anti human cd19 pe cy7  (Cytek Biosciences)

Journal: Frontiers in Immunology

Article Title: AIP56, an AB toxin secreted by Photobacterium damselae subsp. piscicida , has tropism for myeloid cells

doi: 10.3389/fimmu.2024.1527088

Figure Lengend Snippet: Antibodies used for flow cytometry.

Article Snippet: , Mouse anti-human CD19 , PE-Cy7 , HIB19 , Invitrogen/25-0199-42.

Techniques: Cytometry

Journal: Mucosal immunology

Article Title: Single-cell analysis of human nasal mucosal IgE antibody secreting cells reveals a newly minted phenotype

doi: 10.1016/j.mucimm.2023.02.008

Figure Lengend Snippet: B cell and plasma cell composition of NPs. A, primary population gating strategy for fluorescence-activated cell sorted CD19 + ASCs, CD19 − ASCs, and B cell subsets. One representative patient sample was selected for display (AFRS 1); B, B cell subset frequency of total CD19 + B cells in NPs; C, histograms of CD138 expression in naïve B cells from one donor (AFRS 1) and in ASCs from AFRS 1, 2, and 3 (left). CD19 + CD138 + cell frequency of total CD19 + ASCs (right); D, relative proportions of immunoglobulin isotype subclasses as a percentage of total VDJ sequences isolated ( n = 5907); E, stacked barplots showing the absolute number of ASCs per polyp (left) and the relative composition of each NP VDJ repertoire (right). AFRS = allergic fungal rhinosinusitis; ASC = antibody secreting cell; CD = cluster of differentiation; DN = double-negative B cells; Ig = immunoglobulin; NP = nasal polyp; SWM = switched memory; USM = unswitched memory; VDJ = immunoglobulin gene.

Article Snippet: An antibody cocktail was then added to the single-cell suspension, which included the following: normal mouse serum, FITC anti-human IgD (BD, clone IA6-2), Brilliant Violet 711 anti-human CD3 (Biolegend, clone OKT3), Brilliant Violet 711 anti-human CD14 (Biolegend, clone M5E2), PE/Cy7 anti-human CD19 (BD, clone SJ25C1), V450 anti-human CD38 (BD, clone HIT2), APC anti-human CD138 (Miltenyi Biotec, clone 44F9), APC-eFluor 780 anti-human CD27 (eBiosciences, clone O323), and LiveDead Aqua (Invitrogen).

Techniques: Fluorescence, Expressing, Isolation

Journal: Mucosal immunology

Article Title: Single-cell analysis of human nasal mucosal IgE antibody secreting cells reveals a newly minted phenotype

doi: 10.1016/j.mucimm.2023.02.008

Figure Lengend Snippet: NP VDJ repertoire features. A, relative lineage size in the CD19 + and CD19 − compartments, stratified by polyp. IgE members of lineages are denoted by red spots above their respective lineage; B, violin plot showing IGHV mutation frequencies across isotype subclasses; C, BASELINe analysis showing cumulative selection pressures on VDJ complementarity determining regions (solid line) and framework regions (dashed line) across isotype subclasses; D, alluvial plots showing connectivity between CD19 + ASCs and CD19 − ASCs within each polyp; E, relative VDJ repertoire diversity within the CD19 + and CD19 − compartments as measured by the inverse Simpson index. Statistical significance was determined using (B) Kruskal-Wallis testing with Dunn multiple comparisons testing between groups or (E) paired Student’s t test. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001. AFRS = allergic fungal rhinosinusitis; ASC = antibody secreting cell; CD = cluster of differentiation; Ig = immunoglobulin; IGHV = IgE ASCs featured both high and low heavy chain V region; lins = number of lineages; NP = nasal polyp; ns = not significant; Seqs = number of sequences; VDJ = immunoglobulin gene.

