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:

Article Title: Inhibition of Human Immunodeficiency Virus Type 1 Env-Mediated Fusion by DC-SIGN

doi: 10.1128/JVI.77.9.5313-5323.2003

Figure Lengend Snippet: DC-SIGN enhances viral infection. (A) Surface expression of DC-SIGN in immature DCs, in HeLa CD4+ DC-SIGN+ (P4 DC3 and P4 DC4 clones) cells, and in P4C5 (HeLa CD4+ CCR5+) cells expressing WT (DCWT) or an LL mutant (DCLL) of DC-SIGN. Cells were stained with anti-DC-SIGN antibodies and were analyzed with flow cytometry. An isotypic MAb was used as a negative control (dotted lines). (B) Enhancement of HIV-1 infection by DC-SIGN. HeLa CD4+ (P4 clone) and P4 DC3 and P4 DC4 cells, which both carry an integrated HIV LTR-lacZ cassette, were infected with the indicated doses of HIVNL43 or HIV(VSV), an HIV-1 variant with a deletion of Env pseudotyped with the VSV-G envelope. Similar results were obtained with higher viral inputs of HIV(VSV) (not shown). (C) Enhancement of infection of various X4 and R5 HIV strains. P4C5 cell derivatives were infected with the indicated viral strain (5 ng of p24). Infection was assessed by measuring β-Gal activity in cell extracts 24 h later. Data are means plus or minus standard deviations of triplicates and are representative of at least three independent experiments. OD, optical density.

Article Snippet: Cells were stained with anti-DC-SIGN (MAb 161-PE; R&D Systems) and anti-CD4 MAbs (SK3-PE; Becton Dickinson) and were processed for flow cytometry as described earlier ( 31 ).

Techniques: Infection, Expressing, Clone Assay, Mutagenesis, Staining, Flow Cytometry, Negative Control, Variant Assay, Activity Assay

Journal:

Article Title: Inhibition of Human Immunodeficiency Virus Type 1 Env-Mediated Fusion by DC-SIGN

doi: 10.1128/JVI.77.9.5313-5323.2003

Figure Lengend Snippet: DC-SIGN inhibits syncytium formation between HIV-1 Env+ and CD4+ cells. (A) Syncytium formation assay. HeLa243Env cells, which express gp120/gp41 complexes and Tat, were mixed (at a 1:5 ratio) with the indicated P4 cell clones. After a 6-h incubation at 37°C, cells were fixed and stained for β-Gal detection. Giant multinucleated cells express β-Gal after Tat-induced transactivation of the HIV LTR-lacZ cassette. Representative fields are shown. (B) Quantitative analysis of the inhibitory activity of DC-SIGN. The indicated P4 cell clones (104 cells per point) were cocultivated with 103 or 104 HeLa243Env+ cells. After 6 h, β-Gal activity was measured in cell extracts. P4 LL15 and P4 LL17 are two independent clones expressing the DC-SIGN LL/AA mutant. Data are means plus or minus standard deviations of triplicates and are representative of three independent experiments. O.D., optical density.

Article Snippet: Cells were stained with anti-DC-SIGN (MAb 161-PE; R&D Systems) and anti-CD4 MAbs (SK3-PE; Becton Dickinson) and were processed for flow cytometry as described earlier ( 31 ).

Techniques: Tube Formation Assay, Clone Assay, Incubation, Staining, Activity Assay, Expressing, Mutagenesis

Journal:

Article Title: Inhibition of Human Immunodeficiency Virus Type 1 Env-Mediated Fusion by DC-SIGN

doi: 10.1128/JVI.77.9.5313-5323.2003

Figure Lengend Snippet: High CD4 levels balance the antifusogenic activity of DC-SIGN. (A) Surface expression of CD4 in P4 and P4 DC3 cells. Cells were transiently transduced with a lentiviral vector encoding CD4, yielding P4-CD4high and P4 DC3-CD4high cells. Cells were then analyzed for CD4 expression by flow cytometry. An isotypic MAb was used as a negative control (dotted lines). (B) Syncytium formation assay. The indicated P4 derivatives (104 cells) were cocultivated with 103 or 104 HeLa243Env+ cells. After 6 h, β-Gal activity was measured in cell extracts. Data are means plus or minus standard deviations of triplicates and are representative of two independent experiments. OD, optical density.

Article Snippet: Cells were stained with anti-DC-SIGN (MAb 161-PE; R&D Systems) and anti-CD4 MAbs (SK3-PE; Becton Dickinson) and were processed for flow cytometry as described earlier ( 31 ).

Techniques: Activity Assay, Expressing, Transduction, Plasmid Preparation, Flow Cytometry, Negative Control, Tube Formation Assay