# Comparative evaluation of 5 combination adjuvants on immunogenicity and efficacy of approved seasonal influenza vaccines

**Authors:** Jenny Hernandez-Davies, Jiin Felgner, Erwin Strahsburger, Jacob Laster, Aarti Jain, Timothy Yates, Emily Silzel, Rafael Assis, Rie Nakajima, Algimantas Jasinskas, Andriy Yeromin, Egest J. Pone, Sharon Jan, Luis M. de la Maza, Li Liang, Philip Felgner, Lisa E. Wagar, Anthony E. Gregory, D. Huw Davies

PMC · DOI: 10.1038/s41541-025-01339-y · 2026-01-27

## TL;DR

This study compares five new vaccine adjuvants with seasonal flu vaccines in mice to see how well they boost immunity and protection.

## Contribution

The study provides a direct comparison of five novel adjuvants with the same vaccines and model system.

## Key findings

- All adjuvants were immunogenic and protective against H1N1 challenge.
- T-VANT, TRAC-478, and IVAX-1/-3 induced strong Th1 responses and systemic cytokines.
- Males showed greater morbidity after H1N1 challenge compared to females.

## Abstract

The benefits of adjuvants for enhancing vaccine immunogenicity and efficacy are well known. Numerous novel adjuvants are at advanced levels of characterization, including some in clinical trials. However, understanding the relative benefits of each is hindered by a lack of comparative studies between adjuvants within the same study. To address this, we have performed a side-by-side comparison of 5 novel combination adjuvants (Alhydroxyquim-II, T-VANT, TRAC-478, IVAX-1 and IVAX-3) by formulating each with two approved seasonal influenza vaccines, Flublok® and Fluzone HD®, and assessing immunogenicity and efficacy in female and male C57Bl/6 mice. Although all tested adjuvants were immunogenic and protective against H1N1 challenge, T-VANT, TRAC-478 and IVAX-1 and -3 were associated with systemic inflammatory cytokines and robust Th1 responses, while Alhydroxyquim-II elicited lower levels of inflammatory cytokines and a Th2 response. Greater morbidity after challenge was also detected in males compared to females. Side-by-side comparisons of existing and novel adjuvants with the same antigen and model system will help inform rational adjuvant selection and guide vaccine development for influenza and other infectious diseases.

## Linked entities

- **Diseases:** influenza (MONDO:0005812)

