antibody against human l sign  (R&D Systems)

Journal: PLoS ONE

Article Title: Contribution of Corneal Neovascularization to Dendritic Cell Migration into the Central Area during Human Corneal Infection

doi: 10.1371/journal.pone.0109859

Figure Lengend Snippet: DCs were observed under a light microscope. Primary antibodies against CD1a (MTB1; mouse IgG1, κ; 1∶30 dilution; Novocastra; Newcastle-upon-Tyne, UK), DC-SIGN (CD209; H-200; rabbit polyclonal; 1∶400; Santa Cruz Biotechnology; Santa Cruz, CA, USA), langerin (CD207; 12D6; mouse IgG2b; 1∶100; Ylem S.R.L.; Rome, Italy), and CD83 (1H4b; mouse IgG1, κ; 1∶40; Novocastra) were used to observe DCs. A. Case 17; infected endophthalmitis. CD1a + DCs were observed mainly in the epithelium and the subepithelial space of an inflamed cornea. B. Case 17; infected endophthalmitis. Langerin + DCs were observed in the epithelium. C. Case 18; infected keratitis. DC-SIGN + DCs were observed in the stroma. D. Case 22; corneal perforation, post PKP. CD83 + DCs were observed in the subepithelial space together with lymphocytic infiltration.

Article Snippet: IHC analyses were performed using paraffin-embedded corneal tissues and primary antibodies against CD1a (MTB1; mouse IgG1, κ; 1∶30 dilution; Novocastra; Newcastle-upon-Tyne, UK), DC-SIGN (CD209; H-200; rabbit polyclonal; 1∶400; Santa Cruz Biotechnology; Santa Cruz, CA, USA), langerin (CD207; 12D6; mouse IgG2b; 1∶100; Ylem S.R.L.

Techniques: Light Microscopy, Infection

Journal: PLoS ONE

Article Title: Contribution of Corneal Neovascularization to Dendritic Cell Migration into the Central Area during Human Corneal Infection

doi: 10.1371/journal.pone.0109859

Figure Lengend Snippet: A. Case 18; infected keratitis. Blood vessel formation was observed in the upper stroma. Blood vessels were determined by staining with anti-CD31 antibodies (JC70A; mouse IgG1,κ; 1∶50; Abcam). B. Case 17; infected endophthalmitis. Lymphatic vessel formation was assessed using anti-D2-40 antibodies (mouse IgG1, κ, 1∶50, Dako). Lymphatic vessel formation was observed in the subepithelial space. C. Case 18; infected keratitis. Lymphatic vessel was determined using anti-D2-40 (mouse IgG1, κ, 1∶50, Dako) antibodies, and immature DCs were identified using anti-DC-SIGN antibodies (CD209; H-200; rabbit polyclonal; 1∶400; Santa Cruz Biotechnology; Santa Cruz, CA, USA). A merged image of immunofluorescence staining for D2-40 (green), DC-SIGN (red), and DAPI (blue) captured using a fluorescence microscope. DC-SIGN + DCs (arrows) were observed around a lymphatic vessel (*). D. Case 17; infected endophthalmitis. Blood vessels were determined using anti-von Willebrand factor staining (factor VIII (rabbit polyclonal; 1∶1000, Dako; Carpinteria, California, USA), and mature DCs were identified using anti-CD83 antibodies (1H4b; mouse IgG1, κ; 1∶40; Novocastra). A merged image of immunofluorescence staining for factor 8 (green), CD83 (red), and DAPI (blue) was captured using a fluorescence microscope. CD83 + DCs (arrow) were observed around the blood vessels (*).

Article Snippet: IHC analyses were performed using paraffin-embedded corneal tissues and primary antibodies against CD1a (MTB1; mouse IgG1, κ; 1∶30 dilution; Novocastra; Newcastle-upon-Tyne, UK), DC-SIGN (CD209; H-200; rabbit polyclonal; 1∶400; Santa Cruz Biotechnology; Santa Cruz, CA, USA), langerin (CD207; 12D6; mouse IgG2b; 1∶100; Ylem S.R.L.

Techniques: Infection, Staining, Immunofluorescence, Fluorescence, Microscopy

Journal: PLoS ONE

Article Title: Contribution of Corneal Neovascularization to Dendritic Cell Migration into the Central Area during Human Corneal Infection

doi: 10.1371/journal.pone.0109859

Figure Lengend Snippet: Multiple regression analysis.

