15 research outputs found
Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes
10.1186/1476-8518-6-2Journal of Immune Based Therapies and Vaccines6
Use of Ultraviolet Light Irradiated Multiple Myeloma Cells as Immunogens to generate Tumor Specific Cytolytic T Lymphocytes
Background:
As the eradication of tumor cells in vivo is most efficiently performed by cytolytic Tlymphocytes (CTL), various methods for priming tumor-reactive lymphocytes have been developed. In this study, a method of priming CTLs with ultraviolet (UV)-irradiated tumor cells, which results in termination of tumor cell proliferation, apoptosis, as well as upregulation of heat shock proteins (HSP) expression is described.
Methods:
Peripheral blood mononuclear cells (PBMC) were primed weekly with UV-irradiated or mitomycin-treated RPMI 8226 multiple myeloma cells. Following three rounds of stimulation over 21 days, the lymphocytes from the mixed culture conditions were analyzed for anti-MM cell reactivity.
Results:
By day 10 of cultures, PBMCs primed using UV-irradiated tumor cells demonstrated a higher percentage of activated CD8+/CD4- T lymphocytes than non-primed PBMCs or PBMCs primed using mitomycin-treated MM cells. Cytotoxicity assays revealed that primed PBMCs were markedly more effective (p \u3c 0.01) than non-primed PBMCs in killing RPMI 8226 MM cells. Surface expression of glucose regulated protein 94 (Grp94/Gp96) and Grp78 were both found to be induced in UV-treated MM cells.
Conclusion:
Since, HSP-associated peptides are known to mediate tumor rejection; these data suggest that immune-mediated eradication of MM cells could be elicited via a UV-induced HSP process. The finding that the addition of 17-allylamide-17-demethoxygeldanamycin (17AAG, an inhibitor of HSP 90-peptide interactions) resulted in decreased CTL-induced cytotoxicity supported this hypothesis. Our study, therefore, provides the framework for the development of anti-tumor CTL cellular vaccines for treating MM using UV-irradiated tumor cells as immunogens
Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes-4
2% polyacrylamide gel. Western immunoblotting was performed with anti-Grp78 (A) and anti-Grp94 (B) antibodies and data is representative to two independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes"</p><p>http://www.jibtherapies.com/content/6/1/2</p><p>Journal of Immune Based Therapies and Vaccines 2008;6():2-2.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2383894.</p><p></p
Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes-8
A synthesis using standard H-TdR (0.25 μCi/well) incorporation assays 3 hr after UV-irradiation. Three triplicate experiments were performed and data is shown as box plots. Statistical differences were determined using the Mann-Whitney test.<p><b>Copyright information:</b></p><p>Taken from "Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes"</p><p>http://www.jibtherapies.com/content/6/1/2</p><p>Journal of Immune Based Therapies and Vaccines 2008;6():2-2.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2383894.</p><p></p
Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes-3
cell immunogens for 28 days in 'mixed-culture conditions'. Non-primed, mito-primed, or UV-primed and PBMCs (effector cells) were then co-cultured with viable H-TdR-labeled RPMI 8226 MM cells (target cells) in a re-directed CTL assay. A standard H-TdR cytotoxicity assays was performed at E:T ratios of 1:1, 3:1, 6:1, 12:1 or 25:1, 50:1 and 100:1.<p><b>Copyright information:</b></p><p>Taken from "Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes"</p><p>http://www.jibtherapies.com/content/6/1/2</p><p>Journal of Immune Based Therapies and Vaccines 2008;6():2-2.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2383894.</p><p></p
Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes-5
R using actin and grp78 (A) or grp94 (B) primers was performed and data is expressed as calibrator corrected and actin normalized ratios.<p><b>Copyright information:</b></p><p>Taken from "Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes"</p><p>http://www.jibtherapies.com/content/6/1/2</p><p>Journal of Immune Based Therapies and Vaccines 2008;6():2-2.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2383894.</p><p></p
Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes-7
cell immunogens for 28 days in 'mixed-culture conditions'. Non-primed, mito-primed, or UV-primed and PBMCs (effector cells) were then co-cultured with viable H-TdR-labeled RPMI 8226 MM cells (target cells) in a re-directed CTL assay. 17-AAG was added at 10 uM to the UV-irradiated RPMI cells for four hours and then washed away before the cells were added to the CTL culture. A standard H-TdR cytotoxicity assays was performed at E:T ratio of 50:1. Experiments were performed triplicate and expressed as the mean ± 2SEM.<p><b>Copyright information:</b></p><p>Taken from "Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes"</p><p>http://www.jibtherapies.com/content/6/1/2</p><p>Journal of Immune Based Therapies and Vaccines 2008;6():2-2.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2383894.</p><p></p
Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes-0
A synthesis using standard H-TdR (0.25 μCi/well) incorporation assays 3 hr after UV-irradiation. Three triplicate experiments were performed and data is shown as box plots. Statistical differences were determined using the Mann-Whitney test.<p><b>Copyright information:</b></p><p>Taken from "Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes"</p><p>http://www.jibtherapies.com/content/6/1/2</p><p>Journal of Immune Based Therapies and Vaccines 2008;6():2-2.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2383894.</p><p></p
Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes-1
E first irradiated with 120 mJ/cm2 of UV light and serially (0, 2, 4, 6 and 8 hr post UV-irradiation) analyzed for the onset of early apoptosis (annexin V+PI-) or established apoptosis (annexin V+PI+) using annexin V-FITC/PI dual staining and fluorescence flow cytometric analysis. All experiments were performed triplicate and expressed as mean values.<p><b>Copyright information:</b></p><p>Taken from "Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes"</p><p>http://www.jibtherapies.com/content/6/1/2</p><p>Journal of Immune Based Therapies and Vaccines 2008;6():2-2.</p><p>Published online 28 Apr 2008</p><p>PMCID:PMC2383894.</p><p></p