Journal: Cell Reports Medicine

Article Title: Response and recurrence correlates in individuals treated with neoadjuvant anti-PD-1 therapy for resectable oral cavity squamous cell carcinoma

doi: 10.1016/j.xcrm.2021.100411

Figure Lengend Snippet:

Article Snippet: ICOS (C398.4A) , Fluidigm , Cat# 3168024B, RRID: AB_2858237.

Techniques: Recombinant, Isolation, Mass Cytometry, Software

Journal: Cell reports

Article Title: Molecular Signatures of Dengue Virus-Specific IL-10/IFN-γ Co-producing CD4 T Cells and Their Association with Dengue Disease

doi: 10.1016/j.celrep.2019.11.098

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Article Snippet: CD278/ICOS, 148Nd , Fluidigm , Cat#3148019B; Clone C398.4A; RRID: AB_2756435.

Techniques: Recombinant, Staining, Activation Assay, Software

Journal: Cell Reports

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

doi: 10.1016/j.celrep.2021.108860

Figure Lengend Snippet:

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

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

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MiR-155 -targeted IcosL controls tumor rejection.

doi: 10.1073/pnas.2408649121

Figure Lengend Snippet: Fig. 1. MiR-155 impairs T cell antitumor response at least in part by targeting ICOSL transcripts. (A) Detection of miR-155 (dark blue) in mice with miR-155 ON and OFF using ISH. Left, miR-155 detection (dark blue) when this microRNA is turned ON in Nestin-Cre x miR-155LSLtTAoffspring; Right, Lack of miR-155 expression when this microRNA has been turned OFF in the same mice. (B) Tumor infiltrates in mice with miR-155 ON for 3 mo (Top panels) then back OFF for 1 wk (Bottom panels). Top panels: Left: H&E staining of the kidney showing infiltrates (dark blue color); Middle: B220 (red signal) and CD4 (brown signal) staining of the infiltrate from the kidney indicated by the square); Right: B220 (red signal) and CD4 (brown signal) staining of a skeletal muscle. Bottom panels, tumor infiltrates in kidneys. From left to right: H&E staining at low (first panel) and high magnification (second panel); B220 staining (red, third panel, notice lack of B220-positive cells); CD4 staining (brown, fourth panel). (C) Top panels: Left: IHC for CD4 (brown) showing T cell infiltration surrounding a lymph node. Right: Magnification of a part of the image on the Left. Bottom panels: IHC for Icos (brown) on lymphomas with miR-155 ON (Left) and then turned back OFF (Right). (D) Dual-Luciferase reporter assays performed in Raw264.7 macrophages cotransfected with a Renilla luciferase reporter vector containing the human ICOSL-3′-UTR with either the WT miR-155 binding site or its mutated version, along with either a miR-Control RNA, a human miR-155 RNA, or a human miR-155-Inhibitor RNA, as indicated. n = 6 replicates/each experimental setting; P = 5.09182E-07. (E) Raw264.7 cells transfected with either a miR-Control RNA, a mouse miR-155 RNA, or a mouse miR-155- Inhibitor RNA, as indicated, were treated with either LPS (100 ng/mL) or the vehicle 24 h after transfection. Two days posttransfection, cell lysates were analyzed by western blot for IcosL expression. GAPDH was used as a loading control. (F) Two days after transfection with either a miR-Control RNA or human-miR-155 RNA, BL cell lines BJAB and NAMALWA were analyzed for ICOSL expression by western blotting. α-tubulin and GAPDH were used as loading control. (G) Inverse correlation between miR-155 and ICOSL transcripts as measured by qRT-PCR in CLL (MEC1 and MEC2) and BL (NAMALWA, BJAB, RAJI, PH3R1, DAUDI) cell lines.

Article Snippet: The specific antibodies used (source and catalog numbers) were as follows: ICOS (ProSci; 8685), ICOSL (ProSci; 8687), T cell (CD3) (Abcam; ab16669), CD163 (Abcam; ab182422), CD31 (Abcam; ab28364); and CD20 B cells (Biogenex; AM537GP).

