Journal: bioRxiv

Article Title: Specific induction of double negative B cells during protective and pathogenic immune responses

doi: 10.1101/2020.09.08.285148

Figure Lengend Snippet: Distribution of B cell subsets in the peripheral blood of patients with neuro-inflammatory disorders or following vaccination with influenza or tick borne encephalitis virus. B cell subsets are presented as a percentage of total CD19 + B cells and defined as follows: naïve B cells CD19 + CD20 + CD27 − CD38 + IgD + , memory B cells CD19 + CD20 + CD27 + CD38 + , double negative B cells (DN B cells) CD19 + CD20 low CD27 − IgD − , plasmablasts CD19 + CD20 low CD27 + CD38 high IgD − . ( A ) Peripheral blood B cell subsets were quantified from patients with neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis, meningitis / encephalitis (mening./enceph.), Guillain-Barré syndrome (GBS), multiple sclerosis (MS), non-inflammatory neurological diseases (NIND) and patients with rheumatic diseases. Red dots highlight samples obtained from rituximab-treated patients. Clinical groups were compared using a Kruskal-Wallis test with correction for multiple comparisons. Peripheral blood B cell subsets following vaccination against ( B ) influenza virus (n = 22) or ( C ) tick-borne encephalitis virus (n = 6). Statistical measurements were performed using a repeated measures ANOVA with Dunnett’s post-hoc correction. ( D ) Staining of different B cell populations was visualized by ImageStream. Lines in the graphs indicate median values and asterisks describe significance values as follows: * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: For flow cytometric analyses, the following antibodies were used for all analyses: CD38 FITC (BD), IgD PE (Biozol), CD19 ECD (Beckman Coulter), CD3 PeCy7 (Beckman Coulter), CD45 VM (BD), CD27 APC (BD), CD20 APC Cy7 (BD).

Techniques: Staining

Journal: JCI Insight

Article Title: Differential transcriptome and development of human peripheral plasma cell subsets

doi: 10.1172/jci.insight.126732

Figure Lengend Snippet: (A) Top panels divide the CD19+ and CD19– fractions. Lower panels represent subsets of CD19–IgD– (left) and CD19+IgD– (right) fractions. (B) Morphology of blood ASC subsets (×100 magnification) by Wright-Giemsa stain. Left column: Sorted blood ASC subsets on day 7 after tetanus vaccination. ASC populations (pops) 1 to 5 and naive B cells are shown. Right column: Percentage of intracellular BLIMP-1 staining per subset is shown in blue histograms (naive controls in red). (C) Percentage of each ASC subset and naive B cells (N) expressing IgG, IgA, or IgM isotypes after peak vaccination. (D) Quantification of each blood ASC subset (pops 1 to 5) in cells/ml (top) and percentage of PBMCs (bottom). (E) Quantitative RNA expression of 5,000 sorted ASC subsets and naive and memory B cells for Pax5 (top), BLIMP-1 (middle), and Xbp-1 (lower), normalized to GAPDH in blood. Relative mRNA expression is expressed in arbitrary units.

Article Snippet: CD3 – CD14 – cell fractions were stained with the following anti-human antibody staining reagents: IgD-PE (catalog, 555779), CD19–PE-Cy7 (catalog, 557835), CD38–Pacific Blue (catalog, 561378) (BD Pharmingen); CD3–PE-Cy5.5 (catalog MHCD0318), CD14–PE-Cy5.5 (catalog MHCD1418) (Invitrogen); CD138-APC (Miltenyi Biotec, catalog 130-091-250); and CD27–APC-eFluor 780 (eBioscience, catalog 47-0279).

Techniques: Giemsa Stain, Staining, Expressing, RNA Expression

Journal: JCI Insight

Article Title: Differential transcriptome and development of human peripheral plasma cell subsets

doi: 10.1172/jci.insight.126732

Figure Lengend Snippet: (A) CD20, surface Ig (kappa and lamda), and CD27 staining for blood ASC subsets and naive B cells (CD19+IgD+CD27–) illustrated in blue relative to controls in gray (also shown in right-hand panel). (B) HLA-DR and Ki-67 staining for blood ASC subsets. Far right: CD14+ peripheral blood monocytes served as controls for HLA-DR staining and naive B cells for Ki-67. (C and D) Frequency of CXCR4, CD28, IL-6R, FCGR2B, and BCMA in blood ASC subsets (pops 2 to 5) and naive B cells. Respective numbers of subjects are listed in Table 1.

Article Snippet: CD3 – CD14 – cell fractions were stained with the following anti-human antibody staining reagents: IgD-PE (catalog, 555779), CD19–PE-Cy7 (catalog, 557835), CD38–Pacific Blue (catalog, 561378) (BD Pharmingen); CD3–PE-Cy5.5 (catalog MHCD0318), CD14–PE-Cy5.5 (catalog MHCD1418) (Invitrogen); CD138-APC (Miltenyi Biotec, catalog 130-091-250); and CD27–APC-eFluor 780 (eBioscience, catalog 47-0279).

Techniques: Staining