percp anti cd8a  (Miltenyi Biotec)

Journal: Npj Imaging

Article Title: Reactive oxygen species-related oxidative changes are associated with splenic lymphocyte depletion in Ebola virus infection

doi: 10.1038/s44303-025-00079-x

Figure Lengend Snippet: a From left to right, the number and frequencies of total and apoptotic populations of CD8 + T lymphocytes, CD4 + T lymphocytes, B lymphocytes, and CD11b + myeloid cells are presented over the course of the experiments. b Correlation matrix between imaging and immunohistochemistry and flow cytometry of the spleens (Spearman ranks). * indicates statistical significance ( p < 0.05). 4-HNE 4-hydroxy-2-nonenal; MPO myeloperoxidase; MRI magnetic resonance imaging; non-exp non-exposed; 3 dpe 3 d post-exposure; dt day terminal.

Article Snippet: Blocked samples were stained in total volumes of 200 µL with a mixture of conjugated primary antibodies including CD11b BV510 (clone M1/70; Biolegend, San Diego, CA, USA), CD25 Alexa Fluor 647 (clone 7D4; BD Biosciences, Franklin Lakes, NJ, USA), HLA-DR BV650 (clone L243; Biolegend), F4/80 BV711 (clone BM8; Biolegend), CD4 FITC (clone 02; Sino Biological US Inc., Wayne, PA, USA), CD11a PE (clone 2D7; Biolegend), and CD8a PerCP (clone Lyt2; Sino Biological US Inc.), amine-reactive dye (Live Dead Blue; Thermo Fisher Scientific, Waltham, MA, USA) for dead cell exclusion, and Brilliant Stain Buffer Plus (BD Biosciences) for 20–30 min on ice.

Techniques: Imaging, Immunohistochemistry, Flow Cytometry, Magnetic Resonance Imaging

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Immunophenotyping of PBMCs from pregnant SARS-CoV-2 infected and recovered patients. (a) Malaysian Cohort 1: Description of PBMCs used for immunophenotyping collected from pregnant healthy controls (Preg-HC, blue), SARS-CoV-2 infected (Preg-INF, orange) and COVID-19 recovered (Preg-R, pink). (b) Unsupervised clustering of immune cells based on 14-color flow cytometry panel. Each PBMC sample from Preg-HC, Preg-INF, and Preg-R groups were concatenated bioinformatically using the FlowJo software. Two major clusters for lymphocytes (pink) and monocytes (blue) population were identified and presented on a UMAP plot. The combined cell population is shown in gray. (c) UMAP plot characterising 7 identified immune subsets; monocytes (cyan), CD4 + T cells (navy), CD8 + T cells (red), NKT cells (lilac), CD19 + B cells (green), NK cells (rose), and Tregs (yellow). (d) Overlay of UMAP to show the different immune cell landscape; Preg-HC (blue), Preg-INF (orange), and Preg-R (pink) patient groups. The combined cell population is shown in gray. (e) Delineation of all CD8 + T subsets. UMAP overlay showing the central memory (blue), early (red), intermediate (orange), late effector (bright green), and Naïve (dark green) CD8 + T cells. The combined cell population is shown in rose. (f) UMAP overlay depicting the various CD8 + T cells based on the different patient groups. Preg-HC (cyan), Preg-R (pink), and Preg-D (orange). The combined cell population is shown in gray. (g) Original FACS plots of cytotoxic CD8 + T cells. Identification of Naïve, memory, and effector memory CD8 + T cells based on CD45RA and CCR7 markers. FACS plots show the CD45RA + CCR7 + naïve, CD45RA - CCR7 + CM cells, CD45RA +high CCR7 - late EM, CD45RA +mid CCR7 + intermediate EM, CD45RA - CCR7 - early EM cells in the different patient groups. (h) The percentage of Naïve (upper left), intermediate (EM II, right) and late EM III (bottom left) cells are shown as violin plots with each dot represents an individual sample. P-values show the significance among Preg-HC, Preg-INF and Preg-R groups and compared using Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05).

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Infection, Flow Cytometry, Software

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Increased effector CD4 + T cells and NKT cells in infected and recovered pregnant women. (a) UMAP analysis of total CD4 + T cells into different CD4 + T subsets including Naïve T cells (green), central memory T cells (CM; blue), early effector T cells (EMI; red), late effector cells (EMII; orange), and Tregs (yellow). The combined cell population is shown in gray. (b) UMAP for different CD4 + T subsets showing Preg-HC (cyan), Preg-R (pink), and Preg-INF (orange). The combined cell population is shown in gray. (c) Original FACS plots identifies CD4 + T cells subsets based on CCR7 and CD45RA markers. Classification of naïve, memory and effector memory CD4 + T cells based on CD45RA and CCR7 markers. FACS plots show the CD45RA + CCR7 + Naïve, CD45RA - CCR7 + CM cells, CD45RA +high CCR7 - late EM, CD45RA +mid CCR7 + intermediate EM, CD45RA - CCR7 - early EM cells. (d) The percentage of Naïve, early, and late EM cells are shown in violin plots. P-values show the significance among Preg-HC, Preg-INF, and Preg-R groups and compared using Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05). (e) UMAP plots showing the distribution of NKT cells (blue) in the CD3 + T cell compartment. (f) UMAP displaying the overlay with different patient groups. Preg-HC (cyan), Preg-R (pink), and Preg-INF (orange) for NKT cells. Combined samples are shown in gray. (g) Original FACS plots presenting the expression of CD56 on CD3 + CD8 + T cells with antibody markers staining for CD8 and CD56. (h) The percentage of NKT cells displayed by violin plots among Preg-HC, Preg-INF, and Preg-R groups. Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test was used to determine significance (*p ≤0.05).

