lyve 1 blocking experiments mice  (R&D Systems)

Journal: PLoS Pathogens

Article Title: Rapid Lymphatic Dissemination of Encapsulated Group A Streptococci via Lymphatic Vessel Endothelial Receptor-1 Interaction

doi: 10.1371/journal.ppat.1005137

Figure Lengend Snippet: A-D: Adhesion of encapsulated M18 GAS untransfected and LYVE-1 lentivirus-transfected HDLECs as indicated above each panel. (A and B) Left to right; numbers of adherent GAS were determined by quantitative culture and representative fluorescence microscopy of adherent GAS (30 min incubation) in the presence of control mAb or LYVE-1 blocking mAb. Scale bars (20 μm). (C) Dose-dependent inhibition of GAS adhesion to HDLEC by a soluble human LYVE-1 ectodomain fragment. (D) Dose-dependent inhibition of GAS adhesion to lentivirus transfected HDLECs in the presence of high (HMWHA, solid line) and low (LMWHA, dashed line) molecular weight HA. (A-D: n = 4, data represent mean+/-SD) (Mann Whitney U;* = p<0.05). (E) High resolution confocal microscopy of M18 GAS following infection of HDLEC. Scale bars (10 μm). Red = LYVE-1, green = GAS, blue = nuclei. Arrows indicate encapsulated GAS adhering to clusters of LYVE-1.

Article Snippet: For LYVE-1 blocking experiments mice were injected intra-peritoneally with 0.5 mg anti mLYVE-1 mAb2125 (R&D) or control rat IgG (isotype control or polyclonal) (R&D) 24 hours prior to infection.

Techniques: Transfection, Fluorescence, Microscopy, Incubation, Blocking Assay, Inhibition, Molecular Weight, MANN-WHITNEY, Confocal Microscopy, Infection

Journal: PLoS Pathogens

Article Title: Rapid Lymphatic Dissemination of Encapsulated Group A Streptococci via Lymphatic Vessel Endothelial Receptor-1 Interaction

doi: 10.1371/journal.ppat.1005137

Figure Lengend Snippet: Adhesion of M18 GAS to MDLECs. Left to right; numbers of adherent GAS were determined by quantitative culture (n = 4; Data represent mean+/-SD) (Mann Whitney U;* = p<0.05) and representative fluorescence microscopy of adherent GAS (30 min incubation) in the presence of control mAb or LYVE-1 blocking mAb. Scale bars (20 μm). Red = LYVE-1, green = GAS, blue = nuclei. Arrows indicate encapsulated GAS adhering to clusters of LYVE-1.

Article Snippet: For LYVE-1 blocking experiments mice were injected intra-peritoneally with 0.5 mg anti mLYVE-1 mAb2125 (R&D) or control rat IgG (isotype control or polyclonal) (R&D) 24 hours prior to infection.

Techniques: MANN-WHITNEY, Fluorescence, Microscopy, Incubation, Blocking Assay

Journal: PLoS Pathogens

Article Title: Rapid Lymphatic Dissemination of Encapsulated Group A Streptococci via Lymphatic Vessel Endothelial Receptor-1 Interaction

doi: 10.1371/journal.ppat.1005137

Figure Lengend Snippet: A-C: Confocal microscopic images of murine ear skin 6–24 hours after local intradermal inoculation. (A) Low magnification images of lymphatic vessels in skin and surrounding infected tissue 24 hours post infection (scale bar 20 μm). (B) 3D rendered images and (C) orthogonal views of the same z-stacks at 6 hours post infection. Arrows indicate individual streptococci interacting with LYVE-1 dense regions of lymphatic vessel endothelium (scale bar 5 μm). D-E: Epifluorescence microscopic images of frozen sections of draining cervical nodes 6 hours post infection at low power (100X magnification)(D) and high power (400X)(E) (Scale bar 50 μm and 20 μm respectively). Red = LYVE-1, green = GAS, blue = nuclei. Arrows indicate encapsulated GAS adhering to clusters of LYVE-1.

Article Snippet: For LYVE-1 blocking experiments mice were injected intra-peritoneally with 0.5 mg anti mLYVE-1 mAb2125 (R&D) or control rat IgG (isotype control or polyclonal) (R&D) 24 hours prior to infection.

Techniques: Infection

Journal: PLoS Pathogens

Article Title: Rapid Lymphatic Dissemination of Encapsulated Group A Streptococci via Lymphatic Vessel Endothelial Receptor-1 Interaction

doi: 10.1371/journal.ppat.1005137

Figure Lengend Snippet: Dissemination of M18 GAS in murine soft-tissue infection in constitutive LYVE-1 -/- mice (n = 7/group). Numbers of GAS recovered at site of infection (A), ipsilateral draining LN (B), spleen (C) and blood (D) 3 hours post-infection. Lines depict median values in each case (Mann Whitney U;* = p<0.05, ** = p<0.01). (E-I) Confocal micrographs of frozen sections of representative draining inguinal lymph nodes from wildtype (n = 5) (E-G) and LYVE-1 -/- (n = 4) (H and I) mice that were resected 3 hours post infection. Original magnification was 100X (E and H) or 400X (F, G and I). Scale bars are 100, 50, 20, 100 and 50 μm for E, F, G, H and I respectively.

Article Snippet: For LYVE-1 blocking experiments mice were injected intra-peritoneally with 0.5 mg anti mLYVE-1 mAb2125 (R&D) or control rat IgG (isotype control or polyclonal) (R&D) 24 hours prior to infection.

Techniques: Infection, MANN-WHITNEY

Journal: PLoS Pathogens

Article Title: Rapid Lymphatic Dissemination of Encapsulated Group A Streptococci via Lymphatic Vessel Endothelial Receptor-1 Interaction

doi: 10.1371/journal.ppat.1005137

Figure Lengend Snippet: Dissemination of M18 GAS in murine soft-tissue infection following LYVE-1 mAb blockade (n = 22/group). Quantitative culture of GAS at site of infection (A), ipsilateral draining LN (B), spleen (C) and blood (D) 3 hours post-infection. Lines depict median values in each case (Mann Whitney U;* = p<0.05, ** = p<0.01). (Control antibody group: circles indicate isotype control antibody and triangles indicate polyclonal control IgG).

Article Snippet: For LYVE-1 blocking experiments mice were injected intra-peritoneally with 0.5 mg anti mLYVE-1 mAb2125 (R&D) or control rat IgG (isotype control or polyclonal) (R&D) 24 hours prior to infection.

Techniques: Infection, MANN-WHITNEY