Journal: PLOS Pathogens

Article Title: C/EBP-β-regulated complement hyperactivation in spleen of SFTSV-infected mice: A clue to targeted complement therapy

doi: 10.1371/journal.ppat.1014144

Figure Lengend Snippet: (A and B) Serum C3a (A) and C5a (B) levels of IFNAR −/− mice infected with SFTSV for 3 days or mock-infected. Data are presented as the mean ± SD. (C) Immunohistochemical staining for C3aR and C5aR in the spleen. The deposition of C3aR and C5aR significantly increased 3 days after SFTSV infection. Scale bars are indicated in the images. (D and E) C3ar1 (D) and C5ar1 (E) mRNA expression levels in the spleen 3 days post-infection. Data are presented as the mean ± SD. (F) Volcano plot indicating DEGs in mock-infected and SFTSV-infected spleens. Upregulated and downregulated genes with FDR < 0.05 and fold change > 2 are colored red and green, respectively. (G) Gene ontology (GO) analysis based on DEGs in SFTSV-infected spleens (FDR < 0.05 and fold change > 2). (H) Gene set enrichment analysis (GSEA) plot for the complement and coagulation cascades pathway. (I) Heatmap showing the expression profiles of genes involved in the complement and coagulation cascades pathway, with complement genes highlighted in red. (J) Circos graph displaying the correlation network between complement genes and the top 25 DEGs associated with the most enriched biological processes in (G) ; the Gapdh gene was added as a reference. Each sector of the circle represents one gene, and the color of each link represents the Spearman correlation coefficient between the expressions of genes. Two-sided p -values, examined by Student’s t-test (A, B, D and E) , are shown. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: IFNAR −/− C57BL/6 mice (6–8 weeks old) were divided into three groups (n = 5 per group for tissue collection and n = 7 per group for survival rate): mock-infected, SFTSV + vehicle group, and SFTSV + C3aR antagonist (C3aRA, SB290157, TOPSCIENCE) group.

Techniques: Infection, Immunohistochemical staining, Staining, Expressing, Coagulation

Journal: PLOS Pathogens

Article Title: C/EBP-β-regulated complement hyperactivation in spleen of SFTSV-infected mice: A clue to targeted complement therapy

doi: 10.1371/journal.ppat.1014144

Figure Lengend Snippet: C) Intracellular viral RNA (A) and the mRNA expression levels of cytokines IL-1β (B) and IL-6 (C) in SFTSV-infected FRCs treated with or without the C3aR antagonist (SB290157). (D) Schematic diagram of 40-kDa FITC-dextran extravasation analysis using a co-culture of FRCs and the endothelial cell line HMEC-1 (left) and the calculated permeability index in co-cultures treated with SFTSV or SFTSV plus the C3aR antagonist for 24 hpi. Data are presented as the mean ± SD. Two-sided p -values, examined Dunnett’s multiple comparisons test after one-way ANOVA, are shown. ** p < 0.01, *** p < 0.001. (E) A heatmap showing the DEGs of SFTSV-infected FRCs in the presence of the C3aR antagonist or the vehicle. The top 10 upregulated and downregulated genes are highlighted. (F) Significantly enriched pathways analyzed by Metacore using the DEGs of SFTSV-infected FRCs in the presence of the C3aR antagonist or the vehicle.

Article Snippet: IFNAR −/− C57BL/6 mice (6–8 weeks old) were divided into three groups (n = 5 per group for tissue collection and n = 7 per group for survival rate): mock-infected, SFTSV + vehicle group, and SFTSV + C3aR antagonist (C3aRA, SB290157, TOPSCIENCE) group.

Techniques: Expressing, Infection, Co-Culture Assay, Permeability

Journal: PLOS Pathogens

Article Title: C/EBP-β-regulated complement hyperactivation in spleen of SFTSV-infected mice: A clue to targeted complement therapy

doi: 10.1371/journal.ppat.1014144

Figure Lengend Snippet: IFNAR −/− mice were challenged with SFTSV and then administered the C3aR antagonist (C3aRA) or the vehicle control. Viral load in the spleen 3 days after challenge in the SFTSV + vehicle group (n = 5) and the SFTSV + C3aR antagonist group (n = 5). (B) Representative images of the histological analysis of the spleen tissues from mock-infected or SFTSV-infected mice administered the C3aRA or the vehicle 3 days post-infection. Scale bars are indicated in the images. (C-E) The mRNA expression levels of the cytokines IL-6, TNF-α, and IFN-γ in the spleen tissues 3 days post-infection (n = 5 per group). Data are presented as the mean ± SD. Two-sided p-values, examined by Student’s t-test, are shown. * p < 0.05, ** p < 0.01, *** p < 0.001. (F) Kaplan-Meier survival curves of SFTSV-infected IFNAR −/− mice treated with the C3aR antagonist or the vehicle (n = 6 per group). Representative data are shown from two independent experiments. ** p < 0.01 (log-rank test).

