vivo monoclonal antibody  (Bio X Cell)

Journal: Journal of translational medicine

Article Title: Non-invasive imaging with ICOS-targeting monoclonal antibody for preclinical diagnosis of rheumatoid arthritis in a humanized mouse model.

doi: 10.1186/s12967-024-05899-w

Figure Lengend Snippet: Fig. 4 A Scheme of Cy7 NHS ester conjugation with anti-human ICOS monoclonal antibody (mAb). B The characterization of labeled antibody at differ ent concentrations. Relative fluorescence unit (RFU). C Cell uptake of Cy7-ICOS mAb in PMA/Iono activated, blocked, and resting T cells in huPBMCs at 1-hour incubation (n = 3). D MFI was measured to assess cellular uptake of the ICOS mAb labeled with Cy7 fluorophore at a concentration of 100nM. All values represent the mean ± SEM unless otherwise specified. One-way ANOVA and t test was conducted for statistical significance. ****, p < 0.0001; ***, p < 0.001; **, p < 0.01; *, p < 0.05; ns p > 0.05

Article Snippet: The supplier of the in vivo monoclonal antibody (BE0353, clone: C398.4 A) against human ICOS was BioXCell, located in West Lebanon, USA.

Techniques: Conjugation Assay, Labeling, Fluorescence, Incubation, Concentration Assay

Journal: Journal of translational medicine

Article Title: Non-invasive imaging with ICOS-targeting monoclonal antibody for preclinical diagnosis of rheumatoid arthritis in a humanized mouse model.

doi: 10.1186/s12967-024-05899-w

Figure Lengend Snippet: Fig. 5 A Reference of 4-view Regions of Interest (ROI) drawing was implemented, encompassing perspectives from the TOP, Bottom, Left, and Right orientations. B 4-view NIRF images at all time points (24 h, 48 h, 72 h, 96 h) between the huPBMC-AIA (n = 6) and control groups (n = 4). C X-ray and photographs images merge NIRF imaging following ICOS mAb-Cy7 injection at 6 h, 24 h, 48 h, 72 h and 96 h. D Fluorescence Intensity (FI) of four distinct views was systematically assessed at various time points. E ROI quantification of RP and the RP/LP ratio at all time points examined between huPBMC-AIA and control. F Correlation between paw thickness and RP/LP ratio in NIRF. Significant correlation was observed in both huPBMC-AIA and control group. All values represent the mean ± SEM unless otherwise specified. One-way ANOVA and t test was conducted for statistical significance. ****, p < 0.0001; ***, p < 0.001; **, p < 0.01; *, p < 0.05; ns p > 0.05

Article Snippet: The supplier of the in vivo monoclonal antibody (BE0353, clone: C398.4 A) against human ICOS was BioXCell, located in West Lebanon, USA.

Techniques: Control, Imaging, Injection, Fluorescence

Journal: Journal of translational medicine

Article Title: Non-invasive imaging with ICOS-targeting monoclonal antibody for preclinical diagnosis of rheumatoid arthritis in a humanized mouse model.

doi: 10.1186/s12967-024-05899-w

Figure Lengend Snippet: Fig. 6 A Ex vivo imaging in major organs (1 heart, 2 liver, 3 lung, 4 spleen, 5 kidney, 6 intestine, 7 right paw (RP), 8 left paw (LP), 9 bone, 10 muscle, 11 skin, and 12 blood) at day 7 right after the last in vivo NIRF scan (X-ray and photographs images). B Fluorescence intensity (FI) statistics were analyzed for both paw tissues and major organs following the intravenous administration of ICOS mAb labeled with Cy7 fluorophore at the 96 h time point. C Intra-group correlation analysis, the uptake and metabolism of the anti-human ICOS-Cy7 were evaluated. Row/column order determined by unsupervised hierarchi cal clustering. Side bars represent log10 (FI of organs) at 96 h. D Correlogram depicting r2 Pearson correlation between FI measured in the indicated ROI, compared with the log10 (fold-change RP/LP)at various time points. E Principal Component Analysis (PCA) involved the transformation of the data into principal components, enabling a comprehensive exploration of patterns and variations within the normalized ICOS NIRF signals. F ROC analysis showing sensitivity against 100%specificity for distinguishing the huPBMC-AIA from control group based on FI fold change imaged at 24 h and 96 h. All values represent the mean ± SEM unless otherwise specified. One-way ANOVA and t test was conducted for statistical significance. ****, p < 0.0001; ***, p < 0.001; **, p < 0.01; *, p < 0.05; ns p > 0.05

Article Snippet: The supplier of the in vivo monoclonal antibody (BE0353, clone: C398.4 A) against human ICOS was BioXCell, located in West Lebanon, USA.

