scholarly journals Extracellular adenosine triphosphate increases cation permeability of chronic lymphocytic leukemic lymphocytes

Blood ◽  
1989 ◽  
Vol 73 (5) ◽  
pp. 1316-1323 ◽  
Author(s):  
JS Wiley ◽  
GR Dubyak
Keyword(s):  
Adenosine Triphosphate ◽  
Fold Increase ◽  
Normal Subjects ◽  
Peripheral Blood Lymphocytes ◽  
Extracellular Atp ◽  
Total Cell ◽  
Lymphocytic Leukemia ◽  
Maximal Response ◽  
Extracellular Adenosine ◽  
Cation Permeability

Abstract Extracellular adenosine triphosphate (ATP) is known to reversibly increase the cation permeability of a variety of freshly isolated and cultured cell types. In this study the effects of extracellular ATP were studied using peripheral blood lymphocytes (PBL) isolated from both normal subjects and from patients with chronic lymphocytic leukemia (CLL). Changes in the permeability to Na+, Rb+, and Li+ ions were measured using conventional isotope and flame photometry techniques. In addition, changes in cytosolic (Ca2+) were fluorimetrically monitored to assess possible changes in net Ca2+ influx. ATP produced a 12-fold increase in 22Na+ influx into CLL cells but only a 3.5-fold increase in this flux in PBL cells. A maximal response was produced by 0.1 mmol/L ATP in the absence of Mg2+, while a twofold molar excess of Mg2+ over ATP abolished the response. ATP had no effect on the passive (ouabain-insensitive) 86Rb+ influx into PBL cells but stimulated this flux by fivefold in the CLL cells. Li+ influx into CLL cells was also stimulated threefold by ATP. Under these same conditions ATP also produced a net increase in total cell Na and a decrease in total cell K in the CLL cells. Exclusion of two normally impermeable dyes, trypan blue and ethidium bromide, was not altered in the ATP-treated CLL cells. Finally, extracellular ATP (3 mmol/L) produced no significant change in the cytosolic (Ca2+) of normal, monocyte-depleted populations of PBL. Conversely, this same concentration of ATP produced a very rapid (complete within 30 seconds) and a significant (an average threefold peak change) increase in the cytosolic (Ca2+) of cell preparations derived from five out of nine CLL patients. In these latter CLL cells, the ATP-induced elevation in cytosolic (Ca2+) appeared to be due to a net increase in Ca2+ influx, since no elevations were observed when the extracellular (Ca2+) was reduced to less than 0.1 mmol/L. These actions of ATP were specific in that equimolar concentrations of other nucleotides were without effect. These data indicate that treatment of CLL lymphocytes with extracellular ATP4 produces large increases in cation permeability. In contrast, there is less or no ATP-induced permeabilization of normal PBL.

scholarly journals Extracellular adenosine triphosphate increases cation permeability of chronic lymphocytic leukemic lymphocytes

Blood ◽  
1989 ◽  
Vol 73 (5) ◽  
pp. 1316-1323
Author(s):  
JS Wiley ◽  
GR Dubyak
Keyword(s):  
Adenosine Triphosphate ◽  
Fold Increase ◽  
Normal Subjects ◽  
Peripheral Blood Lymphocytes ◽  
Extracellular Atp ◽  
Total Cell ◽  
Lymphocytic Leukemia ◽  
Maximal Response ◽  
Extracellular Adenosine ◽  
Cation Permeability

