Journal: International Journal of Molecular Sciences

Article Title: Pathological Analysis of Ocular Lesions in a Murine Model of Sjögren’s Syndrome

doi: 10.3390/ijms18061209

Figure Lengend Snippet: Immune cell infiltration of target organs of the SS model mice. CD4, CD8, CD19, and F4/80 expressing immune cells were observed by confocal microscopy. ( a ) CD4 + T cells, ( b ) CD8 + T cells, ( c ) CD19 + B cells, and ( d ) F4/80 + macrophages in the lacrimal glands of the SS model mice were detected by Alexa-546-conjugated antibodies. Nuclei were stained with 4′,6-diamdino-2-phenylindole dihydrochloride (DAPI). The results are representative of five mice. Scale bar: 50 μm; ( e ) Flow cytometry analysis using lacrimal gland tissues from the SS model mice. CD4 and CD8 T cell subsets ware shown, and the result is representative of 5 mice; and ( f ) Cell numbers of CD4 + , CD8 + T cells, CD19 + B cells, and F4/8 + macrophages were calculated by flow cytometery analysis. Data are means ± SD of 5 mice.

Article Snippet: Lymphocytes from lacrimal gland tissues were stained with antibodies against PE-Cy7-conjugated anti-mouse CD4 mAb (TONBO Biosciences, San Diego, CA, USA), fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD8 mAb (eBioscience, San Diego, CA, USA), PE-Cy5.5-conjugated anti-mouse CD19 mAb (TONBO Biosciences), and APC-Cy7-conjugated anti-mouse F4/80 mAb (BioLegend, San Diego, CA, USA).

Techniques: Expressing, Confocal Microscopy, Staining, 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: Frontiers in Immunology

Article Title: Canagliflozin reverses Th1/Th2 imbalance and promotes podocyte autophagy in rats with membranous nephropathy

doi: 10.3389/fimmu.2022.993869

Figure Lengend Snippet: Canagliflozin regulates the imbalance in Th1/Th2 in MN rats. (A) Representative images showing DAPI (blue) and SGLT2 (green) staining in vitro T lymphocytes. (B–D) The number of CD4+ T-cells and CD8+ T-cells in peripheral blood PBMCs of rats detected by flow cytometry analysis. (E–H) The number of Th1 cells and Th2 cells in peripheral blood PBMCs of rats detected by flow cytometry analysis. Data are shown as mean ± SEM values. One-way ANOVA followed by Tukey’s test for multiple comparisons was used. *P < 0.05, **P < 0.01.

Article Snippet: Flow cytometry was used to determine the phenotype of each group of rat T-cells, B-cells, and Th1 and Th2 cells using various combinations of the following fluorochrome-conjugated monoclonal antibodies: perCP/Cyanine5.5-conjugated anti-rat CD3 (201417; Biolegend, USA), FITC-conjugated anti-rat CD4 (130-107-667; Miltenyi, Germany), eFluor 450–binding anti-rat CD8 (48-0084-80; Thermo Fisher Scientific, USA), PE-CYN7–binding anti-rat CD45R (25-0460-80; Thermo Fisher Scientific, USA), Alexa Fluor ® 647-binding anti-rat interferon (IFN)-γ (507809; Biolegend, USA), and PE-conjugated anti-rat interleukin (IL)-4 (511905; Biolegend, USA) for multicolor immunofluorescence (IF).

Techniques: Staining, In Vitro, Flow Cytometry

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Flow cytometry antibodies used.

Article Snippet: CD4-VioBright FITC , Miltenyi , 130-113-791 , REA623 , .

Techniques: Flow Cytometry, In Vivo, In Vitro

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Murlentamab opsonization of SKOV3-R2 + activates an effective anti-tumor T cell immune response. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). After 3 days of co-culture, activated T cells coming from the same healthy donor were added in the culture well for 4 more days. ( A ) The CD4 + Th1/Th2 polarization profile, ( B ) the proportion of CD3 + CD4 + CD25 + regulatory T cells and ( C ) the activation of T CD8 + cells were determined by flow cytometry after four days of co-culture. Data shown (boxplots) are the results from two different experiments (performed with two different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test.

Article Snippet: CD4-VioBright FITC , Miltenyi , 130-113-791 , REA623 , .

Techniques: Labeling, Cell Culture, Derivative Assay, Co-Culture Assay, Activation Assay, Flow Cytometry

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Murlentamab/pembrolizumab combination accentuates the anti-tumoral effect of murlentamab monotherapy through the enhancement of T cell activation. ( A – C ) SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors stimulated with M-CSF and IL-10 (TAMs). After 3 days of co-culture, activated T cells coming from the same healthy donor were added in the culture well for 4 more days. Pembrolizumab was added into co-culture wells everyday from day 3 to day 10. ( A ) Opsonized-SKOV3-R2 + cell number was determined by flow cytometry after one and two days of co-culture with TAMs. Data shown (mean ± SEM) are the results from three different experiments (performed with one healthy donors). ** p < 0.01 compared 3C23K-FcKO vs. Murlentamab. # p < 0.05; ## p < 0.01 compared 3C23K-FcKO + anti-PD-1 vs. Murlentamab + anti-PD-1 as determined using one-way ANOVA analysis followed by Dunnett’s multiple comparisons test. ( B , C ) The CD4 + Th1/Th2 polarization profile and the activation of T CD8 + cells were determined by flow cytometry after four days of co-culture. Data shown (mean ± SEM) are the results from three different experiments (performed with one healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test. ( D , E ) 10 × 10 6 COV434-R2 + ovarian tumor cells were transplanted subcutaneously into humanized GM-CSF/IL3/IL4 hu-NOG (NOD/Shi-scid/IL2Rγ null ) mice (Taconic). After 35 days, when tumors were big enough, mice were i.p treated or not with murlentamab (5 mg/kg) +/− pembrolizumab (25 mg/kg) twice a week for 4 weeks. ( D ) Quantification of circulating CD86 + and CD163 + cells by flow cytometry from blood of tumor-bearing mice before treatment and after 24 days of treatment with murlentamab (5 mg/kg) or pembrolizumab (25 mg/kg) as single agents or murlentamab/pembrolizumab combo-therapy. Data are represented as boxplots. *** p < 0.001, **** p < 0.0001 in comparison to baseline. ( E ) In vivo tumor growth. Data are represented as mean + SEM.

Article Snippet: CD4-VioBright FITC , Miltenyi , 130-113-791 , REA623 , .

Techniques: Activation Assay, Labeling, Cell Culture, Derivative Assay, Co-Culture Assay, Flow Cytometry, Comparison, In Vivo