Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: DC-SIGN is a receptor for human herpesvirus 8 on dendritic cells and macrophages.

doi: 10.4049/jimmunol.176.3.1741

Figure Lengend Snippet: FIGURE 3. Infection of DCs with HHV-8 is blocked by anti DC-SIGN mAb. A, Immunofluorescence results on DCs that were left untreated or treated with either anti-DC-SIGN mAb (clone 120507) or anti-CD11a mAb, infected with HHV-8, incubated, and stained with anti-K8.1A/B mAb (red) at 24 h. Uninfected DCs were used as controls. B, Immunoflu- orescence results on DCs that were either treated with anti-DC-SIGN mAb (clone 120507) or left untreated, infected, and stained at 24 h with anti- DC-SIGN mAb (green) and anti-ORF 59 mAb (red). The overlay of com- bined colors for anti-DC-SIGN and ORF 59 is shown. Cells were coun- terstained with DAPI (blue) (600). Data are from one experiment representative of eight independent experiments.

Article Snippet: For blocking studies, cells were pretreated with 20 g/ml anti-human-DC-SIGN mAb (clone 120507; R&D Systems, or clone DCN46; BD Biosciences), anti-CD11a mAb (BD Biosciences), mouse IgG (Sigma-Aldrich), or 100 g/ml mannan (Sigma-Aldrich), for 1 h at 4°C before exposure to HHV-8.

Techniques: Infection, Incubation, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: DC-SIGN is a receptor for human herpesvirus 8 on dendritic cells and macrophages.

doi: 10.4049/jimmunol.176.3.1741

Figure Lengend Snippet: FIGURE 4. DC-SIGN expression renders resistant cells susceptible to HHV-8 infection. A, Immunofluorescence results on K562 and K562-DC- SIGN cells that were infected with HHV-8 and stained with anti-K8.1A/B mAb at 24 h (red). B, Immunofluorescence results on B-LCL and B-LCL DC-SIGN that were infected with HHV-8 and stained after 24 h with anti- K8.1A/B mAb (red). Cells were counterstained with DAPI (600). Data are from one experiment representative of five independent experiments.

Article Snippet: For blocking studies, cells were pretreated with 20 g/ml anti-human-DC-SIGN mAb (clone 120507; R&D Systems, or clone DCN46; BD Biosciences), anti-CD11a mAb (BD Biosciences), mouse IgG (Sigma-Aldrich), or 100 g/ml mannan (Sigma-Aldrich), for 1 h at 4°C before exposure to HHV-8.

Techniques: Expressing, Infection, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: DC-SIGN is a receptor for human herpesvirus 8 on dendritic cells and macrophages.

doi: 10.4049/jimmunol.176.3.1741

Figure Lengend Snippet: FIGURE 5. HHV-8 binds to DC-SIGN. A, Inhibition of binding of radio- actively labeled HHV-8 by treatment of target cells with anti-DC-SIGN mAb. DCs or B-LCL-DC-SIGN were pretreated with anti-DC-SIGN mAb (clone 120507) or mannan, or left untreated. Each bar represents the mean percent of binding inhibition ( SE) (compared with untreated cells) from two duplicate determinations. B, Inhibition of binding of radioactively labeled HHV-8 by treatment of virus with soluble DC-SIGN. Results are the mean ( SE) per- centage of inhibition of binding of soluble DC-SIGN-treated HHV-8 com- pared with binding of radiolabeled untreated virus to each cell type from two determinations. C, Dose response of inhibition of virus binding to DCs by treatment with anti-DC-SIGN mAb. Each bar represents the mean of duplicate reactions ( SE) from duplicate determinations. Data are from one experiment representative of three independent experiments.

Article Snippet: For blocking studies, cells were pretreated with 20 g/ml anti-human-DC-SIGN mAb (clone 120507; R&D Systems, or clone DCN46; BD Biosciences), anti-CD11a mAb (BD Biosciences), mouse IgG (Sigma-Aldrich), or 100 g/ml mannan (Sigma-Aldrich), for 1 h at 4°C before exposure to HHV-8.

Techniques: Inhibition, Binding Assay, Labeling, Virus

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: DC-SIGN is a receptor for human herpesvirus 8 on dendritic cells and macrophages.

doi: 10.4049/jimmunol.176.3.1741

Figure Lengend Snippet: FIGURE 6. HHV-8 infection of IL-13-activated macrophages is related to DC-SIGN expression. A, Flow cytometric analysis showing expression of DC-SIGN on HHV-8-infected (empty histogram, broken line) or un- infected (empty histogram, solid line) IL-13-activated macrophages. Full histogram, isotype controls. B, Im- munofluorescence results on IL-13-activated macro- phages that were infected with HHV-8 for 24 h and stained for ORF 59 (red) and DC-SIGN (green). The overlay of combined colors for anti-DC-SIGN and ORF59 is shown. C, Immunofluorescence results on IL- 13-activated macrophages that were pretreated with anti- DC-SIGN mAb (clone 120507) or mouse IgG, infected with HHV-8 for 24 h and stained for anti-K8.1 mAb (red). Cells were counterstained with DAPI (600). Data are from one experiment representative of four in- dependent experiments.

Article Snippet: For blocking studies, cells were pretreated with 20 g/ml anti-human-DC-SIGN mAb (clone 120507; R&D Systems, or clone DCN46; BD Biosciences), anti-CD11a mAb (BD Biosciences), mouse IgG (Sigma-Aldrich), or 100 g/ml mannan (Sigma-Aldrich), for 1 h at 4°C before exposure to HHV-8.

Techniques: Infection, Expressing, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: DC-SIGN is a receptor for human herpesvirus 8 on dendritic cells and macrophages.

doi: 10.4049/jimmunol.176.3.1741

Figure Lengend Snippet: FIGURE 7. Effect of HHV-8 in- fection of DCs on expression of DC- SIGN and costimulatory molecules. A, DC-SIGN expression on unin- fected or HHV-8 infected DCs. Data are mean MFI (SE) from seven in- dependent experiments. B, Expres- sion of HLA-ABC, HLA-DR, CD83, and DC-SIGN on infected DCs. Blue histogram, HHV-8-infected DCs; yellow histogram, uninfected DCs; empty histogram, fine line, and bro- ken line isotype controls for the infected and uninfected DCs, respec- tively. Data are from one experiment representative of 10 independent ex- periments. C, Confocal microscopy of HHV-8-infected DCs stained with anti-DC-SIGN (green) and anti-ORF 59 (red) mAbs at 24 h (left panel) and 48 h (center panel) after infection. Uninfected DCs served as controls (right panel). Data are from one ex- periment representative of two inde- pendent experiments.

Article Snippet: For blocking studies, cells were pretreated with 20 g/ml anti-human-DC-SIGN mAb (clone 120507; R&D Systems, or clone DCN46; BD Biosciences), anti-CD11a mAb (BD Biosciences), mouse IgG (Sigma-Aldrich), or 100 g/ml mannan (Sigma-Aldrich), for 1 h at 4°C before exposure to HHV-8.

Techniques: Expressing, Infection, Confocal Microscopy, Staining

Journal: bioRxiv

Article Title: Antibodies against SARS-CoV-2 control complement-induced inflammatory responses to SARS-CoV-2

doi: 10.1101/2023.05.29.542735

Figure Lengend Snippet: (A-F) Human monocyte-derived DCs were exposed to SARS-CoV-2 isolate (hCoV-19/Italy-WT, 1000TCID/mL), complement-opsonized SARS-CoV-2 (hCoV-19/Italy-WT, 1000TCID/mL) and LPS (100 ng/mL) in presence or absence of anti-CD32 for 6 h. mRNA levels of APOBEC3G (A), IRF7 (B), CXCL10 (C), IL-6 (D), IL-10 (E) and IL-12p35 (F) were determine with qPCR after 6h (n=14 donors). Data show the mean values and error bars are the SEM. Statistical analysis was performed using (A-F) 2-way ANOVA with Dunnett’s multiple-comparison test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 (n=14 donors).

Article Snippet: The following antibodies were used (all anti-human): CD86 (2331 (FUN-1), BD Pharmingen), CD80 (L307.4, BD Pharmingen), PE-conjugated mouse IgG1 CR3/CD11b (101208, Biolegend), LEAF purified CR3/CD11b mouse IgG1, LEAF purified CR4/CD11c mouse IgG1, CR3/CD11b (M1/70), CR4/CD11c (S-HCL-3), CD32 (FUN-2), DC-SIGN (FAB161F, R&D systems), CD16, CD32 CD64 (all BD Pharmingen) and, viability dye (Ghost DyeTM Violet 510, Tonbo Biosciences, San Diego, USA).

Techniques: Derivative Assay, Comparison

Journal: bioRxiv

Article Title: Antibodies against SARS-CoV-2 control complement-induced inflammatory responses to SARS-CoV-2

doi: 10.1101/2023.05.29.542735

Figure Lengend Snippet: (A-F) SARS-CoV-2 was pre-incubated with patient isolated mAb COVA1-18 and COVA1-27 (10µg/mL) for 30 min at 37°C. Human monocyte-derived DCs were exposed to SARS-CoV-2 isolate (hCoV-19/Italy-WT, 1000TCID/mL) alone or with blocks, to complement-opsonized SARS-CoV-2 (hCoV-19/Italy-WT, 1000TCID/mL) and LPS (100 ng/mL) in presence or absence of anti-CD32 for 2 h and 6 h. mRNA levels of IFNβ (A), APOBEC3G (B), IRF7 (C), CXCL10 (D), IL-6 (E) and IL-10 (F) were determined by qPCR (n=6 donors (A) and (n=4 donors) (B-F). Data show the mean values and error bars are the SEM. Statistical analysis was performed using (A-F) 2-way ANOVA with Tukey’s multiple-comparison test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, (A) (n=6donors) and (B-F) (n=4 donors).

Article Snippet: The following antibodies were used (all anti-human): CD86 (2331 (FUN-1), BD Pharmingen), CD80 (L307.4, BD Pharmingen), PE-conjugated mouse IgG1 CR3/CD11b (101208, Biolegend), LEAF purified CR3/CD11b mouse IgG1, LEAF purified CR4/CD11c mouse IgG1, CR3/CD11b (M1/70), CR4/CD11c (S-HCL-3), CD32 (FUN-2), DC-SIGN (FAB161F, R&D systems), CD16, CD32 CD64 (all BD Pharmingen) and, viability dye (Ghost DyeTM Violet 510, Tonbo Biosciences, San Diego, USA).

Techniques: Incubation, Isolation, Derivative Assay, Comparison

Journal: bioRxiv

Article Title: Antibodies against SARS-CoV-2 control complement-induced inflammatory responses to SARS-CoV-2

doi: 10.1101/2023.05.29.542735

Figure Lengend Snippet: (A) SARS-CoV-2 pseudovirus opsonization patterns with mild and severe COVID-19 patient sera was determined by ELISA (p24 pg/mL) using anti-human C3c and C3d, for iC3b recognition, and anti-human IgG, for immunoglobulins detection. (B) Human monocyte-derived DCs were exposed to SARS-CoV-2 isolate (hCoV-19/Italy-WT, 1000TCID/mL), to complement-opsonized SARS-CoV-2 (hCoV-19/Italy-WT, 1000TCID/mL), COVID-19 patient serum (mild or severe) and antibody/complement-opsonized SARS-CoV-2 (hCoV-19/Italy-WT, 1000TCID/mL) in presence or absence of anti-CD32 for 2 h and 6 h. mRNA levels for IFNβ (B) were determined after 2 h and mRNA levels of IRF7 (C) and IL-6) (D) after 6 h by qPCR (n=6 donors) (B) and (n=8 donors) (C-D). Data show the mean values and error bars are the SEM. Statistical analysis was performed using (B-D) ordinary one-way ANOVA with Tukey’s multiple-comparison test. *p ≤ 0.05, **p ≤ 0.01 (B) (n=6 donors) and (C-D) (n=8 donors).