Extracellular adenosine triphosphate (ATP) is known to reversibly increase the cation permeability of a variety of freshly isolated and cultured cell types. In this study the effects of extracellular ATP were studied using peripheral blood lymphocytes (PBL) isolated from both normal subjects and from patients with chronic lymphocytic leukemia (CLL). Changes in the permeability to Na+, Rb+, and Li+ ions were measured using conventional isotope and flame photometry techniques. In addition, changes in cytosolic (Ca2+) were fluorimetrically monitored to assess possible changes in net Ca2+ influx. ATP produced a 12-fold increase in 22Na+ influx into CLL cells but only a 3.5-fold increase in this flux in PBL cells. A maximal response was produced by 0.1 mmol/L ATP in the absence of Mg2+, while a twofold molar excess of Mg2+ over ATP abolished the response. ATP had no effect on the passive (ouabain-insensitive) 86Rb+ influx into PBL cells but stimulated this flux by fivefold in the CLL cells. Li+ influx into CLL cells was also stimulated threefold by ATP. Under these same conditions ATP also produced a net increase in total cell Na and a decrease in total cell K in the CLL cells. Exclusion of two normally impermeable dyes, trypan blue and ethidium bromide, was not altered in the ATP-treated CLL cells. Finally, extracellular ATP (3 mmol/L) produced no significant change in the cytosolic (Ca2+) of normal, monocyte-depleted populations of PBL. Conversely, this same concentration of ATP produced a very rapid (complete within 30 seconds) and a significant (an average threefold peak change) increase in the cytosolic (Ca2+) of cell preparations derived from five out of nine CLL patients. In these latter CLL cells, the ATP-induced elevation in cytosolic (Ca2+) appeared to be due to a net increase in Ca2+ influx, since no elevations were observed when the extracellular (Ca2+) was reduced to less than 0.1 mmol/L. These actions of ATP were specific in that equimolar concentrations of other nucleotides were without effect. These data indicate that treatment of CLL lymphocytes with extracellular ATP4 produces large increases in cation permeability. In contrast, there is less or no ATP-induced permeabilization of normal PBL.

scholarly journals Visualization of tubulin in lymphocytes. I. Comparison of normal and chronic lymphocytic leukemia (CLL) lymphocytes

Blood ◽  
1978 ◽  
Vol 51 (6) ◽  
pp. 1031-1037 ◽  
Author(s):  
G Dighiero ◽  
E Karsenti ◽  
JY Follezou ◽  
M Bornens
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Peripheral Blood ◽  
Normal Subjects ◽  
Peripheral Blood Lymphocytes ◽  
Lower Number ◽  
Lymphocytic Leukemia ◽  
Blood Lymphocytes ◽  
Cell Structures ◽  
Immunoperoxidase Assay

Abstract Cell structures containing tubulin were studied in peripheral blood lymphocytes from 8 normal donors and 11 patients with CLL using specific antitubulin antibodies revealed by immunoperoxidase assay. The centriole and microtubules were clearly visible in both groups. A “nucleus-associated tubulin-containing structure” was revealed by antitubulin antibodies and was found in virtually all lymphocytes of normal subjects but in a considerably lower number of CLL lymphocytes. The nature of this structure and its relationship to other cell structures are discussed.

Role of Extracellular Adenosine Triphosphate in Human Skin

2004 ◽  
Vol 8 (2) ◽  
pp. 90-96 ◽  
Author(s):  
Aton M. Holzer ◽  
Richard D. Granstein
Keyword(s):  
Human Skin ◽  
Adenosine Triphosphate ◽  
Current Knowledge ◽  
Extracellular Atp ◽  
Cell Type ◽  
Skin Injury ◽  
Direct Role ◽  
Extracellular Adenosine ◽  
Proliferation And Apoptosis

Background: The nucleotide adenosine triphosphate (ATP) has long been known to drive and participate in countless intracellular processes. Extracellular ATP and its metabolite adenosine have also been shown to exert a variety of effects on nearly every cell type in human skin. Knowledge of the sources and effects of extracellular ATP in human skin may help shape new therapies for skin injury, inflammation, and numerous other cutaneous disorders. Objective: The objective of this review is to introduce the reader to current knowledge regarding the sources and effects of extracellular ATP in human skin and to outline areas in which further research is necessary to clarify the nature and mechanism of these effects. Conclusion: Extracellular ATP seems to play a direct role in triggering skin inflammatory, regenerative, and fibrotic responses to mechanical injury, an indirect role in melanocyte proliferation and apoptosis, and a complex role in Langerhans cell-directed adaptive immunity.

Adenosine Triphosphate–Induced Shedding of CD23 and L-Selectin (CD62L) From Lymphocytes Is Mediated by the Same Receptor but Different Metalloproteases

Blood ◽  
1998 ◽  
Vol 92 (3) ◽  
pp. 946-951 ◽  
Author(s):  
B. Gu ◽  
L.J. Bendall ◽  
J.S. Wiley
Keyword(s):  
Adenosine Triphosphate ◽  
Phorbol Ester ◽  
P2x7 Receptor ◽  
Transmembrane Protein ◽  
Lymphocytic Leukemia ◽  
P2x7 Receptors ◽  
Extracellular Adenosine ◽  
Zinc Chelator ◽  
High Concentrations ◽  
American Society

