Structural basis for the proinflammatory cytokine activity of high mobility group box 1.
High mobility group box 1 (HMGB), a ubiquitous DNA-binding protein, has been implicated as a proinflammatory cytokine and late mediator of lethal endotoxemia. HMGB1 is released by activated macrophages. It amplifies and extends the inflammatory response by inducing cytokine release and mediating acute lung injury, anorexia, and the inflammatory response to tissue necrosis. The kinetics of HMGB1 release provide a wide therapeutic window for endotoxemia because extracellular levels of HMGB1 begin to increase 12 to 24 h after exposure to inflammatory stimuli. Here, we demonstrate that a DNA-binding domain of HMGB1, the B box, recapitulates the cytokine activity of full length HMGB1 and efficiently activates macrophages to release tumor necrosis factor (TNF) and other proinflammatory cytokines. Truncation of the B box revealed that the TNF-stimulating activity localizes to 20 amino acids (HMGB1 amino acids 89 to 108). Passive immunization of mice with antibodies raised against B box conferred significant protection against lethal endotoxemia or sepsis, induced by cecal perforation. These results indicate that a proinflammatory domain of HMGB1 maps to the highly conserved DNA-binding B box, making this primary sequence a suitable target in the design of therapeutics.
Li
Jianhua
J
Laboratory of Biomedical Science, North Shore-Long Island Jewish Research Institute, 350 Community Drive, Manhasset, NY 11030, USA.
Kokkola
Riikka
R
Tabibzadeh
Siamak
S
Yang
Runkuan
R
Ochani
Mahendar
M
Qiang
Xiaoling
X
Harris
Helena E
HE
Czura
Christopher J
CJ
Wang
Haichao
H
Ulloa
Luis
L
Wang
Hong
H
Warren
H Shaw
HS
Moldawer
Lyle L
LL
Fink
Mitchell P
MP
Andersson
Ulf
U
Tracey
Kevin J
KJ
Yang
Huan
H
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D024202:Electrophoretic Mobility Shift Assay; D019446:Endotoxemia / Q000517:prevention & control; D005982:Glutathione Transferase / Q000378:metabolism; D024243:HMGB1 Protein / Q000737:chemistry* / Q000502:physiology*; D007116:Immunization, Passive; D008070:Lipopolysaccharides / Q000494:pharmacology; D008262:Macrophage Activation; D008264:Macrophages / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011817:Rabbits; D018805:Sepsis / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D016207:Cytokines; D017931:DNA Primers; D024243:HMGB1 Protein; D008070:Lipopolysaccharides; D010446:Peptide Fragments; D014409:Tumor Necrosis Factor-alpha; D005982:Glutathione Transferase