The effect of tumour necrosis factor-α (TNF-α) muteins on human neutrophils in vitro
Tumour necrosis factor-α (TNF-α) has been implicated as an important inflammatory mediator. In vitro, TNF-α is reported to activate human polymorphonuclear neutrophils (PMN), inducing responses such as phagocytic activity, degranulation and oxidative metabolism. Biological responses to TNF-α are ini...
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| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
1993-01-01
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| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/S0962935193000055 |
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| author | H. Tchorzewski K. Zeman J. Kantorski E. Paleolog M. Kahan M. Feldmann M. Kwinkowski P. Guga B. Szymanska P. Parniewski A. Wilk J. Jarosz |
| author_facet | H. Tchorzewski K. Zeman J. Kantorski E. Paleolog M. Kahan M. Feldmann M. Kwinkowski P. Guga B. Szymanska P. Parniewski A. Wilk J. Jarosz |
| author_sort | H. Tchorzewski |
| collection | DOAJ |
| description | Tumour necrosis factor-α (TNF-α) has been implicated as an important inflammatory mediator. In vitro, TNF-α is reported to activate human polymorphonuclear neutrophils (PMN), inducing responses such as phagocytic activity, degranulation and oxidative metabolism. Biological responses to TNF-α are initiated by its binding to specific cell surface receptors, and various studies have shown that the major TNF receptor species on PMN is the 75 kDa receptor. To verify the suggestion that the receptor binding domain includes the region close to the N-terminus of the TNF-α molecule, four TNF-α derivatives termed muteins were constructed, using a synthetic cDNA fragment substituting the N-terminal 3–7 selected hydrophilic or hydrophobic amino acids in the original TNF-α genomic DNA. Binding of muteins to PMN was assessed using monoclonal antibodies recognizing either the 55 kDa (p55) or the 75 kDa (p75) TNF receptor subtypes. Blocking by muteins of anti-p75 antibody binding to PMN was as expected from their N-terminal amino acid composition and hydrophilic properties. Hydrophilic muteins competed well with anti-TNF receptor antibodies for binding to the p75 receptor. In contrast, hydrophobic muteins were unable to block anti-p75 binding. Similarly, degranulation, chemiluminescence or enhancement of the PMN response to specific stimuli by the muteins correlated with the hydrophilic properties of the muteins. The significance of these observations in relation to the molecular structure of TNF-α is discussed. |
| format | Article |
| id | doaj-art-29485d65bd9f48d69b286d1afe214df6 |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 1993-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-29485d65bd9f48d69b286d1afe214df62025-02-03T05:47:02ZengWileyMediators of Inflammation0962-93511466-18611993-01-0121414810.1155/S0962935193000055The effect of tumour necrosis factor-α (TNF-α) muteins on human neutrophils in vitroH. Tchorzewski0K. Zeman1J. Kantorski2E. Paleolog3M. Kahan4M. Feldmann5M. Kwinkowski6P. Guga7B. Szymanska8P. Parniewski9A. Wilk10J. Jarosz11Department of Pathophysiology and Immunology, Institute of Basic Medical Sciences, Military Medical Academy, Lodz, PolandDepartment of Pathophysiology and Immunology, Institute of Basic Medical Sciences, Military Medical Academy, Lodz, PolandDepartment of Pathophysiology and Immunology, Institute of Basic Medical Sciences, Military Medical Academy, Lodz, PolandKennedy Institute of Rheumatology, Sunley Division, London, UKKennedy Institute of Rheumatology, Sunley Division, London, UKKennedy Institute of Rheumatology, Sunley Division, London, UK Department of Bioorganic Chemistry, Polish Academy of Sciences, Lodz, Poland Department of Bioorganic Chemistry, Polish Academy of Sciences, Lodz, Poland Department of Bioorganic Chemistry, Polish Academy of Sciences, Lodz, Poland Department of Bioorganic Chemistry, Polish Academy of Sciences, Lodz, Poland Department of Bioorganic Chemistry, Polish Academy of Sciences, Lodz, Poland Department of Bioorganic Chemistry, Polish Academy of Sciences, Lodz, PolandTumour necrosis factor-α (TNF-α) has been implicated as an important inflammatory mediator. In vitro, TNF-α is reported to activate human polymorphonuclear neutrophils (PMN), inducing responses such as phagocytic activity, degranulation and oxidative metabolism. Biological responses to TNF-α are initiated by its binding to specific cell surface receptors, and various studies have shown that the major TNF receptor species on PMN is the 75 kDa receptor. To verify the suggestion that the receptor binding domain includes the region close to the N-terminus of the TNF-α molecule, four TNF-α derivatives termed muteins were constructed, using a synthetic cDNA fragment substituting the N-terminal 3–7 selected hydrophilic or hydrophobic amino acids in the original TNF-α genomic DNA. Binding of muteins to PMN was assessed using monoclonal antibodies recognizing either the 55 kDa (p55) or the 75 kDa (p75) TNF receptor subtypes. Blocking by muteins of anti-p75 antibody binding to PMN was as expected from their N-terminal amino acid composition and hydrophilic properties. Hydrophilic muteins competed well with anti-TNF receptor antibodies for binding to the p75 receptor. In contrast, hydrophobic muteins were unable to block anti-p75 binding. Similarly, degranulation, chemiluminescence or enhancement of the PMN response to specific stimuli by the muteins correlated with the hydrophilic properties of the muteins. The significance of these observations in relation to the molecular structure of TNF-α is discussed.http://dx.doi.org/10.1155/S0962935193000055 |
| spellingShingle | H. Tchorzewski K. Zeman J. Kantorski E. Paleolog M. Kahan M. Feldmann M. Kwinkowski P. Guga B. Szymanska P. Parniewski A. Wilk J. Jarosz The effect of tumour necrosis factor-α (TNF-α) muteins on human neutrophils in vitro Mediators of Inflammation |
| title | The effect of tumour necrosis factor-α (TNF-α) muteins on human neutrophils in vitro |
| title_full | The effect of tumour necrosis factor-α (TNF-α) muteins on human neutrophils in vitro |
| title_fullStr | The effect of tumour necrosis factor-α (TNF-α) muteins on human neutrophils in vitro |
| title_full_unstemmed | The effect of tumour necrosis factor-α (TNF-α) muteins on human neutrophils in vitro |
| title_short | The effect of tumour necrosis factor-α (TNF-α) muteins on human neutrophils in vitro |
| title_sort | effect of tumour necrosis factor α tnf α muteins on human neutrophils in vitro |
| url | http://dx.doi.org/10.1155/S0962935193000055 |
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