Journal: Journal of Hematology & Oncology

Article Title: Intratumoral anti-HuD immunotoxin therapy for small cell lung cancer and neuroblastoma

doi: 10.1186/s13045-014-0091-3

Figure Lengend Snippet: Western blot analysis of HuD proteins in SCLC, neuroblastoma, and leukemia/lymphoma cell lines. Protein extracts from cell lysates were subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a NuPAGE™ 4-12% Bis-Tris gel, transferred to nitrocellulose membrane. The membrane was probed with anti-HuD mAb (16A11) and a HRP-conjugated goat anti-mouse IgG detection system. The neuroblastoma (Neuro-2a) and SCLC (NCI-H69, DSM79) extracts demonstrated strong bands at 40 kD, whereas two control cell line (BW5147, K562) extracts were devoid of signal.

Article Snippet: Human SCLC (NCI-H69, DMS-79), human erythroleukemia (K-562), mouse NB (neuro-2a) and mouse T lymphoma (BW5147) cell lines were purchased from the American Type Culture Collection (ATCC; Rockville, MD).

Techniques: Western Blot, Polyacrylamide Gel Electrophoresis, SDS Page, Membrane, Control

Journal: Journal of Hematology & Oncology

Article Title: Intratumoral anti-HuD immunotoxin therapy for small cell lung cancer and neuroblastoma

doi: 10.1186/s13045-014-0091-3

Figure Lengend Snippet: Immunotoxin BW-2 efficiently kills SCLC and neuroblastoma cancer cells. BW-2 kills (A) HuD-positive H69 cells and (B) neuro-2a, whereas no cytotoxicity occurred in the control (C) HuD-negative K562 cells. Log dilutions of BW-2 (0–5 μg/ml) were added to the cell culture wells (5 × 10 4 cells/well) and incubated at 37°C for 72 hr. Exposure to equal molar concentrations of streptavidin-saporin (Streptavidin-SP) alone or mAb (16A11) alone served as controls. BW-2 efficiently killed the targeted H69 and neuro-2a cancer cells with great potency ( P < 0.005 ). SAP-streptavidin alone or 16A11 mAb alone induced only mild non-specific killing at higher concentrations. Data represents the mean of three tests in a single representative experiment repeated 3 times.

Article Snippet: Human SCLC (NCI-H69, DMS-79), human erythroleukemia (K-562), mouse NB (neuro-2a) and mouse T lymphoma (BW5147) cell lines were purchased from the American Type Culture Collection (ATCC; Rockville, MD).

Techniques: Control, Cell Culture, Incubation

Journal: Journal of Hematology & Oncology

Article Title: Intratumoral anti-HuD immunotoxin therapy for small cell lung cancer and neuroblastoma

doi: 10.1186/s13045-014-0091-3

Figure Lengend Snippet: Nude mouse model of human SCLC and pathology. Viable H69 SCLC cells (1x10 7 /100 μl PBS) were subcutaneously implanted into the flank of nude mice. (A) Subcutaneous tumor nodules developed and progressed rapidly after s.c. implantation and multiple organ cancer metastases occurred when the s.c. tumor reached a certain size (~1000 mm 3 ). (B) The metastasized cancer cells were easily detected by immunoreaction with biotinylated anti-HuD mAb and Cy3 fluorescent detection.

Article Snippet: Human SCLC (NCI-H69, DMS-79), human erythroleukemia (K-562), mouse NB (neuro-2a) and mouse T lymphoma (BW5147) cell lines were purchased from the American Type Culture Collection (ATCC; Rockville, MD).

Techniques:

Journal: Journal of Hematology & Oncology

Article Title: Intratumoral anti-HuD immunotoxin therapy for small cell lung cancer and neuroblastoma

doi: 10.1186/s13045-014-0091-3

Figure Lengend Snippet: Intratumorally injected immunotoxin BW-2 inhibits growth in HuD-positive H69 SCLC tumor xenografted mice. SCLC tumor xenografted nude mice (n = 6) were treated with one time intratumoral immunotoxin, or an equivalent dose of mAb alone, saporin toxin alone, or saline. A) Tumor size in BW-2 treated mice remained static or actually regressed for ~3 weeks. B) The control mice exhibited rapidly progressive increases in tumor volume and were overwhelmed by large tumor burden as well as metastases.

Article Snippet: Human SCLC (NCI-H69, DMS-79), human erythroleukemia (K-562), mouse NB (neuro-2a) and mouse T lymphoma (BW5147) cell lines were purchased from the American Type Culture Collection (ATCC; Rockville, MD).

Techniques: Injection, Saline, Control