role of apoptosis of lymphocytes in pediatric patients with chronic renal failure and end stage renal disease

ABSTRACT

Apoptosis is an essential form of physiological cell suicide in which the cell itself initiates, regulates and executes cell death pathway using a highly structured arsenal of specific proteins. Patients with chronic kidney disease [CKD] have an impairment of the immune response. Apoptosis of lymphocytes can contribute to this immune defect. The aim of this study is to study the effect of apoptosis on lymphocytes of chronic renal failure and end stage renal disease pediatric patients. This study was conducted on 7 pediatric patients with chronic renal failure under conservative treatment, 7 pediatric cases with end stage renal disease [ESRD] under hemodialysis and 8 healthy control subjects. Lymphocytes were isolated by ficoll-Hypaque method. Apoptosis of lymphocytes was assessed by Annexin V FITC assay kit. Percentage of apoptotic lymphocytes [annexin V FITC +ve and propidium iodide -ve] was determined by flow cytometer. Extraction of DNA from lymphocytes was done using DNeasy prep-kit. DNA electrophoresis was done on 2% agarose gel and stained with ethidium bromide. The bands are seen through UV trans-illuminator. Mean positive apoptotic lymphocytes were 35%, 27%, and 4% in end stage renal disease, chronic renal failure patients and control personnel respectively. Apoptosis was significantly higher in end stage renal disease patients in comparison to control personnel [Chi Square and p value, 28. 7 and <0.0001 respectively]. Apoptosis was significantly higher in chronic renal failure patients in comparison to control personnel [Chi Square and p value, 18.5 and <0.0001 respectively]. Apoptosis was more in lymphocytes of end stage renal disease patients than chronic renal failure patients but this difference is not significant [Chi Square and p value, 1.145 and 0.2845 respectively]. By DNA electrophoresis, 7 cases of ESRD and 6 out of 7 CRF cases give the DNA ladder characteristic to apoptosis while no control gives this ladder pattern. This study concludes -that apoptosis significantly affects lymphocytes in chronic renal failure and end stage disease renal disease pediatric patients so it can contribute to immune deficiency seen in these patients