Tumor necrosis factor inhibitor: purification, NH2-terminal amino acid sequence and evidence for anti-inflammatory and immunomodulatory activities.
The urine of some febrile patients has been shown to contain a tumor necrosis factor-alpha-inhibiting activity (TNF-alpha INH) when tested in a cytotoxicity assay using the TNF-susceptible cell line L-929. The inhibitor was purified to homogeneity using a simple three-step procedure which included a TNF-alpha affinity column, cation exchange and reverse-phase chromatography. The NH2-terminal amino acid sequence of the inhibitor showed no sequence similarity with proteins in the data bases used. Using gel filtration, it was shown that TNF-alpha and the inhibitor form a stable complex which eluted with a molecular weight of about 75,000. This value corresponds to the sum of the inhibitor (approximately 30,000) and TNF-alpha (approximately 45,000-50,000) molecular weight. The TNF-alpha INH blocked prostaglandin E2 production by dermal fibroblasts in a dose-dependent manner, providing evidence for antiinflammatory activity. TNF-alpha INH also blocked class I antigen expression in a dose-dependent manner as measured using the human Colo 205 tumor cell line. Furthermore, TNF-alpha INH affected TNF-alpha synergism with IFN-gamma-induced HLA-DR antigen expression but had no effect on IFN-gamma activity. The data presented demonstrate that TNF-alpha bioactivity can be regulated at the protein level.
Seckinger
P
P
Division of Immunology and Allergy (Hans Wilsdorf Laboratory), Department of Medicine, Hôpital Cantonal Universitaire, Geneva, Switzerland.
Vey
E
E
Turcatti
G
G
Wingfield
P
P
Dayer
J M
JM
D000276:Adjuvants, Immunologic / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000302:isolation & purification*; D002460:Cell Line; D015232:Dinoprostone / Q000096:biosynthesis; D005347:Fibroblasts / Q000378:metabolism; D006680:HLA Antigens / Q000096:biosynthesis; D006681:HLA-D Antigens / Q000096:biosynthesis; D006801:Humans; D007371:Interferon-gamma / Q000502:physiology; D008969:Molecular Sequence Data; D011506:Proteins / Q000302:isolation & purification* / Q000502:physiology; D012867:Skin / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors*
D000276:Adjuvants, Immunologic; D000894:Anti-Inflammatory Agents, Non-Steroidal; D006680:HLA Antigens; D006681:HLA-D Antigens; D011506:Proteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D015232:Dinoprostone