Journal: iScience

Article Title: CIITA promoter polymorphism impairs monocytes HLA-DR expression in patients with septic shock

doi: 10.1016/j.isci.2022.105291

Figure Lengend Snippet: Monocyte HLA-DR expression in 203 patients with septic shock Circulating monocyte HLA-DR expression is expressed as number of antibodies bound per cell (AB/C). Results are presented as median with 95% CI and by genotype of CIITA rs12596540 and G-286A∗rs3087456 in a dominant model. ∗p< 0.03 vs. basal (Wilcoxon signed rank test).

Article Snippet: PE-Cy™7 Mouse Anti-Human HLA-DR , BD Biosciences , Cat#560651; RRID: AB_1727528.

Techniques: Expressing

Journal: iScience

Article Title: CIITA promoter polymorphism impairs monocytes HLA-DR expression in patients with septic shock

doi: 10.1016/j.isci.2022.105291

Figure Lengend Snippet: Ex vivo HLA-DR expression in monocytes of healthy controls Results are presented as violin plot with median and quartiles lines, and by genotype of CIITA G-712A∗rs12596540 and G-286A∗rs3087456 in an A-dominant model (results for the co-dominant model presented in Figure S2 ). p values were calculated using Mann-Whitney Test. ∗∗∗∗p< 0.0001 and ∗p< 0.05.

Article Snippet: PE-Cy™7 Mouse Anti-Human HLA-DR , BD Biosciences , Cat#560651; RRID: AB_1727528.

Techniques: Ex Vivo, Expressing, MANN-WHITNEY

Journal: iScience

Article Title: CIITA promoter polymorphism impairs monocytes HLA-DR expression in patients with septic shock

doi: 10.1016/j.isci.2022.105291

Figure Lengend Snippet:

Article Snippet: PE-Cy™7 Mouse Anti-Human HLA-DR , BD Biosciences , Cat#560651; RRID: AB_1727528.

Techniques: Fluorescence, Quantitation Assay, Binding Assay, Recombinant, Luciferase, Transfection, Software, Plasmid Preparation, Mutagenesis

Journal: JCI Insight

Article Title: Dectin-1 genetic deficiency predicts chronic lung allograft dysfunction and death

doi: 10.1172/jci.insight.133083

Figure Lengend Snippet: Following 4 hours of stimulation with control media or zymosan, monocytes were assessed by immunophenotyping those from CLEC7A variants (AC, n = 8) and wild-type CLEC7A (AA, n = 8). (A) Monocytes were defined by forward scatter, side scatter, size, viability, and staining for CD14 and HLA-DR. (B) Surface dectin-1 receptors were measured on monocytes by MFI. Box-and-whisker plots show dectin-1 and HLA-DR monocyte surface expression, with boxes defining the 25th and 75th percentile of data, bisecting line depicting the median value, and whiskers capturing minimum and maximum values. (C) In control media, surface dectin-1 receptor MFI was increased on wild-type monocytes (AA) relative to Y238X variants (AC, P = 0.006). Following zymosan stimulation, this difference was maintained between AA and AC genotypes (P = 0.05). There was no difference in dectin-1 receptor MFI on wild-type or variant-genotype monocytes in control media or following stimulation. (D) HLA-DR MFI was also measured. There was significantly decreased HLA-DR MFI on CLEC7A-variant monocytes (AC) compared with wild-type monocytes (AA, P = 0.03) in control media. The HLA-DR MFI decreased on AA-genotype monocytes (P = 0.01) but not on AC-genotype monocytes following stimulation. Compared to wild-type CLEC7A genotypes, there was no difference in HLA-DR MFI on Y238X variant-genotype monocytes following zymosan stimulation. Differences were assessed using 2-tailed Student’s t test and the experiment was conducted twice.

Article Snippet: Samples were pretreated with saturating concentrations of human aggregated immunoglobulin to prevent nonspecific binding and then labeled with the following reagent and antibody combinations to define monocytes: viability dye (eBioscience, 65-0863-18), fluorescein isothiocyanate–conjugated (FITC-conjugated) anti-CD14 (clone HCD14, VWR International, 325604), allophycocyanin-cyanine 7 (APC-Cy7)–conjugated anti–HLA-DR (clone L243, VWR International, 307617), R-phycoerythrin (PE)–conjugated anti–dectin-1 (clone 259931, Thermo Fisher Scientific, FAB1859P025).

Techniques: Staining, Whisker Assay, Expressing, Variant Assay