igg2 isotype control  (Bio X Cell)

Journal: bioRxiv

Article Title: Adenovirus phagocytosis by neutrophils triggers a pro-inflammatory response

doi: 10.1101/2025.09.02.673675

Figure Lengend Snippet: (A, B) Alexa-488 (A-488) labeled Ad5 or A-488 Ad3 were incubated at 37°C with or without human serum (HS), heat-inactivated (HI-HS) or IgG-depleted HS. Then PMNs were exposed to Ads at 4°C to allow cell binding (MOI 10 4 vp/cell) and analysed by flow cytometry. The results are presented as an association index ± SEM ( i.e . mean fluorescence multiplied by percentage of positive cells) (n ≥ 3). (A) Ad association index without serum. (B) Ad association index in the different conditions described above. Mann-Whitney tests: *, p < 0.05; **, p < 0.01. (C, D) A PMN model cell line, PLB-985 cells, was untreated or incubated with blocking antibodies against CD16 and/or CD32 receptors, or with control isotypes. Then cells were incubated with HS-pre-incubated A-488 Ad5 (C) or Ad3 (D) at 4°C to allow binding. Results show the percentage of binding inhibition (± SEM) for the different Ad + HS conditions (five independent experiments) relative to untreated cells exposed to HS-opsonized Ad: Mann-Whitney test * p < 0.05; ** p < 0.01.

Article Snippet: For Fcγ receptor-blocking experiments, the same protocol was used except that cells were also pre-incubated for 20 min at 4°C with the following blocking antibodies : anti-CD16 [3G8] clone antibody (ARG42244, ArigoBio) 5 μg/mL and/or anti-CD32 IV.3 clone antibody (BioXCell BE0224) 10 μg/mL, or control isotypes IgG1 (ARG20767, ArigoBio) 5μg/mL and IgG2 (MPC-11 clone, BioXCell BE0086) 10 μg/mL.

Techniques: Labeling, Incubation, Binding Assay, Flow Cytometry, Fluorescence, MANN-WHITNEY, Blocking Assay, Control, Inhibition

Journal: bioRxiv

Article Title: Adenovirus phagocytosis by neutrophils triggers a pro-inflammatory response

doi: 10.1101/2025.09.02.673675

Figure Lengend Snippet: IgG-coated Ads bind to PMNs via CD32 receptor recognition. This first step triggers Ad internalization in a CD63 + Rab5 + phagosome. Simultaneously, an Ad-specific transcriptional program is induced while NOX2 produces ROS, and an extracellular calcium entry occurs via calcium (or cationic) channels. These events lead, after a few hours, to the PMN lytic death with NET emission, and IL-8 release. This cell death depends on extracellular calcium and RIPK3 (created with BioRender).

Article Snippet: For Fcγ receptor-blocking experiments, the same protocol was used except that cells were also pre-incubated for 20 min at 4°C with the following blocking antibodies : anti-CD16 [3G8] clone antibody (ARG42244, ArigoBio) 5 μg/mL and/or anti-CD32 IV.3 clone antibody (BioXCell BE0224) 10 μg/mL, or control isotypes IgG1 (ARG20767, ArigoBio) 5μg/mL and IgG2 (MPC-11 clone, BioXCell BE0086) 10 μg/mL.

Techniques:

Journal: Journal of Biomedical Science

Article Title: Unraveling Cathepsin S regulation in interleukin-7-mediated anti-tumor immunity reveals its targeting potential against oral cancer

doi: 10.1186/s12929-025-01154-6

Figure Lengend Snippet: Overexpression of CTSS in HNC tissue is inversely correlated with CD8 + T-cell infiltration. A & B The dot graphs show the mRNA expression levels of CTSS in the head and neck cancer (HNC) datasets from ( A ) TCGA and ( B ) GEO ( GSE6791 ). C The representative photos of the CTSS immunohistochemical (IHC) staining on the HNC tissue array. The dot graphs summarize the quantitative score of each sample by stratification. D & E The representative photos of IHC staining for ( D ) CTSS expression and ( E ) CD8 + T-cell infiltration of the in-house oral cancer (OC) samples were shown (N = 70). The score of each sample is plotted by different parameters on the right panels. F & G The correlation between the IHC staining score of CTSS and CD8 in the in-house OC samples was studied. The results are shown by dot graph plots for ( F ) the entire cohort (N = 70) and ( G ) samples with T1 and T2 Stage (N = 38). H The tumor volume of the subcutaneously-inoculated NHRI-HN1 cell is plotted. Mice were implanted with tumors carrying either sh Ctss or shControl, and were grouped by treatment with αCD8 or IgG antibody (N = 6 for each group). Arrowhead indicates days for antibody administration. I The representative photos of the IHC staining for CTSS and CD8 + T cells for each treatment group