CD23 is a transmembrane protein expressed on the surface of B-lymphocytes that binds IgE, CD21, CD11b, and CD11c. High concentrations of soluble CD23 and L-selectin are found in the serum of patients with B-chronic lymphocytic leukemia (B-CLL). Because extracellular adenosine triphosphate (ATP) causes shedding of L-selectin via activation of P2Z/P2X7 receptors expressed on B-CLL lymphocytes, we studied the effect of ATP on shedding of CD23. ATP-induced shedding of CD23 at an initial rate of 12% of that for L-selectin, whereas the EC50 for ATP was identical (35 μmol/L) for shedding of both molecules. Furthermore, benzoylbenzoyl ATP also produced shedding of CD23 and L-selectin with the same agonist EC50 values for both (10 μmol/L). Inactivation of the P2Z/P2X7 receptor by preincubation with oxidized ATP abolished ATP-induced shedding of both molecules. Moreover, KN-62, the most potent inhibitor for the P2Z/P2X7 receptor, inhibited ATP-induced shedding of both CD23 and L-selectin with the same IC50 (12 nmol/L). Ro 31-9790, a membrane permeant zinc chelator that inhibits the phorbol-ester-stimulated shedding of L-selectin, also inhibited shedding of CD23 from B-CLL lymphocytes. However, the IC50 for this inhibition by Ro31-9790 was different for L-selectin and CD23 (83 v 6 μmol/L, respectively). Although L-selectin was completely shed by incubation of cells with phorbol-ester, CD23 was not lost under these conditions. The data show that extracellular ATP induces shedding of L-selectin and CD23 from B-CLL lymphocytes by an action mediated by the P2Z/P2X7 receptor. However, different membrane metalloproteases seem to mediate the shedding of L-selectin and CD23. © 1998 by The American Society of Hematology.

scholarly journals Modulation of platelet function by extracellular adenosine triphosphate

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 984-993 ◽  
Author(s):  
G Soslau ◽  
J Parker
Keyword(s):  
Platelet Aggregation ◽  
Adenosine Triphosphate ◽  
Platelet Function ◽  
Surface Membrane ◽  
Adenosine Diphosphate ◽  
Extracellular Atp ◽  
Surface Proteins ◽  
Intracellular Camp ◽  
Extracellular Adenosine ◽  
Camp Levels

Abstract A potential physiologic role of extracellular adenosine triphosphate (ATP) on platelet function is proposed in this report. It is widely accepted that ATP competitively inhibits adenosine diphosphate (ADP)- induced platelet aggregation. Our observations of platelet aggregation with the agonists, collagen, epinephrine, and ADP in the presence of 180 mumol/L ATP could support this competitive nature of ATP. However, the disaggregation of maximally aggregated platelets induced by ATP, theophylline, or ATP plus theophylline indicates that additional mechanisms of ATP action may be present. Extracellular gamma-32P-ATP (7 pmol) labels surface-membrane proteins in intact platelets as demonstrated by several criteria. The reaction is Ca++-dependent. Stimulation by calcium occurs in the physiologic range of 1 to 5 mmol/L. Significant levels of phosphorylation occur within one minute with near maximal levels reached by five minutes. Platelet cyclic AMP (cAMP) levels were elevated in a dose-dependent fashion in cells incubated for four minutes with increasing amounts of extracellular ATP (18 to 540 nmol). The addition of ATP plus theophylline resulted in a synergistic stimulation of cAMP levels. ATP was not being hydrolyzed to adenosine by plasma nucleotidases, as demonstrated by the lack of effect of ten U of adenosine deaminase. The phosphorylation of surface proteins by extracellular ATP released from activated platelets may modulate platelet responsiveness to agonists at distances removed from the site of vascular injury. Phosphorylation may also play a role in signal transduction to regulate the levels of intracellular cAMP, which further inhibits platelet activation.

scholarly journals Mitogen stimulation of chronic lymphocytic leukemic lymphocytes: defective phytohemagglutinin stimulation independent of immunologic cell surface markers

Blood ◽  
1980 ◽  
Vol 56 (2) ◽  
pp. 256-261 ◽  
Author(s):  
S Davis ◽  
P Rambotti
Keyword(s):  
Cell Surface ◽  
Peripheral Blood Lymphocytes ◽  
Lymphocytic Leukemia ◽  
Pokeweed Mitogen ◽  
Maximal Response ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Surface Immunoglobulin ◽  
Pha Response ◽  
Stimulation Of

