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Transfer of Invitro CD4 + T Cells with Hypomethylation of Perforin Promoter into Rats' Abdomens Causes Autoimmune
Jia-Jia Liu1,2, Lin Liu1, Hong-Hong Mu1
1Department of Respiratory Medicine, Guizhou Provincial People's Hospital, Guiyang, Guizhou, China.
Abstract:
Our previous study suggested that hypomethylation of perforin promoter of CD4 + T cells might be involved in the pathogenesis of autoimmune emphysema of rats. Whether transfer of this kind of cells hypomethylated in vitro into naive immunocompetent rats also results in emphysema is unknown yet. To test the hypothesis above, thirty Sprague Dawley (SD) rats were randomly divided into three groups: a model group (n = 10), a normal control group (n = 10) and a sham operation group (n = 10). In the model group, spleen-derived CD4 + T cells of normal rats were treated with 5-azacytidine (5-Aza), complete Freund's adjuvant and Phosphate Buffered Saline (PBS), then transferred into naive immunocompetent rats. The normal control group was injected with CD4 + T lymphocytes from spleens of normal rats and the same amount of adjuvant and PBS as above. In sham operation group, normal rats were injected intraperitoneally with complete Freund's adjuvant and PBS. Histopathological evaluations (mean linear Intercept (MLI) and mean alveolar numbers (MAN)), anti-endothelial cell antibodies (AECA) in serum and bronchoalveolar lavage fluid (BALF), lung vascular endothelial growth factor (VEGF)), the apoptotic index (AI) of alveolar septal cells and the methylation levels of perforin promoter of CD4 + T cells were investigated. The levels of the methylation above and MAN were lower in the model group than in the control and the sham operation group, while the AECA in serum and BALF, VEGF, MLI and the AI were greater (all p < 0.05). The methylation levels of perforin promoter were positively correlated with the MAN (r = 0.747, p < 0.05) and negatively correlated with AI, AECA, MLI, and VEGF (r was -0.789, -0.746, -0.743, -0.660, respectively, all p < 0.05). This study suggests that transfer of invitro CD4 + T cells with hypomethylation of perforin promoter into rats causes autoimmune emphysema, possibly by increasing expression of VEGF and promoting alveolar septal cell apoptosis.
Insights
Transferring CD4+ T cells with a hypomethylated perforin promoter into rats induced autoimmune emphysema. This condition was linked to increased vascular endothelial growth factor (VEGF) and alveolar cell apoptosis.
Area of Science:
- Immunology
- Pulmonology
- Epigenetics
Background:
- Previous research indicated perforin promoter hypomethylation in CD4+ T cells may contribute to autoimmune emphysema in rats.
- The direct effect of transferring *in vitro* hypomethylated CD4+ T cells on emphysema development remained uninvestigated.
Purpose of the Study:
- To determine if transferring *in vitro* hypomethylated CD4+ T cells induces autoimmune emphysema in naive rats.
- To investigate the underlying mechanisms, including anti-endothelial cell antibodies (AECA), vascular endothelial growth factor (VEGF), and apoptosis.
Main Methods:
- Thirty Sprague Dawley rats were divided into model, normal control, and sham groups.
- The model group received spleen-derived CD4+ T cells treated with 5-azacytidine (5-Aza) and transferred *in vitro*.
- Histopathological analysis (MLI, MAN), AECA, VEGF levels, apoptotic index (AI), and perforin promoter methylation were assessed.
Main Results:
- The model group exhibited lower perforin promoter methylation and mean alveolar numbers (MAN) compared to controls (p<0.05).
- Conversely, the model group showed increased serum/BALF AECA, VEGF, MLI, and AI (p<0.05).
- Perforin promoter methylation strongly correlated with MAN, AI, AECA, MLI, and VEGF (p<0.05).
Conclusions:
- Transferring *in vitro* hypomethylated CD4+ T cells can induce autoimmune emphysema in rats.
- This process may involve increased VEGF expression and promotion of alveolar septal cell apoptosis.
- Perforin promoter methylation status in CD4+ T cells is a key factor in autoimmune emphysema pathogenesis.
