Journal: PLOS Pathogens

Article Title: Liver stage P. falciparum antigens highly targeted by CD4 + T cells in malaria-exposed Ugandan children

doi: 10.1371/journal.ppat.1012943

Figure Lengend Snippet: A. Protocol for in vitro T cell expansion and testing. B. Response against the 39-peptide pool measured by IFN-γ ELISPOT on day 14 of T cell expansion. Each point represents a T cell line generated from an individual donor using the protocol outlined in A. C. Correlation between total IFN-γ-producing cells measured by ELISPOT and the frequency of IFN-γ + CD4 + T cells measured by flow cytometry in response to the 39-peptide pool. Robust linear regression fit of the data, with Spearman’s ρ and p-value, shown in blue. D. Proportion of donors responding to pooled peptides, grouped by recency of their last malaria episode. SFUPMC: spot-forming units per 10 6 input cells.

Article Snippet: ELISPOT screenings were performed using the ImmunoSpot human IFN-γ ELISPOT kit (CTL, Shaker Heights, OH, USA) following the protocol provided by the manufacturer.

Techniques: In Vitro, Enzyme-linked Immunospot, Generated, Flow Cytometry

Journal: PLOS Pathogens

Article Title: Liver stage P. falciparum antigens highly targeted by CD4 + T cells in malaria-exposed Ugandan children

doi: 10.1371/journal.ppat.1012943

Figure Lengend Snippet: A . Response against individual peptides in deconvolution IFN-γ ELISPOT experiments. Left: allele haplotypes for each of the donors tested (A. donor is homozygous for DRB1 allele on the horizontal axis; A. donor carries the DRB1 on the horizontal axis and another DRB1 allele). Right: points indicate positive IFN-γ responses. B. Magnitude of IFN-γ response against peptides measured by flow cytometry. The gating strategy used to analyze this data is detailed in S2 Fig. In A and B , peptides (x axis) are grouped by their antigen of origin (top ribbons). C. pMHC tetramer staining of CD4+ T cell lines from malaria-exposed children and their IFN-γ secretion in response to stimulation with peptides L.9.30 or L.9.27. FMO: fluorescence-minus-one control.

Article Snippet: ELISPOT screenings were performed using the ImmunoSpot human IFN-γ ELISPOT kit (CTL, Shaker Heights, OH, USA) following the protocol provided by the manufacturer.

Techniques: Enzyme-linked Immunospot, Flow Cytometry, Staining, Fluorescence, Control

Journal: Frontiers in Immunology

Article Title: Local Innate Markers and Vaginal Microbiota Composition Are Influenced by Hormonal Cycle Phases

doi: 10.3389/fimmu.2022.841723

Figure Lengend Snippet: Cytokine and chemokine expression in the plasma of female cynomolgus macaques (n = 9). (A) Heatmap representing the mean fold change in the expression of cytokines and chemokines in the plasma of each animal (n = 9). The fold change was calculated based on the mean expression of each cytokine/chemokine for all females and time points. One way ANOVA was performed to compare the total fold change value to those of each animal for each cytokine. Asterisks indicate p values considered to be statistically significant (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001). (B) Heat map representing the mean fold change in the expression of cytokines and chemokines in the plasma for all animals according to time (n = 9). The fold change was calculated based on the mean expression of each cytokine/chemokine in each female. Numbers 1 to 14 refers to weeks. Red arrows represent menstruation.

Article Snippet: Pro- and anti-inflammatory cytokines, as well as chemokines, were quantified in cervicovaginal fluids and plasma using a 23plex assay for the detection of G-CSF, GM-CSF, IFNγ, IL-1β, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12/23(p40), IL-13, IL-15, IL-17A, CCL2, CCL3, CCL4, sCD40L, TGFα, TNFα, VEGF, and IL-18 (NHP cytokine magnetic bead panel kit; Merck Millipore; Germany), according to the manufacturer’s instructions.

Techniques: Expressing, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: Local Innate Markers and Vaginal Microbiota Composition Are Influenced by Hormonal Cycle Phases

doi: 10.3389/fimmu.2022.841723

Figure Lengend Snippet: Cytokine and chemokine expression in cervicovaginal fluids of female cynomolgus macaques (n = 9). (A) Heatmap representing the mean fold change in the expression of cytokines and chemokines in the cervicovaginal fluids of each animal (n = 9). The fold change was calculated based on the mean expression of each cytokine/chemokine for all females. One way ANOVA was performed to compare the total fold change value to those of each animal for each cytokine. (B) Heat map representing the mean fold change in the expression of cytokines and chemokines in the cervicovaginal fluids of all animals (n = 9). The fold change was calculated based on the mean expression of each cytokine/chemokine for each female. Numbers 1 to 14 refers to weeks. Red arrows represent menstruation (C) Samples clustered into three groups based on the progesterone level or menstruation and each cytokine/chemokine concentration was plotted. A Kruskal-Wallis test with Dunn’s test to adjust the p value was performed. Asterisks indicate p values considered to be statistically significant (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001).

Article Snippet: Pro- and anti-inflammatory cytokines, as well as chemokines, were quantified in cervicovaginal fluids and plasma using a 23plex assay for the detection of G-CSF, GM-CSF, IFNγ, IL-1β, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12/23(p40), IL-13, IL-15, IL-17A, CCL2, CCL3, CCL4, sCD40L, TGFα, TNFα, VEGF, and IL-18 (NHP cytokine magnetic bead panel kit; Merck Millipore; Germany), according to the manufacturer’s instructions.

Techniques: Expressing, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Local Innate Markers and Vaginal Microbiota Composition Are Influenced by Hormonal Cycle Phases

doi: 10.3389/fimmu.2022.841723

Figure Lengend Snippet: Heatmap representing bacterial taxa (family level), neutrophil subpopulations, and cytokine/chemokine levels that varied during the hormonal cycle. The heatmap represents the log10 fold change for each parameter (cytokines, neutrophil subpopulations, bacterial taxa, indicated on the right). Increased values are shown in red and decreased in blue. Each animal is represented by a color code, as well as the hormonal phases (high progesterone in green, low progesterone in orange, menstruation in red). Hierarchical clustering divided the samples (sample ID in the X-axis) into two large clusters (A, B) . Each cluster was then separated into subclusters (A1, A2 and B1, B2).

Article Snippet: Pro- and anti-inflammatory cytokines, as well as chemokines, were quantified in cervicovaginal fluids and plasma using a 23plex assay for the detection of G-CSF, GM-CSF, IFNγ, IL-1β, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12/23(p40), IL-13, IL-15, IL-17A, CCL2, CCL3, CCL4, sCD40L, TGFα, TNFα, VEGF, and IL-18 (NHP cytokine magnetic bead panel kit; Merck Millipore; Germany), according to the manufacturer’s instructions.

Techniques: