scholarly journals Властивості Na+ , K+ -активованої, Mg2+ -залежної АТФ-гідролази лімфоцитів крові у хворих на реактивний артрит

2013 ◽  
Vol 4 (2) ◽  
pp. 57-62
Author(s):  
O.V. Melnyk ◽  
O.P. Kornijchuk ◽  
O.I. Pershyn ◽  
Z.D. Vorobets
Keyword(s):  
Atpase Activity ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Immunocompetent Cells ◽  
Affinity Constant ◽  
Kinetic Properties ◽  
Blood Lymphocytes ◽  
Healthy Donors ◽  
Immunological Mechanisms ◽  
The Impact

A significant role in the development and course of reactive arthritis (ReA) is played by T-lymphocytes as their development and systemic manifestations are based on immunological mechanisms. Additionally, the pathogenesis of many diseases is linked to changes in the structure and function of ion-transporting systems. Therefore, the aim of the study was to find out the kinetic properties of ATP-hydrolysis reaction involving Na+,K+-ATPase of peripheral blood lymphocytes of healthy individuals and patients with ReA. We used the current methodological approaches to the study of ATPase activity in saponin permeabilized cells. We conducted an analysis of the kinetic properties of ouabainsensitive Na+,K+-ATPase activity of saponin-perforated peripheral blood lymphocytes of healthy donors and patients with rheumatoid arthritis (RеA). We found out that in peripheral blood lymphocytes of patients with RеA primary active transport of Na+, K+ ions is slower and less intensive, though characterised by the same capacity, as in healthy donors. The affinity constant for ATP in peripheral blood lymphocytes in patients with RеA is greater by 2.9 times than its value in comparison with healthy donors. We established that in conditions of rheumatic pathology in immunocompetent cells, inhibition of Na+,K+-ATPase activity is not caused by reduction of speed of enzyme work, but by increase of affinity of ouabainsensitive Na+,K+-ATPase to ATP. At the same time, the Mg2+-binding center of Na+,K+-ATPase in patients with RеA is endogenous. We also found that affinity Na+,K+-ATPase to the ions K+ in peripheral blood lymphocytes of healthy donors is 2.4 times higher than in patients with RеA. We observed that Na+,K+-ATPase of peripheral blood lymphocytes of patients with RеA retains its endogenous receptor properties – sensitivity to ouabain does not change. It is assumed that under conditions of rheumatic pathology the impact on the Na+,K+-ATPase structure occurs both externally and on the cytoplasmic membrane surface. The above experimental data can be used for further clarification of the membrane mechanisms of ion exchange in immunocompetent cells of patients suffering from autoimmune diseases.

scholarly journals Antagonistic Effects of CAPE (a Component of Propolis) on the Cytotoxicity and Genotoxicity of Irinotecan and SN38 in Human Gastrointestinal Cancer Cells In Vitro

Molecules ◽  
2020 ◽  
Vol 25 (3) ◽  
pp. 658 ◽  
Author(s):  
Gabriela Gajek ◽  
Beata Marciniak ◽  
Jarosław Lewkowski ◽  
Renata Kontek
Keyword(s):  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Caffeic Acid Phenethyl Ester ◽  
Peripheral Blood Lymphocytes ◽  
Neoplastic Cells ◽  
Blood Lymphocytes ◽  
Human Peripheral Blood Lymphocytes ◽  
Apoptotic Activity ◽  
The Impact

