Raw data of Multiepitope mRNA vaccine against Fasciola hepatica confers T-cell-mediated protection in mice [Dataset]

Abstract

[EN]This dataset contains raw spectral flow cytometry files (FCS format) generated in a mouse immunization and challenge study evaluating an mRNA–solid lipid nanoparticle (mRNA-SLN) vaccine candidate encoding three MHC class II T-cell epitopes from Fasciola hepatica fused to eGFP (eGFP-Fh3Tq). BALB/c mice (n = 33) were allocated into four experimental groups: Untreated (n = 9), Infection control (n = 6), eGFP (n = 9), and eGFP-Fh3Tq (n = 9). Groups eGFP and eGFP-Fh3Tq received a prime–boost immunization regimen (21 days apart) with the corresponding mRNA-SLN formulation. Peripheral blood was collected from three groups (Untreated, eGFP, and eGFP-Fh3Tq) at day 1 post-vaccination (innate panel) and day 42 post-vaccination (adaptive panel); the Infection control group was not differentiated from the Untreated group at these time points and was therefore not sampled separately. For blood panels, samples from two mice were pooled and barcoded with anti-CD45 conjugated to either PerCP or BUV496 before acquisition, yielding three FCS files per group (6 mice/group). At day 42, spleens from three mice per group (Untreated, eGFP, eGFP-Fh3Tq) were harvested for intracellular cytokine staining (ICS) after ex vivo restimulation with the synthetic peptides T14, T15, and T16, PMA/ionomycin (positive control), or left unstimulated (negative control). For ICS, individual splenocyte samples from different conditions were barcoded with anti-CD45 and combined in pairs within each acquisition tube. Where sufficient cells were available, stimulation conditions were performed in duplicate. Remaining mice were orally challenged with 7 F. hepatica metacercariae for survival and protection assessment. All samples were acquired on a Cytek Aurora 5L spectral flow cytometer equipped with five lasers (355, 405, 488, 561, 640 nm).