mouse c5a receptor  (Hycult Biotech)

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: The Journal of Immunology Author Choice

Article Title: Endoplasmic Reticulum Stress of Neutrophils Is Required for Ischemia/Reperfusion–Induced Acute Lung Injury

doi: 10.4049/jimmunol.1500073

Figure Lengend Snippet: C5a interacting with C5aR induces ER stress in neutrophils. (A) Neutrophils prepared from mouse peripheral blood or murine myeloid cell line 32Dcl3 were lysed for immunoblotting with C5aR Ab. Images shown are representative of three experiments. Band intensities were determined with ImageJ. (B) Murine myeloid cells (32Dcl3) were transiently transfected with siC5aR overnight and then lysed. Lysates were analyzed by immunoblotting with C5aR Ab. Images shown are representatives of three experiments. (C) Murine myeloid cells (32Dcl3) were transiently transfected with siC5aR overnight, followed by treatment with C5a and/or C5a neutralizing Ab for 4 h and then lysed. Lysates were analyzed by immunoblotting with indicated Abs. Images shown are representatives of three experiments. (D) Supernatants from cultured neutrophils described as in (C) were collected, and MPO was measured using ELISA. Data are expressed as mean ± SD of three independent experiments. *p < 0.05.

Article Snippet: The anti-mouse complement 5a (C5a) receptor (C5aR) Ab was obtained from OriGene (Rockville, MD).

Techniques: Western Blot, Transfection, Cell Culture, Enzyme-linked Immunosorbent Assay