Flow cytometric analysis of expression of interleukin-2 receptor beta chain (p70-75) on various leukemic cells

Blood ◽  
1990 ◽  
Vol 76 (4) ◽  
pp. 767-774 ◽  
Author(s):  
S Hoshino ◽  
K Oshimi ◽  
M Tsudo ◽  
M Miyasaka ◽  
M Teramura ◽  
...  
Keyword(s):  
T Cell ◽  
Lymphoblastic Leukemia ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Lymphocytic Leukemia ◽  
Leukemic Cells ◽  
Beta Chain ◽  
Alpha Chain ◽  
Interleukin 2 Receptor ◽  
Flow Cytometric

Abstract We analyzed the expression of the interleukin-2 receptor (IL-2R) beta chain (p70–75) on various leukemic cells from 44 patients by flow cytometric analysis using the IL-2R beta chain-specific monoclonal antibody (MoAb), designated Mik-beta 1, which has been recently developed. Flow cytometric analysis demonstrated the expression of the IL-2R beta chain on granular lymphocytes (GLs) from all eight patients with granular lymphocyte proliferative disorders (GLPDs), on adult T- cell leukemia (ATL) cells from all three patients with ATL, and on T- cell acute lymphoblastic leukemia (T-ALL) cells from one of three patients with T-ALL. Although GLs from all the GLPD patients expressed the IL-2R beta chain alone and not the IL-2R alpha chain (Tac-antigen: p55), ATL and T-ALL cells expressing the beta chain coexpressed the alpha chain. In two of seven patients with common ALL (cALL) and in both patients with B-cell chronic lymphocytic leukemia, the leukemic cells expressed the alpha chain alone. Neither the alpha chain nor the beta chain was expressed on leukemic cells from the remaining 28 patients, including all 18 patients with acute nonlymphocytic leukemia, five of seven patients with cALL, all three patients with multiple myeloma, and two of three patients with T-ALL. These results indicate that three different forms of IL-2R chain expression exist on leukemic cells: the alpha chain alone; the beta chain alone; and both the alpha and beta chains. To examine whether the results obtained by flow cytometric analysis actually reflect functional aspects of the expressed IL-2Rs, we studied the specific binding of 125I-labeled IL-2 (125I-IL-2) to leukemic cells in 18 of the 44 patients. In addition, we performed 125I-IL-2 crosslinking studies in seven patients. The results of IL-2R expression of both 125I-IL-2 binding assay and crosslinking studies were in agreement with those obtained by flow cytometric analysis. These results indicate that flow cytometric analysis using MoAbs, anti-Tac, and Mik-beta 1 is useful for detecting the expression of the IL-2R chains.

scholarly journals Flow cytometric analysis of expression of interleukin-2 receptor beta chain (p70-75) on various leukemic cells

Blood ◽  
1990 ◽  
Vol 76 (4) ◽  
pp. 767-774 ◽  
Author(s):  
S Hoshino ◽  
K Oshimi ◽  
M Tsudo ◽  
M Miyasaka ◽  
M Teramura ◽  
...  
Keyword(s):  
T Cell ◽  
Lymphoblastic Leukemia ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Lymphocytic Leukemia ◽  
Leukemic Cells ◽  
Beta Chain ◽  
Alpha Chain ◽  
Interleukin 2 Receptor ◽  
Flow Cytometric

