Monosodium urate, hydroxyapatite, and calcium pyrophosphate crystals induce tumor necrosis factor-alpha expression in a mononuclear cell line.
OBJECTIVE
Tumor necrosis factor-alpha (TNF-alpha) is thought to be important in chronic inflammation of joints characteristic of crystal induced arthritis. Monocytes are instrumental in maintaining that inflammation. We investigated production of mRNA and protein for TNF-alpha in vitro in a murine mononuclear cell line, after exposure to relevant crystal types.


METHODS
Using the cell line designated RAW 264.7, cells were grown in standard medium and exposed to varying amounts of monosodium urate (MSU), hydroxyapatite (HA), and calcium pyrophosphate dihydrate (CPPD) crystals for differing time periods. Analysis of TNF-alpha mRNA induced by such exposure was by Northern hybridization; analysis of TNF-alpha protein was by ELISA:


RESULTS
RNA analyses of cells treated with various levels of MSU, HA, and CPPD crystals showed strong induction of TNF-alpha transcripts. ELISA on culture supernatants confirmed high level TNF-alpha peptide secretion resulting from that transcriptional upregulation. Time course studies showed peak accumulation of TNF-alpha mRNA 1-6 h post-treatment. Study of the signalling pathway involved in TNF-alpha transcriptional upregulation indicated that increased phospholipase A2 activity was required.


CONCLUSIONS
These observations suggest that crystals in joints can directly stimulate production of TNF-alpha, and that the source of that cytokine may be the monocytes known to be present and playing an important role in chronic joint disease.
Meng
Z H
ZH
Arthritis-Immunology Center, Medical Research Service, Philadelphia DVA Medical Center, PA 19104, USA.
Hudson
A P
AP
Schumacher
H R
HR
Baker
J F
JF
Baker
D G
DG
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D002131:Calcium Pyrophosphate / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D003460:Crystallization; D017886:Durapatite / Q000737:chemistry / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D051379:Mice; D009000:Monocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D010003:Osteoarthritis / Q000276:immunology / Q000378:metabolism; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000378:metabolism; D054467:Phospholipases A2; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology; D014158:Transcription, Genetic / Q000187:drug effects / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000378:metabolism; D014527:Uric Acid / Q000737:chemistry / Q000494:pharmacology*
D000098:Acetophenones; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D014527:Uric Acid; D017886:Durapatite; D010741:Phospholipases A; D054467:Phospholipases A2; C006821:4-bromophenacyl bromide; D002131:Calcium Pyrophosphate