il 5  (R&D Systems)

Journal: Science advances

Article Title: Inhalable SPRAY nanoparticles by modular peptide assemblies reverse alveolar inflammation in lethal Gram-negative bacteria infection.

doi: 10.1126/sciadv.ado1749

Figure Lengend Snippet: Fig. 2. The N-terminal modification enables optimal SPRAY to potentiate the M2-type macrophage polarization. (A) The flow cytometry analyzing the CD206 and CD86 expression levels demonstrates that BLKR rather than LKR induces M2 macrophage. (B) The quantification of M2 macrophage percentage (CD11b+F4/80+CD206+ cells) in (A), n = 3, mean ± SD. (C) Representative CLSM images of BMDMs treated by indicated peptide (16 μM) for 24 hours (green, CD206; red, F-actin; blue, DAPI). Scale bar, 20 μm. (D) Real-time qPCR analysis of mRNA expression of M2-related marker (Arg-1, CD206, YM-1, and Fizz-1), n = 3, mean ± SD. (E) The ELISA test of the TH2 cytokines levels (IL-10, left; IL-4, right) in the BMDM supernatant, n = 3, mean ± SD. (F) The GO enrichment analysis in BLKR group compared with LKR group. (G) KEGG pathway enrichment analysis for BLKR versus LKR group. (H) Heatmap plotting for clustering 33 differentially expressed genes under LKR and BLKR treatments in IL-17, MAPK, and TLR pathways. n = 3. (I) Schematic representation of the IL-17, MAPK, and TLR signaling cascade regulated by BLKR to trigger M2 macrophage polarization and inhibit M1 phenotype polarization. IRF-7, interferon regulatory factor-7; FOS, Fos proto-oncogene; CD14, cluster of differentiation 14 for LPS binding; p38, p38 mitogen-activated protein kinase, CCL17, C-C motif chemokine ligand 17; CCL2, C-C motif chemokine ligand 2; CD25, interleukin 2 receptor subunit alpha; CCL5, C-C motif chemokine ligand 5. Statistical analysis in (B), (D), and (E) was conducted using one-way analysis of variance (ANOVA).

Article Snippet: The anti- MMR/ CD206 (no. AF 2535, R&D Systems) was applied at the final concentration of 0.25 μg for 106 cells and incubated overnight at 4°C.

Techniques: Modification, Flow Cytometry, Expressing, Marker, Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: Science advances

Article Title: Inhalable SPRAY nanoparticles by modular peptide assemblies reverse alveolar inflammation in lethal Gram-negative bacteria infection.

doi: 10.1126/sciadv.ado1749

Figure Lengend Snippet: Fig. 4. SPRAY nanoparticles are promising to reverse the inflammation in LPS-induced ALI via M2 macrophage polarization. (A) Scheme illustrating the medica- tion schedule for ALI mice. (B) Body weight monitoring. (C to E) Hematology monitoring of plasma white blood cell (WBC) counts (C), percentage of NEU in plasma (D) and BALF (E), n = 3. WBC, white blood cells; NEU, neutrophils. (F) Biodistribution of BLKR in injured lung at 1 hour after administration. Scale bar, 100 μm. Red, CD68; green, FITC-BLKR; blue, DAPI. (G) Uptake distribution of SPRAY-BLKR-NPs in various lung immune cell types, n = 3. AlvMs, alveolar macrophages; DCs, dendritic cells; Eos, eosino- phils. (H) Representative images of lung H&E staining on day 6 (wide view, scale bar, 2.5 mm; enlarged region, scale bar, 50 μm). (I) Quantification of inflammatory cells in alveoli, min to max; +, mean value. (J) Representative immunofluorescence staining of pulmonary macrophages on day 6. Wide view, scale bar, 400 μm; enlarged region, scale bar, 10 μm (red, CD86; green, CD206; blue, DAPI). (K and L) Percentage of CD86+ (K) and CD206+ macrophages (L) in total pulmonary cells. (M) Representative im- munohistochemistry staining of SFTPC (20× field) on day 6. Scale bar, 100 μm. SFTPC, surfactant protein C. (N) Percentage of SFTPC+ cells in total pulmonary cells, n = 3. (O and P) The IL-6 (O) and IL-4 (P) concentration in BALF on day 6, n = 5. Data in (B) to (E), (G), (I), (K), (L), and (N) to (P) are presented as means ± SD. Statistical analysis was conducted using one-way ANOVA.

Article Snippet: The anti- MMR/ CD206 (no. AF 2535, R&D Systems) was applied at the final concentration of 0.25 μg for 106 cells and incubated overnight at 4°C.

Techniques: Clinical Proteomics, Staining, Immunofluorescence, Concentration Assay

Journal: Journal of leukocyte biology

Article Title: Influenza A virus directly modulates mouse eosinophil responses

doi: 10.1002/JLB.4MA0320-343R

Figure Lengend Snippet: Anti-mouse fluorochrome-labeled Abs used in this study

Article Snippet: CD125 (IL-5R α ) , FITC , T21 , BD Biosciences.

Techniques: