Suppressive oligodeoxynucleotides inhibit CpG-induced inflammation of the mouse lung.
OBJECTIVE
To examine the effect of suppressive oligodeoxynucleotides (ODNs) on the pulmonary inflammation induced by immunostimulatory CpG DNA.


METHODS
Prospective, randomized, controlled study.


METHODS
Research laboratories.


METHODS
RAW 264.7 murine macrophage-like cell line and BALB/c mice.


METHODS
RAW 264.7 cells were incubated with bacterial DNA or CpG ODN, alone or combined with suppressive ODN. The in vivo effect of suppressive ODN was determined using an acute lung injury model. CpG ODN alone or combined with suppressive ODN was instilled into the mouse lung.


RESULTS
Production of tumor necrosis factor (TNF)-alpha and macrophage inflammatory protein (MIP)-2 by RAW 264.7 cells were measured by enzyme-linked immunosorbent assay (ELISA), whereas their messenger RNA levels were determined by reverse transcriptase-polymerase chain reaction. Synthetic ODN containing CpG motifs (CpG ODN) mimicked the ability of bacterial DNA to stimulate the production of TNF-alpha and MIP-2. Suppressive ODN significantly inhibited the activation of RAW 264.7 cells by both bacterial DNA and CpG ODN. In the lung injury model, production of proinflammatory cytokines (TNF-alpha and IL-6) and chemokines (MIP-2 and KC) in bronchoalveolar lavage (BAL) fluids was measured by ELISA. Neutrophil accumulation in the alveolar spaces was also evaluated. Instillation of CpG ODN into the lungs of normal mice triggered the synthesis of TNF-alpha, IL-6, MIP-2, and KC. Suppressive ODN significantly blocked the production of these proinflammatory cytokines and chemokines and also reduced neutrophil mobilization into the alveolar spaces by CpG DNA.


CONCLUSIONS
Proinflammatory cytokines and chemokines are up-regulated by CpG motifs in bacterial DNA. Suppressive ODN significantly inhibits the inflammatory response induced by CpG DNA in murine macrophages and the lung. This study supports the use of suppressive ODN to reduce the deleterious inflammatory responses induced by bacterial DNA.
Department of Anesthesiology, Yokohama City University, School of Medicine, Japan. hyamada@bb.mbn.or.jp
Hiroshi
H
Yamada
Ken J
KJ
Ishii
Dennis M
DM
Klinman
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000276:immunology; D002460:Cell Line; D054426:Chemokine CXCL2; D002634:Chemotaxis, Leukocyte / Q000187:drug effects / Q000276:immunology; D018899:CpG Islands / Q000187:drug effects / Q000276:immunology*; D004269:DNA, Bacterial / Q000276:immunology* / Q000494:pharmacology; D008168:Lung / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D023421:Models, Animal; D015846:Monokines / Q000096:biosynthesis; D009504:Neutrophils; D009838:Oligodeoxyribonucleotides / Q000276:immunology / Q000494:pharmacology*; D011014:Pneumonia / Q000276:immunology*; D011446:Prospective Studies; D012128:Respiratory Distress Syndrome / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D054426:Chemokine CXCL2; D004269:DNA, Bacterial; D015846:Monokines; D009838:Oligodeoxyribonucleotides; D014409:Tumor Necrosis Factor-alpha