Expression of P2X7purinoceptors on human lymphocytes and monocytes: evidence for nonfunctional P2X7receptors

2000 ◽  
Vol 279 (4) ◽  
pp. C1189-C1197 ◽  
Author(s):  
B. J. Gu ◽  
W. Y. Zhang ◽  
L. J. Bendall ◽  
I. P. Chessell ◽  
G. N. Buell ◽  
...  
Keyword(s):  
B Lymphocytes ◽  
Adhesion Molecule ◽  
Human Lymphocytes ◽  
Normal Subjects ◽  
Cell Types ◽  
Surface Expression ◽  
Lymphocytic Leukemia ◽  
Functional Responses ◽  
Intracellular Expression ◽  
Selective Channel

Lymphocytes from normal subjects and patients with B-chronic lymphocytic leukemia (B-CLL) show functional responses to extracellular ATP characteristic of the P2X7receptor (previously termed P2Z). These responses include opening of a cation-selective channel/pore that allows entry of the fluorescent dye ethidium and activation of a membrane metalloprotease that sheds the adhesion molecule L-selectin. The surface expression of P2X7receptors was measured in normal leucocytes, platelets, and B-CLL lymphocytes and correlated with their functional responses. Monocytes showed four- to fivefold greater expression of P2X7than B, T, and NK lymphocytes, whereas P2X7expression on neutrophils and platelets was weak. All cell types demonstrated abundant intracellular expression of this receptor. All 12 subjects with B-CLL expressed lymphocyte P2X7at about the same level as B lymphocytes from normal subjects. P2X7function, measured by ATP-induced uptake of ethidium, correlated closely with surface expression of this receptor in normal and B-CLL lymphocytes and monocytes ( n = 47, r = 0.70; P< 0.0001). However, in three patients the ATP-induced uptake of ethidium into the malignant B lymphocytes was low or absent. The lack of P2X7function in these B lymphocytes was confirmed by the failure of ATP to induce Ba2+uptake into their lymphocytes. This lack of function of the P2X7receptor resulted in a failure of ATP-induced shedding of L-selectin, an adhesion molecule that directs the recirculation of lymphocytes from blood into the lymph node.

scholarly journals Both Freshly Prepared and Frozen-Stored Amniotic Membrane Cells Express the Complement Inhibitor CD59

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Ágnes Füst ◽  
Éva Pállinger ◽  
Adrienn Stündl ◽  
Eszter Kovács ◽  
László Imre ◽  
...  
Keyword(s):  
Cell Surface ◽  
Amniotic Membrane ◽  
Ocular Surface ◽  
Cell Types ◽  
Surface Expression ◽  
Mesenchymal Cells ◽  
Cell Surface Expression ◽  
Complement Inhibitor ◽  
Becton Dickinson ◽  
Intracellular Expression

Amniotic membrane proved to be very effective tool in the treatment of a number of ocular surface diseases. The amniotic membrane, however, has to be stored before its transplantation onto the ocular surface followed by mandatory serologic control in order to exclude the transmission of certain viruses. Therefore it is most important to study if cryopreservation of the membrane affects cell surface expression of the molecules. We measured cell surface expression of CD59, a membrane-bound complement inhibitor on the cells of freshly prepared and cryopreserved amniotic membrane. Cells of amniotic membrane were separated mechanically. Epithelial and mesenchymal cells were identified by the intracellular expression of nanog and the cell surface ICAM1 positivity, respectively. Multicolor flow cytometric immunophenotyping was used for determination of the CD59 expression. CellQuest-Pro software program (Becton Dickinson) was used both for measurements and analysis. CD59-positive cells could be detected in all investigated samples and in all investigated cell types, although the expression level of CD59 differed. CD59 was expressed both on freshly prepared and frozen-stored samples. Higher level of CD59 was detected on ICAM1+ mesenchymal cells than on nanog+ epithelial cells. Our findings indicate that amniotic membranes maintain their complement inhibiting capacity after cryopreservation.

