anti l sign antibodies  (R&D Systems)

Journal: PLOS Pathogens

Article Title: N-glycosylation of viral glycoprotein is a novel determinant for the tropism and virulence of highly pathogenic tick-borne bunyaviruses

doi: 10.1371/journal.ppat.1012348

Figure Lengend Snippet: (a) Nucleotide (position 115 to 123)/amino acid (position 33 to 35) sequences of the original (recOri) and Hp50-4 (recOri(U123A)) M segment/GP are shown. A nucleotide mutation observed in the Hp50-4 strain within the region and deduced amino acids are shown in orange. (b) Schematic diagram of the SFTS virus Gn and Gc with five potential N-linked glycosylation sites (black bars) are shown. (c) Western blotting analysis of lysates of Vero cells infected with recOri or recOri(U123A) recombinants were performed with (+) or without (-) glycosidase treatment and anti-Gn antibody. (d) Jurkat cells expressing a control molecule or one of the human C-type lectins (DC-SIGN, DC-SIGNR, and LSECtin) and Vero cells were inoculated with either recOri or recOri(U123A) strain at an MOI of 0.025. Ratios of viral antigen positivity in Jurkat cells to positivity in Vero cells are shown. Data shown are the means and standard deviations (n = 3). Statistical comparisons were performed between control recOri(U123A) and the others indicated (Dunnett’s test).

Article Snippet: Mouse IgG1 isotype control, anti-DC-SIGN (#120507), anti-DC-SIGNR (#120604), and anti-DC-SIGN/DC-SIGNR (DC28) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Mutagenesis, Virus, Western Blot, Infection, Expressing, Control

Journal: PLOS Pathogens

Article Title: N-glycosylation of viral glycoprotein is a novel determinant for the tropism and virulence of highly pathogenic tick-borne bunyaviruses

doi: 10.1371/journal.ppat.1012348

Figure Lengend Snippet: (a) Human monocyte-derived THP-1 cells were cultured in the presence or absence of phorbol 12-myristate 13-acetate (PMA). The expressions of C-type lectins were analyzed by flow cytometry with antibody clones #120507 (DC-SIGN-specific), #120604 (DC-SIGNR-specific), DC28 (DC-SIGN/DC-SIGNR-dual specific), and SOTO-1 (LSECtin-specific). (b) PMA-treated (PMA+) and untreated (PMA-) THP-1 cells were inoculated with infectious viral particle (iVLP) carrying the original GP (iVLP-Ori) and reporter expression was analyzed by flow cytometry (FCM). (c) PMA-treated and untreated THP-1 cells were inoculated with iVLP carrying the GP from the original or recOri(123A) strains (iVLP-Ori or iVLP-Ori(U123A), respectively). Ratios of reporter positivity in treated cells to those in untreated cells are shown. Statistical comparisons were performed (Welch’s t -test). (d) PMA-treated THP-1 cells were pretreated or untreated with human IgG followed by the addition of the indicated concentration of Ab10. The cells were further inoculated with iVLP-Ori or iVLP-Ori(U123A). Reporter expression was analyzed by flow cytometry. All data shown are the means and standard deviations (n = 3).

Article Snippet: Mouse IgG1 isotype control, anti-DC-SIGN (#120507), anti-DC-SIGNR (#120604), and anti-DC-SIGN/DC-SIGNR (DC28) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Derivative Assay, Cell Culture, Flow Cytometry, Clone Assay, Expressing, Concentration Assay

Journal: PLOS Pathogens

Article Title: N-glycosylation of viral glycoprotein is a novel determinant for the tropism and virulence of highly pathogenic tick-borne bunyaviruses

doi: 10.1371/journal.ppat.1012348

Figure Lengend Snippet: Based on the genome sequence of the Heartland virus (HRTV) MO-4-NIID strain, two HRTV recombinants, HRTVrec and HRTVΔ1stNgly, were produced by reverse genetics. HRTVrec had no mutation but HRTVΔ1stNgly had an asparagine-to-glutamine substitution at position 35 of GP, resulting in the destruction of the N-linked glycosylation motif. (a) AG129 mice were subcutaneously inoculated with 10 2 focus forming units of HRTV MO4, HRTVrec, and HRTVΔ1stNgly (nine or ten mice per group) and observed for 14 days. Survival curves are shown. (b) Jurkat cells expressing control molecule or one of the human C-type lectins (DC-SIGN, DC-SIGNR, and LSECtin) and Vero cells were inoculated at a multiplicity of infection of 0.025. Ratios of viral antigen positivity in Jurkat cells to viral antigen positivity in Vero cells are shown. Data shown are the means and standard deviations (n = 3). Statistical comparisons were performed between control HRTVΔ1stNgly and the others indicated (Dunnett’s test).

Article Snippet: Mouse IgG1 isotype control, anti-DC-SIGN (#120507), anti-DC-SIGNR (#120604), and anti-DC-SIGN/DC-SIGNR (DC28) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Sequencing, Virus, Produced, Mutagenesis, Expressing, Control, Infection