Journal: The Journal of General Virology

Article Title: MERS-CoV infection causes brain damage in human DPP4-transgenic mice through complement-mediated inflammation

doi: 10.1099/jgv.0.001667

Figure Lengend Snippet: Anti-C5aR1 antibody treatment decreased inflammation and viral spread in brains of hDPP4-transgenic mice. Representative images of immunohistochemical staining of cleaved caspase-3 ( a, b ), phosphorylated P38 ( c, d ), IBA-1 ( e, f ), and antiviral NP ( g, h ) in similar regions of cerebral cortex in brains of hDPP4 transgenic mice 7 days after infection with MERS-CoV and treatment with anti-C5aR1 monoclonal antibody. Fewer immunopositive cells were detected in the anti-C5aR1 antibody treatment group ( n =3 per group).

Article Snippet: Mice were treated intravenously (600 µg kg −1 ) with a monoclonal antibody (mAb) to the mouse C5a receptor (C5aR1, HM1076; Hycult Biotech, PB Uden, The Netherlands) for complement inhibition immediately after virus challenge or with the same volume of isotype antibody (HI4041, Hycult Biotech, PB Uden, The Netherlands) as a control.

Techniques: Transgenic Assay, Immunohistochemical staining, Staining, Infection

Journal: The Journal of General Virology

Article Title: MERS-CoV infection causes brain damage in human DPP4-transgenic mice through complement-mediated inflammation

doi: 10.1099/jgv.0.001667

Figure Lengend Snippet: Anti-C5aR1 antibody treatment decreased brain damage in hDPP4 transgenic mice. ( a, b ) Representative images of H&E staining of brain sections of hDPP4-transgenic mice 7 days after infection with MERS-CoV and treatment with anti-C5aR1 or sham control. The representative image of the brains in anti-C5aR1 treatment mice showed less oedema, fewer infiltrating inflammatory cells, especially around vessels in the cerebellum compared to those receiving sham treatment. ( c, d ) Evans blue staining of mice brain on day 7. The brain of a MERS-CoV-infected mouse appeared blue compared with that of a mouse treated with anti-C5aR1 antibody. ( e–h ) Representative images of immunohistochemical staining for neutrophil infiltration ( e, f ) and NF-κB localization ( g, h ). ( i, j ) Semiquantitative analysis of brain damage via H&E scores ( i ) and neutrophil infiltration ( j ). #, Undetectable; ** P <0.01 (Student’s t -test with Welch’s correction)

Article Snippet: Mice were treated intravenously (600 µg kg −1 ) with a monoclonal antibody (mAb) to the mouse C5a receptor (C5aR1, HM1076; Hycult Biotech, PB Uden, The Netherlands) for complement inhibition immediately after virus challenge or with the same volume of isotype antibody (HI4041, Hycult Biotech, PB Uden, The Netherlands) as a control.

Techniques: Transgenic Assay, Staining, Infection, Immunohistochemical staining

Journal: The Journal of General Virology

Article Title: MERS-CoV infection causes brain damage in human DPP4-transgenic mice through complement-mediated inflammation

doi: 10.1099/jgv.0.001667

Figure Lengend Snippet: Diagram illustrating damage to brain tissues in human DPP4-transgenic mice. Neurons infected by MERS-CoV secrete complement components which could activate microglia, which, in turn, could also secrete complement components in brain. Excessive complement activation could activate the endothelial cells of BBB, enhancing the infiltration of inflammatory cells, such as neutrophils and macrophages, into brain parenchyma. The infiltrated inflammatory cells secrete proinflammatory cytokines which could further enhance neuronal damage. However, the inhibition of C5a-C5aR1 interaction could inhibit BBB damage and decrease second damage owing to the excessive inflammatory response.

Article Snippet: Mice were treated intravenously (600 µg kg −1 ) with a monoclonal antibody (mAb) to the mouse C5a receptor (C5aR1, HM1076; Hycult Biotech, PB Uden, The Netherlands) for complement inhibition immediately after virus challenge or with the same volume of isotype antibody (HI4041, Hycult Biotech, PB Uden, The Netherlands) as a control.