scholarly journals Distribution of complement receptors on human normal and malignant mononuclear cells

Blood ◽  
1980 ◽  
Vol 56 (5) ◽  
pp. 792-797
Author(s):  
RB Slease ◽  
JE Gadek ◽  
MM Frank ◽  
I Scher
Keyword(s):  
Fluorescence Intensity ◽  
Mononuclear Cells ◽  
Human Subjects ◽  
Normal Subjects ◽  
Cell Types ◽  
Lymphocytic Leukemia ◽  
Homogeneous Distribution ◽  
Cell Leukemia ◽  
Peripheral Blood Mononuclear ◽  
Heterogeneous Distribution

Mononuclear cells from normal human subjects and patients with chronic lymphocytic leukemia (CLL), chronic lymphosarcoma cell leukemia (LCL), and hairy cell leukemia (HCL) were labeled with fluoresceinated, purified human C3b (FI-C3b) and analyzed using the fluorescence- activated cell sorter (FACS). FI-C3b labeled 17.6% +/- 6.0% of peripheral blood mononuclear cells (PBM) from 20 normal subjects, which, when separated by the FACS, consisted of B lymphocytes and approximately 5% monocytes. Analyses in which either monocytes or B lymphcoytes were excluded from consideration demonstrated that both these cell types were labeled by the FI-C3b with a heterogeneous distribution of fluorescence intensity, indicating either heterogeneity of CR density or variable avidity of individual CR for the FI-C3b. FACS profiles of PBM ( < 5% monocytes) from 14 of 15 patients with CLL showed a homogeneous distribution of very low fluorescence intensity, with > 60% of the cells being slightly more fluorescent than unlabeled controls. This low, homogeneous distribution of fluorescence is strikingly similar to profiles of CLL cells labeled with anti-Ig reagents and suggests homogeneity of low CR density and/or avidity. Similarly, CR+ mononuclear cells from five patients with HCL and three patients with LCL displayed more homogeneous FI-C3b labeling than normal CR+ PBM. Homogeneity of FI-C3b binding to CLL, LCL, and HCL cells further supports the concept for a clonal origin for these disorders.

scholarly journals Human lymphocyte motility: normal characteristics and anomalous behavior of chronic lymphocytic leukemia cells

Blood ◽  
1976 ◽  
Vol 48 (5) ◽  
pp. 717-729 ◽  
Author(s):  
SC Jarvis ◽  
R Snyderman ◽  
HJ Cohen
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Human Lymphocyte ◽  
Human Lymphocytes ◽  
Surface Membrane ◽  
Cell Types ◽  
Anomalous Behavior ◽  
Lymphocytic Leukemia ◽  
Boyden Chamber ◽  
Surface Receptor ◽  
Lymphocyte Motility

Abstract The characteristics of human lymphocyte motility and its relationship to the redistribution of surface membrane antigens (capping) are poorly defined. Since chronic lymphocytic leukemia (CLL) cells cap poorly when compared with normal human lymphocytes, this study was undertaken to compare the motility of these two cell types. A modification of the Boyden chamber system was employed to quantify lymphocyte motility by placing lymphocyte suspensions on 8-mum convoluted-pore nitrocellulose filters and measuring the depth of migration of the cells into the filter at 37 degrees C. After 3 hr of incubation, CLL cells migrated significantly less into the filter than normal cells. Incubation in the presence of sodium azide or at 4 degrees C abolished all motility, indicating the active nature of the process. The relative motility of individual CLL patients' cells correlated best with the proportion of abnormal cells present as determined by surface receptor assays. The possibility that decreased cell motility in CLL was a reflection of enrichment by a “bone marrow-derived” (B cell) population was eliminated by the finding that normal B cells purified by gradient separation of rosetted cells migrated faster than normal T cells and considerably faster than CLL cells. Motility of normal and CLL lymphocytes was decreased by cytochalasin B and increased by colchicine, vincristine, and vinblastine. Thus, human lymphocyte motility appears to be dependent on microfilament integrity but not to require the colchicine-sensitive cytoskeleton. The decreased motility of CLL cells is the result of an intrinsic cell abnormality, but this finding cannot fully explain the decreased capping, since in human lymphocytes the latter is not prevented by an inhibitor of motility.

scholarly journals The surface morphology of human B lymphocytes as revealed by immunoelectron microscopy.