Article Snippet: IFNAR −/− C57BL/6 mice (6–8 weeks old) were divided into three groups (n = 5 per group for tissue collection and n = 7 per group for survival rate): mock-infected, SFTSV + vehicle group, and SFTSV + C3aR antagonist (C3aRA, SB290157, TOPSCIENCE) group.

Techniques: Control, Infection, Expressing

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: C3aR is localized to the paranodal region. Longitudinal and cross‐sections of human sural nerves were analyzed by immunofluorescence using an anti‐C3aR antibody (red, A, D). The paranodal region was stained with an anti‐Caspr antibody (green, B), and the axons were stained with an anti‐peripherin antibody (green, E). Double‐stained images are merged (C, F). Teased mouse sciatic fibers were stained for C3aR (red, G, J, M), Caspr (green, H, K), and peripherin (green, N). The white arrow points to the Schmidt–Lanterman incisures (G). Double‐stained images are merged (I, L, O). Caspr‐contactin‐associated protein; n = 10 mice; Scale bar, 20 μM.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Immunofluorescence, Staining

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: mRNA and protein expression in the mouse sciatic nerve. Quantitative PCR (qPCR) analysis and western blotting were performed on sciatic nerves from 13‐week‐old mice ( n = 6 nerves). Intrinsic expression of the complement receptors and factors is seen (A). The outcomes are presented relative to hypoxanthine guanine phosphoribosyltransferase expression using the 2 −ΔCT method calculation method. Proteins C3aR and C5aR1 are present in mouse sciatic nerves ( n = 6 nerves) (B, C). HPRT‐hypoxanthine guanine phosphoribosyltransferase; ** p < 0.01.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: TLQP‐21 C3aR agonist‐induced neuronal sensitivity. Representative recruitment curves from a control nerve (A) and a nerve treated with the C3aR agonist (B) are shown. TLQP‐21‐treated nerves responded to a lower stimulation current compared to control nerves, prevented by the application of the antagonist (C). EC50 is presented as the EC50 plus 95% CI (profile likelihood) (D). Control n = 10; TLQP‐21 n = 13; JR14a n = 7; TLQP‐21 JR14a n = 10. **** p < 0.0001.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Control

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: The C3aR agonist modulates excitatory neuronal activity. Panel (A) shows a representative example of the refractory response in a control nerve (black, top) compared with a nerve treated with the C3aR agonist (green, bottom). The application of the C3aR agonist led to a reduction in the refractory period, which was prevented by the application of an antagonist (B). In Panel (C), representative examples of the response to high‐frequency stimulation (600–1000 Hz) are presented, comparing a control nerve (black, top) with a nerve following C3aR agonist application (green, bottom). In panel (D), representative examples of the slope at 1000 Hz are presented for a control nerve (black, left) compared with a C3aR agonist‐treated nerve (green, right). The slope in the C3aR agonist‐treated nerves was less steep than that in the control nerves, with the antagonist preventing this excitatory response (E). In panel (F), representative examples of the compound action potential response to 1000 Hz are shown (control: black, top; C3aR agonist: green, bottom). The difference between consecutive compound action potentials was greater in the control nerves than in the C3aR agonist‐treated nerves (G). Control n = 9, TLQP‐21 n = 13, and TLQP‐21 + JR14a n = 10, **** p < 0.0001.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Activity Assay, Control

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: C3aR neuronal hyperexcitability through potassium channel mediation. The slope of the response to high frequencies was similar for low KCl and low KCl + TLQP‐21 (A) Control = white; TLQP‐21 = green. A decreased response to high‐frequency firing rates was observed in the TEA and TEA‐TLQP‐21 nerves (B). Control n = 9; TLQP‐21 n = 12; Low KCl n = 4; Low KCl + TLQP‐21 n = 5; TEA n = 4; TEA + TLQP‐21 n = 6; TEA‐ tetraethyl ammonium chloride; ** p < 0.01.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Control