Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: A Novel Mode of Action for a Microbial-Derived Immunotoxin: The Cytolethal Distending Toxin Subunit B Exhibits Phosphatidylinositol 3,4,5-Triphosphate Phosphatase Activity 1

doi:

Figure Lengend Snippet: Assessment of CdtB mutants for their ability to induce G2 arrest in Jurkat cells. Jurkat cells were exposed to medium alone (A) or 10 ng/ml each of CdtA and CdtC in the presence of 4 ng/ml CdtBWT (B) or CdtBH160Q (C), CdtBH274Q (D), CdtBR117A (E), or CdtBD199S (F). Cells were analyzed for cell cycle distribution 18 h after exposure to toxin subunits using flow cytometric analysis of propidium iodide fluorescence (27). The numbers in each panel represent the percentages of cells in G0/G1, S, and G2/M. Cells exposed to only 10 ng/ml each of CdtA and CdtC exhibited 14.7% G2 cells (data not shown). Results are representative of three experiments.

Article Snippet: The human leukemic T cell lines Jurkat, CEM, and Molt were maintained in RPMI 1640 supplemented with 10% FCS, 2 mM glutamine, 10 mM HEPES, 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Fluorescence

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: A Novel Mode of Action for a Microbial-Derived Immunotoxin: The Cytolethal Distending Toxin Subunit B Exhibits Phosphatidylinositol 3,4,5-Triphosphate Phosphatase Activity 1

doi:

Figure Lengend Snippet: Relationship between exposure to Cdt, toxicity, and cellular content of PI-3,4,5-P3. A, Jurkat cells were incubated in the presence of 50 pg/ml CdtABC for varying periods of time. Cells were then harvested, phospholipids were extracted, and PI-3,4,5-P3 levels determined by ELISA. Data are plotted as PI-3,4,5-P3 content (pM/107 cells; mean ± SD) vs time. B, PI3K inhibitors were used to lower Jurkat cell PI-3,4,5-P3 and thereby alter susceptibility to CdtABC. Jurkat cells were preincubated in medium, 250 nM wortmannin, or 40 µM LY290004. The cells were then treated with medium or 40 pg/ml CdtABC and assessed for cell cycle distribution by flow cytometry. The numbers in each panel represent the percentages of cells in G0/G1, S, and G2/M.

Article Snippet: The human leukemic T cell lines Jurkat, CEM, and Molt were maintained in RPMI 1640 supplemented with 10% FCS, 2 mM glutamine, 10 mM HEPES, 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: A Novel Mode of Action for a Microbial-Derived Immunotoxin: The Cytolethal Distending Toxin Subunit B Exhibits Phosphatidylinositol 3,4,5-Triphosphate Phosphatase Activity 1

doi:

Figure Lengend Snippet: Comparison of lymphoid cell line susceptibility to Cdt-induced G2 arrest. Jurkat cells (●), Hut78 cells (▼), Molt cells (■), and CCRF-CEM cells (♦) were treated with varying amounts of CdtABC and incubated for 18 h. The cells were then harvested, stained with propidium iodide, and analyzed by flow cytometry for cell cycle distribution. Data are plotted as percent G2 cells vs CdtABC concentration. Results represent the mean ± SD of three experiments.

Article Snippet: The human leukemic T cell lines Jurkat, CEM, and Molt were maintained in RPMI 1640 supplemented with 10% FCS, 2 mM glutamine, 10 mM HEPES, 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Comparison, Incubation, Staining, Flow Cytometry, Concentration Assay

Journal: Retrovirology

Article Title: In vivo expression of the HBZ gene of HTLV-1 correlates with proviral load, inflammatory markers and disease severity in HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP)

doi: 10.1186/1742-4690-6-19

Figure Lengend Snippet: HTLV-1 mRNA load, proviral load and mRNA/DNA ratio in HTLV-1 – infected individuals and T-cell lines.

Article Snippet: We could not detect any HTLV-1 tax and HBZ mRNA expression in any of the 20 NCs and 3 uninfected human leukemic T-cell lines (Jurkat, MOLT-4, and CEM) tested (data not shown).

Techniques: Infection