Journal: Journal of leukocyte biology

Article Title: Blockade of the CCR3 receptor reduces neutrophil recruitment to the lung during acute inflammation.

doi: 10.1093/jleuko/qiae203

Figure Lengend Snippet: Fig. 1. SB328437 reduces neutrophil recruitment to the lung in the LPS-induced lung injury model. LPS-induced respiratory distress model was induced in C57/BL6 mice. After 24 h of LPS administration, BALF and BM were obtained. A) Expression of CCR3 on neutrophils from BALF and BM was determined by flow cytometry. Representative histograms of CCR3 expression on neutrophils (left panel) and percentage of CCR3+ neutrophils in BM and BALF (right panel) are presented. Mice were treated as indicated, and 24 h after treatment, BALF, lung, blood, and BM were obtained. B) The total number of cells per milliliter of BALF was determined. C) Cells from BALF were recovered and analyzed t by flow cytometry to determine the presence of neutrophils; representative contour plots (left panel) and neutrophils per milliliter of BALF (right panel) are presented. The percentage of neutrophils from CD45+ cells was determined by flow cytometry in D) lung parenchyma, E) blood, and F) BM. G) H&E-stained lung sections from one representative animal from each group. The data shown comprise three independent experiments. Bars represent mean ± SE; each circle represents a mouse in each group. Data were analyzed using the Mann–Whitney test A) or one-way ANOVA B–E). Significant changes are indicated by *P ≤ 0.05, **P ≤ 0.01, ****P ≤ 0.0001.

Article Snippet: Then, cells were stained with the following monoclonal antibodies: PerCP/Cy5.5 antimouse CD45 (clone 30-F11), FITC antimouse Ly6G (clone 1A8), Pacific Blue antimouse CD11b (clone M1/70) from BioLegend, PE antimouse CCR3, and APC antimouse CCR3 (clone FAB729P) from R&D Systems.

Techniques: Expressing, Flow Cytometry, Staining, MANN-WHITNEY

Journal: Journal of leukocyte biology

Article Title: Blockade of the CCR3 receptor reduces neutrophil recruitment to the lung during acute inflammation.

doi: 10.1093/jleuko/qiae203

Figure Lengend Snippet: Fig. 2. CCR3 specific antagonist, SB-328437, reduces neutrophil recruitment to the lung during influenza A infection. C57/BL6 mice were infected intranasally with 1 × 106 PFUs of H1N1 pmd9 influenza, and the control group was administrated with PBS. At Days 2, 4, and 7, postinfection BALF was obtained, and A) the total number of cells per milliliter was determined. B) Cells from BALF were recovered and analyzed by flow cytometry to determine the presence of neutrophils, and the number of neutrophils per milliliter of BALF was calculated. C) Cells from BALF were analyzed by flow cytometry to determine the percentage of CCR3+ neutrophils. D) Schematic representation of the experimental design. E) Mice were weighed every day, and the weight change percentage was determined. At Days 2, 4, and 7, postinfection BALF, lung, and blood were obtained. F) The total number of cells per milliliter of BALF was determined. G, H) Cells from BALF were recovered and analyzed by flow cytometry to determine the presence of neutrophils; representative contour plots at Day 4 postinfection (pi) G) and H) neutrophils per milliliter of BALF at 2, 4, and 7 d pi are presented. The percentage of neutrophils from CD45+ cells was determined by flow cytometry in the lung I) and blood J) at 2, 4, and 7 d pi. K) H&E-stained lung sections from one representative animal at 4 d pi from each group. Bars represent mean ± SE; each circle represents a mouse in each group. The data shown comprise two independent experiments. Data were analyzed by one-way ANOVA C) or t-test comparing influenza vs. influenza + SB-328437 groups at the indicated time points E, F, and H–J). Significant changes are indicated by *P ≤ 0.05, **P ≤ 0.01. ns = not significant.

Article Snippet: Then, cells were stained with the following monoclonal antibodies: PerCP/Cy5.5 antimouse CD45 (clone 30-F11), FITC antimouse Ly6G (clone 1A8), Pacific Blue antimouse CD11b (clone M1/70) from BioLegend, PE antimouse CCR3, and APC antimouse CCR3 (clone FAB729P) from R&D Systems.

Techniques: Infection, Control, Flow Cytometry, Staining

Journal: Journal of leukocyte biology

Article Title: Blockade of the CCR3 receptor reduces neutrophil recruitment to the lung during acute inflammation.

doi: 10.1093/jleuko/qiae203

Figure Lengend Snippet: Fig. 3. Neutrophils acquire expression of CCR3 in the alveolar space during lung inflammation. Flow cytometry analysis of expression of CCR3 on neutrophils from A) lung parenchyma and BALF of mice treated with LPS and LPS + SB-328437, representative histograms of CCR3 expression on neutrophils (left panel) and percentage of CCR3+ neutrophils in BALF and lung parenchyma are presented (right panel). B, C) Expression of CCR3 on neutrophils from lung parenchyma and BALF of mice infected with influenza or influenza + SB-328437 at 2, 4, and 7 d pi, representative histograms of CCR3 expression on neutrophils at Day 4 pi B) and percentage of CCR3+ neutrophils in BALF and lung parenchyma at 2, 4, and 7 d pi are presented C). MIF of CD11b on neutrophils from BALF of LPS treated or D) influenza-infected mice at Day 4 pi E), representative histograms of CD11b expression and MIF of CD11b (left panels) and MIF of CD11b (right panels) are presented. A, C–E) Bars represent mean ± SE, each circle represents a mouse in each group, and data were analyzed by one-way ANOVA. Significant changes are indicated by *P ≤ 0.05, ***P ≤ 0.001, ****P ≤ 0.0001.

Article Snippet: Then, cells were stained with the following monoclonal antibodies: PerCP/Cy5.5 antimouse CD45 (clone 30-F11), FITC antimouse Ly6G (clone 1A8), Pacific Blue antimouse CD11b (clone M1/70) from BioLegend, PE antimouse CCR3, and APC antimouse CCR3 (clone FAB729P) from R&D Systems.

Techniques: Expressing, Flow Cytometry, Infection