Journal: Retrovirology

Article Title: Construction of doxycyline-dependent mini-HIV-1 variants for the development of a virotherapy against leukemias

doi: 10.1186/1742-4690-3-64

Figure Lengend Snippet: Replication and cell killing capacity of dox-inducible viruses in SupT1 cells . ( Left ) Virus replication of LAI (△), HIV-rtTA (▲), HIV-rtTAΔ6 A (○), HIV-rtTAΔ6 B (◆) and HIV-rtTA without dox (×) was determined by measuring of the supernatant CA-p24 concentration after infection with virus (20 ng CA-p24) in a 5 mL SupT1 culture. ( Right ) The number of cells in each culture was determined by a 30 sec time limit FACS analysis. The cell killing capacity of the viruses was determined as the ratio of SupT1 cells present in the infected culture versus the uninfected control culture.

Article Snippet: The human T lymphocyte cell line SupT1 [ATCC CRL-1942, [ ]] was cultured in RPMI 1640 (Gibco BRL, Gaithersburg, MD) supplemented with 10% (v/v) FCS, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37°C and in 5% CO 2 .

Techniques: Concentration Assay, Infection

Journal: Retrovirology

Article Title: Construction of doxycyline-dependent mini-HIV-1 variants for the development of a virotherapy against leukemias

doi: 10.1186/1742-4690-3-64

Figure Lengend Snippet: Dox regulated replication of the mini-rtTA virus rtTAΔ6 A . SupT1 cells were infected with rtTAΔ6 A virus (1 ng CA-p24). The culture was split and the cells were cultured with dox (upper panels) or without (lower panels). Virus replication was monitored by CA-p24 Elisa on the culture supernatant. At day 9 post infection, both cultures were washed and each culture split into one culture with dox (left panels) and one without (right panels). Filled triangles indicate cultures without dox and open triangles indicate cultures with 1000 ng/mL dox.

Article Snippet: The human T lymphocyte cell line SupT1 [ATCC CRL-1942, [ ]] was cultured in RPMI 1640 (Gibco BRL, Gaithersburg, MD) supplemented with 10% (v/v) FCS, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37°C and in 5% CO 2 .

Techniques: Infection, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Retrovirology

Article Title: Construction of doxycyline-dependent mini-HIV-1 variants for the development of a virotherapy against leukemias

doi: 10.1186/1742-4690-3-64

Figure Lengend Snippet: Selective SupT1 killing in SupT1/PBMC co-cultures by mini-rtTA viruses . Infections were started with virus corresponding to 40 ng CA-p24 or mock infected. The FACS dot plot of the initial PBMC + SupT1 cell mixture is in the lower left corner, and the separate cultures are shown above. The gates for CD4+ PBMC (blue), SupT1 (red) and CD8+ PBMC (green) are indicated. The composition of the PBMC + SupT1 culture was followed over time upon infection with the indicated viruses.

Article Snippet: The human T lymphocyte cell line SupT1 [ATCC CRL-1942, [ ]] was cultured in RPMI 1640 (Gibco BRL, Gaithersburg, MD) supplemented with 10% (v/v) FCS, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37°C and in 5% CO 2 .

Techniques: Infection

Journal: Retrovirology

Article Title: HIV-1 latency in actively dividing human T cell lines

doi: 10.1186/1742-4690-5-37

Figure Lengend Snippet: Overview of SupT1 clones with integrated HIV-rtTA provirus

Article Snippet: The human T lymphocytic cell lines SupT1 (ATCC CRL-1942) [ ], MT2, C8166 and Jurkat (ATCC TIB-152) were cultured in advanced RPMI 1640 medium (Gibco BRL, Gaithersburg, MD) supplemented with 1% (v/v) FCS, 40 U/mL penicillin, and 40 μg/mL streptomycin at 37°C and 5% CO 2 .

Techniques: Clone Assay

Journal: Retrovirology

Article Title: HIV-1 latency in actively dividing human T cell lines

doi: 10.1186/1742-4690-5-37

Figure Lengend Snippet: Northern blot analysis of shRNA production in rtTA-shNef SupT1 cell lines . RNAs from the different HIV-rtTA-shNef SupT1 clones were isolated and separated on an agarose gel. After blotting the membrane was probed with a LNA probe against the shNef. Lane M, marker; lane 1, shNef clone B9; lane 2, clone C10 (a negative control cell line without a provirus); lane 3, shNef clone D2; lane 4, shNef clone D11; lane 5, shNef clone F7; lane 6, shNef clone F8; lane 7 and 8 contain positive controls from a transfection with the F-shNef - and F-shNef + plasmid [31]; lane 9 contains a negative empty-vector control; lane 10 in vitro produced shNef RNA.

