Journal:

Article Title: Induction of Specific Cytotoxic Lymphocytes in Mice Vaccinated with Brucella abortus RB51

doi: 10.1128/IAI.69.9.5502-5508.2001

Figure Lengend Snippet: Different roles of CD4+ and CD8+ T cells in the cytotoxic lysis and release of IFN-γ. The two T-cell populations were isolated by the MACS magnetic kit and showed more than 92% purity by flow cytometry. The individual T-cell populations were cocultured at various concentrations with either strain RB51-infected or normal J774.A1 cells. The cytotoxic activity (A) and the amount of IFN-γ released into the supernatants (B) were measured. The data are means for triplicate estimations, and standard deviations did not exceed 20% of the means. Cocultures of CD4+ T cells with noninfected target macrophages (solid squares) and RB51-pulsed target macrophages (solid triangles) and CD8+ T cells with noninfected target cells (solid diamonds) and RB51-pulsed target cells (solid circles) were tested.

Article Snippet: Briefly, live T cells isolated by Histopaque column purification were incubated with MACS magnetic MicroBeads to which monoclonal antibodies against CD4 molecule (clone GK1.5; isotype, rat IgG2b) or CD8 molecule (clone 53-6.7; isotype, rat IgG2a) had been coupled (Miltenyi Biotec, Auburn, Calif.) at the concentration of 10 μl of MicroBeads per 10 7 total cells for 15 min in a refrigerator at 4°C (see Fig. ).

Techniques: Lysis, Isolation, Flow Cytometry, Infection, Activity Assay

Journal:

Article Title: Induction of Specific Cytotoxic Lymphocytes in Mice Vaccinated with Brucella abortus RB51

doi: 10.1128/IAI.69.9.5502-5508.2001

Figure Lengend Snippet: Specific phenotype analysis of effector cells by flow cytometry a

Article Snippet: Briefly, live T cells isolated by Histopaque column purification were incubated with MACS magnetic MicroBeads to which monoclonal antibodies against CD4 molecule (clone GK1.5; isotype, rat IgG2b) or CD8 molecule (clone 53-6.7; isotype, rat IgG2a) had been coupled (Miltenyi Biotec, Auburn, Calif.) at the concentration of 10 μl of MicroBeads per 10 7 total cells for 15 min in a refrigerator at 4°C (see Fig. ).

Techniques: Flow Cytometry

Journal: Nature Communications

Article Title: Fasting mimicking diet in mice delays cancer growth and reduces immunotherapy-associated cardiovascular and systemic side effects

doi: 10.1038/s41467-023-41066-3

Figure Lengend Snippet:

Article Snippet: Anti-mouse CD4, VioBright FITC (REA604) , Miltenyi Biotec , 130-118-692.

Techniques: In Vivo

Journal: Science Advances

Article Title: Altered X-chromosome inactivation predisposes to autoimmunity

doi: 10.1126/sciadv.adn6537

Figure Lengend Snippet: ( A ) Representative flow cytometry analysis of spontaneously activated B220 + CD69 + B cells in spleen from 1-year-old WT and Ftx −/− females. Percentages in leucocytes are shown on the graphs beneath. Each triangle represents a mouse. Median values are shown (Mann-Whitney test, * P < 0.05 and ** P < 0.01). ( B ) Same as (A) for spontaneously activated CD4 + CD69 + T cells (Mann-Whitney test, * P < 0.05 and ** P < 0.01). ( C ) Total IgG, IgM, IgG2b, and IgG2c natural antibody levels in sera of 3-month-, 1-year, and >1.5-year-old WT or Ftx −/− females measured by ELISA. Each circle represents a mouse. Mean values are shown (Mann-Whitney test, * P < 0.05 and ** P < 0.01). ( D ) Cytokines levels in the blood analyzed with cytometric bead array assays on sera from 3-month-, 1-year-, or 2-year-old WT and Ftx −/− females. Each triangle represents a mouse. Median values are shown ( t test, * P < 0.05).