Article Snippet: An antibody cocktail was then added to the single-cell suspension, which included the following: normal mouse serum, FITC anti-human IgD (BD, clone IA6-2), Brilliant Violet 711 anti-human CD3 (Biolegend, clone OKT3), Brilliant Violet 711 anti-human CD14 (Biolegend, clone M5E2), PE/Cy7 anti-human CD19 (BD, clone SJ25C1), V450 anti-human CD38 (BD, clone HIT2), APC anti-human CD138 (Miltenyi Biotec, clone 44F9), APC-eFluor 780 anti-human CD27 (eBiosciences, clone O323), and LiveDead Aqua (Invitrogen).

Techniques: Mutagenesis, Selection

Journal: Mucosal immunology

Article Title: Single-cell analysis of human nasal mucosal IgE antibody secreting cells reveals a newly minted phenotype

doi: 10.1016/j.mucimm.2023.02.008

Figure Lengend Snippet: Connectivity of V(D)J region of nasal polyp ASCs with sorted B cell populations. A, connectivity plots for CD19 + sorted ASCs from polyp AFRS 1 with bulk VDJ sequencing data from four sorted B cell subsets. Red dots denote IgE ASCs; B, connectivity plots for CD19 − sorted ASCs from polyp AFRS 1 with bulk VDJ sequencing data from four sorted B cell subsets. Red dots denote IgE ASCs; C, connectivity of VDJ regions of CD19 + sorted ASCs (left) and CD19 − ASCs (right) with four sorted B cell populations. Statistical significance was determined using paired Student’s t test. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001. AFRS = allergic fungal rhinosinusitis; ASC = antibody secreting cell; CD = cluster of differentiation; DN = double-negative B cells; Ig = immunoglobulin; ns = not significant; SWM = switched memory; USM = unswitched memory; VDJ = immunoglobulin gene.

Article Snippet: An antibody cocktail was then added to the single-cell suspension, which included the following: normal mouse serum, FITC anti-human IgD (BD, clone IA6-2), Brilliant Violet 711 anti-human CD3 (Biolegend, clone OKT3), Brilliant Violet 711 anti-human CD14 (Biolegend, clone M5E2), PE/Cy7 anti-human CD19 (BD, clone SJ25C1), V450 anti-human CD38 (BD, clone HIT2), APC anti-human CD138 (Miltenyi Biotec, clone 44F9), APC-eFluor 780 anti-human CD27 (eBiosciences, clone O323), and LiveDead Aqua (Invitrogen).

Techniques: Sequencing

Journal: Mucosal immunology

Article Title: Single-cell analysis of human nasal mucosal IgE antibody secreting cells reveals a newly minted phenotype

doi: 10.1016/j.mucimm.2023.02.008

Figure Lengend Snippet: IgE ASC-specific transcriptional features. A, heatmap showing the gene expression of the top 10 DEGs identified per cluster. The red box indicates the 10 DEGs upregulated in cluster 5; B, volcano plot of DEGs identified by comparing IgE ASCs against all other CD19 + ASCs; C, dot plot indicating the number of genes and average log2fold change of genes differentially expressed in pathways enriched in IgE ASCs. DEGs detected by comparing IgE versus all other CD19 + NP ASCs; D, dot plot showing IgE specific genes, including MHC Class I and MHC Class II genes, across sorted ASC NPs grouped by isotypes; E, sorted violin plots for the log normalized gene expression for genes encoding BAFFR, TACI and BCMA. ASC = antibody secreting cell; CD = cluster of differentiation; DEGs = differentially expressed genes; HLA = ; Ig = immunoglobulin; MHC = ; NP = nasal polyp; scRNA-seq = single-cell RNA sequence.

Article Snippet: An antibody cocktail was then added to the single-cell suspension, which included the following: normal mouse serum, FITC anti-human IgD (BD, clone IA6-2), Brilliant Violet 711 anti-human CD3 (Biolegend, clone OKT3), Brilliant Violet 711 anti-human CD14 (Biolegend, clone M5E2), PE/Cy7 anti-human CD19 (BD, clone SJ25C1), V450 anti-human CD38 (BD, clone HIT2), APC anti-human CD138 (Miltenyi Biotec, clone 44F9), APC-eFluor 780 anti-human CD27 (eBiosciences, clone O323), and LiveDead Aqua (Invitrogen).

Techniques: Expressing, Sequencing