title: Molecular mechanisms of paraptosis induction: implications for a non-genetically modified tumor vaccine.
abstract: Paraptosis is the programmed cell death pathway that leads to cellular necrosis. Previously, rodent and human monocytes/macrophages killed glioma cells bearing the membrane macrophage colony stimulating factor (mM-CSF) through paraptosis, but the molecular mechanism of this killing process was never identified. We have demonstrated that paraptosis of rat T9 glioma cells can be initiated through a large potassium channel (BK)-dependent process initiated by reactive oxygen species. Macrophage mediated cytotoxicity upon the mM-CSF expressing T9-C2 cells was not prevented by the addition of the caspase inhibitor, zVAD-fmk. By a combination of fluorescent confocal and electron microscopy, flow cytometry, electrophysiology, pharmacology, and genetic knock-down approaches, we demonstrated that these ion channels control cellular swelling and vacuolization of rat T9 glioma cells. Cell lysis is preceded by a depletion of intracellular ATP. Six-hour exposure to BK channel activation caused T9 cells to over express heat shock proteins (Hsp 60, 70, 90 and gp96). This same treatment forced HMGB1 translocation from the nuclear region to the periphery. These last molecules are "danger signals" that can stimulate immune responses. Similar inductions of mitochondrial swelling and increased Hsp70 and 90 expressions by BK channel activation were observed with the non-immunogenic F98 glioma cells. Rats injected with T9 cells which were killed by prolonged BK channel activation developed immunity against the T9 cells, while the injection of x-irradiated apoptotic T9 cells failed to produce the vaccinating effect. These results are the first to show that glioma cellular death induced by prolonged BK channel activation improves tumor immunogenicity; this treatment reproduces the vaccinating effects of mM-CSF transduced cells. Elucidation of strategies as described in this study may prove quite valuable in the development of clinical immunotherapy against cancer.
authors.affiliation: Diagnostic and Molecular Medicine Healthcare Group, Veterans Affairs Medical Center, Long Beach, California, United States of America.
authors.forename: Neil
authors.initials: N
authors.lastname: Hoa
authors.forename: Michael P
authors.initials: MP
authors.lastname: Myers
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Douglass
authors.forename: Jian Gang
authors.initials: JG
authors.lastname: Zhang
authors.forename: Christina
authors.initials: C
authors.lastname: Delgado
authors.forename: Lara
authors.initials: L
authors.lastname: Driggers
authors.forename: Linda L
authors.initials: LL
authors.lastname: Callahan
authors.forename: Gerald
authors.initials: G
authors.lastname: VanDeusen
authors.forename: Jimmy T H
authors.initials: JT
authors.lastname: Pham
authors.forename: Nirav
authors.initials: N
authors.lastname: Bhakta
authors.forename: Lisheng
authors.initials: L
authors.lastname: Ge
authors.forename: Martin R
authors.initials: MR
authors.lastname: Jadus
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D019496:Cancer Vaccines / Q000008:administration & dosage* / Q000276:immunology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D055785:Gene Knockdown Techniques; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008928:Mitochondria / Q000378:metabolism; D008933:Mitochondrial Swelling; D010455:Peptides / Q000494:pharmacology; D010693:Phloretin / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D019496:Cancer Vaccines; D051036:Large-Conductance Calcium-Activated Potassium Channels; D010455:Peptides; D017382:Reactive Oxygen Species; C064719:iberiotoxin; D000255:Adenosine Triphosphate; D010693:Phloretin