A 63 kDa tumor necrosis factor inhibitor released from a human monocytic leukemia cell line, THP-1.
A human monocytic cell line, THP-1-S, was cultured in a serum-free medium. The effect of the culture supernatant of THP-1-S on the cytotoxicity of rTNF-alpha to three kinds of cell lines and the binding of rTNF to its receptor were tested. The supernatant inhibited the cytotoxicity of rTNF-alpha when tested by the neutral red uptake method. In addition, the supernatant blocked the binding of 125I-rTNF-alpha to its receptor. Furthermore, following precipitation with PEG we detected complexes between rTNF-alpha and the inhibitory factor which formed during incubation with the culture supernatant from THP-1-S cells. However, the supernatant did not bind to or down-regulate the receptor for TNF-alpha on the cell surface of L-M-2d6 cells. This factor eluted with an apparent molecular mass of 63,000 Da by gel filtration and did not react with antibodies against p55 and p75 TNF receptors. These data suggest that human monocytic cells are capable of releasing an inhibitory factor against rTNF-alpha in serum-free culture conditions.
Aso
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Department of Animal Physiology, National Institute of Animal Industry, Tsukuba, Ibaraki, Japan. asosan@niai.affrc.go.jp
Yoshite
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O
Tomioka
Y
Y
Rose
M T
MT
Mizugaki
M
M
D002470:Cell Survival; D002850:Chromatography, Gel; D016895:Culture Media, Serum-Free; D006367:HeLa Cells; D006801:Humans; D007948:Leukemia, Monocytic, Acute / Q000473:pathology; D009000:Monocytes / Q000166:cytology / Q000378:metabolism*; D009499:Neutral Red / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor / Q000096:biosynthesis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000378:metabolism
D016895:Culture Media, Serum-Free; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D009499:Neutral Red