Article Snippet: The following antibodies were used (all anti-human): CD86 (2331 (FUN-1), BD Pharmingen), CD80 (L307.4, BD Pharmingen), PE-conjugated mouse IgG1 CR3/CD11b (101208, Biolegend), LEAF purified CR3/CD11b mouse IgG1, LEAF purified CR4/CD11c mouse IgG1, CR3/CD11b (M1/70), CR4/CD11c (S-HCL-3), CD32 (FUN-2), DC-SIGN (FAB161F, R&D systems), CD16, CD32 CD64 (all BD Pharmingen) and, viability dye (Ghost DyeTM Violet 510, Tonbo Biosciences, San Diego, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Comparison

Journal: International journal of molecular sciences

Article Title: Analysis of CD1a-Positive Monocyte-Derived Cells in the Regional Lymph Nodes of Patients with Gallbladder Cancer.

doi: 10.3390/ijms252312763

Figure Lengend Snippet: Figure 2. Representative images of immunohistochemistry of CD209 (DC-SIGN) ((a) original mag- nification ×100, (b) original magnification ×200). (a) Many CD209-positive cells are found at the sinuses of LNs. (b) In metastatic LNs, CD209-positive cells are found at stroma surrounding cancer cells, some of them having dendritic shapes. (c) Image of immunohistochemistry of CD1a, almost the same area of (b). Dendritic-shaped CD1a-positive cells are found at stroma surrounding cancer cells (original magnification ×200).

Article Snippet: The following primary antibodies were used: the mouse monoclonal anti-CD1a antibody (clone 010; IS06930–2; prediluted; Dako, Glostrup, Denmark); the rabbit polyclonal antiS100 antibody (GA50461–2 J; prediluted; Dako); and the mouse monoclonal anti-CD209 antibody (clone 120507; R&D systems, Minneapolis, MN, USA).

Techniques: Immunohistochemistry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Internalizing antibodies to the C-type lectins, L-SIGN and DC-SIGN, inhibit viral glycoprotein binding and deliver antigen to human dendritic cells for the induction of T cell responses.

doi: 10.4049/jimmunol.176.1.426

Figure Lengend Snippet: FIGURE 1. Screening of phage-displayed Fabs for L-SIGN reactivity. A, Ninety-six Fab phage clones selected from three rounds of pan- ning on recombinant DC-SIGN (negative selec- tion) and L-SIGN (positive selection) proteins were screened for binding to K562 (negative control) and K562/L-SIGN-transfected cells by flow cytometry using goat anti-mouse IgG PE conjugate for detection. Only clones showing at least a 5-fold higher binding to K562/L-SIGN cells are shown. B, To select Fab phage clones uniquely reactive with L-SIGN, but not DC- SIGN, K562/L-SIGN cell-reactive clones were screened for reactivity with DC-SIGN-Fc and L- SIGN-Fc fusion proteins in a capture ELISA. mAb162 (L-SIGN specific), mAb1621, and mAb16211 (DC-SIGN/L-SIGN cross-reactive) were used as positive controls. Data are repre- sentative of two independent experiments.

Article Snippet: Before their use in assays, RBC were lysed and any remaining dead cells were further depleted using a dead cell removal kit (Miltenyi Biotec; catalog 130-090-101), per the manufacturer’s instructions. mAbs mAb162 (reactive only with L-SIGN), mAb1621, and mAb16211 (cross-reactive with DC-SIGN and L-SIGN) were purchased from R&D Systems.

Techniques: Clone Assay, Recombinant, Selection, Binding Assay, Negative Control, Transfection, Cytometry, Enzyme-linked Immunosorbent Assay