Elevated levels of TNF in the joints of adjuvant arthritic rats.
The primary purpose of this study was to determine whether local levels of tumor necrosis factor (TNF) were elevated in chronically inflamed joints in rats with adjuvant-induced arthritis (AA). We also wished to develop methodology for the quantitative measurement of joint TNF, and to examine the effects of known anti-inflammatory agents on joint TNF levels. TNF levels were measured in joints from AA rats taken during the systemic phase (day 20) of arthritic disease. Using the L929 bioassay, joint extracts from AA rats had significantly greater TNF levels (1054 +/- 147 pg/g tissue) than joint extracts from normal rats (110 +/- 42 pg/g tissue). Administration of ibuprofen failed to significantly inhibit TNF levels in the joint at a time point when paw swelling was significantly reduced. The immunomodulating agents, methotrexate, cyclosporin A (CSA) and HWA486 profoundly inhibited both joint TNF levels and paw swelling. The specificity of this assay for TNF was supported by studies with a polyclonal rabbit anti-mouse TNF antibody which neutralized 78-87% of the TNF activity in the joint extracts. Our studies demonstrate a quantitative increase in local TNF expression in adjuvant arthritis and support a role for TNF in chronic inflammation.
Smith-Oliver
T
T
Department of Cellular Biochemistry, Glaxo Inc., Research Triangle Park, North Carolina 27709.
Noel
L S
LS
Stimpson
S S
SS
Yarnall
D P
DP
Connolly
K M
KM
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D001169:Arthritis, Experimental / Q000378:metabolism*; D001681:Biological Assay; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology; D004487:Edema / Q000517:prevention & control; D005620:Freund's Adjuvant; D007052:Ibuprofen / Q000494:pharmacology; D007555:Isoxazoles / Q000494:pharmacology; D007596:Joints / Q000187:drug effects / Q000378:metabolism*; D007739:L Cells; D000077339:Leflunomide; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008727:Methotrexate / Q000494:pharmacology; D051379:Mice; D009169:Mycobacterium tuberculosis; D011817:Rabbits; D051381:Rats; D011917:Rats, Inbred Lew; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000378:metabolism*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D007555:Isoxazoles; D014409:Tumor Necrosis Factor-alpha; D016572:Cyclosporine; D005620:Freund's Adjuvant; D000077339:Leflunomide; D007052:Ibuprofen; D008727:Methotrexate