1975 ◽  
Vol 141 (2) ◽  
pp. 392-410 ◽  
Author(s):  
R J Lejonc ◽  
M F Gourdin ◽  
P Mannoni ◽  
B Dreyfus ◽  
F Reyes
Keyword(s):  
B Lymphocytes ◽  
Immunoelectron Microscopy ◽  
Binding Capacity ◽  
Human Lymphocytes ◽  
Lymphocytic Leukemia ◽  
Surface Appearance ◽  
Normal Individuals ◽  
Villous Surface ◽  
Intermediate Cells ◽  
Immunoglobulin Binding

Surface immunoglobulins (sIg) were detected on human lymphocytes by immunoelectron microscopy with peroxidase-conjugated antibodies. Blood, marrow, and thymus cells from normal individuals and patients with lymphoproliferative disorders were examined. Samples were fixed before exposure to specific reagents. Normal lymphocyts with detectable sIg, i.e. B lymphocytes, were characterized by a villous surface; nonlabeled blood lymphocytes and thymocytes were smooth cells. Intermediate cells were also found which in sections appeared moderately villous and labeled, thus identified as B lymphocytes. Further evidence for a relationship between villous surface and sIg was given by the finding of a few lymphocytes with polar concentration of labeled microvilli. In chronic lymphocytic leukemia patients, most cells exhibited a villous surface with parallel variations of the number of microvilli and of anti-immunoglobulin-binding capacity. However, some labeled smooth blastic cells were also observed. On the other hand, abnormal lymphocytes from Sézary's syndrome which could exhibit segments of villous membrane had no detectable sIg. This study confirms that in most cases human B lymphocytes have a characteristic surface appearance and that the detection of sIg in normal lymphocytes correlates with the presence of microvilli.

scholarly journals Abnormal levels of the α chain of the CD22 adhesion molecule may account for low CD22 surface expression in chronic lymphocytic leukemia

Leukemia ◽  
2006 ◽  
Vol 20 (5) ◽  
pp. 877-878 ◽  
Author(s):  
B Payelle-Brogard ◽  
G Dumas ◽  
C Magnac ◽  
A I Lalanne ◽  
G Dighiero ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Adhesion Molecule ◽  
Surface Expression ◽  
Lymphocytic Leukemia ◽  
Α Chain

scholarly journals Microspectrophotometric Estimation of the Desoxyribonucleic Acid (DNA) Content of Individual Normal and Leukemic Human Lymphocytes

Blood ◽  
1953 ◽  
Vol 8 (10) ◽  
pp. 905-915 ◽  
Author(s):  
NICHOLAS L. PETRAKIS
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Dna Content ◽  
Acute Lymphocytic Leukemia ◽  
Human Lymphocytes ◽  
Normal Subjects ◽  
Lymphocytic Leukemia ◽  
Desoxyribonucleic Acid ◽  
Dna Values ◽  
Circulating Lymphocytes ◽  
Normal Human

Abstract The DNA content, in arbitrary units, of individual circulating lymphocytes from nine normal subjects, nine patients with chronic lymphocytic leukemia, and five patients with acute lymphocytic leukemia was estimated microspectrophotometrically with the Feulgen dye. Normal circulating lymphocytes were found to contain twice the average DNA content of normal human spermatids, corroborating their diploid chromosome number. Lymphocytes from normal and leukemic lymph node and bone marrow were frequently found possessing four times the average spermatid DNA value. Three out of nine patients with chronic lymphocytic leukemia, and four of five patients with acute lymphocytic leukemia had significantly increased numbers of circulating lymphocytes containing DNA values which were elevated above the normal diploid value. The patients demonstrating cells with elevated DNA values were in a clinically exacerbated phase of their disease. The significance of these findings is discussed.

