Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Analysis of an IL-10 polymorphism in idiopathic pulmonary fibrosis

Abstract

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive fibrotic disorder of the lung parenchyma. We have demonstrated changes in IL-10 protein production by alveolar macrophages (AMs) from patients with IPF, which we hypothesise could be because of an IL-10 gene polymorphism. We have screened the coding sequence and 3′ untranslated region of IL-10 for polymorphisms using single-standard conformational polymorphism analysis. A novel polymorphism was identified resulting in a G to A substitution of +43 nucleotides from the start codon changing glycine to arginine at amino acid 15 of the signal peptide sequence. We have introduced the signal peptide mutation into the IL-10 gene and compared secretion of the mutant and wild-type forms after transient transfection of COS-7 cells. Our studies showed that the signal peptide mutation did not have a significant effect on secretion at 24 h post-transfection (P=0.4529 by Mann-Whitney test). However, by 48 h there are significantly lower levels of mutant IL-10 (P=0.0515). There were no differences in the level of cell-associated IL-10 at either 24 or 48 h (P=0.9296 and 0.4268). We suggest that the mutation could affect the efficiency of protein translocation and signal peptide cleavage resulting in lower levels of IL-10 protein secretion.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Nathan CF . Secretory products of macrophages. J Clin Invest 1987; 79: 319–326.

    Article  CAS  Google Scholar 

  2. Tracey KJ, Cerami A . Tumour necrosis factor: a pleiotropic cytokine and therapeutic target. Annu Rev Med 1994; 45: 491–503.

    Article  CAS  Google Scholar 

  3. Piguet PF, Grau GE, Vassalli P . Subcutaneous perfusion of tumour necrosis factor induces local proliferation of fibroblasts, capillaries and epidermal cells or massive tumour necrosis. Am J Pathol 1990; 136: 103–110.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Wanidworanun C, Strober W . Predominant role of tumour necrosis factor-alpha in human monocyte IL-10 synthesis. J Immunol 1993; 151: 6853–6861.

    CAS  PubMed  Google Scholar 

  5. Armstrong L, Jordan N, Millar A . Interleukin 10 (IL-10) regulation of tumour necrosis factor alpha (TNF-alpha) from human alveolar macrophages and peripheral blood monocytes. Thorax 1996; 51: 143–149.

    Article  CAS  Google Scholar 

  6. Martinez JA, King TE, Brown K et al. Increased expression of the interleukin-10 gene by alveolar macrophages in interstitial lung disease. Am J Physiol 1997; 273: L676–L683.

    Article  CAS  Google Scholar 

  7. Zhang Y, Lee TC, Guillemin B, Yu M, Rom WN . Enhanced IL-10 and tumour necrosis factor-a release and messenger RNA expression in macrophages from idiopathic fibrosis or after asbestos exposure. J Immunol 1993; 150: 4188–4196.

    CAS  PubMed  Google Scholar 

  8. Eskdale J, Gallagher G, Verweij CL, Keijsers V, Westendorp RG, Huizinga TW . Interleukin 10 secretion in relation to IL-10 locus haplotypes. Proc Natl Acad Sci USA 1998; 95: 9465–9470.

    Article  CAS  Google Scholar 

  9. Turner DM, Williams DM, Sankaran D, Lazarus M, Sinnott PJ, Hutchinson IV . An investigation of polymorphism in the interleukin-10 gene promoter. Eur J Immunogenet 1997; 24: 1–8.

    Article  CAS  Google Scholar 

  10. Donger C, Georges JL, Nicaud V et al. New polymorphisms in the interleukin-10 gene-relationships to myocardial infarction. Eur J Clin Invest 2001; 31: 9–14.

    Article  CAS  Google Scholar 

  11. Martoglio B, Dobberstein B . Signal sequences: more than just greasy peptides. Trends Cell Biol 1998; 8: 410–415.

    Article  CAS  Google Scholar 

  12. Awad MR, El-Gamel A, Hasleton P, Turner DM, Sinnott PJ, Hutchinson IV . Genotypic variation in the transforming growth factor-beta 1 gene: association with transforming growth factor-beta 1 production, fibrotic lung disease, and graft fibrosis after lung transplantation. Transplantation 1998; 66: 1014–1020.

    Article  CAS  Google Scholar 

  13. Ito M, Oiso Y, Murase T et al. Possible involvement of inefficient cleavage of preprovassopressin by signal peptidase as a cause for familial central diabetes insipidus. J Clin Invest 1993; 91: 2565–2571.

    Article  CAS  Google Scholar 

  14. Racchi M, Watzke HH, High KA, Lively MO . Human coagulation factor X deficiency caused by a mutant signal peptide that blocks cleavage by signal peptidase but not targeting and translocation to the endoplasmic reticulum. J Biol Chem 1993; 268: 5735–5740.

    CAS  PubMed  Google Scholar 

  15. Zschenker O, Jung N, Rethmeier J et al. Characterization of lysosomal acid lipase mutations in the signal peptide and mature polypeptide region causing Wolman disease. J Lipid Res 2001; 42: 1033–1040.

    CAS  PubMed  Google Scholar 

  16. Karaplis AC, Lim S, Baba H, Arnold A, Kronenberg HM . Inefficient membrane targeting, translocation and proteolytic processing by signal peptidase of a mutant preproparathyroid hormone protein. J Biol Chem 1995; 270: 1629–1635.

    Article  CAS  Google Scholar 

  17. Schwacha MG, Schneider CP, Bland KI, Chaudry IH . Resistance of macrophages to the suppressive effect of interleukin-10 following thermal injury. Am J Physiol Cell Physiol 2001; 281: C1180–C1187.

    Article  CAS  Google Scholar 

  18. Avdiushko R, Hongo D, Lake-Bullock H, Kaplan A, Cohen D . IL-10 receptor dysfunction in macrophages during chronic inflammation. J Leuk Biol 2001; 70: 624–632.

    CAS  Google Scholar 

  19. Demedts M, Costabel U . ATS/ERS international multidisciplinary consensus classification of the idiopathic interstitial pneumonias. Eur Respir J 2002; 19: 794–796.

    Article  CAS  Google Scholar 

  20. Gustincich S, Manfioletti G, Del Sal G, Schneider C, Carninci P . A fast method for high-quality genomic DNA extraction from whole human blood. Biotechniques 1991; 11: 298–300.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to A B Millar.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Whittington, H., Freeburn, R., Godinho, S. et al. Analysis of an IL-10 polymorphism in idiopathic pulmonary fibrosis. Genes Immun 4, 258–264 (2003). https://doi.org/10.1038/sj.gene.6363959

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gene.6363959

Keywords

This article is cited by

Search

Quick links