Synthesis and maturation of recombinant human tumor necrosis factor in eukaryotic systems

Müller, R; Marmenout, Anne; Fiers, Walter

doi:10.1016/0014-5793(86)80306-4

Cited by 25 publications

(20 citation statements)

References 20 publications

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“…3). This small increase immediately after UV-B was reproducible and may represent transcription-independent processing of internalized preformed TNFa precursors (18,19). The levels then rose in epidermis and, to a lesser extent, dermis, at 12 h and declined significantly by 24 h (Fig.…”

mentioning

confidence: 72%

Regulation of TNFα production and release in human and mouse keratinocytes and mouse skin after UV‐B irradiation

Yarosh

Both

Kibitel

et al. 2000

Photoderm Photoimm Photomed

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TNFalpha is a primary cytokine responsible for inflammatory and immunosuppressive responses in skin. After UV-B irradiation of cultured human keratinocytes, we found that TNFalpha was released into the media, as monitored by ELISA, and was bound to cells, as observed by immunofluorescence microscopy. The release of TNFalpha into cell culture supernatant during the 24 h after UV-B irradiation was augmented by the addition of IL-1alpha to the cells. Further, we found this secretion was unaffected by rapamycin, and therefore independent of FRAP DNA-protein kinase mediated signal transduction. However, UV-B also induced expression of membrane-bound TNFalpha, and this was dependent on FRAP signaling. In wild type mice, TNFalpha bound to skin increased immediately after irradiation, declined at 6 h, and then rose again at 12 h before falling by 24 h. This pattern of induction was confirmed by RT-PCR of TNFalpha mRNA message in cultured epidermal cells. Induction of membrane-bound TNFalpha was also found in c-fos gene knockout mice deficient in the AP-1 transcription factor, suggesting that, although AP-1 containing c-fos signaling is required for some UV responses, AP-1 containing c-fos is not required for this TNFalpha activation. However, in homozygous p53 knockout mice the basal level of TNFalpha bound to the epidermis was greatly elevated without UV irradiation. This level declined and remained constant following irradiation. This implies that p53 directly or indirectly represses TNFalpha gene expression and that modification of p53 mRNA stability or phosphorylation of p53 protein after UV may be responsible for TNFalpha induction in the membrane. Overexpression of the immunosuppressive cytokine TNFalpha in this locale may contribute to the carcinogen-susceptibility of p53 knockout mice.

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confidence: 72%

Regulation of TNFα production and release in human and mouse keratinocytes and mouse skin after UV‐B irradiation

Yarosh

Both

Kibitel

et al. 2000

Photoderm Photoimm Photomed

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“…TNF-a is a 17 kDa cytokine, consisting of 157 amino acids and exhibiting a variety of immunobiological functions. 11 Its proinflammatory and antitumor activity was first noticed more than 100 y ago, but the protein was only identified in 1975. 12 Its gene was cloned in 1984, 13 and it was mapped to the short arm of chromosome 6, 14 near the HLA-B focus in the MHC.…”

Section: Introductionmentioning

confidence: 99%

TNF-α protects dendritic cells from prostate cancer-induced apoptosis

Pirtskhalaishvili

Shurin

Esche

et al. 2001

Prostate Cancer Prostatic Dis

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We have recently shown that human prostate cancer (PCa) cells induced apoptotic death of the most potent antigen-presenting cells, dendritic cells (DC), which are responsible for the induction of specific antitumor immune responses. Here we have evaluated the effect of murine PCa cells RM-1 on the survival of immature and tumor necrosis factor-a (TNF-a)-stimulated mature DC. PCa cells and DC were co-incubated for 24 -48 h and DC apoptosis was assessed by morphologic criteria, Annexin V assay, and TUNEL staining. We have shown that co-incubation of RM-1 cells with DC is accompanied by an increased level of DC apoptosis, which was mediated by decreased expression of anti-apoptotic protein Bcl-2. Stimulation of DC maturation by TNF-a resulted in increased resistance of DC to PCa-induced apoptosis. In TNF-a treated mature DC, but not in immature DC, the expression of Bcl-2 was not blocked after exposure to RM-1-derived factors. Thus, these data suggest that TNF-a-induced maturation of DC increases their resistance to PCa induced apoptosis. This is likely to be due to the stabilizing of the expression of anti-apoptotic protein Bcl-2. The difference in the sensitivity of mature and immature DC to PCa-induced cell death should be considered during the design of DC-based clinical trials for PCa patients. Prostate Cancer and Prostatic Diseases (2001) 4, 221-227.

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“…It is initially synthesized and expressed as a functional transmembrane prohormone with a type II orientation (12,16). The pathophysiological effects of TNF may be mediated by juxtacrine interactions of the prohormone and/or autocrine-paracrine interactions of the mature, secreted molecule.…”

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confidence: 99%

“…However, deletion of residues Ϫ55 to Ϫ34 resulted in blocked translocation and secretory mechanisms. This result was rationalized on the basis of inadequate hydrophobicity of the remaining transmembrane domain (16,26). However, since the latter mutant also lacked the Arg-49-Arg-48 doublet, a role for cytoplasmic positive charge in effective translocation of human pro-TNF might also exist.…”

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confidence: 99%

Human Pro-Tumor Necrosis Factor: Molecular Determinants of Membrane Translocation, Sorting, and Maturation

Utsumi

Akimaru

Kawabata

et al. 1995

Molecular and Cellular Biology

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Human pro-tumor necrosis factor (pro-TNF) is a type II transmembrane protein with a highly conserved 76-residue leader sequence. We have analyzed the behavior, both in a microsomal translocational system and by transfection, of a series of mutants with deletions from the cytoplasmic, transmembrane, and linking domains. Cytoplasmic deletions included the Arg doublet at ؊49 and ؊48 and/or the Lys doublet at ؊58 and ؊57; additional mutants included deletion of residues ؊73 to ؊55 and ؊73 to ؊55, ؊49, and ؊48. The transmembrane and linking domain mutants included deletions in the ؊42 to ؊35 region, combined with the deletion of residues ؊32 to ؊1. Two hybrid mutants combined the cytoplasmic deletions with the deletion of residues ؊32 to ؊1. All of the cytoplasmic deletion mutants were properly translocated, as were the transmembrane deletion mutants with deletions up to residues ؊36, ؊35, ؊32 to ؊1, although the last one exhibited reduced efficiency; further incremental deletions, including deletions of residues ؊38 to ؊35 and ؊32 to ؊1, completely blocked translocation. Both hybrid mutants were effectively translocated; furthermore, transfection analysis revealed competent expression and maturation of both the cytoplasmic and hybrid mutants. Thus, proper expression and maturation of human pro-TNF can be accomplished with as few as ϳ12 of the 26 residues of the native transmembrane domain and with a net negative charge in the cytoplasmic domain flanking the transmembrane region.

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Synthesis and maturation of recombinant human tumor necrosis factor in eukaryotic systems

Cited by 25 publications

References 20 publications

Regulation of TNFα production and release in human and mouse keratinocytes and mouse skin after UV‐B irradiation

Regulation of TNFα production and release in human and mouse keratinocytes and mouse skin after UV‐B irradiation

TNF-α protects dendritic cells from prostate cancer-induced apoptosis

Human Pro-Tumor Necrosis Factor: Molecular Determinants of Membrane Translocation, Sorting, and Maturation

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