Digestive Diseases and Sciences

, Volume 56, Issue 12, pp 3517–3524 | Cite as

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

  • Cathrine Jespersgaard
  • Peder Fode
  • Marianne Dybdahl
  • Ida Vind
  • Ole Haagen Nielsen
  • Claudio Csillag
  • Pia Munkholm
  • Ben Vainer
  • Lene Riis
  • Margarita Elkjaer
  • Natalia Pedersen
  • Elisabeth Knudsen
  • Paal Skytt Andersen
Original Article


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.


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.


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).


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.


Crohn’s disease Colonic tissue Copy number variation Defensin Genetic association Real-time PCR 



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


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.

Supplementary material

10620_2011_1794_MOESM1_ESM.doc (211 kb)
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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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Cathrine Jespersgaard
    • 1
  • Peder Fode
    • 2
  • Marianne Dybdahl
    • 1
  • Ida Vind
    • 3
  • Ole Haagen Nielsen
    • 3
  • Claudio Csillag
    • 3
  • Pia Munkholm
    • 3
  • Ben Vainer
    • 5
  • Lene Riis
    • 3
  • Margarita Elkjaer
    • 3
  • Natalia Pedersen
    • 3
  • Elisabeth Knudsen
    • 6
  • Paal Skytt Andersen
    • 4
  1. 1.Department of Clinical Biochemistry and ImmunologyStatens Serum InstitutCopenhagenDenmark
  2. 2.Laboratory for Microbial Pathogenesis and Host Susceptibility, Department for Microbiological Surveillance and ResearchStatens Serum InstitutCopenhagenDenmark
  3. 3.Department of Gastroenterology, Medical SectionHerlev HospitalHerlevDenmark
  4. 4.Laboratory for Microbial Pathogenesis and Host Susceptibility, Department for Microbiological Surveillance and ResearchStatens Serum InstitutCopenhagen SDenmark
  5. 5.Department of Pathology, RigshospitaletUniversity of CopenhagenCopenhagenDenmark
  6. 6.Department of GastroenterologyGlostrup University HospitalGlostrupDenmark

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