htb-4 uroepithelial cell line  (Future Medicine Ltd)

Journal: FEMS Immunology & Medical Microbiology

Article Title: Highly-purifiedHelicobacter pyloriLPS preparations induce weak inflammatory reactions and utilize Toll-like receptor 2 complex but not Toll-like receptor 4 complex

doi: 10.1111/j.1574-695x.2007.00288.x

Figure Lengend Snippet: Fig. 1. IL-8 induction by LPS derived from Heli- cobacter pylori and Escherichia coli in the human uroepithelial cell line T24 and in human gastric carcinoma lines MKN28 and MKN45. Cells were treated with various concentrations of LPS for 24 h. Concentrations of IL-8 in the resulting culture supernatant were determined by ELISA. The LPSs used were derived from Helicobacter pylori strain GU2 (), DU2 (), CA2 (n), CA6 (m), CG10 (&), and DU8 (’), and Escherichia coli O111 : B4 wild-type () and Re mutant (1) strains. The data represent results for the con- ventional LPS preparations. Similar results were obtained when the highly purified LPS prepara- tion was used.

Article Snippet: The human uroepithelial cell line T24 and the human monocytic cell line THP-1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Mutagenesis

Journal: FEMS Immunology & Medical Microbiology

Article Title: Highly-purifiedHelicobacter pyloriLPS preparations induce weak inflammatory reactions and utilize Toll-like receptor 2 complex but not Toll-like receptor 4 complex

doi: 10.1111/j.1574-695x.2007.00288.x

Figure Lengend Snippet: Fig. 2. Expression of (a) mRNA levels (by real-time RT-PCR) of TLR2, TLR4, TLR1, TLR6, CD14 and MD2, and (b) cell surface protein levels (by flow cytometry) of TLR2, TLR4 and CD14 in MKN28, MKN45 and T24 cells. THP-1 cells were used as positive controls. The relative amounts of mRNA levels in relation to THP-1 cells (expression levels in THP-1 cells taken as 1) were calculated. (b) Hatched lines indicate cells incubated with isotype-matched control antibodies.

Article Snippet: The human uroepithelial cell line T24 and the human monocytic cell line THP-1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, Quantitative RT-PCR, Cytometry, Incubation, Control

Journal: FEMS Immunology & Medical Microbiology

Article Title: Highly-purifiedHelicobacter pyloriLPS preparations induce weak inflammatory reactions and utilize Toll-like receptor 2 complex but not Toll-like receptor 4 complex

doi: 10.1111/j.1574-695x.2007.00288.x

Figure Lengend Snippet: Fig. 3. Effect of overexpression of dominant negative mutants of TLR2 or TLR4 on LPS- induced IL-8 induction in T24 cells. Expression plasmids encoding dominant negative mutants of TLR2 (m) or TLR4 (’), or empty plasmid (), were transfected into T24 cells. After 24 h of transfection, cells were treated with Helicobacter pylori LPS derived from strain GU2 (a), strain CA2 (b) or strain CG10 (c), or LPS from Escherichia coli O111 : B4 (d) or Escherichia coli Re mutant (e). Concentrations of IL-8 in the resulting culture supernatants were determined by ELISA. Experi- ments were done in triplicate. The results were expressed as fold induction (mean values SD), relative to the value obtained from control ex- periments without LPS stimulation. P o 0.01 compared to LPS-untreated control. The data represent results for the highly purified LPS pre- parations. Similar results were obtained when the conventional LPS preparations were used.

Article Snippet: The human uroepithelial cell line T24 and the human monocytic cell line THP-1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Over Expression, Dominant Negative Mutation, Expressing, Plasmid Preparation, Transfection, Derivative Assay, Mutagenesis, Enzyme-linked Immunosorbent Assay, Control