Journal: bioRxiv

Article Title: Designed nanofiber immunogens prime antibody responses to diverse conformations of the HIV-1 fusion peptide

doi: 10.64898/2026.10.02.756244

Figure Lengend Snippet: (a) Schematic of FP-Coil29 nanofibers based on the structure of Coil29 (PDB 3j89) capped with size-controlling Coil29Caps (dark grey), FP fusions (pink) and glycomimetic peptides (gold). (b) Structure of FP-Coil29 peptides with helical Coil29 motif and helical EAAAK spacers (grey) with N-terminally linked FP sequence AVIGIAVFL (pink). (c) Antibody titers of C57BL/6 mice immunized with FP-Coil29 with or without helical EAAAK spacers. Mice were immunized at 0, 2, and 4 weeks. IgG titers to FP were measured at 5 weeks after primary immunization. Animals immunized with FP-(EAAAK) 2 -Coil29 constructs showed significantly higher anti-FP IgG titers (p=0.04, unpaired t-test with Welch’s correction). (d) Glycomimetic peptides (gold) displayed adjacent to the Coil29 sequence (grey) with either a linear peptide linker (Glyco-Coil29) or after branching lysine resides (Glyco 4 -Coil29). (e) Circular Dichroism of unmodified fusion peptide, and fusion peptide linked to Coil29 via a SGSG- or EAAAK-linker (left). Circular Dichroism of soluble glycomimetic peptides and its fusions to Coil29 in linear monomeric and branched tetrameric format (right). (f) AFM images of nanofiber formulations assembled in PBS. Images displayed above are representative height maps. (scale bar = 800 nm) (g) Viscosity of FP-Coil29 nanofibers formulated with capping and glycomimetic peptides was measured using a viscometry shear stress ramp from 0.1 to 1000 Hz . (h) Uptake of co-assembled and separately-assembled linear and branched peptides by immune cells in lavage fluid after intraperitoneal injection compared by one-way ANOVA with Fisher’s LSD post-hoc analysis (*p<0.05, ***p<0.001).

Article Snippet: To study immune responses to FP-Coil29 and FP-KLH immunogens, 8-week-old female C57BL/6 mice (Envigo) were injected subcutaneously at the tail base.

Techniques: Sequencing, Construct, Circular Dichroism, Viscosity, Viscometric Assay, Shear, Injection, Analysis

Journal: Frontiers in Immunology

Article 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

doi: 10.3389/fimmu.2026.1929726

Figure Lengend Snippet: αGC in mRNA-LNPs activates iNKT cells and primes antigen-specific CD8 + T cells in a dose-dependent manner (A) Schematic representation of C12–200 mRNA-LNPs encapsulating m1Ψ-modified truncated ovalbumin (tOVA) mRNA encoding tOVA flanked by the LAMP1 signal peptide (SP) and LAMP3 trafficking motifs (TM) formulated as mRNA-LNPS without iNKT agonist, αGC (blue) or with (green). (B) C57BL/6 mice received a single intramuscular injection with various doses of the αGC-adjuvanted LNPs (0.5,1,2 and 5µg/mice), 5 µg of identical unadjuvanted mRNA-LNPs or Tris buffer. iNKT cell frequencies and activation status were assessed 72h later by flow cytometry. Frequency of iNKT cells in (C) draining lymph node (dLN) and (D) spleen. (E) Histograms of programmed cell death protein 1 (PD-1) expression (MFI) on iNKT cells as a measure for iNKT activation in dLN and spleen comparing unvaccinated mice (grey) with mice vaccinated with 5µg mRNA-LNP (blue) and 5µg αGC-adjuvanted mRNA-LNPs (green). (F, G) MFI of PD-1 expression on iNKT cells in dLN and spleen of mice vaccinated with 5µg of unadjuvanted mRNA-LNPs (blue) and increasing doses of αGC-adjuvanted mRNA-LNPs (green). (H) C57BL/6 mice received a prime and boost intramuscular injection with increasing doses (0.5,1,2.5 or 5µg/mice) of unadjuvanted (blue) or αGC-adjuvanted mRNA-LNPs (green) with a 2-week interval. Two weeks post boost the spleen was collected and analyzed with flow cytometry. (for statistic information see ) (I) Weights were monitored every 24 hours for a period of 72 hours after prime injection. (J) Frequency of antigen specific CD8 + T cells (H-2Kb/SIINFEKL tetramer) in the spleen. Each individual point represents one animal. Results given as mean ± SD, n=5 per group. P values were determined by a one-way (C, D, F, G, J) or two-way (I) ANOVA followed by Tukey’s post-hoc test. See Supplementary Figures 1 and for flow gating strategy.

