Hypomethylation of perforin regulatory elements in CD4+ T cells from rat spleens contributes to the development of autoimmune emphysema

Respirology. 2014 Apr;19(3):376-81. doi: 10.1111/resp.12240. Epub 2014 Feb 10.

Abstract

Background and objective: It is widely accepted that perforin regulatory elements are hypomethylated in CD4+ T cells from patients with active lupus, but whether this is the case in autoimmune emphysema is not known.

Methods: Twenty rats were randomly divided into a normal control group and an emphysema group. Rat models of emphysema were established by intraperitoneal injection with xenogeneic endothelial cells. The levels of tumour necrosis factor-α, interleukin-8 and matrix metalloproteinase (MMP)-9 in bronchoalveolar lavage fluid (BALF) were measured, lung mean linear intercept and destructive index measured. Mean methylation of perforin gene promoter in CD4+ T cells and the expression of perforin were investigated. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling methods were used to examine the percentage of apoptotic cells in the alveolar septa.

Results: The levels of MMP-9 in BALF were higher in emphysema group than in control group (P < 0.05). The mean linear intercept and destructive index were higher in emphysema group than in control group (P < 0.05). The mean perforin gene promotor methylation of emphysema group was significantly decreased as compared with control group, while the expression levels of perforin gene were relatively higher (P < 0.05). There were more terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling-positive cells in the alveolar septa in control group than in emphysema group.

Conclusions: Hypomethylation of perforin regulatory elements in CD4+ T cells may result in the lung septal cell apoptosis associated with the development of experimental autoimmune emphysema. MMP-9 may play an important role in the pathogenesis of this kind of disease.

Keywords: CD4+ T cell; autoimmune emphysema; hypomethylation; perforin regulatory element; rat.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autoimmune Diseases / genetics*
  • Autoimmune Diseases / metabolism
  • Bronchoalveolar Lavage Fluid
  • CD4-Positive T-Lymphocytes / metabolism*
  • Cell Culture Techniques
  • DNA Methylation*
  • Disease Models, Animal
  • In Situ Nick-End Labeling
  • Interleukin-8 / metabolism
  • Male
  • Matrix Metalloproteinase 9 / metabolism
  • Pore Forming Cytotoxic Proteins / genetics*
  • Pore Forming Cytotoxic Proteins / metabolism
  • Pulmonary Emphysema / genetics*
  • Pulmonary Emphysema / metabolism
  • Pulmonary Emphysema / pathology
  • Rats
  • Rats, Sprague-Dawley
  • Real-Time Polymerase Chain Reaction
  • Regulatory Elements, Transcriptional / genetics*
  • Sequence Analysis, DNA
  • Spleen / cytology*
  • Spleen / metabolism
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Interleukin-8
  • Pore Forming Cytotoxic Proteins
  • Tumor Necrosis Factor-alpha
  • perforin, rat
  • Matrix Metalloproteinase 9