Journal: Gene therapy

Article Title: Tumor cells expressing a fusion protein of MULT1 and Fas are rejected in vivo by apoptosis and NK cell activation.

doi: 10.1038/gt.2008.77

Figure Lengend Snippet: Figure 3 Mouse UL16-binding protein-like transcript 1 E (MULT1E)/FasTI induces apoptosis. A total of 1 106 cells of TC-1 and clones L-5, L-7 and L-10 were treated with 1 mg/ml NKG2D/Fc for 16 h. The cells were then analyzed for apoptosis and necrosis using Annexin V assay (a–c) or caspase-3 assay (d) according to the manufacturers’ protocols. (a) An example of the fluorescence-activated cell sorting (FACS) data. (b, c) Summaries of data from three separate experiments. The statistical analyses were conducted between the controls (open bars) and NKG2D/Fc-treated cells (solid bars) using two-way analysis of variance (ANOVA). The difference between NKG2D/Fc-treated L-5 cells and NKG2D/Fc-treated L-10 cells was also compared using Student’s t-test. *Po0.05; **Po0.01 and ***Po0.001.

Article Snippet: Induction of apoptosis in cells expressing the fusion protein To determine if cells expressing the fusion protein can be induced to undergo apoptosis, one million cells of TC-1 and clones L-5, L-7 and L-10 were treated with 1 mg/ml of NKG2D/Fc for 16 h. Apoptosis of the cells was measured using two systems: a TACS Annexin V-FITC Apoptosis Kit (R&D Systems) and a caspase-3 fluorometric assay (R&D Systems).

Techniques: Binding Assay, Clone Assay, Annexin V Assay, Caspase-3 Assay, FACS

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Immune impairment in patients with terminal cancers: influence of cancer treatments and cytomegalovirus infection

doi: 10.1007/s00262-009-0753-0

Figure Lengend Snippet: Altered T cell differentiations in cancer patients. The expression of the phenotypic markers CD45RA and CCR7 was used to define the stages of T cell differentiation. The proportion of naïve T cells (CD45RA+CCR7+) (a) (p = 0.0296, one-way ANOVA) and central memory T cells (CD45RA−CCR7+) (b) (p = 0.0312, one-way ANOVA) in the heavy-treated patients were markedly lower than normal. The numbers of effector memory T cells (CD45RA−CCR7−) (c) (p = 0.0104, one-way ANOVA) in the heavy-treated patients were higher than those of the normal groups. However, the proportion of effector T cells (CD45RA+CCR7−) (d) did not differ among the three groups (e) Overview of the three subpopulations in cancer patients. The proportion of early differentiated subpopulations, naïve or central memory cells, gradually declined from the normal group to the heavy-treated group. Asterisks indicate statistically significant differences between the normal and heavy-treated groups (p < 0.05). N normal, TN treatment-naive, and HT heavy-treated

Article Snippet: Flow cytometry Peripheral blood mononuclear cells were stained with antibodies against CD8 (FITC, clone HIT8a), CD27 (FITC, clone M-T271), CD28 (PE and APC, clone CD28.2), CD45RA (APC, clone HI100), and CD127 (PE, clone hIL-7R-M21) (BD PharmingenTM, San Jose, CA, USA); CD4 (PE and PerCP, clone SK3) and CD8 (PerCP, clone SK1) (BD, San Jose, CA, USA); and CCR7 (FITC, clone 150503) (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Cell Differentiation

Journal: Immunology

Article Title: Lentiviral calnexin-modified dendritic cells promote expansion of high-avidity effector T cells with central memory phenotype.

doi: 10.1111/j.1365-2567.2009.03067.x

Figure Lengend Snippet: Figure 6. Increased CCR7 expression in calnexin–dendritic cell (CNX-DC)-activated T cells. (a) Flow cytometry analysis of memory markers in T cells after coculture with lentiviral vector (LV)-modi- fied DCs. T cells were cocultured with different LV-modified DCs as indicated, restimulated for 7 days and stained with GLC-pentamer together with anti-CD8 antibody and antibodies against CCR7, CD62L, CD28 or CD69. GLC pentamer-positive T cells were CD8- gated and analysed for the expression of differentiation markers. The percentages of different cell population are indicated in the flow graphs. (b) Quantitative analysis of CCR7+ cells in pentamer-positive T cells. Data are representative of four experiments.

Article Snippet: Other fluorochromeconjugated antibodies used in this study included FITC-CCR7 (R&D Systems Inc., Minneapolis, MN), FITC-conjugated human leucocyte antigen DR (HLA-DR) (Caltag Laboratories, Invitrogen, Carlsbad, CA), APC-DCSIGN and APC-CD28 (eBioscience, San Diego, CA).

Techniques: Expressing, Flow Cytometry, Plasmid Preparation, Staining