scholarly journals Adenosine and adenosine analogues are more toxic to chronic lymphocytic leukemia than to normal lymphocytes

Blood ◽  
1983 ◽  
Vol 62 (1) ◽  
pp. 75-80 ◽  
Author(s):  
S Bajaj ◽  
J Insel ◽  
F Quagliata ◽  
R Hirschhorn ◽  
R Silber
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Lymphocytes ◽  
Proliferative Response ◽  
Differential Susceptibility ◽  
Normal Subjects ◽  
Lymphocytic Leukemia ◽  
Leucine Incorporation ◽  
Adherent Cells ◽  
Adenosine Analogues ◽  
Residual Inhibition

Abstract We compared the effect of adenosine and adenosine analogues on the phytohemagglutinin-induced proliferative response of blood lymphocytes from normal subjects and patients with chronic lymphocytic leukemia. As measured by the inhibition of thymidine or leucine incorporation, adenosine was more toxic to chronic lymphocytic leukemia (CLL) than to normal lymphocytes. This difference was not affected by the removal of adherent cells. The patients' B lymphocytes were more susceptible to adenosine toxicity than normal B lymphocytes. Similar responses were noted in T lymphocytes from both sources. Differential susceptibility was also observed with deoxyadenosine and adenosine analogues, including 5′deoxyadenosine. Uridine rescue from adenosine toxicity was observed for normal and CLL lymphocytes. In the presence of uridine, there was no difference in the residual inhibition of CLL as compared to normal lymphocytes. Intact CLL lymphocytes metabolized 14C-adenosine at a much lower rate than normal lymphocytes. While it appears that the greater toxicity of adenosine to CLL lymphocytes reflects the impaired catabolism of this nucleoside by these cells, evidence is presented that this is not the only mechanism underlying the differential susceptibility. These results may serve as the basis for further pharmacologic investigations of adenosine and adenosine deaminase inhibitors in chronic lymphocytic leukemia.

scholarly journals Adenosine and adenosine analogues are more toxic to chronic lymphocytic leukemia than to normal lymphocytes

Blood ◽  
1983 ◽  
Vol 62 (1) ◽  
pp. 75-80 ◽  
Author(s):  
S Bajaj ◽  
J Insel ◽  
F Quagliata ◽  
R Hirschhorn ◽  
R Silber
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Lymphocytes ◽  
Proliferative Response ◽  
Differential Susceptibility ◽  
Normal Subjects ◽  
Lymphocytic Leukemia ◽  
Leucine Incorporation ◽  
Adherent Cells ◽  
Adenosine Analogues ◽  
Residual Inhibition

We compared the effect of adenosine and adenosine analogues on the phytohemagglutinin-induced proliferative response of blood lymphocytes from normal subjects and patients with chronic lymphocytic leukemia. As measured by the inhibition of thymidine or leucine incorporation, adenosine was more toxic to chronic lymphocytic leukemia (CLL) than to normal lymphocytes. This difference was not affected by the removal of adherent cells. The patients' B lymphocytes were more susceptible to adenosine toxicity than normal B lymphocytes. Similar responses were noted in T lymphocytes from both sources. Differential susceptibility was also observed with deoxyadenosine and adenosine analogues, including 5′deoxyadenosine. Uridine rescue from adenosine toxicity was observed for normal and CLL lymphocytes. In the presence of uridine, there was no difference in the residual inhibition of CLL as compared to normal lymphocytes. Intact CLL lymphocytes metabolized 14C-adenosine at a much lower rate than normal lymphocytes. While it appears that the greater toxicity of adenosine to CLL lymphocytes reflects the impaired catabolism of this nucleoside by these cells, evidence is presented that this is not the only mechanism underlying the differential susceptibility. These results may serve as the basis for further pharmacologic investigations of adenosine and adenosine deaminase inhibitors in chronic lymphocytic leukemia.

