Abstract
Introduction: SLE is a systemic autoimmune disorder. One of the mechanisms by which elimination of autoreactive lymphocytes takes place is apoptosis. Adefect in apoptosis may thus contribute to the development of autoimmune disease. Patients and methods: The study included 34 Russian children patients with SLE in age between 6-17 years old. Lymphocytes were isolated from venous blood by method of gradient centrifugation of all the blood through a Ficoll-pak solution. The quantity apoptotic cells was determined in leukocytes by flow cytometry Epics XL-2 (“Beckman Coulter”, USA). Analysis of lymphocyte subpopulations was carried by using two fluorescent labels: FITC conjugated monoclonal antibodies (CD-25 and CD-20) and phycoerythrin conjugated monoclonal antibody (CD-3, CD-4 and CD-8). Results: After in vitro incubation in CO2 incubator for 36hr; the percentage of apoptotic lymphocytes from patients with SLE was significantly higher than from healthy donors (7.2* ± 4.7 % vs. 5.5 ± 3.2% respectively). This was found to be depending upon the SLE disease activity. Moreover, observed correlations between level apoptosis lymphocyte in SLE and the titer of antibodies to dsDNA (r = 0.5) and dosage prednisolone (r = 0.6). By analyzing lymphocyte phenotypes; there was no significant difference in percentage of CD3+T-cells from SLE compared to normal donors (62.4±7.2 vs. 68.7±9.9.); the same had been observed in CD8+ T-cells (20.9±5.5 vs. 25.3±5.6); and in CD20+ B-cells (9.7±8.9 vs. 13.7±8.0). However, the percentage of CD4+T cell was significantly decreased (P = 0,001) in patients with SLE compared to normal donors (25.5*±7.3 vs. 40.6±8.7). The immunoregulatory index was decreased in SLE compared to normal donors (1.2 vs 1.6 respectively). This phenotypes disbalance was associated with significant increased in the level of activated CD25+ lymphocytes in SLE compared to normal healthy donors. Keywords: SLE, apoptosis, lymphocyte phenotypes, prednisolone, dsDNA autoantibodies, flow cytometry Sudan Journal of Medical Sciences Vol. 1(1) 2006: 14-19
Highlights
Percentages of apoptosis in granulocytes from patients with Systemic lupus erythematosus (SLE) and healthy donors both in freshly intact (3.8 ± 3.1% vs. 3.6 ± 1.4% respectively), and after being cultured in culturing medium at 37C0 and 5 % CɈ2 (14.1 ± 8.4% vs. 15.3 ± 5.6% respectively) ; no significant difference was reveal in level of apoptosis (Fig.1)
Apoptosis of lymphocytes and SLE disease activity Depending upon the SLE disease activity, the level apoptosis of lymphocytes after in vitro culturing was raised (Table 1)
Our data shows that significant increased in percentage apoptosis in lymphocyte subsets CD4, CD8, and CD20 lymphocyte in patients with SLE compared to healthy donors; CD4 T cell in SLE underwent the much apoptosis than other lymphocyte subsets (Table 2)
Summary
One of the mechanisms by which elimination of autoreactive lymphocytes takes place is apoptosis. Adefect in apoptosis may contribute to the development of autoimmune disease. The study included 34 Russian children patients with SLE in age between 6-17 years old. Lymphocytes were isolated from venous blood by method of gradient centrifugation of all the blood through a Ficoll-pak solution. The quantity apoptotic cells was determined in leukocytes by flow cytometry Epics XL-2 (“Beckman Coulter”, USA). Analysis of lymphocyte subpopulations was carried by using two fluorescent labels: FITC conjugated monoclonal antibodies (CD-25 and CD-20) and phycoerythrin conjugated monoclonal antibody (CD-3, CD-4 and CD-8).
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