Journal: The Journal of allergy and clinical immunology

Article Title: Regulatory effects of Interleukin (IL)-15 on allergen-induced airway obstruction

doi: 10.1016/j.jaci.2017.05.025

Figure Lengend Snippet: ELISA analysis of IL-4, IL-5, IL-13, IL-10 IFNγ and IgE levels in control and rIL-15 treated saline or Aspergillus-challenge mice are shown (A-F). BALF eosinophil and PAS+ goblet cell numbers in control and rIL-15 treated saline or Aspergillus-challenge mice are shown (G, H). Airway hyperreactivity (AHR), airway resistance (RI) and compliance (Cydn) in response to methacholine were measured in saline challenged, rIL-15 pretreated saline, Aspergillus challenged alone and rIL-15 pretreated Aspergillus challenged groups of mice are shown (I, J, K). Data is expressed as mean ± SD, n=12 mice/group. *p<0.05, ** p<0.001

Article Snippet: ELISA analysis Cytokine levels in BALF supernatants and lung homogenates were measured using ELISA kits (IL-4, IL-5, IL-10, IL-13, IL-15 and IFN-γ from eBiosciences, San Diego, CA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: The inflammatory microenvironment of the lung at the time of infection governs innate control of SARS-CoV-2 replication

doi: 10.1101/2024.03.27.586885

Figure Lengend Snippet: (A) Left: WT mice were administered PBS, CpG or Pm3 intrapharyngeally (i.ph.). Lungs were collected at seven days post treatment and homogenates were assayed for mouse ACE2 by ELISA. Right: K18-hACE2 Tg mice were administered PBS, CpG or Pm3 i.ph., lungs were collected at 10 days post-treatment and homogenates were assayed for human ACE2 by ELISA, n= 3 – 8, data combined from 1 – 2 independent experiments. (B) Trem2 −/− or (C) Ccr2 −/− mice were given either PBS or 10μg CpG i.ph. seven days prior to being i.n. infected with 3.5×10 4 TCID 50 SCV2 (B.1.351), mice were euthanized 3 days later. Viral loads in lung are shown as measured by TCID 50 on Vero E6 cells, n= 3 – 8, data combined from 1 – 2 independent experiments. Geometric mean, significance determined by two-tailed Mann Whitney test, LD= limit of detection, n.s.= not significant.

Article Snippet: Mouse and human ACE2 protein levels were quantified from lung homogenates by ELISA (R&D Systems #DY3437, #DY933).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Two Tailed Test, MANN-WHITNEY

Journal: Inflammopharmacology

Article Title: Anti-inflammatory and antioxidant activities of Gymnema Sylvestre extract rescue acute respiratory distress syndrome in rats via modulating the NF-κB/MAPK pathway

doi: 10.1007/s10787-022-01133-5

Figure Lengend Snippet: HAEGS attenuated the inflammatory cytokine levels and oxidative stress mediators in LPS-stimulated lung tissue homogenates by modulating the Nrf2 pathway. After 24 h of treatment, BALF samples were collected and lungs were isolated. Further, BALF ( A ) and tissue samples ( B – D ) were subjected to ELISA for the specified protein estimations. A part of the tissues was homogenized and subjected to nitric oxide, for immunoblot and immunohistochemistry analysis for the specified antibodies. E Nitric oxide estimation. F , G Representative graphs of relative expression (immunohistochemistry score) on NRF-2 and HO-1 normalized to area. H , I Representative immunohistochemistry images of NRF-2 and HO-1. J Representative images of western blot analysis. K and L Graph represents densitometric quantification of the specified proteins. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. the LPS control group. Data represented as mean ± SEM, n = 8. One-way ANOVA was performed for statistical analysis

Article Snippet: IL-6, CCL-2 and IL-1β protein levels were measured in cell culture supernatants, BALF and lung tissue homogenates using ELISA kits (R&D Biosystems, Minneapolis, MN, USA) as per the manufacturer’s instructions.