Techniques: Expressing, Staining, Luciferase, Plasmid Preparation, Binding Assay, Control, Transfection, Western Blot, Quantitative RT-PCR

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MiR-155 -targeted IcosL controls tumor rejection.

doi: 10.1073/pnas.2408649121

Figure Lengend Snippet: Fig. 2. Turning ON the expression of miR-155 in lymphocytes of Nestin-Cre x miR-155LSLtTA mice leads to development of B cell lymphomas lacking IcosL expression, while turning miR-155 back OFF allows the formation of B (IcosL) - T (Icos) cells synapses mostly located around blood vessels. (A) Flow cytometry analyses for B220 and IcosL expression in lymph nodes of littermates with miR-155 OFF (Left) or ON (Right) for 2 mo. Representative images. (B) First row: IcosL (green; Left) and CD20 (red; Middle) staining in a WT spleen. Colocalization (yellow; Right). Second row: Lack of IcosL (green; Left) expression by CD20-positive cells (red; Middle) in lymphomas with miR-155 ON (no yellow signal, Right). IcosL-positive cells in the left are distinct from malignant B cells. Third row: IcosL-positive cells (green; Left) in lymphomas with miR-155 ON are CD31-positive (red, Middle) with colocalization (yellow; Right). Fourth row: A few benign B cells present in the mice with miR-155 ON then OFF for 1 wk had started to re-express IcosL. CD20 (green; Left); IcosL (red; Middle); colocalization (yellow; Right). Panels are representative of staining of 3 mice/group. (C) IHC for B220 (red; Top Left), CD3 (green; Top Right) in infiltrates from mice with miR-155 ON for 10 wk then back OFF for 4 d. Scattered B220-CD3 synapses are seen as yellow fluorescence in the pictures at the Bottom Left after merging B220-CD3 signals; Bottom Right is a magnification of the square in the left picture. (D) IHC for Icos (red; Top) and IcosL (green; Middle) and coexpression (yellow; Bottom) in infiltrates from mice with miR-155 ON for 2 to 3 mo then OFF for 4 d. The immunological synapses between B cells (IcosL) and T cells (Icos) are located mostly around blood vessels (ovals). (Scale bar, 100 µm.)

Article Snippet: The specific antibodies used (source and catalog numbers) were as follows: ICOS (ProSci; 8685), ICOSL (ProSci; 8687), T cell (CD3) (Abcam; ab16669), CD163 (Abcam; ab182422), CD31 (Abcam; ab28364); and CD20 B cells (Biogenex; AM537GP).

Techniques: Expressing, Flow Cytometry, Staining, Fluorescence

Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet:

Article Snippet: 151Eu‐conjugated hamster anti‐human ICOS , Fluidigm , Clone DX29; Cat# 3151020B.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Marker, Infection, Purification, Flow Cytometry, Blocking Assay, Generated, Recombinant, Saline, Staining, Software, Cytometry

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: Journal of Immunology Research

Article Title: Implementation of Mass Cytometry for Immunoprofiling of Patients with Solid Tumors

doi: 10.1155/2019/6705949

Figure Lengend Snippet: Panel of antibodies for mass cytometry.

Article Snippet: 25 , CD278 (ICOS) , 143Nd , DX29 , Surface , Fluidigm.

Techniques: Cytometry, Staining

Journal: Scientific Reports

Article Title: Identification of m6A methyltransferase-related WTAP and ZC3H13 predicts immune infiltrates in glioblastoma

doi: 10.1038/s41598-025-88671-4

Figure Lengend Snippet: Relative expression of genes expression in GBM tissues. Expression levels of WTAP and ZC3H13 genes were compared with those of the control group on GBM tissues, 10 GBM and adjacent tissues, respectively ( A ). The expression levels of siRNA WTAP and overexpression ZC3H13 genes ( B and C ). The expression levels of CD27, CD70, CD80, CD86, ICOS, CTLA4, and LAG3 in siRNA WTAP and overexpression ZC3H13 were demonstrated in Fig. 9D and F, respectively. The result were determined by qRT-PCR. The data are mean ± standard deviation of six replicate experiments. * P < 0.05, **P < 0.01 and ***P < 0.001 compared with the corresponding control values.

Article Snippet: After blocking with 5% nonfat milk, the membranes were immunoblotted with the primary antibodies: WTAP (A04296-2, Boster Biotech, Wuhan, China), CD27 (A01148-2, Boster Biotech), CD70 (A02853-2, Boster Biotech), CD80 (A00196-3, Boster Biotech), CD86(BM4121, Boster Biotech), ICOS (A00291-2, Boster Biotech), CTLA4 (A00020-1, Boster Biotech), LAG3 (M02869-2, Boster Biotech), Beta-actin (BM3873, Boster Biotech).

Techniques: Expressing, Control, Over Expression, Quantitative RT-PCR, Standard Deviation