Journal: Journal of clinical & cellular immunology

Article Title: The Anti-inflammatory Effect of Alpha-1 Antitrypsin in Rhinovirus-infected Human Airway Epithelial Cells

doi: 10.4172/2155-9899.1000475

Figure Lengend Snippet: Increased IL-8 production in cultured COPD brushed airway epithelial cells. Brushed airway epithelial cells from COPD patients (n=6) and normal subjects (n=6) were cultured under air-liquid interface (ALI) condition for 10 days. After 24 hours of air exposure, IL-8 was measured by ELISA. Data are expressed as means ± SEM.

Article Snippet: Protein levels of human IL-8, or mouse KC were determined by using a human IL-8 or a mouse KC DuoSet ELISA kit (R&D Systems, Minneapolis, MN) as per manufacturer’s instruction.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Journal of Immunology Research

Article Title: Tumor-Derived CXCL1 Promotes Lung Cancer Growth via Recruitment of Tumor-Associated Neutrophils

doi: 10.1155/2016/6530410

Figure Lengend Snippet: Expression and secretion of CXCL1 in vitro and in vivo . (a) Levels of CXCL1 in the serum of naive mice ( n = 11) or 3LL tumor-bearing mice ( n = 27) were measured by ELISA at 14 days after tumor inoculation. Each dot represents an individual mouse. (b) CXCL1 protein levels in the 3LL culture supernatants were determined by ELISA. (c) mRNA expression of CXCL1 in 3LL cells was measured by quantitative RT-PCR. (d) ELISA analysis of CXCL1 production in Lewis lung carcinoma cell line 3LL, mouse epithelial cell line MLE, and human non-small lung carcinoma cell line A549. (e) Ly6G + MPO + neutrophils in 3LL tumor tissues were determined by IF and IHC. (f) Quantitative RT-PCR analysis of CXCL1 expression in 3LL tumor cells and TANs (sorted from 3LL tumor tissues) stimulated with or without LPS (100 ng/mL) for 6 hours. ∗∗ p < 0.01; data are shown as mean ± SD.

Article Snippet: Mouse and human CXCL1 ELISA kit (R&D systems) were used to determine the concentrations of CXCL1 in cell culture supernatant or serum from tumor-bearing mice according to the manufacturer's instructions.

Techniques: Expressing, In Vitro, In Vivo, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Journal of Immunology Research

Article Title: Tumor-Derived CXCL1 Promotes Lung Cancer Growth via Recruitment of Tumor-Associated Neutrophils

doi: 10.1155/2016/6530410

Figure Lengend Snippet: Ablation of CXCL1 in 3LL cells limits tumor growth in vivo . (a) 3LL cells were stable transfected with negative control shRNA (NC) or specific CXCL1 shRNA (shCXCL1); CXCL1 mRNA expression was measured by quantitative RT-PCR. (b) Protein levels of CXCL1 in the 3LL/NC and 3LL/shCXCL1 supernatants were measured by ELISA. (c) Serum levels of CXCL1 from 3LL/NC or 3LL/shCXCL1 bearing mice were measured by ELISA. Each dot represents an individual mouse. (d) Primary tumors excised from 3LL/NC bearing mice and 3LL/shCXCL1 bearing mice at day 16 after tumor inoculation. (e) Growth of NC and shCXCL1 tumors was monitored for 16 days after cell inoculations. (f) CXCL1 mRNA expression in tumor tissues derived from 3LL/NC or 3LL/shCXCL1 tumor-bearing mice was examined by quantitative RT-PCR. ∗ p < 0.05, ∗∗ p < 0.01. Results are shown as mean ± SD.

Article Snippet: Mouse and human CXCL1 ELISA kit (R&D systems) were used to determine the concentrations of CXCL1 in cell culture supernatant or serum from tumor-bearing mice according to the manufacturer's instructions.

Techniques: In Vivo, Transfection, Negative Control, shRNA, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: Journal of Immunology Research

Article Title: Tumor-Derived CXCL1 Promotes Lung Cancer Growth via Recruitment of Tumor-Associated Neutrophils

doi: 10.1155/2016/6530410

Figure Lengend Snippet: Knockdown of CXCL1 expression in 3LL cells decreases the number of neutrophils in vivo . 5 × 10 5 3LL/NC or 3LL/shCXCL1 cells were inoculated subcutaneously (s.c.) into C57BL/6J mice. At day 16, mice were scarified. The CD11b + Ly6G + neutrophils in peripheral blood were detected as described in the materials and methods. Representative flow cytometric graph (a) and percentage of CD11b + Ly6G + neutrophils in peripheral blood (b) were analyzed. Each dot plot represents an individual mice; horizontal lines are shown as mean of each group. (c) Flow cytometric analysis of the frequency of CD11b + Ly6G + neutrophils in bone marrow from mice inoculated with 3LL/NC or 3LL/shCXCL1. (d) Flow cytometric analysis of the frequency of CD11b + Ly6G + neutrophils in spleen from mice inoculated with 3LL/NC or 3LL/shCXCL1 tumor cells. (e, f, g, and h) IHC staining of Ly6G + or MPO + cells in the tumor tissues from mice inoculated with 3LL/NC or 3LL/shCXCL1 tumor cells. IHC staining quantification of Ly6G + (f) or MPO + (h) cells (per high-power field, HPF). Scale bars, 20 μ m or 40 μ m. (i) Neutrophils were isolated from spleen or peripheral blood derived from tumor-bearing mice and the chemoattracted CD11b + Ly6G + neutrophils were analyzed towards the condition supernatant from NC and shCXCL1 tumor cells by in vitro transwell assay. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Mouse and human CXCL1 ELISA kit (R&D systems) were used to determine the concentrations of CXCL1 in cell culture supernatant or serum from tumor-bearing mice according to the manufacturer's instructions.

Techniques: Knockdown, Expressing, In Vivo, Immunohistochemistry, Isolation, Derivative Assay, In Vitro, Transwell Assay