Consequently, secondary antibodies (anti-human IgG Q800, Grace Biolabs, #110,635), were applied at a dilution of 1 1:250 and incubated for 2?h. Dot storyline graphs display the relation between the estimated quantity of pre/post-immunization GMZ2 stimulated cytokine producing CD4+ T cells (top side), the number of and B cells subsets (bottom side) regarding medical malaria status after CHMI. Monotone increase of parasitemia with symptoms (Malaria) is definitely represented by black places. Low oscillating parasitemia with no symptoms (Control) plus individuals with neither parasitemia nor symptoms (Guarded) are displayed by open circles. Comparison of the cell number of GMZ2 stimulated CD4+ T cells, of CD20+ B cells and the GMZ2-specific B subsets was performed using Mann-Whitney (for T cells) or unpaired t-tests (for B cells). Data are from a single experiment after several optimization checks, and individual volunteers were measured in separate experiments. Symbols represent individual samples. Number S5. Post-immunization cell frequencies and the time to treatment after CHMI. Graphs display the time to 1st malaria treatment concerning the portion of specific triple and double positive CD4+ T cells, total B Thapsigargin cells and the CD27+/- cluster subsets of GMZ2-specific within CD20+IgG+ B cells (a), or the number of GMZ2-stimulated triple and double positive CD4+ T cells, and the number of total B cells and different GMZ2+B cells (b) at D84. Ideals above the median are displayed in reddish whereas data below the median are demonstrated in blue. The Log-rank test was used to compare the two curves. p value lower than 0.05 is considered significant. Number S6. Correlation between the estimated quantity of B cell phenotypes Thapsigargin and the anti-GMZ2 IgG concentration at baseline. The association between the estimated quantity of CD20+IgG+ B cells, the estimated quantity of GMZ2-specific B cells and the anti-GMZ2 IgG concentration, was performed on D0 data using Pearsons correlation after log transformation. Data are from a single experiment after several optimization checks, and individual volunteers were measured in separate experiments. Symbols represent individual samples. A p-value less than 0.05 is considered as statistically significant. Number S7. B cell phenotypes rate of recurrence following Thapsigargin immunization concerning vaccine treatment. Frequencies of CD20+IgG positive, CD20+IgG+GMZ2-specific and GMZ2-specific CD27+/- B cells between D0 and D84 are compared for those volunteers concerning vaccine treatment. Vaccinated subjects with Rabies control vaccine are displayed with opened squares. GMZ2 vaccinated discriminate vaccinees receiving 100g GMZ2-Alhydrogel (opened dots), 30g GMZ2-CAF01 (gray dots), and 100g GMZ2-CAF01 (black dots). Wilcoxon test following by Bonferroni correction for multiple assessment is performed to test statistical significance. p value below 0.05 is considered statistically significant. Data are from a single experiment after several optimization checks, and individual volunteers were measured in separate experiments. Symbols represent individual samples. Red lines symbolize the median ideals with interquartile range. p value lower than 0.05 is considered significant. Table S1. Cox proportional analysis post-immunization 12936_2022_4169_MOESM1_ESM.docx (5.0M) GUID:?5FC1DC26-18E5-4B1A-B435-AB33075F83A2 Additional file 2: Table S2. Complete list of antigens noticed on the protein microarray. 12936_2022_4169_MOESM2_ESM.xlsx (34K) GUID:?3B06DA23-0EF1-4B22-A213-A93D3D41AA9B Additional file 3: Table S3. The antibody response against Thapsigargin the vaccine antigen (GMZ2) or any of the main parts (MSP3, GLURP), estimated before vaccination, was tested for correlation with any of the antibody reactions, as estimated from the ENOX1 protein microarray before CHMI. The log-transformed ELISA data and the protein microarray data were tested usind Student’s t-test for correlation. Antigens with correlations resulting in p-values < 0.05 are shown. 12936_2022_4169_MOESM3_ESM.xlsx (22K) GUID:?F46F47F8-9C70-4C34-92C3-C93D37754D51 Data Availability StatementThe datasets generated during and/or analysed during the current study are available from.