Article Snippet: The human T lymphocytic cell lines SupT1 (ATCC CRL-1942) [ ], MT2, C8166 and Jurkat (ATCC TIB-152) were cultured in advanced RPMI 1640 medium (Gibco BRL, Gaithersburg, MD) supplemented with 1% (v/v) FCS, 40 U/mL penicillin, and 40 μg/mL streptomycin at 37°C and 5% CO 2 .

Techniques: Northern Blot, shRNA, Clone Assay, Isolation, Agarose Gel Electrophoresis, Marker, Negative Control, Transfection, Plasmid Preparation, In Vitro, Produced

Journal: Retrovirology

Article Title: HIV-1 latency in actively dividing human T cell lines

doi: 10.1186/1742-4690-5-37

Figure Lengend Snippet: Silencing and reactivation in HIV-rtTA SupT1 cell lines . The HIV-rtTA SupT1 cell lines F7 and B9 were induced with dox or with dox in combination with the indicated activators. DMSO is the solvent for genistein and therefore used as an additional control. Statistical significance was determined with a two-tailed student's T test for each combination versus dox alone (GraphPad Prism). Significant changes ( P -value < 0.05) are indicated with asterisks.

Article Snippet: The human T lymphocytic cell lines SupT1 (ATCC CRL-1942) [ ], MT2, C8166 and Jurkat (ATCC TIB-152) were cultured in advanced RPMI 1640 medium (Gibco BRL, Gaithersburg, MD) supplemented with 1% (v/v) FCS, 40 U/mL penicillin, and 40 μg/mL streptomycin at 37°C and 5% CO 2 .

Techniques: Two Tailed Test

Journal: Retrovirology

Article Title: HIV-1 latency in actively dividing human T cell lines

doi: 10.1186/1742-4690-5-37

Figure Lengend Snippet: Latent HIV-1 infection in SupT1 T cells . TNFα-induced reactivation of silent HIV-1 proviruses in SupT1 cells was analyzed by determining the percentage of CA-p24 producing cells by intracellular FACS analysis. (A) After a single round infection with the LAI isolate, the culture was split and one culture activated for 24 h with TNFα and the other used as a control. (B) The control culture was maintained for one week and split again into a TNFα treated and control culture. Statistical significance was determined with a two-tailed student's T test (GraphPad Prism).

Article Snippet: The human T lymphocytic cell lines SupT1 (ATCC CRL-1942) [ ], MT2, C8166 and Jurkat (ATCC TIB-152) were cultured in advanced RPMI 1640 medium (Gibco BRL, Gaithersburg, MD) supplemented with 1% (v/v) FCS, 40 U/mL penicillin, and 40 μg/mL streptomycin at 37°C and 5% CO 2 .

Techniques: Infection, Two Tailed Test

Journal: Retrovirology

Article Title: HIV-1 latency in actively dividing human T cell lines

doi: 10.1186/1742-4690-5-37

Figure Lengend Snippet: Latent HIV-1 and HIV-rtTA infection in SupT1 T cells . TNFα-induced reactivation of silent HIV-1 proviruses in SupT1 cells was analyzed by determining the percentage of CA-p24 producing cells by intracellular FACS analysis. After a single round infection with HIV-1 (LAI isolate) or the HIV-rtTA-Tat wt , each culture was split and either activated for 24 h with TNFα or not. The fold TNFα activation is the percentage CA-p24 positive cells in the culture with TNFα divided by the percentage of CA-p24 positive cells in the control culture.

Article Snippet: The human T lymphocytic cell lines SupT1 (ATCC CRL-1942) [ ], MT2, C8166 and Jurkat (ATCC TIB-152) were cultured in advanced RPMI 1640 medium (Gibco BRL, Gaithersburg, MD) supplemented with 1% (v/v) FCS, 40 U/mL penicillin, and 40 μg/mL streptomycin at 37°C and 5% CO 2 .

Techniques: Infection, Activation Assay