Abstract Peripheral blood lymphocytes (PBL) from 107 untreated patients with chronic lymphocytic leukemia (CLL) were analyzed for the presence of surface immunoglobulin (Ig) and the ability to form rosettes with sheep erythrocytes (SRBC). Four groups were identified based on the cell surface markers: (1) 81 patients' PBL expressed primarily IgM kappa or IgM lambda, 4 further patients' PBL expressed IgM with equal percentages of kappa and lambda surface markers; (2) 13 patients had equal percentages of PBL expressing lg and SRBC receptors; (3) 6 patients' PBL primarily formed rosettes with SRBCs, and (4) in 3 patients and the majority of cells had no detectable markers (null cells). Lymphocytes from all patients within each group were tested for their ability to respond to phytohemagglutinin (PHA) and pokeweed mitogen (PWM). The maximum response in PHA-stimulated normal cell cultures appeared at 2--3 days; for PWM-stimulated cultures, maximal response was at 3--5 days. CLL cultures from all patients in each of the four groups required 5--7 days to develop a maximal PHA response. The response of CLL lymphocytes in all groups to PWM stimulation was similar to normal lymphocytes. Thus, the abnormal PHA response of CLL lymphocytes was independent of the presence or pattern of cell surface markers.

scholarly journals Mitogen stimulation of chronic lymphocytic leukemic lymphocytes: defective phytohemagglutinin stimulation independent of immunologic cell surface markers

Blood ◽  
1980 ◽  
Vol 56 (2) ◽  
pp. 256-261
Author(s):  
S Davis ◽  
P Rambotti
Keyword(s):  
Cell Surface ◽  
Peripheral Blood Lymphocytes ◽  
Lymphocytic Leukemia ◽  
Pokeweed Mitogen ◽  
Maximal Response ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Surface Immunoglobulin ◽  
Pha Response ◽  
Stimulation Of

Peripheral blood lymphocytes (PBL) from 107 untreated patients with chronic lymphocytic leukemia (CLL) were analyzed for the presence of surface immunoglobulin (Ig) and the ability to form rosettes with sheep erythrocytes (SRBC). Four groups were identified based on the cell surface markers: (1) 81 patients' PBL expressed primarily IgM kappa or IgM lambda, 4 further patients' PBL expressed IgM with equal percentages of kappa and lambda surface markers; (2) 13 patients had equal percentages of PBL expressing lg and SRBC receptors; (3) 6 patients' PBL primarily formed rosettes with SRBCs, and (4) in 3 patients and the majority of cells had no detectable markers (null cells). Lymphocytes from all patients within each group were tested for their ability to respond to phytohemagglutinin (PHA) and pokeweed mitogen (PWM). The maximum response in PHA-stimulated normal cell cultures appeared at 2--3 days; for PWM-stimulated cultures, maximal response was at 3--5 days. CLL cultures from all patients in each of the four groups required 5--7 days to develop a maximal PHA response. The response of CLL lymphocytes in all groups to PWM stimulation was similar to normal lymphocytes. Thus, the abnormal PHA response of CLL lymphocytes was independent of the presence or pattern of cell surface markers.

scholarly journals Visualization of tubulin in lymphocytes. I. Comparison of normal and chronic lymphocytic leukemia (CLL) lymphocytes

Blood ◽  
1978 ◽  
Vol 51 (6) ◽  
pp. 1031-1037
Author(s):  
G Dighiero ◽  
E Karsenti ◽  
JY Follezou ◽  
M Bornens
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Peripheral Blood ◽  
Normal Subjects ◽  
Peripheral Blood Lymphocytes ◽  
Lower Number ◽  
Lymphocytic Leukemia ◽  
Blood Lymphocytes ◽  
Cell Structures ◽  
Immunoperoxidase Assay

Cell structures containing tubulin were studied in peripheral blood lymphocytes from 8 normal donors and 11 patients with CLL using specific antitubulin antibodies revealed by immunoperoxidase assay. The centriole and microtubules were clearly visible in both groups. A “nucleus-associated tubulin-containing structure” was revealed by antitubulin antibodies and was found in virtually all lymphocytes of normal subjects but in a considerably lower number of CLL lymphocytes. The nature of this structure and its relationship to other cell structures are discussed.