scholarly journals CDw60 glycolipid antigens of human leukocytes: structural characterization and cellular distribution

Blood ◽  
1993 ◽  
Vol 82 (6) ◽  
pp. 1776-1786
Author(s):  
B Kniep ◽  
WA Flegel ◽  
H Northoff ◽  
EP Rieber
Keyword(s):  
T Lymphocytes ◽  
B Lymphocytes ◽  
Human Leukocyte ◽  
Cell Types ◽  
Surface Expression ◽  
Cellular Distribution ◽  
Human Leukocytes ◽  
Costimulatory Signals ◽  
Thymus Cells ◽  
Derivatives Of

Monoclonal CDw60 antibodies recognize glycolipid antigens with restricted surface expression on human leukocytes. They allow us to define new functional subpopulations of T lymphocytes and are able to induce costimulatory signals. In this report, we describe the molecular composition of CDw60 glycolipid antigens derived from different human leukocyte subpopulations. The glycolipids were isolated and their structures were identified by immunochemical methods. All molecules containing the CDw60 determinant were found in the disialoganglioside fraction. They were O-acetylated derivatives of the gangliosides II3 (Neu5Ac)2-LacCer (GD3), IV3 (Neu5Ac)2-nLc4Cer (DSPG), and VI3 (Neu5Ac)2- nLc6Cer (DSnHC), respectively. The most common CDw60 glycolipid antigen in human leukocytes was 9-O-acetyl GD3. In a comparison of various cell types, the highest concentration of 9-O-acetyl GD3 on a per cell basis was determined in granulocytes and in blood T lymphocytes, whereas B lymphocytes, thymus cells, and monocytes contained considerably smaller amounts of this molecule. Polar CDw60 antigens such as 9-O-acetyl DSPG and 9-O-acetyl DSnHC were only detected in granulocytes.

scholarly journals Glutathione depletion in chronic lymphocytic leukemia B lymphocytes

Blood ◽  
1992 ◽  
Vol 80 (8) ◽  
pp. 2038-2043 ◽  
Author(s):  
R Silber ◽  
CM Farber ◽  
E Papadopoulos ◽  
D Nevrla ◽  
L Liebes ◽  
...  
Keyword(s):  
T Cells ◽  
Chronic Lymphocytic Leukemia ◽  
B Lymphocytes ◽  
B Lymphocyte ◽  
Alkylating Agents ◽  
Normal Subjects ◽  
Lymphocytic Leukemia ◽  
Glutathione Depletion ◽  
Cd8 Cells ◽  
Baseline Values

Abstract Glutathione (GSH) content may be the major determinant of a cell's sensitivity to cytotoxic alkylating agents. In the present study, the GSH concentration was determined in lymphocytes isolated from the blood of normal subjects and patients with chronic lymphocytic leukemia (CLL). Comparable levels were found in both types of cells. Incubation for 20 hours led to a decrease in GSH to 51% of baseline values in CLL B cells. Under the same conditions, normal B- or T-lymphocyte GSH content remained constant. GSH depletion was shown to be a characteristic of the B-CLL B lymphocyte. It was not found in the T cells of patients with B-CLL or in cells from patients with T-CLL. Chlorambucil (CLB) contributes to the decrease in GSH in B-CLL lymphocytes; after incubation with the drug, lower levels of GSH were found than in the normal B or T lymphocytes, B-CLL T cells, or T-CLL (CD4 or CD8) cells. GSH depletion of CLL B lymphocytes may be related to the greater therapeutic efficacy of CLB in B-CLL than in T-CLL.

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