Techniques: Ex Vivo, Imaging, In Vivo, Fluorescence, Labeling, Transformation Assay, Control

Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet: Validation of co-signaling receptor modulation in independent mouse models, related to Figure 2 (A) Experimental setup. 10 6 naive Cor93 T (Cor93 T N ) cells were adoptively transferred into HBV-Tg mice (lineage MUP-core). 24 h later, indicated groups of mice were injected intraperitoneally with PBS or 100 μg of monoclonal antibodies (mAbs) blocking PD-1, LAG-3, or CTLA-4. Livers were collected and analyzed at day 5. (B) Total numbers of intrahepatic leukocytes (IHLs) isolated from the indicated mice. (C) Total numbers of Cor93 T cells in the livers of the indicated mice. (D) Representative density plots of IFN-γ expression among Cor93 T cells in the liver of the indicated mice. Numbers represent the percentage of cells within the indicated gates. (E) Total number of IFN-γ-producing Cor93 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction. Each group was compared with PBS-injected controls. (F) Amount of serum alanine transaminases (sALTs) in the serum of the indicated groups of mice at the indicated time points. (G) Experimental setup. HBV replication-competent transgenic mice (lineage 1.3.32) were injected intraperitoneally with PBS or with 100 μg of agonist mAbs activating OX40 or 4-1BB. Livers were collected and analyzed at day 4. (H) Total numbers of IHL isolated from the indicated mice. (I) Representative micrographs of liver sections from the indicated groups of mice. The upper panels show hematoxylin-eosin (H&E) staining, the middle panels show immunohistochemical staining for cleaved caspase 3 (ΔCas3, brown), and the lower panels show immunohistochemical staining for HBcAg (brown). Scale bar represents 100 μm. (J) HBV DNA quantification by southern blot analysis of liver lysates from the indicated mice. Bands corresponding to the expected size of the integrated transgene (Tg), relaxed circular (RC), double-stranded (DS) linear, and single-stranded (SS) HBV DNAs are indicated. (K) Amount of sALT in the serum of the indicated groups of mice at the indicated time points. (L) Experimental setup. 10 6 Cor93 T N cells were adoptively transferred into HBV-Tg mice (lineage MUP-core). 24 h later, selected groups of mice were injected intraperitoneally with PBS or 100 μg of mAbs activating ICOS, OX40, or 4-1BB. Livers were collected and analyzed at day 5. (M) Total numbers of IHL isolated from the indicated mice. (N) Numbers of Cor93 T cells isolated from the liver of the indicated mice. (O) Representative density plots of IFN-γ expression among Cor93 T cells in the liver of the indicated mice. (P) Total number of IFN-γ-producing Cor93 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with control. (Q) Amount of sALT in the serum of the indicated group of mice at the indicated time points. n = 3–4; two-way ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with control group (simple effect within row). (R) Experimental setup. 10 6 naive Env28 CD8 + TCR transgenic cells (Env28 T N ) were adoptively transferred into HBV replication-competent transgenic mice (lineage 1.3.32; background C57BL/6 × BALB/c H-2 bxd hybrids). 24 h later, selected groups of mice were injected intraperitoneally with PBS or 100 μg of mAbs activating OX40 or 4-1BB. Livers were collected and analyzed at day 5. (S) Numbers of IHL isolated from the indicated mice. (T) Numbers of Env28 T cells isolated from the liver of the indicated mice. (U) Representative density plots of IFN-γ expression among Env28 T cells in the liver of the indicated mice. (V) Percentages of IFN-γ-producing Env28 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction. Each group was compared with PBS-injected controls. (W) Amount of sALT in the serum of the indicated groups of mice at the indicated time points. n = 3–4; two-way ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with PBS-injected controls (simple effect within row). (X) Representative micrographs of liver sections from the indicated groups of mice. The upper panels show staining for HBcAg, and the lower panels show staining for cleaved caspase 3 (ΔCas3). Scale bar represents 100 μm. (Y and Z) Representative histograms (Y) and percentages (Z) of in vitro differentiated Cor93 T effector (Cor93 T E ) or Env28 T effector (Env28 T E ) cells producing IFN-γ upon cognate in vitro peptide stimulation at the indicated concentrations. Results are representative of two independent experiments giving similar results.

Article Snippet: In indicated experiments, recipient mice were injected i.p. with wither 100μg of anti-mouse PD-1 (clone RMP1-14, Biolegend #114114), anti-mouse LAG-3 (clone C9B7W, BioXcell #BE0174), anti-mouse CTLA-4 (clone 9H10, BioXcell #BE0131), anti-mouse ICOS (clone C398.4A, BioXcell #BE0353), anti-mouse OX40 (clone OX-86, BioXcell #BE0031) or anti-mouse 4-1BB (clone 3H3, BioXcell #BE0239) antibodies at saturating concentration one day after T cell transfer, unless otherwise indicated.