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Infection, Expressing, Staining

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Validation of immunophenotyping studies and dysregulated humoral immune response in German cohort 2. (a) German Cohort 2: description of cohort and experimental plan using matched PBMCs and serum from pregnant healthy controls (Preg-HC) and COVID-19 recovered (Preg-R) pregnant women. (b) Reduced percentage of CD14 + monocytes and significantly increased CD14 + CD16 + in Preg-R patients. Box plots show median, interquartile range (IQR), and the whiskers corresponding to the highest and lowest points within 1.5 times of IQR. Each dot represents an individual sample. Wilcoxon rank-sum test was used for p-value significance to compare pregnant healthy control (Preg-HC) and COVID-19 recovered (Preg-R). P ≤0.05 considered significant (*p ≤0.05). (c) Box and whisker plot representing the percentage of Naïve, CM, EM I, and EM II CD8 + T cells in Preg-HC and Preg-R patients. CD8 + Naïve and CD8 + CM T cells were lower in Preg-R. CD8 + EM I and CD8 + EM II T cells were significantly increased Preg-R. Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). (d) Box and whisker plot representing NKT cells in Preg-R compared with Preg-HC. (e) Box and whisker’s plot showing reduced CD56 + NK cells and CD56 + CD16 + NK II cells in Preg-R. Increased levels of CD56 - HLA-DR + lymphoid cells were significantly increased in Preg-R. Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). (f) High levels of Nucleocapsid IgG antibody levels in serum of Preg-R group compared to Preg-HC (left graph). Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). Spike S1 (middle graph) and RBD IgG concentration (ug/mL; right graph) at different collection time-points (CT) points (CT1, 0 day; CT2, 39 days; and CT3, 89 days post infection) during their pregnancy. Decreased levels of Spike S1 and RBD IgG antibodies 89 days post first collection time point in Preg-R group. Each dot represents an individual. Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05). (g) Examination of IL-10, MCP-1, and IL-8 levels in the serum of recovered women up to 89 days post infection. Each dot represents an individual on a box-whisker plot. CT1, 0 day; CT2, 39 days; and CT3, 89 days post infection-during their pregnancy Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05, **p ≤0.01, ***p ≤0.001, ****p ≤0.0001).

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Biomarker Discovery, Control, Whisker Assay, Concentration Assay, Infection

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Single cell atlas and immune cell composition of healthy controls and recovered COVID-19 pregnant women. (a) Integrated UMAP (UMAP-CCA) of 30,394 cells derived from PBMCs. Sc-RNA-seq data were analyzed using Seurat pipeline. UMAP plots shown the different immune cell subsets based on distinct gene expression. Cell types are color-coded as shown in UMAP. (b) Dot plots show the expression level of canonical cell markers used to assign cell subset identification for major cell type including CD4 + T, CD8 + T, and B cells. (c) Feature plots show the key canonical markers used for identification of individual cell types. Green intensity shows increasing expression. (d) Percentage of major cell types in Preg-HC and Preg-R patients. (e) UMAP analysis of individual Preg-HC (blue) and Preg-R (orange) patient groups which highlight a reduced NK I cell population.

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Derivative Assay, RNA Sequencing, Gene Expression, Expressing

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Reduced cytotoxic functions of NKT and NK cells in recovered pregnant women. (a) UMAP plots show the NKT (upper panel) and NK I (lower panel) cells from pregnant healthy controls and recovered patients. (b) Differential gene expression analysis of NKT, NK I, and CD8 + TEMRA cells using volcano plots. (c) GSEA pathway enrichment analysis for NK (I) Selected activated and suppressed pathways are shown on GSEA bubble plots. (d) KEGG pathway analysis of NK I cells. Most significantly pathways are shown on the GSEA plot. (e) Dot plot represents the selected cytotoxic function expressing genes in NKT (C3), NK I (C7), NK II (C8), and NK III (C18) cells. (f) Violin plots show significantly regulated genes related with cytotoxic functions in NK I cells. Each represents one cell. (g) Dot plot representing genes related with cytotoxic functions in CD8 + TEMRA (C2) cells.

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Gene Expression, Expressing