Goblet cells, and the mucins they produce, have been implicated in Th2-mediated defence in mice resistant to a primary infection [82, 83]. mice. (A) MLNs from mice vaccinated with 50 g of pre-mixed of VLPs+T-cell epitopes (HBc-CBD1243-1259, HBc-CBD241-257, HBc-CLSP143-158 Cytarabine and HBc-CLSP398-416), 50 g of VLP (HBc-Ag), 50 g ES/Alum, or PBS on Cytarabine day -20 and boosted on day -10 were stimulated at 5×106/ml with 50 g/ml ES. After 48 hours of stimulation, cell culture supernatants were harvested and assayed by cytometric bead array for IL-4 (A), IL-6 (B), IL-10 (C), IL-17 (D), and TNF- (E) production. Results are shown as mean SEM. n = 11 mice per group. The results presented are from two separated experiment pooled together. (F) Serum mMCPT-1 levels assayed at a 1:100 dilution utilising a mouse mMCPT-1 ELISA kit (Invitrogen) according to manufacturers instructions. (G) Day 14 p.i. sera were titrated against purified anti-mouse IgE to assess serum IgE antibody levels in VLPs+T-cell epitopes, VLP (HBc-Ag), PBS and in ES/Alum vaccinated mice by ELISA (reading at 405 nm, with reference of 570nm subtracted). Statistical analyses were carried out using the Kruskal-Wallis test (multiple comparisons). Significant differences between groups are represented by (*P0.05, **P0.01, ***P0.001) with a line. This experiment was repeated two times, and the ELISA results shown here are representative of the two experiments.(PPTX) ppat.1008243.s002.pptx (323K) GUID:?CF1A2694-B722-477A-8D5D-D509B300869C S1 Table: List of Keywords used to screen for MHC I and II prediction tools. (DOCX) ppat.1008243.s003.docx (25K) GUID:?B6B56D2E-9F39-4C42-9758-AC9956640616 S2 Table: List of MHC class I and II and HLA binders peptides prediction tools. (DOCX) ppat.1008243.s004.docx (36K) GUID:?070E4504-08B7-4778-83C6-835F2B59BA38 S3 Table: MHC-II T-cell epitopes dataset used to evaluate the MHC-II prediction tools. (DOCX) ppat.1008243.s005.docx (59K) GUID:?573CF8B2-81CB-4F8E-8D76-F858501F937C S4 Table: List of MHC-II T-cell epitope sequences and primers used to amplify the target. (DOCX) ppat.1008243.s006.docx (26K) GUID:?F21D531D-3323-4E10-B3CF-2A4AE686A1AB Data Availability StatementAll relevant data are within the manuscript and its Supporting Information files. Abstract is a parasite that infects 500 million people worldwide, leading to colitis, growth retardation and Trichuris dysentery syndrome. There are no licensed vaccines available to prevent infection and current treatments are of Cytarabine limited efficacy. infections are linked to poverty, reducing childrens educational performance and the economic productivity of adults. We employed a systematic, multi-stage process to identify a candidate vaccine against trichuriasis based on the incorporation of selected T-cell epitopes into virus-like particles. We conducted a systematic review to identify the most appropriate prediction tools to predict histocompatibility complex class II (MHC-II) molecule T-cell epitopes. These tools were used to identify candidate MHC-II epitopes from predicted ORFs in the genome, selected using inclusion and exclusion criteria. Selected epitopes were incorporated into Hepatitis B core antigen virus-like particles (VLPs). Bone marrow-derived dendritic cells and bone marrow-derived macrophages responded to VLPs irrespective of whether the VLP also included T-cell epitopes. The VLPs were internalized and co-localized in the antigen CD44 presenting cell lysosomes. Upon challenge infection, mice vaccinated with the VLPs+T-cell epitopes showed a significantly reduced worm burden, and mounted genome-based CD4+ T-cell epitope prediction, combined with VLP delivery, offers a promising pipeline for the development of an effective, safe and affordable helminth vaccine. Author summary The soil transmitted helminth is a major parasite in developing countries; development of a comprehensive vaccine has been elusive. Here we used a systematic approach based on identification of MHC-II T-cell epitopes from genome sequences, their incorporation into a virus-like particle (VLP), characterization of the assemblies and testing in an murine infection model. Animals vaccinated with a.