Tumor necrosis factor up-regulates γ-interferon binding in a human carcinoma cell line

Arthur B. Raitano, Murray Korc

Research output: Contribution to journalArticle

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Abstract

WiDR colorectal carcinoma cells are highly sensitive to the synergistic cytotoxic effects of tumor necrosis factor (TNF) and γ-interferon (IFN-γ). In the present study, we have investigated the effects of recombinant human (rh) TNF and IFN-γ on the binding of both ligands in this cell line. WiDR cells exhibited high affinity binding sites for both 125I-rhTNF (Kd = 1.66 × 10-10 M, 920 sites/cell) and 125I-rhIFN-γ (Kd = 4.15 × 10-10 M, 18,960 sites/cell). Preincubation of the cells with rhTNF (24 h) increased cell-associated 125I-rhIFN-γ radioactivity by 129% when binding was carried out at 37°C, as a result of an increase in both surface bound and internalized 125I-rhIFN-γ. However, rhTNF did not alter the degradation profile of released 125I-rhIFN-γ radioactivity. Scatchard analysis of 125I-rhIFN-γ binding data (4°C) revealed that rhTNF induced a 245% increase in 125I-rhIFN-γ binding sites. Conversely, rhIFN-γ caused a 68% increase in 125I-rhTNF binding sites and a 58% increase in receptor affinity. rhIFN-γ also increased the subsequent binding of 125I-rhIFN-γ, whereas rhTNF increased the subsequent binding of 125I-rhTNF. Furthermore, preincubation of the cells with both rhTNF and rhIFN-γ also resulted in an increase in the binding of both ligands. Actinomycin D and cycloheximide blocked all the effects of rhTNF and rhIFN-γ on ligand binding. However, the basal level of 125I-rhIFN-γ binding was insensitive to either inhibitor, whereas the basal level of 125I-rhTNF binding was decreased by both inhibitors. These data indicate that in some cell types TNF and IFN-γ may induce an increase in their own receptors (homologous up-regulation) and concomitantly increase each other's receptors (heterologous up-regulation) and that these actions are due, in part, to enhanced receptor synthesis.

Original languageEnglish (US)
Pages (from-to)10466-10472
Number of pages7
JournalJournal of Biological Chemistry
Volume265
Issue number18
StatePublished - Jun 25 1990
Externally publishedYes

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Interferons
Up-Regulation
Tumor Necrosis Factor-alpha
Binding Sites
Cells
Radioactivity
Ligands
Carcinoma
Cell Line
Dactinomycin
Cycloheximide
Degradation
Colorectal Neoplasms
human TNF protein

ASJC Scopus subject areas

  • Biochemistry

Cite this

Tumor necrosis factor up-regulates γ-interferon binding in a human carcinoma cell line. / Raitano, Arthur B.; Korc, Murray.

In: Journal of Biological Chemistry, Vol. 265, No. 18, 25.06.1990, p. 10466-10472.

Research output: Contribution to journalArticle

Raitano, Arthur B. ; Korc, Murray. / Tumor necrosis factor up-regulates γ-interferon binding in a human carcinoma cell line. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 18. pp. 10466-10472.
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abstract = "WiDR colorectal carcinoma cells are highly sensitive to the synergistic cytotoxic effects of tumor necrosis factor (TNF) and γ-interferon (IFN-γ). In the present study, we have investigated the effects of recombinant human (rh) TNF and IFN-γ on the binding of both ligands in this cell line. WiDR cells exhibited high affinity binding sites for both 125I-rhTNF (Kd = 1.66 × 10-10 M, 920 sites/cell) and 125I-rhIFN-γ (Kd = 4.15 × 10-10 M, 18,960 sites/cell). Preincubation of the cells with rhTNF (24 h) increased cell-associated 125I-rhIFN-γ radioactivity by 129{\%} when binding was carried out at 37°C, as a result of an increase in both surface bound and internalized 125I-rhIFN-γ. However, rhTNF did not alter the degradation profile of released 125I-rhIFN-γ radioactivity. Scatchard analysis of 125I-rhIFN-γ binding data (4°C) revealed that rhTNF induced a 245{\%} increase in 125I-rhIFN-γ binding sites. Conversely, rhIFN-γ caused a 68{\%} increase in 125I-rhTNF binding sites and a 58{\%} increase in receptor affinity. rhIFN-γ also increased the subsequent binding of 125I-rhIFN-γ, whereas rhTNF increased the subsequent binding of 125I-rhTNF. Furthermore, preincubation of the cells with both rhTNF and rhIFN-γ also resulted in an increase in the binding of both ligands. Actinomycin D and cycloheximide blocked all the effects of rhTNF and rhIFN-γ on ligand binding. However, the basal level of 125I-rhIFN-γ binding was insensitive to either inhibitor, whereas the basal level of 125I-rhTNF binding was decreased by both inhibitors. These data indicate that in some cell types TNF and IFN-γ may induce an increase in their own receptors (homologous up-regulation) and concomitantly increase each other's receptors (heterologous up-regulation) and that these actions are due, in part, to enhanced receptor synthesis.",
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N2 - WiDR colorectal carcinoma cells are highly sensitive to the synergistic cytotoxic effects of tumor necrosis factor (TNF) and γ-interferon (IFN-γ). In the present study, we have investigated the effects of recombinant human (rh) TNF and IFN-γ on the binding of both ligands in this cell line. WiDR cells exhibited high affinity binding sites for both 125I-rhTNF (Kd = 1.66 × 10-10 M, 920 sites/cell) and 125I-rhIFN-γ (Kd = 4.15 × 10-10 M, 18,960 sites/cell). Preincubation of the cells with rhTNF (24 h) increased cell-associated 125I-rhIFN-γ radioactivity by 129% when binding was carried out at 37°C, as a result of an increase in both surface bound and internalized 125I-rhIFN-γ. However, rhTNF did not alter the degradation profile of released 125I-rhIFN-γ radioactivity. Scatchard analysis of 125I-rhIFN-γ binding data (4°C) revealed that rhTNF induced a 245% increase in 125I-rhIFN-γ binding sites. Conversely, rhIFN-γ caused a 68% increase in 125I-rhTNF binding sites and a 58% increase in receptor affinity. rhIFN-γ also increased the subsequent binding of 125I-rhIFN-γ, whereas rhTNF increased the subsequent binding of 125I-rhTNF. Furthermore, preincubation of the cells with both rhTNF and rhIFN-γ also resulted in an increase in the binding of both ligands. Actinomycin D and cycloheximide blocked all the effects of rhTNF and rhIFN-γ on ligand binding. However, the basal level of 125I-rhIFN-γ binding was insensitive to either inhibitor, whereas the basal level of 125I-rhTNF binding was decreased by both inhibitors. These data indicate that in some cell types TNF and IFN-γ may induce an increase in their own receptors (homologous up-regulation) and concomitantly increase each other's receptors (heterologous up-regulation) and that these actions are due, in part, to enhanced receptor synthesis.

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