Journal: Cancers

Article Title: Defining the Immune Checkpoint Landscape in Human Colorectal Cancer Highlights the Relevance of the TIGIT/CD155 Axis for Optimizing Immunotherapy

doi: 10.3390/cancers14174261

Figure Lengend Snippet: In situ assessment of T-cell density and IC expression profile using immunohistochemistry in the IC high/low subgroups of CRC identified via flow cytometry. ( A ) Representative CD3 (left) and CD8 (right) immunostaining of an MSS CRC with an IC high expression profile. Stained cells appear in brown within tumor glands and in the surrounding stroma; nuclei are counterstained in blue. Bars indicate 100 μm. ( B ) Box plots in the lower panels indicate the percentages of CD3 + or CD8 + TILs counted inside tumor glands (intraepithelial TILs) or in the peritumoral stroma using QuPath software, according to IC low or IC high subgroups; Mann–Whitney test. MSS tumors are shown in pale green and MSI tumors in dark blue. ( C ) Representative immunostaining of PD-1, TIGIT and CD94 of the same MSS CRC with an IC high expression profile, as in ( A ). Numerous TIGIT + TILs can be seen within the tumor and in the stroma, and only a few PD-1 + cells and CD94 + TILs. Bars indicate 100 μm. ( D ) Box plots recapitulate the percentages of cells expressing PD-1, TIGIT or CD94, counted inside tumor glands (intraepithelial TILs) or in the peritumoral stroma using QuPath software, according to IC low vs. IC high subgroups; Mann–Whitney test (* p < 0.05; ** p < 0.01). MSS tumors are shown in pale green and MSI tumors in dark blue.

Article Snippet: The following primary antibodies were used: CD3 (Agilent Technologies, polyclonal rabbit anti-human, RRID:AB_2335677), CD8 (Agilent Technologies, monoclonal mouse anti-human clone C8/144B, RRID:AB_2075537), PD-1 (Abcam, Cambridge, UK, monoclonal mouse anti-human clone NAT105, RRID:AB_881954), TIGIT (Cell Signaling Technology, Danvers, MA, USA; monoclonal rabbit anti-human clone ESY1W, RRID:AB_2922806), Tim-3 (Bio-Techne, Minneaoplis, MN, USA, polyclonal goat anti-human, RRID:AB_355235), Lag3 (Novus, Bio-Techne, monoclonal mouse anti-human clone 17B4, RRID:AB_11162489), CD94 (Diaclone, Besançon, France; monoclonal mouse anti-human clone B-D49, RRID:AB_2922808), PD-L1 (Programmed death-ligand 1) (Cell Signaling Technology, monoclonal rabbit anti-human clone E1L3N, RRID:AB_2687655), CD155 (Cell Signaling Technology, monoclonal mouse anti-human clone D8A5G, RRID:AB_2799970), galectin-9 (Abcam, polyclonal rabbit anti-human, RRID:AB_1268942), HLA-DR (Abcam, monoclonal mouse anti-human clone SPM289, RRID:AB_444051) and HLA-E (Bio-Rad, Marnes-la-Coquette, France; monoclonal mouse anti-human clone MEM-E/02, RRID:AB_324025).

Techniques: In Situ, Expressing, Immunohistochemistry, Flow Cytometry, Immunostaining, Staining, Software, MANN-WHITNEY

Journal:

Article Title: Binding of LFA-1 (CD11a) to Intercellular Adhesion Molecule 3 (ICAM-3; CD50) and ICAM-2 (CD102) Triggers Transmigration of Human Immunodeficiency Virus Type 1-Infected Monocytes through Mucosal Epithelial Cells

doi: 10.1128/JVI.76.1.32-40.2002

Figure Lengend Snippet: Presence of HIV proviral DNA in MDM recovered after transmigration through an epithelial cell monolayer. DNA was extracted from MDM recovered from the basal transwell chamber. The 219-bp amplicon of HIV was visualized after migration on an agarose gel of PCR-amplified proviral DNA. Lane 1, DNA extracted from transmigrated MDM; lane 2, DNA extracted from transmigrated HIV-infected MDM at 4°C; lane 3, DNA extracted from transmigrated HIV-infected MDM at 37°C in the absence of proinflammatory stimuli; lanes 4 to 6, DNA extracted from transmigrated HIV-infected MDM under conditions where HEC-1 cells had been preincubated with anti-CD102 (lane 4) or anti-CD50 (lane 5) or both MAb (lane 6) and the assay was performed under proinflammatory conditions. Results are those from one representative experiment out of three that were performed.