Article Snippet: IHC analyses were performed using paraffin-embedded corneal tissues and primary antibodies against CD1a (MTB1; mouse IgG1, κ; 1∶30 dilution; Novocastra; Newcastle-upon-Tyne, UK), DC-SIGN (CD209; H-200; rabbit polyclonal; 1∶400; Santa Cruz Biotechnology; Santa Cruz, CA, USA), langerin (CD207; 12D6; mouse IgG2b; 1∶100; Ylem S.R.L.

Techniques: Infection

Journal: Frontiers in Immunology

Article Title: Proteus mirabilis Targets Atherosclerosis Plaques in Human Coronary Arteries via DC-SIGN (CD209)

doi: 10.3389/fimmu.2020.579010

Figure Lengend Snippet: hDC-SIGN and hLangerin are receptors for the rough P. mirabilis. (a) P. mirabilis invades the CHO cell line that expresses hDC-SIGN and hLangerin. Y. pseudotuberculosis (Y1); E coli K-12 (CS180 and CS1861); P. mirabilis and P. mirabilis- pAY100.1 were examined for their ability to invade CHO/CHO-hDC-SIGN (A) and CHO/CHO-hLangerin cells (B) using the gentamicin protection assay. The number of phagocytosed bacteria was determined by counting the recovered CFUs. The data were pooled from three independent experiments. *** P < 0.001, **** P < 0.0001. b The interactions between P. mirabilis with CHO-hDC-SIGN and CHO-hLangerin were blocked by the anti-hDC-SIGN antibody, by mannan oligosaccharides by shielding the ligand of the LPS core by expressing the O-antigen. (C) P. mirabilis was incubated with CHO-hDC-SIGN for 2 h in the presence or absence of anti-hDC-SIGN and mannan. The phagocytosis rate of P. mirabilis was evaluated by recovering the bacteria from the gentamicin protection assay. E. coli K12 CS180 was used as the control strain to show the core LPS-dependent interaction with hDC-SIGN. (D) Anti-hLangerin and mannan were examined for their ability to inhibit the interaction between CHO-hLangerin and P. mirabilis . The data presented were pooled from three independent experiments. *** P < 0.001, **** P < 0.0001.

Article Snippet: Afterwards, the slides were blocked with serum for 1 h and incubated with primary antibodies against human DC-SIGN (Pharmingen, San Diego, CA, USA) for 1 h. After washing slides thrice with PBS, they were incubated with the corresponding secondary antibody for 1 h. The slides were examined under an ECLIPSE CI microscope (ECLIPSE CI, Nikon, Japan) and analyzed with ImageJ 1.50 (National Institutes of Health, Bethesda, Maryland, USA) software.

Techniques: Expressing, Incubation

Journal: Frontiers in Immunology

Article Title: Proteus mirabilis Targets Atherosclerosis Plaques in Human Coronary Arteries via DC-SIGN (CD209)

doi: 10.3389/fimmu.2020.579010

Figure Lengend Snippet: P. mirabilis interacts with atherosclerotic plaques, which was inhibited by the anti-hDC-SIGN antibody, mannan oligosaccharide, as well as by covering the ligand. A similar approach to Figure 4 , the arteries with atherosclerotic plaques were regarded as CHO-DC-SIGN, respectively. (A) The two sets of bacteria including E. coli K-12 strains (CS180 and CS1861), P. mirabilis , and P. mirabilis- pAY100.1 were incubated with atherosclerotic plaques to determine the adherence ability of the plaques. (B) P. mirabilis was incubated with atherosclerotic plaques for 2 h in the presence or absence of anti-hDC-SIGN, mannan, and core oligosaccharide from P. mirabilis . E. coli K12 CS180 was used as the control strains. The data were pooled from three independent experiments. *** P < 0.001, **** P < 0.0001.

Article Snippet: Afterwards, the slides were blocked with serum for 1 h and incubated with primary antibodies against human DC-SIGN (Pharmingen, San Diego, CA, USA) for 1 h. After washing slides thrice with PBS, they were incubated with the corresponding secondary antibody for 1 h. The slides were examined under an ECLIPSE CI microscope (ECLIPSE CI, Nikon, Japan) and analyzed with ImageJ 1.50 (National Institutes of Health, Bethesda, Maryland, USA) software.

Techniques: Incubation