Journal: Neoplasia (New York, N.Y.)

Article Title: Downregulation and/or Release of NKG2D Ligands as Immune Evasion Strategy of Human Neuroblastoma 1

doi:

Figure Lengend Snippet: MICA, MICB, ULBP-1, -2, and -3 gene expression in primary GD2+ NB cells as assessed by RT-PCR. In panel A, the results obtained with two representative tumors out of the eight analyzed are shown. Neuroblasts were isolated as GD2+ cells by immunomagnetic bead manipulation. From left to right: MW = molecular weight markers; NC = negative control, represented by water in place of cDNA; PC = positive control, represented by 1) the GD2- cell fraction isolated from primary NB tumors for CD45 and DRβ gene expression, 2) HeLa cells for MICA gene expression, 3) U937 cells for MICB gene expression, and 4) 293T cells for ULBP gene expression; Pt 1 = NB cells from patient 1; Pt 2 = NB cells from patient 2. The first upper panel shows the amplification product of the G3PDH housekeeping gene tested as control. On the right side of each panel, the expected MWs of the amplified bands are shown. Panel B summarizes the results obtained from the study of the eight primary NB tumors.

Article Snippet: In some experiments, a 1-ml aliquot of sMICA + NB serum or of ACN cell line supernatant was preincubated with immunomagnetic beads (Immunotech), which had been coated with anti-MICA mAb, for 3 hours at room temperature in continuous rotation before being tested in the above assay.

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Isolation, Molecular Weight, Negative Control, Positive Control, Amplification, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Downregulation and/or Release of NKG2D Ligands as Immune Evasion Strategy of Human Neuroblastoma 1

doi:

Figure Lengend Snippet: Reduction of NKG2D-mediated killing of NB cells by soluble MICA. The surface MICA+ ACN NB cell line was tested as target of normal activated NK cells in a 4-hour chromium release assay. NKG2D-dependent cytolytic activity was evaluated by preincubating effector cells with medium, anti-MICA mAb, sMICA+ serum of a NB patient, sMICA+ serum of the same NB patient pretreated with anti-MICA mAb bound to immunomagnetic beads, or sMICA- normal serum. The figure shows one representative experiment out of the three performed.

Article Snippet: In some experiments, a 1-ml aliquot of sMICA + NB serum or of ACN cell line supernatant was preincubated with immunomagnetic beads (Immunotech), which had been coated with anti-MICA mAb, for 3 hours at room temperature in continuous rotation before being tested in the above assay.

Techniques: Release Assay, Activity Assay

Journal: EBioMedicine

Article Title: Radiotherapy Upregulates Programmed Death Ligand-1 through the Pathways Downstream of Epidermal Growth Factor Receptor in Glioma

doi: 10.1016/j.ebiom.2018.01.027

Figure Lengend Snippet: The effect of PD-L1 blockade on the suppressing ability of tumor cells for CD8 + T cell proliferation. Cell lines (U87 and U251) were treated with RT (5 Gy), PD-L1 Ab (20 μg/mL), RT + PD-L1 Ab, AG490 (10 μM), RT + AG490, or RT + AG490 + PD-L1 Ab, and then co-cultured with CD8 + T cells. At 48 h, CD8 + T cell proliferation was evaluated by FCM. Non-specific Stimulation: anti-CD3/CD28 beads. Representative images and quantitative data are shown. Each column is shown as the means of three separate experiments; bars, SD. Student's t -test; *** P < 0.001. ANOVA; ns, nonsignificant.

Article Snippet: CD8 + T cell immunomagnetic beads positive selection kit was purchased from Stem-Cell Tech (USA).

Techniques: Cell Culture

Journal: EBioMedicine

Article Title: Radiotherapy Upregulates Programmed Death Ligand-1 through the Pathways Downstream of Epidermal Growth Factor Receptor in Glioma

doi: 10.1016/j.ebiom.2018.01.027

Figure Lengend Snippet: The effect of PD-L1 blockade on the cytotoxicity of CD8 + T cells against glioma cells. Cell lines (U87 and U251) were treated with RT (5 Gy), PD-L1 Ab (20 μg/mL), RT + PD-L1 Ab, AG490 (10 μM), RT + AG490, or RT + AG490 + PD-L1 Ab, and then co-cultured with CD8 + T cells. At 48 h, tumor cell apoptosis was evaluated by FCM. Representative images and quantitative data are shown. Each column is shown as the means of three separate experiments; bars, SD. Student's t-test; *** P < 0.001. ANOVA; ns, nonsignificant.

Article Snippet: CD8 + T cell immunomagnetic beads positive selection kit was purchased from Stem-Cell Tech (USA).

Techniques: Cell Culture