Journal: Nature Communications

Article Title: Exposure to pesticides in utero impacts the fetal immune system and response to vaccination in infancy

doi: 10.1038/s41467-020-20475-8

Figure Lengend Snippet: a Cord blood non-naive (Supplementary Fig. ) CD4 T cell frequency in exposed (red, n = 98) and unexposed (blue, n = 68) infants. b Cord blood T reg frequency (CD25 high FoxP3 + CD127 lo ) in exposed (red, n = 98) and unexposed (blue, n = 68) infants. c Myeloid (Lin-2 – HLA-DR + CD11c + CD123 - ) and plasmacytoid (Lin-2 – HLA-DR + CD11c − CD123 + ) dendritic cell counts in whole cord blood, in exposed (red, n = 142) and unexposed (blue, n = 96) infants. d Cord blood plasma cytokine levels (unadjusted p values: TNF-RI ( p < 0.0001), IL-2 ( p < 0.0001), IL-4 ( p = 0.037), IL-5 ( p < 0.0001), IL-7 ( p < 0.0001), MDC ( p < 0.0001), TNF-RI ( p < 0.0001), IP-10 ( p = 0.0009), and MIP-1α ( p = 0.01), ran in duplicate) in exposed (red, n = 114) and unexposed (blue, n = 73) infants. Following Bonferroni adjustment for all cytokines and chemokines measured, all associations indicated above remained significant with the exception of IL-4 and MIP-1α. All comparisons were made by Wilcoxon rank-sum test. Bars indicate median with interquartile range. Two-sided p values were calculated for all test statistics.

Article Snippet: Dendritic cell staining was performed on 238 cord blood samples using FITC Lin-2 (clone MφP9/SJ25C1/SK7/L27/NCAM16.2, anti-CD3, CD14, CD19, CD20, CD56), PE CD123 (clone 9F5), PerCP HLA-DR (clone L243), and APC CD11c (clone S-HCL-3, BD Pharmingen).

Techniques:

Journal: Journal of Translational Medicine

Article Title: First-line chemoimmunotherapy in metastatic breast carcinoma: combination of paclitaxel and IMP321 (LAG-3Ig) enhances immune responses and antitumor activity

doi: 10.1186/1479-5876-8-71

Figure Lengend Snippet: IMP321 increases the numbers of monocytes, NK and activated CD8 T cells in blood (panel A) . Fresh blood samples were collected pre-dosing, at D1, D85 and D170 and directly stained with fluorochrome-conjugated antibodies in tubes containing a precise number of fluorescent control beads. The results show the mean ± sd of the absolute numbers of CD45 + CD14 + (monocytes), CD3 - CD56 + (NK cells) and CD38 + HLA-DR + CD8 + (activated CD8 + cells) cells. The paired non-parametric Wilcoxon signed rank test was used to compare increases observed between D85 or D170 and D1. When significant (< 0.05), p values are indicated. IMP321 increases the percentages of dendritic cells and cytotoxic CD45RA + Effector-Memory CD8 + T cells (EMRA) in PBMCs (panel B). PBMCs cells collected pre-dosing, at D1, D85 and D170 were isolated and stained with fluorochrome-conjugated antibodies and analyzed by flow cytometry. The results show the mean ± sd of the percentages of plasmacytoid dendritic cells (pDC, CD45 + CD14 - CD16 - HLA-DR + CD123 + CD11c - ) and myeloid dendritic cells (mDC, CD45 + CD14 - CD16 - HLA-DR + CD123 - CD11c + ) in CD45 + cells, as well as the percentages of CD45RA + CD45RO - CD62L - in the CD8 + T cell population (CD45RA + EM CD8 T cells or EMRA). The significant Wilcoxon p values are indicated.

Article Snippet: Blood samples were collected pre-dosing at D1, D85 and D170 and directly stained with BD Multitest CD8-FITC/CD38-PE/CD3-PerCP/HLA-DR-APC, with BD Multitest 6-color TBNK (CD3-FITC/CD16-PE+CD56-PE/CD45-PerCPCy5.5/CD4PE-Cy7/CD19-APC/CD8-APC-Cy7), BD Simultest LeucoGate (CD45-FITC/CD14-PE) in tubes containing a precise number of fluorescent control beads (BD Trucount™tubes, BD Biosciences).

Techniques: Staining, Isolation, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: First-line chemoimmunotherapy in metastatic breast carcinoma: combination of paclitaxel and IMP321 (LAG-3Ig) enhances immune responses and antitumor activity

doi: 10.1186/1479-5876-8-71

Figure Lengend Snippet: IMP321 increases the expression of activation markers on blood monocytes . Blood samples were collected pre-dosing, at D1, D85 and D170 and directly stained with fluorochrome-conjugated CD45, CD14, anti-HLA-DR and CD11a, CD11b, CD16, CD35, CD54, CD64, CD80 or CD86 antibodies in tubes containing a precise number of fluorescent control beads. The expression of activation markers on monocytes was directly proportional to the cell-bound fluorescence. The results shown in panel A are the mean ± sd after normalization of the cell-bound fluorescence against the fluorescence of control beads. Statistically significant increases between D85 or D170 and D1 are analyzed using Wilcoxon signed rank test and significant p values (< 0.05) are shown. In panel B, the percentage of patients showing increases in the expression of the indicated numbers of activation markers at D85 or D170 compared to the baseline at D1 was calculated. The number of markers (n) displaying an increase by at least 50% was calculated for each patient in the 1.25 mg (7 patients) and 6.25 mg (12 patients) groups. The pie charts represent the percentages of patients with increases in the indicated number of markers.

Article Snippet: Blood samples were collected pre-dosing at D1, D85 and D170 and directly stained with BD Multitest CD8-FITC/CD38-PE/CD3-PerCP/HLA-DR-APC, with BD Multitest 6-color TBNK (CD3-FITC/CD16-PE+CD56-PE/CD45-PerCPCy5.5/CD4PE-Cy7/CD19-APC/CD8-APC-Cy7), BD Simultest LeucoGate (CD45-FITC/CD14-PE) in tubes containing a precise number of fluorescent control beads (BD Trucount™tubes, BD Biosciences).

Techniques: Expressing, Activation Assay, Staining, Fluorescence