Article Snippet: The following murine monoclonal antibodies (MAb) were purchased from the indicated sources: anti-CD50 (clone HP2/19), anti-CD102 (clone B-T1), anti-CD54 (clone HA58), rhodamine-labeled anti-HIV-1 p24 (RD KC57), Beckman Coulter, Brea, Calif.; anti-CXCR4, anti-CCR5 (clone 2D7), anti-TNF-α, anti-IL-1β, fluorescein-conjugated anti-CD50 (anti-CD50 fluorescein isothiocyanate [FITC]), anti-CD11a FITC (clone H111), anti-CD54 FITC (clone HA58), MAb to human immunoglobulin G (IgG) isotypes, Becton Dickinson, San Diego, Calif., anti-CD102 FITC, Diaclone, Besançon, France.

Techniques: Transmigration Assay, Amplification, Migration, Agarose Gel Electrophoresis, Infection

Journal:

Article Title: Binding of LFA-1 (CD11a) to Intercellular Adhesion Molecule 3 (ICAM-3; CD50) and ICAM-2 (CD102) Triggers Transmigration of Human Immunodeficiency Virus Type 1-Infected Monocytes through Mucosal Epithelial Cells

doi: 10.1128/JVI.76.1.32-40.2002

Figure Lengend Snippet: Expression of CD50 (left), CD102 (middle), and CD54 (right) on the membrane of confluent HEC-1 cells, either unstimulated (top) or stimulated with N. meningitidis LPS (5 ng/ml), IL-1β (2 ng/ml), and TNF-α (10 ng/ml) (bottom). Relevant antibodies were conjugated with FITC, and cells were examined by laser confocal microscopy.

Article Snippet: The following murine monoclonal antibodies (MAb) were purchased from the indicated sources: anti-CD50 (clone HP2/19), anti-CD102 (clone B-T1), anti-CD54 (clone HA58), rhodamine-labeled anti-HIV-1 p24 (RD KC57), Beckman Coulter, Brea, Calif.; anti-CXCR4, anti-CCR5 (clone 2D7), anti-TNF-α, anti-IL-1β, fluorescein-conjugated anti-CD50 (anti-CD50 fluorescein isothiocyanate [FITC]), anti-CD11a FITC (clone H111), anti-CD54 FITC (clone HA58), MAb to human immunoglobulin G (IgG) isotypes, Becton Dickinson, San Diego, Calif., anti-CD102 FITC, Diaclone, Besançon, France.

Techniques: Expressing, Confocal Microscopy

Journal:

Article Title: Binding of LFA-1 (CD11a) to Intercellular Adhesion Molecule 3 (ICAM-3; CD50) and ICAM-2 (CD102) Triggers Transmigration of Human Immunodeficiency Virus Type 1-Infected Monocytes through Mucosal Epithelial Cells

doi: 10.1128/JVI.76.1.32-40.2002

Figure Lengend Snippet: Inhibition of transmigration by preincubation of HEC-1 cells with anti-CD50, anti-CD54, and anti-CD102 MAb. The HEC-1 monolayer was preincubated with anti-CD50, anti-CD54, or anti-CD102 MAb (50 μg/ml) or with a mixture of anti-CD50 and anti-CD102 MAb for 30 min at room temperature prior to performing the transmigration assay under proinflammatory conditions. The results are expressed as in Fig. ​Fig.44.

Article Snippet: The following murine monoclonal antibodies (MAb) were purchased from the indicated sources: anti-CD50 (clone HP2/19), anti-CD102 (clone B-T1), anti-CD54 (clone HA58), rhodamine-labeled anti-HIV-1 p24 (RD KC57), Beckman Coulter, Brea, Calif.; anti-CXCR4, anti-CCR5 (clone 2D7), anti-TNF-α, anti-IL-1β, fluorescein-conjugated anti-CD50 (anti-CD50 fluorescein isothiocyanate [FITC]), anti-CD11a FITC (clone H111), anti-CD54 FITC (clone HA58), MAb to human immunoglobulin G (IgG) isotypes, Becton Dickinson, San Diego, Calif., anti-CD102 FITC, Diaclone, Besançon, France.

Techniques: Inhibition, Transmigration Assay