The incidence of gastrointestinal cancers is increasing every year. Irinotecan (CPT-11), a drug used in the treatment of colorectal cancer and gastric cancer, is metabolized by carboxylesterases to an active metabolite, SN-38, which is more cytotoxic. CAPE (caffeic acid phenethyl ester) is an active component of propolis, which has a high antibacterial, antiviral, and antineoplastic potential. This study analyses the impact of CAPE on the cytotoxic (MTT assay), genotoxic (comet assay) and proapoptotic (caspase-3/7 activity) potential of irinotecan and its metabolite SN-38 in cultures of gastrointestinal neoplastic cells (HCT116, HT29, AGS). Cytotoxicity and genotoxicity activities of these compounds were carried out in comparison with human peripheral blood lymphocytes (PBLs) in vitro. The antioxidant potential of CAPE was investigated in relation H2O2-induced oxidative stress in the both neoplastic cells and PBLs. CAPE expressed cytotoxic, genotoxic, and pro-apoptotic activity against AGS, HCT116, and HT29 tumor cells. CAPE, in the presence of different concentrations of irinotecan or SN38, decreased the cytotoxicity, genotoxicity, and pro-apoptotic activity in these cell lines, but it has no such action on normal human peripheral blood lymphocytes.

scholarly journals The kinetic properties of the ecto-ATPase of human peripheral blood lymphocytes and of chronic lymphatic leukemia cells

Blood ◽  
1983 ◽  
Vol 62 (5) ◽  
pp. 1041-1046
Author(s):  
HR Gutmann ◽  
YM Chow ◽  
RL Vessella ◽  
B Schuetzle ◽  
ME Kaplan
Keyword(s):  
B Cell ◽  
Peripheral Blood ◽  
Specific Activity ◽  
Human Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Kinetic Constants ◽  
Kinetic Properties ◽  
Blood Lymphocytes ◽  
Significant Difference ◽  
Specific Activities

This study examines whether the activity of the Mg2+-dependent ecto- ATPase of the surface membrane of the human lymphocyte is changed in chronic lymphocytic B-cell leukemia (CLL-B) and may be an indicator of malignant transformation. The ecto-ATPase activities of preparations consisting predominantly of T or B cells were compared to each other and to the ecto-ATPase of the CLL peripheral blood lymphocytes (PBL). The specific activities and kinetic constants of the ecto-ATPase of the cell preparations were determined with [gamma-32P] adenosine triphosphate (ATP) as substrate. B-enriched lymphocytes had nearly fourfold greater specific activity and apparent Vmax than T-enriched lymphocytes, while the Km values of both cell types showed no significant difference. The specific activities and kinetic constants of the ecto-ATPase of the CLL PBL were significantly higher than the corresponding values of PBL or of B-enriched lymphocytes. Judging from the kinetic constants the ecto-ATPase of the CLL-B lymphocyte appears to be an enzyme that is distinctly different from that of the normal B cell. On the basis of the kinetic properties, the ecto-ATPase of the B cell appears to be identical with that of the T cell. The differences in the maximal velocities of the hydrolysis of ATP by B and T cells are likely due to a greater number of enzymatic sites on the B cell.

scholarly journals About hematopoietic properties of peripheral blood lymphocytes RNA from patients with polycythemia vera and healthy donors

2015 ◽  
Vol 10 (2) ◽  
pp. 58 ◽  
Author(s):  
A. G. Babaeva ◽  
N. M. Gevorkyan ◽  
N. V. Tishevskaya ◽  
L. L. Golovkina ◽  
Yu. O. Muratova ◽  
...  
Keyword(s):  
Polycythemia Vera ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Healthy Donors

HLA-A*0201 Restricted Killing Activity of WT1-Specific CTL Generated From the Peripheral Blood Lymphocytes of Normal Healthy Donors.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3026-3026
Author(s):  
Deepa Kolaseri Krishnadas ◽  
Mindy Stamer ◽  
Kim Dunham ◽  
Lei Bao ◽  
Kenneth Lucas
Keyword(s):  
T Cells ◽  
Cell Line ◽  
Cd8 T Cells ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Target Cells ◽  
Cd8 Cells ◽  
Blood Lymphocytes ◽  
Healthy Donors ◽  
Ifn Γ

