Journal: Frontiers in Immunology

Article Title: Toll-Like Receptor 7 Enhances Rabies Virus-Induced Humoral Immunity by Facilitating the Formation of Germinal Centers

doi: 10.3389/fimmu.2019.00429

Figure Lengend Snippet: TLR7 is indispensable for optimal antibody production after RABV immunization. TLR7 −/− ( n = 12), TLR3 −/− ( n = 10), or WT mice ( n = 11–12) were immunized i.m. with 10 6 FFU RABV vaccine strain LBNSE in the hind legs. At indicated times post immunization, sera were collected for measuring RABV-specific total IgG (A) and VNA titers (B) . Error bars represent standard error of mean (SEM) ( * P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: Bound Ab was detected using HRP-conjugated Abs (IgG (1:1,000), IgG1 (1:1,500), IgG2a (1:1,500), IgG2b (1:2,000), IgG3 (1:1,500), or IgM (1:2,000) (Boster, Wuhan, China).

Techniques:

Journal: Frontiers in Immunology

Article Title: Toll-Like Receptor 7 Enhances Rabies Virus-Induced Humoral Immunity by Facilitating the Formation of Germinal Centers

doi: 10.3389/fimmu.2019.00429

Figure Lengend Snippet: TLR7 facilitates the formation of GCs. (A,B) TLR7 −/− and WT mice were immunized i.m. with 10 6 FFU LBNSE in the hind legs. Draining LNs were excised, and tissue sections were prepared and stained for GCs (GL-7, red; B220, blue; and IgG, green). Scale bars represent 200 μm. Representative sections are shown in (A) Numbers of GCs (GL-7 positive cell clusters labeled with white arrows) in the draining LNs are calculated and shown in (B) ( n = 3). (C) Blood samples were collected at indicated time points and the concentration of serum CXCL13 was determined by using a commercial ELISA kit ( n = 9). Error bars represent SEM ( * P < 0.05; *** P < 0.001).

Article Snippet: Bound Ab was detected using HRP-conjugated Abs (IgG (1:1,000), IgG1 (1:1,500), IgG2a (1:1,500), IgG2b (1:2,000), IgG3 (1:1,500), or IgM (1:2,000) (Boster, Wuhan, China).

Techniques: Staining, Labeling, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Toll-Like Receptor 7 Enhances Rabies Virus-Induced Humoral Immunity by Facilitating the Formation of Germinal Centers

doi: 10.3389/fimmu.2019.00429

Figure Lengend Snippet: TLR7 facilitates Th1-biased immune responses after RABV vaccination. TLR7 −/− and WT mice were immunized i.m. with 10 6 FFU LBNSE in the hind legs. Spleens were collected at 7 d.p.i. and subjected to ELISpot assay to measure RABV-specific IFN-γ (A,B) and IL-4 (C,D) producing cells ( n = 3 mice/ group). (E–G) Blood samples were collected at the indicated times p.i. for analysis of RABV G-specific IgG1 (E) , IgG2a (F) , and IgG2b (G) ( n = 10). Error bars represent SEM. ND, not detectable. NS, no significantly difference ( * P < 0.05; *** P < 0.001).

Article Snippet: Bound Ab was detected using HRP-conjugated Abs (IgG (1:1,000), IgG1 (1:1,500), IgG2a (1:1,500), IgG2b (1:2,000), IgG3 (1:1,500), or IgM (1:2,000) (Boster, Wuhan, China).

Techniques: Enzyme-linked Immunospot

Journal:

Article Title: Inhibition of IgE-mediated mast cell activation by the paired Ig-like receptor PIR-B

doi:

Figure Lengend Snippet: PIR-B inhibitory activity in FcεRI-mediated Ca2+ mobilization. Indo-1/AM dye preloaded mast cells from wild-type mice (upper panel) and motheaten mice (lower panel) were analyzed by flow cytometry for intracellular Ca2+ levels in the presence or absence (data not shown) of extracellular Ca2+. Cells were stimulated with rat IgE anti-DNP (IgE), F(ab′)2 fragments of rat anti-PIR (anti-PIR), or both mAb’s (IgE + anti-PIR). F(ab′)2 fragments of rabbit anti-rat Ig antibodies were used as the cross-linking reagent.

Article Snippet: Briefly, mast cells were loaded with Indo-1/AM (1 μg/ml; Calbiochem-Novabiochem Corp., San Diego, California, USA) in the presence or absence of rat IgE anti-DNP mAb (1 μg/ml) for 30 minutes at 37°C, resuspended at 10 6 cells/ml either in HBSS containing 5% FCS or Ca 2+ -free HBSS containing 1 mM MgCl 2 and were incubated first with F(ab′) 2 fragments of the rat anti-PIR mAb (10.1 clone; γ2bκ; 20 μg/ml) and then with F(ab′) 2 fragments of rabbit anti-rat Ig antibody (50 μg/ml; Southern Biotechnology Associates) as a cross-linker.

Techniques: Activity Assay, Flow Cytometry

Journal:

Article Title: Inhibition of IgE-mediated mast cell activation by the paired Ig-like receptor PIR-B

doi:

Figure Lengend Snippet: PIR-B inhibitory activity in FcεRI-induced serotonin release. 3H-Serotonin preloaded mast cells from wild-type neonatal spleen (left) and motheaten mutant neonatal spleen (right) were incubated with rat IgE anti-DNP mAb plus various concentrations of F(ab′)2 fragments of rat mAb specific for PIR (filled circles) or unknown cell surface antigen (open circles). Antibody-primed cells were then triggered with F(ab′)2 fragments of rabbit anti-rat Ig antibody as a cross-linker, and the 3H-serotonin release was measured as described in Methods. The standard deviation from the mean (circle) is indicated by bars only for the points at which this value (1 SD) is greater than the circle radius.

Article Snippet: Briefly, mast cells were loaded with Indo-1/AM (1 μg/ml; Calbiochem-Novabiochem Corp., San Diego, California, USA) in the presence or absence of rat IgE anti-DNP mAb (1 μg/ml) for 30 minutes at 37°C, resuspended at 10 6 cells/ml either in HBSS containing 5% FCS or Ca 2+ -free HBSS containing 1 mM MgCl 2 and were incubated first with F(ab′) 2 fragments of the rat anti-PIR mAb (10.1 clone; γ2bκ; 20 μg/ml) and then with F(ab′) 2 fragments of rabbit anti-rat Ig antibody (50 μg/ml; Southern Biotechnology Associates) as a cross-linker.

Techniques: Activity Assay, Mutagenesis, Incubation, Standard Deviation

Journal: iScience

Article Title: TLR7 neo-functionalizes to sense dsRNA and trigger antiviral and antibacterial immunity in non-tetrapod vertebrates

doi: 10.1016/j.isci.2023.108315

Figure Lengend Snippet:

Article Snippet: Goat anti-rabbit Ig-HRP conjugate secondary Ab , ECM Biosciences , , Cat# RS3201; RRID: AB_715256.

Techniques: Virus, Isolation, Recombinant, Plasmid Preparation, Software