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemistry, Expressing

Journal: Respiratory Research

Article Title: Pirfenidone inhibits fibrocyte accumulation in the lungs in bleomycin-induced murine pulmonary fibrosis

doi: 10.1186/1465-9921-15-16

Figure Lengend Snippet: Pirfenidone regulated chemokine production in the lungs of bleomycin (BLM)-treated mice. Chemokine (CC motif) ligand-2 (CCL2), CCL12, and chemokine (CXC) ligand-12 (CXCL12) in lung digests were measured with enzyme-linked immunosorbent assay on day 14 of BLM treatment. (A) CCL2 production was significantly increased in BLM-treated mice compared with that in saline- and pirfenidone-treated mice. Pirfenidone significantly attenuated the production of CCL2 in BLM-treated mice ( P = 0.0003). (B) CCL12 production was also significantly stimulated by BLM treatment, and pirfenidone administration significantly attenuated this increase ( P < 0.0001). (C) CXCL12 production was increased in BLM-treated mice compared with that in saline-treated mice. Although a trend toward attenuation of CXCL12 production by pirfenidone treatment was found, statistical significance was not reached. The level of each chemokine in the supernatant of the lung homogenate was standardised with the wet weight (g) of each lung. PFD, pirfenidone.

Article Snippet: Chemokine (CC motif) ligand-2 (CCL2), CCL12, and chemokine (CXC motif) ligand-12 (CXCL12) levels were measured in lung homogenates using specific ELISA kits from R&D Systems Inc. (Minneapolis, Minnesota, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Saline

Journal: Genes

Article Title: Human Vascular Endothelial Growth Factor A 165 Expression Induces the Mouse Model of Neovascular Age-Related Macular Degeneration

doi: 10.3390/genes9090438

Figure Lengend Snippet: Morphologic changes and transgene expression in the Cre and LacZ injected eyes. ( a ) β-gal expression (arrowhead) after LacZ injection was seen two weeks after gene transfer but not at later time points. ( b ) VEGF-A expression (violet) in ganglion cell layer (black arrowhead), photoreceptors (arrowhead), and neovascular membrane (arrow) in the eye of Cre -injected mouse. ( c ) Glial fibrillary acidic protein (GFAP) immunoreactivity was observed in the nerve fiber layer (arrowhead) and Müller cells (arrow) in the outer retina post- Cre injection. ( d ) In the Cre group, F4/80 positive macrophages were seen in the retina and subretinal layers. ( e , f ) Retinal autofluorescence (yellow) with DAPI nuclear counterstain (blue). In Cre -injected retina (e), drusen-like lipofuscin deposits (arrowhead) and the loss of photoreceptors were seen. Intact photoreceptor layer (f, arrowhead) was observed in LacZ -injected eyes. Scale bar is 100 µm. GCL: Ganglion cell layer.

Article Snippet: Enzyme-linked immunoassay assay (ELISA) was carried out to quantify human VEGF-A 165 protein in plasma samples and in liver and lung homogenate (human VEGF Quantikine ELISA Kit, DVE00, R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Injection, Membrane

Journal: Genes

Article Title: Human Vascular Endothelial Growth Factor A 165 Expression Induces the Mouse Model of Neovascular Age-Related Macular Degeneration

doi: 10.3390/genes9090438

Figure Lengend Snippet: Human VEGF-A 165 messenger RNA (mRNA) and protein expression in the eye and off-targets. ( a ) Real-time polymerase chain reaction (RT-PCR) analysis of the expression of hVEGF-A 165 in whole eyes. Cyclophilin A (PPIA) normalized relative mRNA expression of hVEGF-A 165 was two-fold higher six weeks after Cre injection compared with the two-week time point. ( b ) RT-PCR analysis of the expression of hVEGF-A 165 in liver samples. Ribosomal protein lateral stalk subunit P0 (Rplp0) normalized mRNA expression of hVEGF-A 165 was the highest two weeks after Cre injection. ( c – e ) hVEGF-A 165 protein levels in plasma, liver, and lung homogenate after Cre gene transfer, respectively. The same symbols in each graph represent the same animal.

Article Snippet: Enzyme-linked immunoassay assay (ELISA) was carried out to quantify human VEGF-A 165 protein in plasma samples and in liver and lung homogenate (human VEGF Quantikine ELISA Kit, DVE00, R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Injection, Clinical Proteomics