Journal: JCI Insight

Article Title: Protective role of tissue-resident Tregs in a murine model of beryllium-induced disease

doi: 10.1172/jci.insight.156098

Figure Lengend Snippet: Frequency (left panel) and number (right panel) of CD4 + CD25 + FoxP3 + T cells in the lungs of HLA-DP2 Tg mice sensitized with 1 (1×) or 3 (3×) doses of BeO and harvested at day 12 ( A ) or sensitized with 1 (1×), 3 (3×), or 7 (7×, sensitization/boost) BeO exposures and examined at day 21 ( B ). ( C ) Representative dot plots show CD103 and CD69 expression on CD4 + CD25 + FoxP3 + Tregs derived from the spleen (top panels) and lung (bottom panels) of mice exposed to BeO on 1 (1×), 3 (3×), or 7 (7×) occasions and examined at day 21. ( D and E ) Number of resident effector (RE, CD103 – CD69 + ) and resident memory (RM, CD103 + CD69 + ) T cells among tissue-resident Tregs in HLA-DP2 Tg mice exposed to 1, 3, and 7 doses of BeO. ( F ) Representative histograms show expression of ICOS, Nrp-1, GITR, and CTLA-4 on tissue-specific CD25 + FoxP3 + CD4 + Tregs in mice exposed to 1 (1×, red) or 3 (3×, green) doses of BeO and analyzed at day 12 or exposed to 1 (1×, blue), 3 (3×, purple), or 7 (7×, brown) doses of BeO and analyzed at day 21. ( G – I ) T cell homeostatic chemokines CXCL-10 ( G ), CCL19 ( H ), and CCL21 ( I ) were measured in the lung tissue lysates by ELISA. Data (mean ± SEM) are representative of 3 individual experiments (2–5 mice per group). Significance was determined by 1-way ANOVA ( A , B , D , and E ) and Mann-Whitney U test ( G – I ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: In ICOS or CTLA-4 blocking experiments, HLA-DP2 Tg mice were injected with 300 μg of an anti–CTLA-4 antibody (BioXcell; clone UC10-4F10-11) or an anti-ICOS antibody (BioXcell; clone 7E.17G9) on day –1.

Techniques: Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: JCI Insight

Article Title: Protective role of tissue-resident Tregs in a murine model of beryllium-induced disease

doi: 10.1172/jci.insight.156098

Figure Lengend Snippet: ( A and B ) Plots show the number of CD4 + T cells (left panel) and CD44 + T effector cells (right panel) in the lungs of BeO-sensitized HLA-DP2 Tg mice on day 12 treated i.p. with either an isotype control antibody and anti–CTLA-4 ( A ) or anti-ICOS ( B ) blocking antibodies at day –1. ( C ) Percentage of FoxP3 + Tregs in the lungs of isotype and anti–CTLA-4 (left) or anti-ICOS (right) blocking antibody–treated, BeO-exposed HLA-DP2 Tg mice on day 12. ( D ) At day 12, Teff/Treg ratio was calculated by dividing the total number of tissue-specific Teffs (CD44 + ) by total Tregs (CD25 + FoxP3 + ) in anti–CTLA-4 (left) and anti-ICOS (right) antibody–treated HLA-DP2 Tg mice. ( E ) IFN-γ secretion by CD4 + T cells purified from the lungs of BeO-exposed and anti–CTLA-4 (left) or anti-ICOS (right) blocking antibody–treated HLA-DP2 Tg mice and stimulated with BeSO 4 (100 μM) in the presence of irradiated naive splenocytes. ( F ) Protein in the BALF of mice exposed with BeO (3×) and treated with anti–CTLA-4 (left) or anti-ICOS (right) blocking antibody was examined by ELISA. Data (mean ± SEM) are representative of 2 individual experiments (4–5 mice per group). Significance was determined by Mann-Whitney U test. * P < 0.05.

Article Snippet: In ICOS or CTLA-4 blocking experiments, HLA-DP2 Tg mice were injected with 300 μg of an anti–CTLA-4 antibody (BioXcell; clone UC10-4F10-11) or an anti-ICOS antibody (BioXcell; clone 7E.17G9) on day –1.

Techniques: Control, Blocking Assay, Purification, Irradiation, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Cell Reports

Article Title: COX2 regulates senescence secretome composition and senescence surveillance through PGE 2

doi: 10.1016/j.celrep.2021.108860

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Article Snippet: Rat anti-CD278-176Yb , Fluidigm , # 3176014B.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software, Sequencing

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

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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