soluble icam 1 sicam 1  (Elabscience Biotechnology)

Journal: Frontiers in immunology

Article Title: Anti-inflammatory actions of Pentosan polysulfate sodium in a mouse model of influenza virus A/PR8/34-induced pulmonary inflammation.

doi: 10.3389/fimmu.2023.1030879

Figure Lengend Snippet: FIGURE 8 Measurement of pro-fibrotic and remodeling factors in the lungs of PR8-infected mice treated with PPS at 21 dpi. Mice were infected with PR8 and treated with 3 or 6 mg/kg PPS at Day 0 or Day 5 post-infection, daily for 7 days. Lung concentration of (A) sICAM-1 (B) Complement Factor Bb (CF-Bb) and (C) C5b9. All values normalised to tissue weights. Data shown from two independent experiments, denoted Expt#4 and Expt#5. Mean values for each experiment shown by black (Expt#4) and red (Expt#5) line on plot. Statistically significant differences were assessed on pooled data by a Student’s t-test. Statistically significant differences of pooled values were assessed by one-way ANOVA with a Sidak post-test (ns, not significant; **p<0.01; ***p<0.005; ****p<0.001). (sample size: n=4-6 mice/group).

Article Snippet: Complement proteins were quantified using single ELISA kits (Novus Biologicals Mouse Complement C3a ELISA kit, Cat#NBP2-70037; Thermo Fisher Mouse Complement C5a ELISA kit, Cat#EHMC; Hycult Biotech Mouse Complement C3b ELISA kit, Cat#HK216-01; Biomatik mouse Terminal Complement Complex C5b9 (Cat#EKU10320); Cusabio mouse Factor B (Complement Factor Bb; CF-Bb) Cat#CSB-E04695m); R&D Systems mouse soluble ICAM-1 (sICAM-1) Quantikine Cat#MIC100) according to manufacturer’s instructions.

Techniques: Infection, Concentration Assay

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells

doi: 10.1152/ajplung.00025.2019

Figure Lengend Snippet: Effects of washed red blood cells (RBCs) on human pulmonary artery endothelial cell (HPAEC) inflammatory activation. HPAECs were challenged with vehicle, indicated dilutions of fresh (RBC-F) and old (RBC-O) RBCs, 200 ng/mL LPS or 0.2 ng/mL TNF-α, followed by incubation for 16–20 h. A and B: the conditioned medium was analyzed for soluble ICAM-1 (sICAM-1) (A) and IL-8 (B) using ELISA assay. C: Western blot analysis of ICAM-1 and VCAM-1 expression by HPAECs treated with RBC-F (Rf) and RBC-O (Ro) at indicated dilutions or LPS. Membrane reprobing for β-tubulin was used as normalization control. Bar graphs depict the quantitative densitometry analysis of Western blot data; n = 3. *P < 0.05 vs. nonstimulated control. #P < 0.005.

Article Snippet: Human recombinant TNF-α, IL-6, IL-6-soluble receptor, and ELISA kits for human soluble ICAM-1 (sICAM-1)/IL-8 were obtained from R&D Systems (Minneapolis, MN); lipopolysaccharide (LPS; Escherichia coli O55:B5); and antibodies for human ICAM-1/VCAM-1 were from Santa Cruz Biotechnology (Santa Cruz, CA); VE-cadherin antibody was from Cayman Chemical (Ann Arbor, MI); and antibodies for IκBα and β-tubulin were from Cell Signaling (Danvers, MA).

Techniques: Activation Assay, Incubation, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Membrane

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells

doi: 10.1152/ajplung.00025.2019

Figure Lengend Snippet: Effects of washed red blood cells (RBCs) on LPS- or TNF-α-induced inflammatory responses by human pulmonary artery endothelial cells (HPAECs). HPAEC monolayers were incubated with LPS (A) or TNF-α (B) for 1 h followed by addition of fresh (RBC-F) and old (RBC-O) RBCs (1:100 dilution). The conditioned medium was analyzed for soluble ICAM-1 (sICAM-1) and IL-8 using ELISA assay; n = 5. *P < 0.05 vs. LPS or TNF-α; ND, no statistical difference.