Article Snippet: Antibodies were listed in the following: Polyclonal Armenian rat IgG2 isotype control (10 μg/mice) ( cat# BE0086; BioXcell, Lebanon, NH, USA), Anti-mouse/human IL-7 antibody (αIL-7) (10 μg/mice) ( cat# BE0048; BioXcell), Polyclonal Armenian hamster IgG (100 μg/mice) ( cat# BE0091; BioXcell), αPD-1 (100 μg/mice) ( cat# BE0033-2; BioXcell), Rat IgG1 isotype control, anti-horseradish peroxidase (200 μg/mice) ( cat# BP0088; BioXcell), Anti-mouse CD8b antibody (Lyt3.2) (αCD8) (200 μg/mice) ( cat# BE0223; BioXcell).

Techniques: Over Expression, Expressing, Immunohistochemical staining, Immunohistochemistry

Journal: Journal of Biomedical Science

Article Title: Unraveling Cathepsin S regulation in interleukin-7-mediated anti-tumor immunity reveals its targeting potential against oral cancer

doi: 10.1186/s12929-025-01154-6

Figure Lengend Snippet: CTSS inhibits the CD8 + T-cells infiltration and proliferation by downregulating IL-7. A The expression level of IL-7, IL-10, and MCP1 is plotted in the dot graph by each sample (N = 16 in each group). B The tumor volume of the subcutaneously-inoculated NHRI-HN1 cell is plotted. Mice were implanted with tumors carrying either sh Ctss or shControl, and were grouped by treatment with αIL-7 or IgG antibody (N = 10 for each group). Arrowhead indicates days for antibody administration. C The representative photos of the IHC staining for CTSS, IL-7, and CD8 + T-cells for each treatment group. D Tumor infiltrative leukocytes were isolated and analyzed by FACS, the percentage of CD8 + cells in the target quadrant is plotted in the dot graph by each sample. E The percentage of Ki67 + /CD8 + cells in the target quadrant is plotted in the dot graph by each sample. F The percentage of naïve CD8 + cells, central memory CD8 + cells, effector memory CD8 + cells, and tissue-resident CD8 + cells in the target quadrant is plotted in the dot graph by each sample. G & H Mouse CD8 + T-cells were treated with CM with or without the αIL-7. The CM was obtained from in vitro NHRI-HN1 or MOC-1 that were pretreated with siScramble or si Ctss , or that with co-incubation of the mouse IL-7 recombinant protein (mIL-7). ( G) The result of the WST proliferation test for CD8 + T-cells is shown in the dot-bar graph. ( H ) The indicated scale for CFSE cell proliferation is measured and plotted in the dot-bar graph

Article Snippet: Antibodies were listed in the following: Polyclonal Armenian rat IgG2 isotype control (10 μg/mice) ( cat# BE0086; BioXcell, Lebanon, NH, USA), Anti-mouse/human IL-7 antibody (αIL-7) (10 μg/mice) ( cat# BE0048; BioXcell), Polyclonal Armenian hamster IgG (100 μg/mice) ( cat# BE0091; BioXcell), αPD-1 (100 μg/mice) ( cat# BE0033-2; BioXcell), Rat IgG1 isotype control, anti-horseradish peroxidase (200 μg/mice) ( cat# BP0088; BioXcell), Anti-mouse CD8b antibody (Lyt3.2) (αCD8) (200 μg/mice) ( cat# BE0223; BioXcell).

Techniques: Expressing, Immunohistochemistry, Isolation, In Vitro, Incubation, Recombinant