Journal: American journal of respiratory cell and molecular biology

Article Title: Role of Proinflammatory Cytokines IL-18 and IL-1β in Bleomycin-Induced Lung Injury in Humans and Mice

doi: 10.1165/rcmb.2008-0182OC

Figure Lengend Snippet: Increase of IL-18–producing F4/80-positive macrophages, but not CD3+ T cells, in bronchoalveolar lavage fluid (BALF) from bleomycin-treated, WT B6 mice. Recovered BALF cells were isolated from WT B6 mice treated intraperitoneally with bleomycin or control PBS on Day 4. Isolated BALF cells were stained with PE-Cy7–conjugated anti-mCD3 and PE-F4/80 mAb in the presence of anti-mCD16/CD32 mAb. The cells were then fixed, permeabilized, and stained with FITC anti–mIL-18 (M5). Three-color analysis was performed for analysis of cytoplasmic IL-18 expression in F4/80+ neutrophils and CD3+ T cells.

Article Snippet: For intracellular cytokine staining, cells were stained with FITC anti–mIL-18 (M5) and/or FITC rat IgG2a (Caltag Laboratories), as reported previously ( 11 , 13 ).

Techniques: Isolation, Staining, Expressing

Journal: BMC Cancer

Article Title: Intratumoral delivery of IL-18 naked DNA induces T-cell activation and Th1 response in a mouse hepatic cancer model

doi: 10.1186/1471-2407-7-87

Figure Lengend Snippet: Detection of mIL-18 plasmid expression in DNA treatment sites . BALB/c mice were inoculated with murine CT26 cells in the left liver lobe, and treated intratumorally with varying doses of control plasmid or that expressing IL-18 until day 7 after tumor inoculation. Liver tissue was obtained on day 4 after direct injection of IL-18 plasmid DNA, and ELISA used to evaluate IL-18 protein levels. Representative data determined from two separate experiments.

Article Snippet: The 11 kb mIL-18 DNA expression plasmid vector, pCEP4-mIL18, was constructed using a CMV early enhancer/promoter/EBNA-1-based pCEP4 plasmid vector (Invitrogen, San Diego, CA) with an ampicillin selection gene.

Techniques: Plasmid Preparation, Expressing, Injection, Enzyme-linked Immunosorbent Assay

Journal: BMC Cancer

Article Title: Intratumoral delivery of IL-18 naked DNA induces T-cell activation and Th1 response in a mouse hepatic cancer model

doi: 10.1186/1471-2407-7-87

Figure Lengend Snippet: Immune response is enhanced by a single injection of naked IL-18 plasmid DNA . Each group (consisting of 4 mice) was administered an intratumoral injection of pCEP4 or naked mIL-18 DNA. Spleens of CT26 tumor-bearing mice treated with plasmids were harvested at 7 days after plasmid injection, and cells analyzed by flow cytometry using the corresponding FITC- or PE- or PE-Cy5-labeled antibodies and isotype control.

Article Snippet: The 11 kb mIL-18 DNA expression plasmid vector, pCEP4-mIL18, was constructed using a CMV early enhancer/promoter/EBNA-1-based pCEP4 plasmid vector (Invitrogen, San Diego, CA) with an ampicillin selection gene.

Techniques: Injection, Plasmid Preparation, Flow Cytometry, Labeling

Journal: BMC Cancer

Article Title: Intratumoral delivery of IL-18 naked DNA induces T-cell activation and Th1 response in a mouse hepatic cancer model

doi: 10.1186/1471-2407-7-87

Figure Lengend Snippet: IL-18 gene transfer results in regression of CT26 liver tumors . CT26 cells (1.0 × 10 5 ) were injected into a left lobe of the livers of 6-8-week-old BALB/c mice. pCEP4 or mIL-18 plasmid DNA was injected directly into the tumors on day 7. Tumor growth was measured on days 1, 4, 7, and 14 after plasmid implantation. Data are presented as the mean of tumor weights of 8 mice/group.

Article Snippet: The 11 kb mIL-18 DNA expression plasmid vector, pCEP4-mIL18, was constructed using a CMV early enhancer/promoter/EBNA-1-based pCEP4 plasmid vector (Invitrogen, San Diego, CA) with an ampicillin selection gene.

Techniques: Injection, Plasmid Preparation

Journal: BMC Cancer

Article Title: Intratumoral delivery of IL-18 naked DNA induces T-cell activation and Th1 response in a mouse hepatic cancer model

doi: 10.1186/1471-2407-7-87

Figure Lengend Snippet: Intracellular IFN-γ production by CT26-specific T cells from pmIL-18-treated mice . Splenocytes from CT26-tumor bearing mice treated with three rounds of pCEP4 or mIL-18 were pooled and incubated in medium alone or that containing CT26 lysates. Numbers within the gates in the FACS plots depict the percentage of CD69 + IFN-γ + CD4 + cells. Data are presented from experiments that were repeated at least twice.

Article Snippet: The 11 kb mIL-18 DNA expression plasmid vector, pCEP4-mIL18, was constructed using a CMV early enhancer/promoter/EBNA-1-based pCEP4 plasmid vector (Invitrogen, San Diego, CA) with an ampicillin selection gene.

Techniques: Incubation

Journal: BMC Cancer

Article Title: Intratumoral delivery of IL-18 naked DNA induces T-cell activation and Th1 response in a mouse hepatic cancer model

doi: 10.1186/1471-2407-7-87

Figure Lengend Snippet: Elevated IFN-γ levels after intratumoral injection of nonviral mIL-18 plasmid DNA . Splenocytes from CT26 tumor-bearing mice after gene therapy were collected at 4, 7, and 14 days after gene treatment, and cultured with anti-CD3 and anti-CD28 antibodies for 3 days before performing ELISA (* = 0.006).

Article Snippet: The 11 kb mIL-18 DNA expression plasmid vector, pCEP4-mIL18, was constructed using a CMV early enhancer/promoter/EBNA-1-based pCEP4 plasmid vector (Invitrogen, San Diego, CA) with an ampicillin selection gene.

Techniques: Injection, Plasmid Preparation, Cell Culture, Enzyme-linked Immunosorbent Assay