Journal: medRxiv

Article Title: FcγRIIIb-deficient neutrophils have defects in ROS production, phagocytosis and actin polymerization following stimulation through FcγRs

doi: 10.1101/2025.05.08.25327271

Figure Lengend Snippet: (A) Representative gating in CD14 vs. CD16a density plot of PBMC monocytes. (B) Proportions of monocytes in FcγRIIIb-expressing and FcγRIIIb null individuals (C) Proportions of non-classical FcγRIIIa-expressing monocytes in the peripheral blood from FcγRIIIb-expressing and FcγRIIIb null individuals. (D) MFI of FcγRIIIa present on the surface of non-classical monocytes. Peripheral blood FcγRIIIb-expressing neutrophils (ctrl, green circles). Peripheral blood FcγRIIIb null neutrophils (null, purple squares).

Article Snippet: Briefly, 2×10 5 cells were incubated on ice with Fc block (Miltenyi Biotech, cat. 130-059-901) for 15 min, then stained with titrated concentrations of CD11b-AF700 (BD, cat. 557918), CD15-BV786 (BD, cat. 563838), CD64-BV650 (BD, cat. 745316), CD32-APC (BD, cat. 559769), CD16b-PE (BD, cat. 550868) and CD16a-AF405 (R&D systems, cat. FAB107512V), or titrated concentrations of TLR-2-AF647 (Biolegend, cat. 309714), TLR-4-APC (Biolegend, cat. 312816), TLR-6-PE (Biolegend, cat. 334708) and CD62L-FITC (BD, cat. 555543) for 30 min at 4 °C in the dark.

Techniques: Expressing

Journal: Pathogens

Article Title: Relationship Between Cell Surface Viral Glycoprotein Expression and Resistance of Parainfluenza Virus Persistently Infected Cells to Complement-Mediated Lysis

doi: 10.3390/pathogens14080815

Figure Lengend Snippet: C′ regulators are not significantly upregulated in H1975 and HEp2 PI cells. ( A , B ) RNA samples from H1975 ( A ) and HEp2 ( B ) cells were collected as described in the legend to . Samples were analyzed via RT-qPCR to determine gene expression of three C′ regulators: CD46, CD55, and CFH. Values were normalized to expression of β-actin. C′ inhibitor expression in mock-infected cell cultures was set equal to 1 and expression in AI and PI cel cultures is expressed as fold change relative to mock. ( C ) Mock-infected, AI, and PI HEp2 cells were stained with an anti-CD55 antibody and processed via flow cytometry. MFI; Mean Fluorescence Intensity. ns stands for not significant, **** indicates p -value < 0.0001.

Article Snippet: CD55 surface expression was monitored using an AF405 conjugated anti-human CD55 antibody (R&D Systems, FAB20091V).

Techniques: Quantitative RT-PCR, Gene Expression, Expressing, Infection, Staining, Flow Cytometry, Fluorescence

Journal: Nature Biomedical Engineering

Article Title: Antibody-displaying extracellular vesicles for targeted cancer therapy

doi: 10.1038/s41551-024-01214-6

Figure Lengend Snippet: a , Flow cytometry measurement of uptake by MFI of mNG + EVs (Fc-EVs versus ctrl-EVs versus no EVs) in HER2 positive breast cancer cells (SKBR-3 cells) when incubated with HER2-Ab (trastuzumab), control-Ab (IgG ctrl) or no Ab, showing significant increase in uptake of Fc-EVs by trastuzumab. b , Flow cytometry measurement of uptake (by MFI) of mNG + Fc-EVs decorated with the PD-L1-Ab atezolizumab in PD-L1 expression stimulated (IFNγ) or unstimulated malignant melanoma (B16F10) cells. c , Fluorescence microscopy images of B16F10 cells stained with DAPI (blue), untreated (UT) or treated with mNG + (green) Fc-EVs alone or with the PD-L1-Ab atezolizumab, showing increased uptake of the Fc-EVs when decorated with PD-L1-Ab. Panels a and b are shown as mean ± s.d. n = 3 biological replicates. Statistical significance was calculated using one-way ( b ) or two-way ( a ) ANOVA with Tukey’s post-test compared with each value; P values are indicated above the plots throughout.

Article Snippet: The tissues were processed as described in ref. , with lysed tissue and nLuc + EV input being analysed for nanoluc luminescence or for single-cell suspension for flow cytometry, as described above, but with the HER2 Ab (R&D Systems, FAB9896V-100UG) used for flow cytometry applications.

Techniques: Flow Cytometry, Incubation, Control, Expressing, Fluorescence, Microscopy, Staining

Journal: Nature Biomedical Engineering

Article Title: Antibody-displaying extracellular vesicles for targeted cancer therapy

doi: 10.1038/s41551-024-01214-6

Figure Lengend Snippet: IV injection of Fc-EVs with HER2-Ab (trastuzumab, Fc-EV + HER2-Ab) compared to control-Ab (Fc-EV + IgG-ctrl) in HER2 + breast cancer (SKBR-3) tumour-bearing Swiss nude mice. a , The experimental set-up with inoculation of SKBR-3 cells followed by tumour formation for 2 months before IV injection of nLuc + Fc-EVs with Abs, followed by tissue collection 30 min post injection. b , Fold change in detected EVs (based on luminescence) per gram tumour tissue compared to Fc-EV + IgG-ctrl. c , d , Accumulation of Fc-EV + HER2-Ab (based on luminescence) compared to Fc-EV + IgG-ctrl per gram spleen ( c ) and liver ( d ). All data are shown as mean ± s.d. n = 10 mice. Statistical significance was calculated using two-tailed unpaired t -test analysis compared with each value; P values are indicated above the plots throughout.

Article Snippet: The tissues were processed as described in ref. , with lysed tissue and nLuc + EV input being analysed for nanoluc luminescence or for single-cell suspension for flow cytometry, as described above, but with the HER2 Ab (R&D Systems, FAB9896V-100UG) used for flow cytometry applications.

Techniques: IV Injection, Control, Injection, Two Tailed Test