Adenosine Triphosphate–Induced Shedding of CD23 and L-Selectin (CD62L) From Lymphocytes Is Mediated by the Same Receptor but Different Metalloproteases

Blood ◽  
1998 ◽  
Vol 92 (3) ◽  
pp. 946-951 ◽  
Author(s):  
B. Gu ◽  
L.J. Bendall ◽  
J.S. Wiley
Keyword(s):  
Adenosine Triphosphate ◽  
Phorbol Ester ◽  
P2x7 Receptor ◽  
Transmembrane Protein ◽  
Lymphocytic Leukemia ◽  
P2x7 Receptors ◽  
Extracellular Adenosine ◽  
Zinc Chelator ◽  
High Concentrations ◽  
American Society

Abstract CD23 is a transmembrane protein expressed on the surface of B-lymphocytes that binds IgE, CD21, CD11b, and CD11c. High concentrations of soluble CD23 and L-selectin are found in the serum of patients with B-chronic lymphocytic leukemia (B-CLL). Because extracellular adenosine triphosphate (ATP) causes shedding of L-selectin via activation of P2Z/P2X7 receptors expressed on B-CLL lymphocytes, we studied the effect of ATP on shedding of CD23. ATP-induced shedding of CD23 at an initial rate of 12% of that for L-selectin, whereas the EC50 for ATP was identical (35 μmol/L) for shedding of both molecules. Furthermore, benzoylbenzoyl ATP also produced shedding of CD23 and L-selectin with the same agonist EC50 values for both (10 μmol/L). Inactivation of the P2Z/P2X7 receptor by preincubation with oxidized ATP abolished ATP-induced shedding of both molecules. Moreover, KN-62, the most potent inhibitor for the P2Z/P2X7 receptor, inhibited ATP-induced shedding of both CD23 and L-selectin with the same IC50 (12 nmol/L). Ro 31-9790, a membrane permeant zinc chelator that inhibits the phorbol-ester-stimulated shedding of L-selectin, also inhibited shedding of CD23 from B-CLL lymphocytes. However, the IC50 for this inhibition by Ro31-9790 was different for L-selectin and CD23 (83 v 6 μmol/L, respectively). Although L-selectin was completely shed by incubation of cells with phorbol-ester, CD23 was not lost under these conditions. The data show that extracellular ATP induces shedding of L-selectin and CD23 from B-CLL lymphocytes by an action mediated by the P2Z/P2X7 receptor. However, different membrane metalloproteases seem to mediate the shedding of L-selectin and CD23. © 1998 by The American Society of Hematology.

scholarly journals Modulation of platelet function by extracellular adenosine triphosphate

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 984-993 ◽  
Author(s):  
G Soslau ◽  
J Parker
Keyword(s):  
Platelet Aggregation ◽  
Adenosine Triphosphate ◽  
Platelet Function ◽  
Surface Membrane ◽  
Adenosine Diphosphate ◽  
Extracellular Atp ◽  
Surface Proteins ◽  
Intracellular Camp ◽  
Extracellular Adenosine ◽  
Camp Levels

A potential physiologic role of extracellular adenosine triphosphate (ATP) on platelet function is proposed in this report. It is widely accepted that ATP competitively inhibits adenosine diphosphate (ADP)- induced platelet aggregation. Our observations of platelet aggregation with the agonists, collagen, epinephrine, and ADP in the presence of 180 mumol/L ATP could support this competitive nature of ATP. However, the disaggregation of maximally aggregated platelets induced by ATP, theophylline, or ATP plus theophylline indicates that additional mechanisms of ATP action may be present. Extracellular gamma-32P-ATP (7 pmol) labels surface-membrane proteins in intact platelets as demonstrated by several criteria. The reaction is Ca++-dependent. Stimulation by calcium occurs in the physiologic range of 1 to 5 mmol/L. Significant levels of phosphorylation occur within one minute with near maximal levels reached by five minutes. Platelet cyclic AMP (cAMP) levels were elevated in a dose-dependent fashion in cells incubated for four minutes with increasing amounts of extracellular ATP (18 to 540 nmol). The addition of ATP plus theophylline resulted in a synergistic stimulation of cAMP levels. ATP was not being hydrolyzed to adenosine by plasma nucleotidases, as demonstrated by the lack of effect of ten U of adenosine deaminase. The phosphorylation of surface proteins by extracellular ATP released from activated platelets may modulate platelet responsiveness to agonists at distances removed from the site of vascular injury. Phosphorylation may also play a role in signal transduction to regulate the levels of intracellular cAMP, which further inhibits platelet activation.

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