Techniques: Biomarker Discovery, Injection, Bioprocessing, Blocking Assay, Isolation, Expressing, Ex Vivo, Transgenic Assay, Staining, Immunohistochemical staining, Southern Blot, Control, In Vitro

Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet:

Article Snippet: In indicated experiments, recipient mice were injected i.p. with wither 100μg of anti-mouse PD-1 (clone RMP1-14, Biolegend #114114), anti-mouse LAG-3 (clone C9B7W, BioXcell #BE0174), anti-mouse CTLA-4 (clone 9H10, BioXcell #BE0131), anti-mouse ICOS (clone C398.4A, BioXcell #BE0353), anti-mouse OX40 (clone OX-86, BioXcell #BE0031) or anti-mouse 4-1BB (clone 3H3, BioXcell #BE0239) antibodies at saturating concentration one day after T cell transfer, unless otherwise indicated.

Techniques: Purification, Virus, Recombinant, Staining, Saline, Fluorsave, Sequencing, DNA Labeling, Cell Isolation, Sample Prep, Reverse Transcription, Software, Microscopy

Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet: Validation of co-signaling receptor modulation in independent mouse models, related to Figure 2 (A) Experimental setup. 10 6 naive Cor93 T (Cor93 T N ) cells were adoptively transferred into HBV-Tg mice (lineage MUP-core). 24 h later, indicated groups of mice were injected intraperitoneally with PBS or 100 μg of monoclonal antibodies (mAbs) blocking PD-1, LAG-3, or CTLA-4. Livers were collected and analyzed at day 5. (B) Total numbers of intrahepatic leukocytes (IHLs) isolated from the indicated mice. (C) Total numbers of Cor93 T cells in the livers of the indicated mice. (D) Representative density plots of IFN-γ expression among Cor93 T cells in the liver of the indicated mice. Numbers represent the percentage of cells within the indicated gates. (E) Total number of IFN-γ-producing Cor93 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction. Each group was compared with PBS-injected controls. (F) Amount of serum alanine transaminases (sALTs) in the serum of the indicated groups of mice at the indicated time points. (G) Experimental setup. HBV replication-competent transgenic mice (lineage 1.3.32) were injected intraperitoneally with PBS or with 100 μg of agonist mAbs activating OX40 or 4-1BB. Livers were collected and analyzed at day 4. (H) Total numbers of IHL isolated from the indicated mice. (I) Representative micrographs of liver sections from the indicated groups of mice. The upper panels show hematoxylin-eosin (H&E) staining, the middle panels show immunohistochemical staining for cleaved caspase 3 (ΔCas3, brown), and the lower panels show immunohistochemical staining for HBcAg (brown). Scale bar represents 100 μm. (J) HBV DNA quantification by southern blot analysis of liver lysates from the indicated mice. Bands corresponding to the expected size of the integrated transgene (Tg), relaxed circular (RC), double-stranded (DS) linear, and single-stranded (SS) HBV DNAs are indicated. (K) Amount of sALT in the serum of the indicated groups of mice at the indicated time points. (L) Experimental setup. 10 6 Cor93 T N cells were adoptively transferred into HBV-Tg mice (lineage MUP-core). 24 h later, selected groups of mice were injected intraperitoneally with PBS or 100 μg of mAbs activating ICOS, OX40, or 4-1BB. Livers were collected and analyzed at day 5. (M) Total numbers of IHL isolated from the indicated mice. (N) Numbers of Cor93 T cells isolated from the liver of the indicated mice. (O) Representative density plots of IFN-γ expression among Cor93 T cells in the liver of the indicated mice. (P) Total number of IFN-γ-producing Cor93 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with control. (Q) Amount of sALT in the serum of the indicated group of mice at the indicated time points. n = 3–4; two-way ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with control group (simple effect within row). (R) Experimental setup. 10 6 naive Env28 CD8 + TCR transgenic cells (Env28 T N ) were adoptively transferred into HBV replication-competent transgenic mice (lineage 1.3.32; background C57BL/6 × BALB/c H-2 bxd hybrids). 24 h later, selected groups of mice were injected intraperitoneally with PBS or 100 μg of mAbs activating OX40 or 4-1BB. Livers were collected and analyzed at day 5. (S) Numbers of IHL isolated from the indicated mice. (T) Numbers of Env28 T cells isolated from the liver of the indicated mice. (U) Representative density plots of IFN-γ expression among Env28 T cells in the liver of the indicated mice. (V) Percentages of IFN-γ-producing Env28 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction. Each group was compared with PBS-injected controls. (W) Amount of sALT in the serum of the indicated groups of mice at the indicated time points. n = 3–4; two-way ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with PBS-injected controls (simple effect within row). (X) Representative micrographs of liver sections from the indicated groups of mice. The upper panels show staining for HBcAg, and the lower panels show staining for cleaved caspase 3 (ΔCas3). Scale bar represents 100 μm. (Y and Z) Representative histograms (Y) and percentages (Z) of in vitro differentiated Cor93 T effector (Cor93 T E ) or Env28 T effector (Env28 T E ) cells producing IFN-γ upon cognate in vitro peptide stimulation at the indicated concentrations. Results are representative of two independent experiments giving similar results.

