Antigenic regions of tumor necrosis factor alpha and their topographic relationships with structural/functional domains

Angelo Corti, Giorgio Fassina, Fabrizio Marcucci, Giovanni Cassani

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Abstract

The immunogenic regions of human Tumor Necrosis Factor alpha (huTNF) have been mapped by studying the interaction between various mouse anti-huTNF sera and synthetic huTNF fragments, spanning the entire sequence of huTNF. Three main immunogenic regions were identified within residues 1-23, 95-116 and 137-157 of huTNF and two other less immunogenic regions within residues 117-136 and 37-55. The same huTNF regions were found to contain antigenic sites by binding studies with cognate anti-peptide sera. Competitive binding experiments with shorter synthetic subfragments provided evidence for the location of strong antigenic sites within residues 1-10, 17-23, 104-112 and 137-143. In particular the immunodominant site was found to be located within residues 104-112. huTNF regions corresponding to residues 24-36, 56-75, 76-94, and 147-157 resulted to be not or poorly antigenic. However, treatment of huTNF with Triton X-100 under conditions that partially dissociate the oligomeric quaternary structure resulted in the exposition of sites recognized by sera against peptides huTNF [56-75] and huTNF [76-94], suggesting that antigenic sites not accessible in the oligomeric huTNF are exposed in the dissociated form. The principal antigenic sites in the oligomeric molecule are localized in the flexible N-terminal part and in hydrophilic segments located in the "middle/top" region of the molecule, opposite to the C-terminus. Protein segments of the "bottom" region, close to the C-terminus, were poorly immunoreactive. Neutralization assays of TNF cytolytic activity on L-M cells showed that binding of antibodies to epitopes located in the "middle/top" regions of huTNF does not affect its cytolytic activity, supporting the hypothesis of a receptor binding site location at the "bottom" of TNF trimer.

Original languageEnglish
Pages (from-to)471-479
Number of pages9
JournalMolecular Immunology
Volume29
Issue number4
DOIs
Publication statusPublished - 1992

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Tumor Necrosis Factor-alpha
human TNF protein
Serum
Binding Sites
Immunodominant Epitopes
Peptides
Competitive Binding
Octoxynol
Epitopes
Antibodies

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

Cite this

Antigenic regions of tumor necrosis factor alpha and their topographic relationships with structural/functional domains. / Corti, Angelo; Fassina, Giorgio; Marcucci, Fabrizio; Cassani, Giovanni.

In: Molecular Immunology, Vol. 29, No. 4, 1992, p. 471-479.

Research output: Contribution to journalArticle

Corti, Angelo ; Fassina, Giorgio ; Marcucci, Fabrizio ; Cassani, Giovanni. / Antigenic regions of tumor necrosis factor alpha and their topographic relationships with structural/functional domains. In: Molecular Immunology. 1992 ; Vol. 29, No. 4. pp. 471-479.
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