The Influence of Recombinant Human Erythropoietin on Apoptosis and Cytokine Production of CD4 + lymphocytes from Hemodia
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The Influence of Recombinant Human Erythropoietin on Apoptosis and Cytokine Production of CD4+ lymphocytes from Hemodialyzed Patients Katarzyna A. Lisowska & Alicja Dębska-Ślizień & Aleksandra Jasiulewicz & Agnieszka Daca & Ewa Bryl & Jacek M. Witkowski
Received: 18 September 2012 / Accepted: 8 November 2012 / Published online: 20 November 2012 # The Author(s) 2012. This article is published with open access at Springerlink.com
Abstract Recombinant human erythropoietin (rhEPO) treatment of hemodialyzed (HD) patients normalizes the altered phenotype of CD4+ lymphocytes and restores the balance of Th1/Th2 cytokines. We decided to test how the presence of rhEPO in cell culture modulates cytokine production of CD4+ lymphocytes in HD patients with stable hemoglobin level and expression of activation antigens of stimulated CD4+ lymphocytes similar to those observed in healthy individuals. We also tested whether the presence of rhEPO in cell culture protects stimulated CD4+ lymphocytes of HD patients from apoptosis. Peripheral blood mononuclear cells (PBMC) of HD patients were stimulated with an immobilized anti-CD3 antibody with or without addition of rhEPO. The percentage of apoptotic CD4+ lymphocytes and the level of Th1/Th2 cytokines in culture supernatants were measured with flow cytometry. HD patients showed a decrease in the percentage of apoptotic CD4+ cells after stimulation with the anti-CD3 antibody combined with rhEPO. The level of IFN-g and IL-10 was increased while the level of TNF-α was decreased in the presence of rhEPO in cell culture from HD patients. These results confirm the role of rhEPO signaling in T lymphocytes of HD patients. Keywords CD4+ lymphocytes . apoptosis . cytokines . rhEPO K. A. Lisowska (*) : A. Jasiulewicz : A. Daca : E. Bryl : J. M. Witkowski Department of Pathophysiology, Medical University of Gdańsk, Dębinki 7, 80-211 Gdańsk, Poland e-mail: [email protected] A. Dębska-Ślizień Department of Nephrology, Transplantology and Internal Diseases, Medical University of Gdańsk, Gdańsk, Poland
Introduction Chronic renal failure is accompanied by a deficiency state in both cell-mediated and humoral immunity which is deepened by hemodialysis (HD) [1]. A direct contact of the patient’s peripheral blood mononuclear cells (PBMC) with an artificial membrane increases the production of pro-inflammatory cytokines: tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1 and IL-6 [2, 3]. What’s more, HD patients demonstrate a decreased production of IL-2 and interferon gamma (IFN-g) [4, 5]. The deficient production of cytokines is probably related to changes in phenotype of CD4+ lymphocytes from HD patients, which exhibit decreased expression of the major co-stimulatory CD28 antigen and main activation markers: CD25 and CD69 [6]. Recombinant human erythropoietin (rhEPO) administered to HD patients to correct the anemia state can influence both the phenotype of T lymphocytes and the production of cytokines. Our previous studies have shown that rhEPO treatment normalizes the impai
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