Article Snippet: InVivoMAb anti-mouse/human/rat/monkey ICOS (CD278) , Bio X Cell , Cat# BE0353; RRID: AB_2894772.

Techniques: Biomarker Discovery, Injection, Bioprocessing, Blocking Assay, Isolation, Expressing, Ex Vivo, Transgenic Assay, Staining, Immunohistochemical staining, Southern Blot, Control, In Vitro

Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet:

Article Snippet: InVivoMAb anti-mouse/human/rat/monkey ICOS (CD278) , Bio X Cell , Cat# BE0353; RRID: AB_2894772.

Techniques: Purification, Virus, Recombinant, Staining, Saline, Fluorsave, Sequencing, DNA Labeling, Cell Isolation, Sample Prep, Reverse Transcription, Software, Microscopy

Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B.

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet: Figure 2. Intrahepatically primed, dysfunctional CD8+ T cells are unresponsive to co-inhibitory receptor blockade but can be revived by OX40 and 4-1BB agonism (A) Experimental setup. (B) Numbers of IHL isolated from the indicated mice. (C) Numbers of Cor93 T cells in the livers of the indicated mice. (D) Representative plots of IFN-g expression among Cor93 T cells in the liver of the indicated mice. (E) Number of IFN-g-producing Cor93 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 4–5; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction. Each group was compared with PBS-injected controls.

Article Snippet: Splenectomy was performed according to standard protocols.56 In indicated experiments, recipient mice were injected i.p. with wither 100mg of anti-mouse PD-1 (clone RMP1-14, Biolegend #114114), anti-mouse LAG-3 (clone C9B7W, BioXcell #BE0174), anti-mouse CTLA-4 (clone 9H10, BioXcell #BE0131), anti-mouse ICOS (clone C398.4A, BioXcell #BE0353), anti-mouse OX40 (clone OX-86, BioXcell #BE0031) or anti-mouse 4-1BB (clone 3H3, BioXcell #BE0239) antibodies at saturating concentration one day after T cell transfer, unless otherwise indicated.

Techniques: Isolation, Expressing, Ex Vivo, Injection

Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B.

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet: Figure 5. Upon chronic antigenic stimulation, only 4-1BB but not OX40 stimulation results in significant T cell reinvigoration (A) Heatmap displaying the expression of a selection of genes. Data on the left derived from the scRNA-seq dataset conducted in our study, while the data on the right originates from the dataset presented in Miller et al.26 Specific cell subsets are labeled above the heatmap. Highlighted squares indicate comparisons between subsets identified in both studies: genes common to both the Dys-TRM and TEX subsets (red), genes exclusive to the Dys-TRM subsets (orange), genes shared between the Dys-TSL and TPEX subsets (green), and genes specific to Dys-TSL (blue). Color scale indicates the normalized/scaled expression. (B) UMAP projection (top left) and feature plots of signatures extracted from the re-analysis of the scRNA-seq dataset of Miller et al.,26 showing the enrichment of the TPEX (top right), TEX (bottom left), and effector-like (bottom right) signatures on the UMAP from Figure 3E. Color scale indicates the signature score. (C and D) Violin plots showing the normalized expression profile of Tnfrsf4 (C) and Tnfrsf9 (D) on the subsets described in (A).

Article Snippet: Splenectomy was performed according to standard protocols.56 In indicated experiments, recipient mice were injected i.p. with wither 100mg of anti-mouse PD-1 (clone RMP1-14, Biolegend #114114), anti-mouse LAG-3 (clone C9B7W, BioXcell #BE0174), anti-mouse CTLA-4 (clone 9H10, BioXcell #BE0131), anti-mouse ICOS (clone C398.4A, BioXcell #BE0353), anti-mouse OX40 (clone OX-86, BioXcell #BE0031) or anti-mouse 4-1BB (clone 3H3, BioXcell #BE0239) antibodies at saturating concentration one day after T cell transfer, unless otherwise indicated.

Techniques: Expressing, Selection, Derivative Assay, Labeling