Particle Bombardment for Gene Transfer into Nerve Cell Systems

Yang, Ning‐Sun; Jiao, Shoushu; Luna, Carolyn De

doi:10.1016/b978-0-12-185291-7.50031-x

Cited by 1 publication

(1 citation statement)

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“…1 Therefore, the irradiation of tumor cell lines seems to have a negligible effect on cytokine production, and has been reported by some investigators to actually increase total protein production over time. 7 These assays address a major concern with any tumor vaccine protocol: CML-6M and CML-1 cells were transfected as described previously, and supernatant was collected from separate cultures at 12,24,36,48,60, and 72 hours in a cumulative fashion for both irradiated and nonirradiated samples of each line. Thus, the 12-hour samples represent hGM-CSF production accumulated over 12 hours, 24-hour samples represent accumulation over 24 hours, and so forth.…”

Section: Discussionmentioning

confidence: 99%

Preclinical development of human granulocyte-macrophage colony-stimulating factor-transfected melanoma cell vaccine using established canine cell lines and normal dogs

Jk²,

Culp³

et al. 1999

Cancer Gene Ther

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Tumor vaccines and gene therapy have received significant attention as means of increasing cellular and humoral immune responses to cancer. We conducted a pilot study of seven research dogs to determine whether intradermal injection of canine tumor cells transfected via the Accell particle-mediated gene transfer device with the cDNA for human granulocyte-macrophage colony-stimulating factor (hGM-CSF) would generate biologically relevant levels of protein and result in demonstrable histological changes at sites of vaccination. Tumor cell vaccines of 10 7 irradiated canine melanoma cells were nontoxic, safe, and well tolerated. No significant alterations in blood chemistry values or hematological profiles were detected. A histological review of control vaccine sites revealed inflammatory responses predominated by eosinophils, whereas vaccine sites with hGM-CSF-transfected tumor cells had an influx of neutrophils and macrophages. Enzyme-linked immunosorbent assays of skin biopsies from vaccine sites had local hGM-CSF production (8.68 -16.82 ng/site of injection) at 24 hours after injection and detectable levels (0.014 -0.081 ng/site) for Յ2 weeks following vaccination. Flow cytometric analysis of hGM-CSF-transfected cells demonstrated Յ25% transfection efficiency, and hGM-CSF levels obtained during time-course assays demonstrated biologically relevant levels for both irradiated and nonirradiated samples. These data demonstrate the in vivo biological activity of irradiated hGM-CSF-transfected canine tumor cells and help provide evidence for a valid translational research model of spontaneous tumors.

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