Summary
The bcl-2 gene product has been shown to regulate apoptotic cell death, and its dysregulation has been shown to induce several abnormalities in the immune system. No data exist regarding bcl-2 expression in autoimmune diseases, such as human insulin-dependent diabetes mellitus (IDDM). We investigated bcl-2 protein expression by testing T lymphocytes from 15 newly-diagnosed (<3 weeks) IDDM patients in comparison to 10 age-matched control subjects. The expression of bcl-2 on CD3+ lymphocyte subsets was investigated after membrane permeabilization by two- or three-colour immunofluorescence. When the percentage and mean fluorescence intensity (MFI) of bcl-2+/CD3+ cells from normal individuals and patients were compared, we found that bcl-2 expression within the CD3+ and CD4+ CD45R0+ T-cell populations was reduced significantly in IDDM patients (46.8±15.4 vs 79.6±11.7; 25.7±3.8 vs 47.15±5.7, respectively; p<0.001). To establish whether low bcl-2 expression in T cells from newly-diagnosed patients reflects their susceptibility to death by an apoptotic process, we also evaluated DNA staining with propidium iodide in CD3+ lymphocyte suspension after a (24–72 h) culture period (spontaneous apoptosis). We found that IDDM patients have higher levels of spontaneous apoptosis (mean±SEM: 24 h=4.6±0.8; 48 h=9.9±1; 72 h=12.8±1.1) than control subjects (24 h=1.8±0.4; 48 h=4.6±0.4; 72 h=5.7±0.3; p<0.02–0.001). Our study suggests that recent onset IDDM is characterised by reduced bcl-2 expression, which in turn may be associated with the increased spontaneous apoptosis we observed.
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Abbreviations
- MFI:
-
Mean fluorescence intensity
- mAb:
-
monoclonal antibody
- PMA:
-
phorbol myristate
- PHA:
-
phytoemagglutinin
- Con A:
-
concanavalin-A
- PKC:
-
protein kinase C
- ICA:
-
islet cell autoantibody
- IAA:
-
insulin autoantibody
- PE:
-
phycoerythrin
- PI:
-
propidium iodide
- FSC:
-
forward scatter
- SSC:
-
side scatter
- PBMC:
-
peripheral blood mononuclear cells
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Giordano, C., Stassi, G., Todaro, M. et al. Low bcl-2 expression and increased spontaneous apoptosis in T-lymphocytes from newly-diagnosed IDDM patients. Diabetologia 38, 953–958 (1995). https://doi.org/10.1007/BF00400585
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DOI: https://doi.org/10.1007/BF00400585