Since a dysregulated synthesis of tumor necrosis factor alpha (TNF-alpha) may be involved in the pathogenesis of autoimmune diseases, it was of interest to precisely locate the recently reported NcoI restriction fragment length polymorphism (RFLP) of the TNF-alpha region. However, by mapping of 56.8 kb of overlapping cosmid clones and direct sequencing, we could localize the polymorphic NcoI restriction site within the first intron of the TNF-beta gene and not in the TNF-alpha gene. To study whether regulatory mechanisms are affected by this polymorphism, we analyzed the TNF-alpha/TNF-beta production of phytohemagglutinin-stimulated peripheral blood mononuclear cells of individuals homozygous for the TNF-beta NcoI RFLP by ELISA and concomitant Northern blot analysis. On days 2-4 after stimulation with mitogen, the TNFB*1 allele corresponding to a 5.3-kb NcoI fragment presented with a significantly higher TNF-beta response. A mRNA analysis demonstrated that higher protein levels of TNF-beta correlate also with increased amounts of TNF-beta transcripts. No allelic association was found in respect to TNF-alpha production. To further investigate a possible allelic influence on transcription, we determined the DNA sequence of 2 kb of the 5' portion of our cloned TNFB*2 allele and compared it with the available TNF-beta sequences. By computer-aided recognition motif search of DNA binding factors, we report putative binding sites conserved between mouse and man in the 5' flanking region as well as in intron 1 of the TNF-beta gene, found also in other cytokine promoter sequences. In addition, by polymerase chain reaction amplification and sequencing of 740 bp of the 5' part of TNF-beta of individuals typed homozygously for the NcoI RFLP, we could show that amino acid position 26 is conserved as asparagine in the TNFB*1 and as threonine in the TNFB*2 sequence. A previously reported, EcoRI RFLP in the 3' untranslated region of TNF-beta does not segregate with either of the two alleles. Thus, four TNFB alleles can be defined at the DNA level.
Article|
January 01 1991
Polymorphic structure of the tumor necrosis factor (TNF) locus: an NcoI polymorphism in the first intron of the human TNF-beta gene correlates with a variant amino acid in position 26 and a reduced level of TNF-beta production.
G Messer,
G Messer
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
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U Spengler,
U Spengler
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
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M C Jung,
M C Jung
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
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G Honold,
G Honold
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
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K Blömer,
K Blömer
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
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G R Pape,
G R Pape
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
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G Riethmüller,
G Riethmüller
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
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E H Weiss
E H Weiss
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
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G Messer
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
U Spengler
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
M C Jung
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
G Honold
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
K Blömer
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
G R Pape
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
G Riethmüller
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
E H Weiss
Institute of Immunology, Ludwig-Maximilians University, Munich 2, Federal Republic of Germany.
Online ISSN: 1540-9538
Print ISSN: 0022-1007
J Exp Med (1991) 173 (1): 209–219.
Citation
G Messer, U Spengler, M C Jung, G Honold, K Blömer, G R Pape, G Riethmüller, E H Weiss; Polymorphic structure of the tumor necrosis factor (TNF) locus: an NcoI polymorphism in the first intron of the human TNF-beta gene correlates with a variant amino acid in position 26 and a reduced level of TNF-beta production.. J Exp Med 1 January 1991; 173 (1): 209–219. doi: https://doi.org/10.1084/jem.173.1.209
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