Article Snippet: BM, spleen, blood, and peritoneal cavity cells were stained using the following antibodies: CD3 PerCP-Vio770 (130-119-656, Miltenyi Biotec), CD4-allophycocyanin (APC) (130-123-207, Miltenyi Biotec), CD5-APC-Vio770 (130-120-165, Miltenyi Biotec), CD8-fluorescein isothiocyanate (FITC) (130-118-468, Miltenyi Biotec), CD11b APC (553312, BD Pharmingen), CD11c phycoerythrin (PE)–Vio770 (130-110-840, Miltenyi Biotec), CD19-FITC (557398, BD Pharmingen), CD21-APC-Vio770 (130-111-733, Miltenyi Biotec), CD23-PE-Vio770 (130-118-764, Miltenyi Biotec), CD38-PE (130-123-571, Miltenyi Biotec), CD43-PE (130-112-887, Miltenyi Biotec), CD69-PE (130-115-575, Miltenyi Biotec), CD138 PE-Vio615 (130-108-989, Miltenyi Biotec), F4/80 FITC (130-117-509, Miltenyi Biotec), Ter119 PE (130-112-909, Miltenyi Biotec), SiglecH APC-Vio770 (130-112-299, Miltenyi Biotec), B220-APC (130-110-847, Miltenyi Biotec), B220 VioBlue (130-110-851, Miltenyi Biotec), IgM-VioBlue (130-116-318, Miltenyi Biotec), IgD-PE (130-111-496, Miltenyi Biotec), GL7-PE-Cy7 (144619, BioLegend), Ly6C-FITC (130111-915, Miltenyi Biotec), streptavidin FITC (554060, BD Biosciences), CD138 BV605 (563147, BD-Horizon), CD23 BV605 (101637, BioLegend), I-A/I-E BV711 (107643, BioLegend), CD19 BV786 (563333, BD Horizon), T and B cell activation antigen (GL7) PE (561530, BD Pharmingen), CD95 PE-Cy7 (557653, BD Pharmingen), IgM APC-eFluor 780 (47-5790-82, Invitrogen), CD45R/B220 APC/cyanine7 (103224, BioLegend), CD11b eFluor 450 (48-0112-82, eBioscience), CD267 (TACI) BV421 (742840, BD Biosciences), IgD BUV395 (564274, BD Horizon), streptavidin APC (4317-82, eBioscience), CD3 Biotin (100304, BioLegend), Biotin CD11c (568970, BD Biosciences), CD21 PercP Cy5.5 (562797, BD Biosciences), and Fixable Viability Dye eFluor 506 (65-0866-18, Invitrogen) following the recommendations of the manufacturers.

Techniques: Flow Cytometry, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: DNase I alleviates renal inflammatory injury in MRL/lpr mice by inhibiting NETs formation

doi: 10.3389/fimmu.2025.1656069

Figure Lengend Snippet: DNase I treatment decreased the number of immune inflammatory cells in the kidneys and spleens of mice in the MRL/lpr model. (A, B) The TIMER algorithm was used to assess the proportion of immune cell infiltration in the kidneys of control (Ctr), MRL/lpr (Lpr), and DNase I-treated MRL/lpr (DNase) mice at 21 weeks of age. The percentages of CD3+CD4+ T cell subsets (C, G) , CD3+CD8+ T cell subsets (D, H) , and CD3-CD19+B cell subsets (E, I) in the spleen were quantified using flow cytometry across the normal control, MRL/lpr, and DNase I treatment MRL/lpr group. (F, J) Flow cytometric and quantitative analysis of CD4+CD25+Foxp3+ regulatory T cell (Treg) percentages in the spleens of mice from each experimental group were also conducted. Data were expressed as means ± SD for groups of three mice. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. normal control ( t test). # P < 0.05 vs. MRL/lpr mice (Bonferroni correction; two comparisons were made).

Article Snippet: The following antibodies were utilized for staining: anti-CD3 antibody (E-AB-F1013Q), anti-CD4 antibody (E-AB-F1097S), anti-CD8 antibody (E-AB-F1104J), anti-CD19 antibody (E-AB-F0986E), anti-CD25 antibody (E-AB-F1102D), and anti-Foxp3 antibody (E-AB-F1238E), all sourced from Elabscience (China).

Techniques: Control, Flow Cytometry