Article Snippet: 7-week-old female C57BL/6 or CB6F1/J mice were ordered from Envigo (The Netherlands) or Charles River laboratories (France), respectively.

Techniques: Modification, Injection, Activation Assay, Flow Cytometry, Expressing

Journal: Frontiers in Immunology

Article 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

doi: 10.3389/fimmu.2026.1929726

Figure Lengend Snippet: mRNA-LNPs and αGC-adjuvanted mRNA-LNPs induce Mtb antigen specific (memory) T cells after prime-boost vaccination in draining lymph node and spleen (A) Construct design of mRNA sequences encoding full length Mtb antigens ESAT-6 and truncated Ag85B flanked by sig-lamp signaling elements. Both mRNAs were co-formulated in a (1:1 weight ratio) in C12–200 LNPs (blue) or αGC-adjuvanted C12-200 LNPs (green), (B) Female C57BL/6 mice were vaccinated with 2.5 µg of the vaccines using a prime-boost regimen with a two-week interval. Two weeks after the boost vaccination draining lymph node (dLN) and spleen were collected for flow cytometry analysis. Antigen specific T cells were detected using ESAT-6 and Ag85B-specific I-Ab tetramers in (C) dLN and (D) spleen with frequencies of CD69 + CD62L - , CD69 - CD62L + , CD69 - CD62L - , CD69 + CD62L + subsets in spleen subsets in spleen. Each individual point represents one animal, with bars or diagrams indicating mean, n=6 per group. P values were determined by a one-way ANOVA followed by Tukey’s post-hoc test. See Supplementary Figure 3 for gating strategy.

Article Snippet: 7-week-old female C57BL/6 or CB6F1/J mice were ordered from Envigo (The Netherlands) or Charles River laboratories (France), respectively.

Techniques: Construct, Vaccines, Flow Cytometry, Analysis

Journal: Frontiers in Immunology

Article 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

doi: 10.3389/fimmu.2026.1929726

Figure Lengend Snippet: Impact of αGC inclusion on systemic CD4 + T H responses after vaccination with mRNA-LNPs encoding Mtb antigens ESAT-6 and Ag85B. Female C57BL/6 mice were vaccinated with 2.5 µg mRNA-LNPs with or without the adjuvant αGC encoding full length Mtb antigens ESAT-6 and Ag85B in a prime boost regimen with a 2-week interval. Analysis of T H responses after re-stimulation of splenocytes with ESAT-6 and Ag85B peptide pools: (A) Intracellular cytokine staining to detect T H 1 CD4 + T cells (IFN-γ, TNF-α and IL-2); (B) polyfunctional T H 1cells; (C) T H 2 cytokine (IL-4) and (D) T H 17 responses (IL-17A). (E) Cytokine levels in the supernatant of splenocytes restimulated with full length proteins ESAT-6 and Ag85B. Boolean gating was applied to analyze the co-expression of the polyfunctional cytokine producing CD4 + T cells (Y-axis range: 0–140 pg/mL). Each individual point represents one animal, with bars indicating mean or heatmap, n= 6 per group. P values were determined by one-way (or a two-way in case of polyfunctional T cells and (E) ANOVA followed by Tukey’s post-hoc test. Reported P values were corrected for multiple testing across all comparisons within the full grid using the Tukey adjustment. See Supplementary Figure 4 for gating strategy.

Article Snippet: 7-week-old female C57BL/6 or CB6F1/J mice were ordered from Envigo (The Netherlands) or Charles River laboratories (France), respectively.

Techniques: Adjuvant, Analysis, Staining, Expressing