Techniques: Transgenic Assay, Infection, Activation Assay, Inhibition

Journal: International journal of biological sciences

Article Title: Suppressing MDSC Infiltration in Tumor Microenvironment Serves as an Option for Treating Ovarian Cancer Metastasis.

doi: 10.7150/ijbs.70013

Figure Lengend Snippet: Figure 7. The C5a receptor inhibitor suppresses MDSC infiltration in TME against cancer metastasis and tumorigenesis. A. Schematic diagram of subcutaneous ID8-luciferase tumors in C57BL/6 mice with CCX168 treatment. The ID8-luciferase cells (5x105) were inoculated into 6-week C57BL/6 mice subcutaneously. After inoculation, the CCX168 (10 mg/kg) or normal saline as control was given by oral gavage every day. B-D. Tumor growth curves, post-dissection measured tumor weights, and volumes of C57BL/6 mice treated with CCX168 or normal saline as control. ID8-luciferase cells (5x106) were inoculated s.c. into 6-week C57BL/6 mice (n=6 per group). The C57BL/6 mice were treated with CCX168 (C5aR inhibitor, 10 mg/kg) or by oral gavage every day. E. The flow cytometry analysis showed fraction change of MDSCs (CD11b+, Gr-1+) in ID8 mice tumor microenvironment. F. Bioluminescence imaging of intraperitoneal ID8-luciferase tumors in C57BL/6 mice with CCX168 treatment. The ID8-luciferase cells (5x105) were inoculated i.p. into 6-week C57BL/6 mice. After 9-day inoculation, the CCX168 (10 mg/kg) or normal saline as control was given by oral gavage every day. Statistic plot of total flux in bioluminescence imaging. G. The graphic illustration of LncOVM-mediated PPIP5K2 regulating complement C5 secretion through the Golgi complex and MDSC recruitment in tumor microenvironment.

Article Snippet: 18 https://www.ijbs.com 3706 Besides, a C5aR antibody (α-C5aR, HM1076, HycultBiotech) was also subcutaneously injected into ID8 tumor surrounding surface (0.6 mg/kg, every second day) in C57BL/6 mice (Fig. S7A).

Techniques: Luciferase, Saline, Control, Dissection, Flow Cytometry, Imaging

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: A Dual Role of Complement Activation in the Development of Fulminant Hepatic Failure Induced by Murine-Beta-Coronavirus Infection

doi: 10.3389/fcimb.2022.880915

Figure Lengend Snippet: Involvement of complement after MHV-3 infection. WT mice were euthanized 0, 6, 12, 24, and 48 hrs after MHV-3 challenge. (A) Immunohistochemical staining for the deposition of C3, C5b-9, and for the expression of C3aR and C5aR in liver tissue samples (CV, central vein; cross, inflammatory infiltrates or necrotic areas). (B, C) Levels of C3aR mRNA and C5aR mRNA, as determined by relative quantitative real-time RT-qPCR in liver tissues at the indicated times. (D, E) Serum concentrations of C3a and C5a at the indicated times. These results are representative of three independent experiments with similar results (n = 4–5 per group). * p <0.05, ** p <0.01, *** p <0.001 compared with 0 time. C3 original magnification, ×200; C5b-9, C3aR, and C5aR original magnification, ×400.

Article Snippet: The monoclonal Ab (mAb) against mouse C5aR (HM1076, clone20/70) and the isotype Rat IgG2b (HI4041, clone RTK4530), were purchased from Hycult Biotech, The Netherlands.

Techniques: Infection, Immunohistochemical staining, Staining, Expressing, Quantitative RT-PCR

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: A Dual Role of Complement Activation in the Development of Fulminant Hepatic Failure Induced by Murine-Beta-Coronavirus Infection

doi: 10.3389/fcimb.2022.880915

Figure Lengend Snippet: Attenuated liver damage in mice treated with anti-C5aR antibody after MHV-3 infection. Mice were administered anti-C5aR antibody or the isotype antibody 12 hrs after MHV-3 infection and euthanized, and liver tissue and serum samples were collected. (A) Histological examination of liver tissue, and immunohistochemical staining for viral antigen and neutrophil infiltration in mouse livers 48 h after virus infection (cross, multifocal damage area). (B, C) Semiquantitative assessment of viral antigen and neutrophil infiltration in livers 48 hrs after virus infection. (D) Serum ALT concentrations 48 hrs after virus infection. (E–G) Serum concentrations of proinflammatory cytokines 24 and 48 hrs after MHV-3 challenge. These results are representative of three independent experiments with similar results (n = 3-4 per group). * p <0.05, ** p <0.01 compared with the isotype control. Original magnification, × 200.

Article Snippet: The monoclonal Ab (mAb) against mouse C5aR (HM1076, clone20/70) and the isotype Rat IgG2b (HI4041, clone RTK4530), were purchased from Hycult Biotech, The Netherlands.

Techniques: Infection, Immunohistochemical staining, Staining