Abstract Abstract 3026 Poster Board II-1002 The Wilms' tumor antigen (WT1) is over-expressed on several human leukemia and solid tumors, and thus is considered as a potential target for cancer immunotherapy. Combating leukemia by targeting WT1 expressing leukemic cells using in vitro generated WT1-specific CTL is one potential approach, but it is difficult to generate an immune response against WT1 due to low T cell precursor frequency in normal healthy individuals. Earlier studies have shown the generation of WT1-A*0201 peptide specific CTL from CD8+ T cells by cloning. Another study reported the production of IFN- γ by WT-1 specific CD8+ T cells. However, the cytolytic killing ability of these IFN- γ producing cells was not further characterized. Here, we demonstrate the generation of WT1-A*0201 specific CTL from the peripheral blood lymphocytes (PBL) of normal healthy donors using CD137 selection. The PBL were stimulated once with RMFPNAPYL (WT1-A*0201 peptide) pulsed autologous dendritic cells and twice with WT1-A*0201 peptide pulsed irradiated peripheral blood mononuclear cells (PBMC). Following three stimulations, the PBL were selected for CD137+ expression and rapidly expanded with OKT3 and IL-2. The WT1-A*0201 specific CTL showed killing of target cells and production of IFN-γ in an antigen-specific manner. The percent killing of WT1-A*0201 peptide pulsed T2 cells (TAP−, HLA- A2+) and autologous B blast (BB) were significantly higher when compared with their control targets. T2 cells and BB either pulsed with an irrelevant A*0201 peptide or un-pulsed served as the control. We have observed similar results with WT1-A*0201 specific CTL generated from normal donor CD8+ cells. However, the efficiency of WT1-A*0201 CTL generated from PBL to kill target cells and produce IFN- γ was higher than CTL from CD8+ cells. The CTL generated from PBL killed BA25, a WT1 expressing A2+ leukemia cell line but failed to kill Molt-4, a WT1 expressing A2− cell line, clearly indicating HLA-A2 restricted CTL activity. The specificity of the generated CTL were further confirmed by staining with WT1-HLA-A*0201 tetramer. The percentage of WT1-specific CD3+CD8+Tetramer+ cells either remained same or higher in CTL generated from PBL when compared with those generated from CD8+ cells. CD137 selection leads to the generation of significant number of CTL in a shorter time when compared to conventional cloning methods. In addition, generation of WT1-A*0201 specific CTL from PBL avoids CD8+ selection. Currently, we are aiming to generate WT1-specific CTL using an overlapping WT1 peptide-mix in order to widen our ability to treat patients with different HLA types. This study has implications for cellular immunotherapy in leukemia patients who relapse following allogeneic stem cell transplantation. Disclosures No relevant conflicts of interest to declare.

Splenectomy selectively affects the distribution and mobility of the recirculating lymphocyte pool

Blood ◽  
2000 ◽  
Vol 96 (3) ◽  
pp. 1180-1183 ◽  
Author(s):  
Tim J. Seabrook ◽  
Wayne R. Hein ◽  
Lisbeth Dudler ◽  
Alan J. Young
Keyword(s):  
Lymph Nodes ◽  
Peripheral Blood ◽  
Blood Lymphocyte ◽  
Lymphocyte Subsets ◽  
Immune Surveillance ◽  
Peripheral Blood Lymphocytes ◽  
Immune Competence ◽  
Blood Lymphocytes ◽  
The Impact ◽  
Residency Time

The spleen plays a major role in immune surveillance, but the impact that splenectomy exerts on the immune competence of an individual is not fully resolved. Here we show that neonatal splenectomy in sheep does not abrogate the development of a large, nonrecirculating pool of lymphocytes and that it has no effect on the acquisition of a normal blood lymphocyte profile. Splenectomy did, however, result in a significant decrease in blood residency time of recirculating lymphocytes and in an enhanced accumulation of recirculating lymphocytes in lymph nodes. Furthermore, nonrecirculating peripheral blood lymphocytes were less likely to migrate to the lung, possibly because of saturation of the marginal pool by recirculating lymphocytes. Although splenectomy has little effect on the development or distribution of lymphocyte subsets in blood and lymph, it has marked effects on the rate of recirculation of lymphocytes, which may have significant implications for peripheral immune surveillance in patients who undergo splenectomy.

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