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Alpha-Defensin DEFA1A3 Gene Copy Number Elevation in Danish Crohn’s Disease Patients

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Abstract

Background and Purpose of Study

Extensive copy number variation is observed for the DEFA1A3 gene encoding alpha-defensins 1–3. The objective of this study was to determine the involvement of alpha-defensins in colonic tissue from Crohn’s disease (CD) patients and the possible genetic association of DEFA1A3 with CD.

Methods

Two-hundred and forty ethnic Danish CD patients were included in the study. Reverse transcriptase PCR assays determined DEFA1A3 expression in colonic tissue from a subset of patients. Immunohistochemical analysis identified alpha-defensin peptides in colonic tissue. Copy number of DEFA1A3 and individual alleles, DEFA1 and DEFA3, were compared with those for controls, by use of combined real-time quantitative PCR and pyrosequencing, and correlated with disease location.

Results

Inflammatory-dependent mRNA expression of DEFA1A3 (P < 0.001), and the presence of alpha-defensin peptides, were observed in colonic tissue samples. Higher DEFA1A3 gene copy number (CD: mean copy number, 7.2 vs. controls 6.7; P < 0.001) and individual DEFA1 alleles (CD mean copy number 5.6 vs. controls 5.1; P < 0.01) were associated with CD, with strong association with colonic location (P < 0.001).

Conclusions

Alpha-defensins are involved in the inflammation of CD, with local mRNA and peptide expression. In combination with the findings that a high DEFA1A3 copy number is significantly linked to CD, these results suggest that a high DEFA1A3 copy number might be important in hindering the normal inflammatory response in CD, particularly colonic CD.

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Acknowledgments

We thank Britta Hansen and Ida Haugaard Bøtcher for expert technical assistance. We would also like to thank Dr John Armour (University of Nottingham, UK) for kindly providing us with data for DEFA1A3 controls, and Nathaniel and Anne Hong for carefully revising the manuscript. This study was supported by the Danish Colitis and Crohn Association, the Augustinus Foundation, Aase and Ejner Danielsens Foundation, the Lundbeck Foundation, the LEO-Pharma Foundation, The Idella Foundation, and “Fonden til Lægevidenskabens Fremme.”.

Conflict of interest

None.

Ethical considerations

The study was conducted according to the Declaration of Helsinki and was approved by the Regional Scientific Ethics Committee. All patients included to the study had given their written consent.

Author information

Authors and Affiliations

  1. Department of Clinical Biochemistry and Immunology, Statens Serum Institut, Copenhagen, Denmark

    Cathrine Jespersgaard & Marianne Dybdahl

  2. Laboratory for Microbial Pathogenesis and Host Susceptibility, Department for Microbiological Surveillance and Research, Statens Serum Institut, Copenhagen, Denmark

    Peder Fode

  3. Department of Gastroenterology, Medical Section, Herlev Hospital, Herlev, Denmark

    Ida Vind, Ole Haagen Nielsen, Claudio Csillag, Pia Munkholm, Lene Riis, Margarita Elkjaer & Natalia Pedersen

  4. Laboratory for Microbial Pathogenesis and Host Susceptibility, Department for Microbiological Surveillance and Research, Statens Serum Institut, 5 Artillerivej, 2300, Copenhagen S, Denmark

    Paal Skytt Andersen

  5. Department of Pathology, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark

    Ben Vainer

  6. Department of Gastroenterology, Glostrup University Hospital, Glostrup, Denmark

    Elisabeth Knudsen

Authors
  1. Cathrine Jespersgaard
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  2. Peder Fode
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Corresponding author

Correspondence to Paal Skytt Andersen.

Additional information

Cathrine Jespersgaard, Peder Fode, and Marianne Dybdahl contributed equally to the manuscript.

Electronic supplementary material

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10620_2011_1794_MOESM1_ESM.doc

Reproducibility of the assay. The graph shows the four controls used in each run plotting the expected copy numbers along the X axis and the observed copy numbers along the Y-axis. The plot illustrates 8 runs. For each run linear regression was performed and the equation for the line was calculated. The figure shows the reproducibility of the assay indicating the standard deviation (SD): TT0296 (Expected CN = 9) SD = 0.14; AF0105 (Expected CN = 7) SD = 0.17; AF0111 (Expected CN = 8) SD = 0.25; AF0103 (Expected CN = 6) SD = 0.08 (DOC 211 kb)

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Jespersgaard, C., Fode, P., Dybdahl, M. et al. Alpha-Defensin DEFA1A3 Gene Copy Number Elevation in Danish Crohn’s Disease Patients. Dig Dis Sci 56, 3517–3524 (2011). https://doi.org/10.1007/s10620-011-1794-8

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  • DOI: https://doi.org/10.1007/s10620-011-1794-8

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