPV types, likely mediated by AS04.To investigate mechanisms of cross-neutralizing potential, six monozygotic twins (12 females aged 9-13 years) were vaccinated with either Cervarix or Gardasil-4 (2 doses, 6 months apart). Serum neutralizing antibody titers against HPV 6,16,18,31,33,45,52, and 58 were assessed pre-vaccination and 7 days post-second dose. Multi-omic single cell RNA and ATAC sequencing of PBMCs was performed at the latter timepoint.Results. Cervarix generated higher cross-neutralizing antibody titers than Gardasil-4. Higher frequencies of dendritic cells and memory B cells were observed. Gene Set Enrichment Analysis (GSEA) indicated enhanced pathways related to NOTCH2 signaling in DCs and cell cycling/RNA translation in B cells, correlating positively with cross-neutralizing antibody titers. Increased chromatin accessability in genes related to NOTCH signaling in cDC1 was also observed. Cervarixvaccinated subjects showed increased DC-to-memory B signaling, through upregulation of NOTCH ligands. Engagement of NOTCH was associated to BCL2 expression in memory B cells, supporting an anti-apoptotic state, and resulted in increased IgG4 expression by B memory cells.Increased DC signaling, including NOTCH, through AS04 in Cervarix supports cell survival and sustained RNA translation in memory B cells, 7 days post-vaccination. This may enhance adaptive immune cell maturation, providing a mechanism that can lead to improved cross-reactivity.
Keywords: HPV vaccine, adjuvant, immune response, Dendritic Cells, memory B cell
Received: 05 May 2025; Accepted: 07 Jul 2025.
Copyright: © 2025 D'Onofrio, Santana, Pauwels, Waerlop, Willems, De Boever, Müller, Sehr, Waterboer, Leroux-Roels, Sharma, Sekaly and Leroux-Roels. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Valentino D'Onofrio, Ghent University, Ghent, Belgium
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