Cytolytic cells induce HMGB1 release from melanoma cell lines.
High mobility group box 1 (HMGB1) is one of the recently defined damage-associated molecular pattern molecules, passively released from necrotic cells and secreted by activated macrophage/monocytes. Whether cytolytic cells induce HMGB1 release from tumor cells is not known. We developed a highly sensitive method for detecting intracellular HMGB1 in tumor cells, allowing analysis of the type of cell death and in particular, necrosis. We induced melanoma cell death with cytolytic lymphokine-activated killing (LAK) cells, tumor-specific cytolytic T lymphocytes, TRAIL, or granzyme B delivery and assessed intracellular HMGB1 retention or release to investigate the mechanism of HMGB1 release by cytolytic cells. HMGB1 release from melanoma cells (451Lu, WM9) was detected within 4 h and 24 h following incubation with IL-2-activated PBMC (LAK activity). HLA-A2 and MART1 or gp100-specific cytolytic T lymphocytes induced HMGB1 release from HLA-A2-positive and MART1-positive melanoma cells (FEM X) or T2 cell-loaded, gp100-specific peptides. TRAIL treatment, however, induced HMGB1 release, and it is interesting that this extrinsic pathway-mediated cell death was blocked with the pancaspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone. Conversely, granzyme B delivery did not induce HMGB1 release. HMGB1, along with other intracellular factors released from tumor cells induced by cytolysis, may be important components of the disordered tumor microenvironment. This has important implications for the immunotherapy of patients with cancer. Specifically, HMGB1 may promote healing or immune reactivity, depending on the nature of the local inflammatory response and the presence (or absence) of immune effectors.
Ito
Norimasa
N
Department of Surgery, University of Pittsburgh School of Medicine, Hillman Cancer Center, 5117 Centre Avenue, Room G.21, Pittsburgh, PA 15213, USA.
DeMarco
Richard A
RA
Mailliard
Robbie B
RB
Han
Jie
J
Rabinowich
Hannah
H
Kalinski
Pawel
P
Stolz
Donna Beer
DB
Zeh
Herbert J
HJ
Lotze
Michael T
MT
D021581:Active Transport, Cell Nucleus; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D016923:Cell Death; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003601:Cytotoxicity Tests, Immunologic; D005434:Flow Cytometry; D053804:Granzymes / Q000494:pharmacology; D024243:HMGB1 Protein / Q000378:metabolism*; D006801:Humans; D015979:Killer Cells, Lymphokine-Activated / Q000502:physiology; D008545:Melanoma / Q000378:metabolism*; D013601:T-Lymphocytes / Q000502:physiology*; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000494:pharmacology
D000590:Amino Acid Chloromethyl Ketones; D024243:HMGB1 Protein; D053221:TNF-Related Apoptosis-Inducing Ligand; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D053804:Granzymes