## Full-text entities

- **Genes:** Il2 (interleukin 2) [NCBI Gene 16183] {aka Il-2}, Gapdh (glyceraldehyde-3-phosphate dehydrogenase) [NCBI Gene 14433] {aka Gapd}, Ifna (interferon alpha complex region) [NCBI Gene 111654] {aka Ifa, Ifa8}, Il10 (interleukin 10) [NCBI Gene 16153] {aka CSIF, If2a, Il-10}, Crp (C-reactive protein, pentraxin-related) [NCBI Gene 12944], Csf2 (colony stimulating factor 2 (granulocyte-macrophage)) [NCBI Gene 12981] {aka CSF, Csfgm, GMCSF, Gm-CSf, MGI-IGM}, Igh-V7183 (immunoglobulin heavy chain (V7183 family)) [NCBI Gene 16059] {aka B9-scFv, IgG, IgH, IgVH1(VSG), VH7183, VI24H}, Il1a (interleukin 1 alpha) [NCBI Gene 16175] {aka Il-1a}, Ighv1-9 (immunoglobulin heavy variable 1-9) [NCBI Gene 668478] {aka Gm16697, Igg2a}, Il6 (interleukin 6) [NCBI Gene 16193] {aka Il-6}, Cxcl10 (C-X-C motif chemokine ligand 10) [NCBI Gene 15945] {aka C7, CRG-2, INP10, IP-10, IP10, Ifi10}, IL6 (interleukin 6) [NCBI Gene 3569] {aka BSF-2, BSF2, CDF, HGF, HSF, IFN-beta-2}, Il17a (interleukin 17A) [NCBI Gene 16171] {aka Ctla-8, Ctla8, IL-17, IL-17A, Il17}, Tlr8 (toll-like receptor 8) [NCBI Gene 170744], Tlr7 (toll-like receptor 7) [NCBI Gene 170743], Il4 (interleukin 4) [NCBI Gene 16189] {aka BSF-1, Il-4}, Tlr9 (toll-like receptor 9) [NCBI Gene 81897], Ccl2 (C-C motif chemokine ligand 2) [NCBI Gene 20296] {aka HC11, JE, MCAF, MCP-1, MCP1, SMC-CF}, Fcr (Fc receptor) [NCBI Gene 109615], Ifnb1 (interferon beta 1, fibroblast) [NCBI Gene 15977] {aka IFN-beta, IFNB, If1da1, Ifb}, Il23a (interleukin 23, alpha subunit p19) [NCBI Gene 83430] {aka IL-23, p19}, IFNG (interferon gamma) [NCBI Gene 3458] {aka IFG, IFI, IMD69}, Igha (immunoglobulin heavy constant alpha) [NCBI Gene 238447] {aka IgA, Igh-2}, IL4 (interleukin 4) [NCBI Gene 3565] {aka BCGF-1, BCGF1, BSF-1, BSF1, IL-4}, LOC105243590 (Ig heavy chain Mem5-like) [NCBI Gene 105243590] {aka IgH, Igg1}, Cxcl11 (chemokine (C-X-C motif) ligand 11) [NCBI Gene 56066] {aka Cxc11, H174, I-tac, Ip9, Itac, Scyb11}, Nelfcd (negative elongation factor complex member C/D, Th1l) [NCBI Gene 57314] {aka 2410003I03Rik, NELF-D, Th1, Th1l}, Tlr4 (toll-like receptor 4) [NCBI Gene 21898] {aka Lps, Ly87, Ran/M1, Rasl2-8}, Cd4 (CD4 antigen) [NCBI Gene 12504] {aka L3T4, Ly-4}, Tnf (tumor necrosis factor) [NCBI Gene 21926] {aka DIF, TNF-a, TNF-alpha, TNFSF2, TNFalpha, Tnfa}, Ifng (interferon gamma) [NCBI Gene 15978] {aka IFN-g, If2f, Ifg}, Il1b (interleukin 1 beta) [NCBI Gene 16176] {aka IL-1beta, Il-1b}, Il27 (interleukin 27) [NCBI Gene 246779] {aka IL-27, IL-27-A, IL-27p28, IL27-A, Il30, p28}
- **Diseases:** infection (MESH:D007239), muscle or joint pain (MESH:D063806), infectious diseases (MESH:D003141), toxicity (MESH:D064420), inflammatory cytokines (MESH:D000080424), influenza (MESH:D007251), COVID-19 (MESH:D000086382), headache (MESH:D006261), deaths (MESH:D003643), Weight loss (MESH:D015431), pain (MESH:D010146), fever (MESH:D005334), Inflammatory (MESH:D007249)
- **Chemicals:** TRIzol (MESH:C411644), Lipids (MESH:D008055), FC (MESH:C095424), TPCK (MESH:D014108), TBS (MESH:D013725), squalene (MESH:D013185), Cal09 (-), MF-59 (MESH:C089950), poly-Histidine (MESH:C033223), AddaVax (MESH:C000590912), sodium citrate (MESH:D000077559), Alexa Fluor647 (MESH:C569686), CpG (MESH:C015772), streptomycin (MESH:D013307), Tween 20 (MESH:D011136), penicillin (MESH:D010406), Triton X-100 (MESH:D017830), paraformaldehyde (MESH:C003043), CB (MESH:C063451), MPLA (MESH:C048436), T (MESH:D014316), 3,3',5,5'-tetramethylbenzidine (MESH:C021758), CO2 (MESH:D002245), PBS (MESH:D007854), QS-21 (MESH:C078785), LPS (MESH:D008070), water (MESH:D014867), isoflurane (MESH:D007530), NaCl (MESH:D012965), saponin (MESH:D012503), H2SO4 (MESH:C033158), Alexa Fluor555 (MESH:C000608607)
- **Species:** Yellow fever virus (no rank) [taxon 11089], Burkholderia pseudomallei (species) [taxon 28450], Orthomyxoviridae (family) [taxon 11308], Chlamydia muridarum (agent of mouse pneumonitis, species) [taxon 83560], Papio hamadryas (baboon, species) [taxon 9557], Gallus gallus (bantam, species) [taxon 9031], Coxiella burnetii (species) [taxon 777], Hepatovirus A (no rank) [taxon 12092], Meleagris gallopavo (common turkey, species) [taxon 9103], Severe acute respiratory syndrome coronavirus 2 (no rank) [taxon 2697049], Mustela putorius furo (black ferret, subspecies) [taxon 9669], Mus musculus (house mouse, species) [taxon 10090], Sus scrofa (pig, species) [taxon 9823], Homo sapiens (human, species) [taxon 9606], H1N1 subtype (serotype) [taxon 114727], H5N1 subtype (serotype) [taxon 102793]
- **Mutations:** V08H
- **Cell lines:** MDCK — Canis lupus familiaris (Dog), Spontaneously immortalized cell line (CVCL_0422), HEK293 — Homo sapiens (Human), Transformed cell line (CVCL_0045), /6 — Homo sapiens (Human), Tongue squamous cell carcinoma, Cancer cell line (CVCL_5985), CCL — Mus musculus (Mouse), Undefined cell line type (CVCL_M023), 34 — Mus musculus (Mouse), Hybridoma (CVCL_J663), TRAC-478 — Mus musculus (Mouse), Hybridoma (CVCL_A0EQ)

## Figures

10 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12852915/full.md

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Source: https://tomesphere.com/paper/PMC12852915