The Effects Of Iron Overload On The Regulation Of Lymphocytes In Patients And Mice

Objective : To investigated the effects of iron overload on lymphocytes in patients and mice.Methods:Peripheral blood from patients was harvested to test the effect on the quantity, differentiation of T cells and the reactive oxygen species(ROS) level of lymphocytes was also evaluated. Male C57Bl/6 mice were intraperitoneally injected with iron dextran at a dose of 25 mg/ml every three days for four weeks and the blood of each mouse was collected for analysis. The changes of the peripheral immune system were investigated at four weeks. The chronic effects on quantity,differentiation, proliferation and apoptosis of T cells were evaluated at 4 weeks. The ratio of CD8+T cells and the cytokines of IFN-γ、TNF-α、Granzyme-B、perforin of CD8+T cells were tested by flow cytometer. The CD8+T cells apoptosis were determined by Annexin V/PI double staining. Real-time PCR assay was performed to evaluate the expression of IFN-γ、TNF-α、Granzyme-B、perforin、bcl-2、bax at mRNA level in CD8+T cells.Results:1. Iron overload could increase the level of LIP of lymphocytes and the level of ROS CD3 T cells.2. Iron overload could decreased the ratio of CD3 T cells, increased the ratio of CD4/CD8, decreased the ratio of Th1/Th2 and Tc1/Tc2.3. Iron overload occurred by spleen iron staining and flow cytometer analysis.4. The level of intracellular ROS of PBMCs and CD8 T cells from spleen with iron overload groups was higher than that of the control groups.5. Compared with the control groups, the ratio and the counts of lymphocytes in mice with iron overload was significantly decreased. Compared with the control groups, the ratio of neutrophil cells in mice with iron overload was significantly decreased, but there were no difference of the counts of neutrophil cells between four groups.6. The ratio of CD3 T cells in mice with iron overload was significantly decreased, CD4/CD8 was significantly increased.7. The ratio of Treg cells in mice with iron overload was significantly increased,Th1/Th2 and Tc1/Tc2 was significantly decreased8. CD3+CD8+T and CD3+CD8-T cells apoptosis in iron overload group was significantly higher than that in control.9. Compared with the control groups, the ratio and the counts of lymphocytes in mice with iron overload was significantly decreased(P<0.05).10. The ratio of CD3+T cells in mice with iron overload was significantly decreased and the ratio of CD4/CD8 was significantly increased(P<0.05).11. The ratio of Treg cells in mice with iron overload was significantly increased,Th1/Th2 and Tc1/Tc2 was significantly decreased(P<0.05).12. CD3+CD8+T and CD3+CD8-T cells apoptosis in iron overload group was significantly higher than that in control(P <0.05).13. These effects could be reversed after treating iron-overloaded mice with DFX or NAC.