Article Snippet: Human recombinant TNF-α, IL-6, IL-6-soluble receptor, and ELISA kits for human soluble ICAM-1 (sICAM-1)/IL-8 were obtained from R&D Systems (Minneapolis, MN); lipopolysaccharide (LPS; Escherichia coli O55:B5); and antibodies for human ICAM-1/VCAM-1 were from Santa Cruz Biotechnology (Santa Cruz, CA); VE-cadherin antibody was from Cayman Chemical (Ann Arbor, MI); and antibodies for IκBα and β-tubulin were from Cell Signaling (Danvers, MA).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells

doi: 10.1152/ajplung.00025.2019

Figure Lengend Snippet: Effects of red blood cell (RBC) supernatants (SUP) on inflammatory response by human pulmonary artery endothelial cells (HPAECs). HPAECs were challenged with RBC supernatants from fresh and old stored RBCs at 1:50 and 1:100 dilutions, or cells were stimulated with LPS as a positive control. A–C: after 16–20 h, the cultured medium was examined by ELISA for inflammatory markers: soluble ICAM-1 (sICAM-1) (A), IL-8 (B), and IL-6 (C); n = 5. *P < 0.05 and #P < 0.01 vs. nonstimulated control. D: Western blot analysis of VCAM-1 and ICAM-1 expression in total lysates of ECs stimulated with RBC supernatants or LPS. Sf, fresh supernatant, less than 14 days; So, old supernatant, more than 28 days; Sm, supernatant between 14 and 28 days of storage. RBC supernatants alone did not induce inflammatory response. Membrane reprobing for β-tubulin was used as normalization control. Bar graphs depict the quantitative densitometry analysis of Western blot data; n = 4, *P < 0.05 vs. nonstimulated control.

Article Snippet: Human recombinant TNF-α, IL-6, IL-6-soluble receptor, and ELISA kits for human soluble ICAM-1 (sICAM-1)/IL-8 were obtained from R&D Systems (Minneapolis, MN); lipopolysaccharide (LPS; Escherichia coli O55:B5); and antibodies for human ICAM-1/VCAM-1 were from Santa Cruz Biotechnology (Santa Cruz, CA); VE-cadherin antibody was from Cayman Chemical (Ann Arbor, MI); and antibodies for IκBα and β-tubulin were from Cell Signaling (Danvers, MA).

Techniques: Positive Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Membrane

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells

doi: 10.1152/ajplung.00025.2019

Figure Lengend Snippet: Effects of red blood cell (RBC) supernatants (SUP) on LPS or TNF-α-induced inflammatory response of human pulmonary artery endothelial cells (HPAECs). A and B: HPAECs were treated with LPS (A) or TNF-α (B) for 1 h followed by addition of fresh RBC supernatants (1:100 dilution) for 16 to 20 h and ELISA analysis of the markers of inflammation: soluble ICAM-1 (sICAM-1), IL-8, and IL-6; n = 3. *P < 0.05 vs. LPS or TNF-α alone. C: Western blot analysis of ICAM-1 and VCAM-1 expression was performed in HPAEC challenged with LPS or TNF-α with or without posttreatment with fresh RBC supernatants. D: effects of fresh and old RBC supernatants on VCAM-1 and ICAM-1 expression caused by LPS or TNF-α. Membrane reprobing for β-tubulin was used as normalization control. Sf, fresh supernatant, less than 14 days; So, old supernatant, more than 28 days Bar graphs depict the quantitative densitometry analysis of Western blot data; n = 3. *P < 0.05.

Article Snippet: Human recombinant TNF-α, IL-6, IL-6-soluble receptor, and ELISA kits for human soluble ICAM-1 (sICAM-1)/IL-8 were obtained from R&D Systems (Minneapolis, MN); lipopolysaccharide (LPS; Escherichia coli O55:B5); and antibodies for human ICAM-1/VCAM-1 were from Santa Cruz Biotechnology (Santa Cruz, CA); VE-cadherin antibody was from Cayman Chemical (Ann Arbor, MI); and antibodies for IκBα and β-tubulin were from Cell Signaling (Danvers, MA).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Membrane