We analyzed the expression of the interleukin-2 receptor (IL-2R) beta chain (p70–75) on various leukemic cells from 44 patients by flow cytometric analysis using the IL-2R beta chain-specific monoclonal antibody (MoAb), designated Mik-beta 1, which has been recently developed. Flow cytometric analysis demonstrated the expression of the IL-2R beta chain on granular lymphocytes (GLs) from all eight patients with granular lymphocyte proliferative disorders (GLPDs), on adult T- cell leukemia (ATL) cells from all three patients with ATL, and on T- cell acute lymphoblastic leukemia (T-ALL) cells from one of three patients with T-ALL. Although GLs from all the GLPD patients expressed the IL-2R beta chain alone and not the IL-2R alpha chain (Tac-antigen: p55), ATL and T-ALL cells expressing the beta chain coexpressed the alpha chain. In two of seven patients with common ALL (cALL) and in both patients with B-cell chronic lymphocytic leukemia, the leukemic cells expressed the alpha chain alone. Neither the alpha chain nor the beta chain was expressed on leukemic cells from the remaining 28 patients, including all 18 patients with acute nonlymphocytic leukemia, five of seven patients with cALL, all three patients with multiple myeloma, and two of three patients with T-ALL. These results indicate that three different forms of IL-2R chain expression exist on leukemic cells: the alpha chain alone; the beta chain alone; and both the alpha and beta chains. To examine whether the results obtained by flow cytometric analysis actually reflect functional aspects of the expressed IL-2Rs, we studied the specific binding of 125I-labeled IL-2 (125I-IL-2) to leukemic cells in 18 of the 44 patients. In addition, we performed 125I-IL-2 crosslinking studies in seven patients. The results of IL-2R expression of both 125I-IL-2 binding assay and crosslinking studies were in agreement with those obtained by flow cytometric analysis. These results indicate that flow cytometric analysis using MoAbs, anti-Tac, and Mik-beta 1 is useful for detecting the expression of the IL-2R chains.

scholarly journals Maturation of acute T-lymphoblastic leukemia cells after CD2 ligation and subsequent treatment with interleukin-2

Blood ◽  
1994 ◽  
Vol 84 (4) ◽  
pp. 1182-1192 ◽  
Author(s):  
F Mentz ◽  
F Ouaaz ◽  
A Michel ◽  
C Blanc ◽  
P Herve ◽  
...  
Keyword(s):  
T Cell ◽  
Tyrosine Phosphorylation ◽  
Lymphoblastic Leukemia ◽  
Interleukin 1 ◽  
Interleukin 2 ◽  
Subsequent Treatment ◽  
Leukemic Cells ◽  
Cell Phenotype ◽  
Cell Functions

Abstract In this study, we have investigated the ability of various cytokines to induce the maturation of acute lymphoblastic leukemia (T-ALL) cells with early T-cell phenotype. Leukemic blasts from 17 untreated T-ALL patients were assayed for their ability to acquire mature T-cell markers, CD3/T-cell receptor (TCR) in particular, after incubation with one or a combination of recombinant human interleukin-1 (IL-1), IL-2, IL-4, IL-7, and CD2-specific monoclonal antibody (MoAb). IL-7 or IL-2 induced the proliferation of some leukemic cells, whereas sequential cell treatment with CD2-MoAb and then IL-2 promoted CD3/TCR expression on nearly all CD2+ cells (15 of 16), except for 1 T-ALL that developed into CD3-CD16+CD56+ cells. Differentiation of T-ALL cells was also evidenced through the downregulation of CD34 precursor cell antigen, the generation of CD4+ and CD8+ cells from CD4+ CD8+ precursors, and the acquisition of mature T-cell functions. CD2 ligation induced a progressive increase of surface expression of IL-2 receptor alpha (IL- 2R alpha) and IL-2R beta and an accelerated in vitro death of leukemic cells. The ligation of IL-2R by IL-2 rescued T-ALL cells from death and promoted their progression toward more mature cells expressing extracellular CD3/TCR alpha beta complexes. Intracellular analysis indicates that TCR alpha transcription and membrane translocation of both TCR alpha and TCR beta were promoted in these conditions. Analysis of intracellular signals transduced during T-ALL differentiation indicated that CD2-ligation induced Ca2+ influx and that the ligation of CD2 and IL-2R induced distinct tyrosine phosphorylation patterns. The addition of inhibitors of tyrosine phosphorylation abolished T-ALL cell differentiation, which suggests the involvement of tyrosine kinases in this phenomenon. Together, we showed the constant maturation of leukemic early T cells after stimulation of surface CD2 and the high- affinity IL-2R.

scholarly journals Interleukin-2 Receptor β-Chain (p70-75) Expressed on Leukemic Cells from Adult T Cell Leukemia Patients

1990 ◽  
Vol 81 (9) ◽  
pp. 902-908 ◽  
Author(s):  
Taiichi Kodaka ◽  
Takashi Uchiyama ◽  
Takayuki Ishikawa ◽  
Masanori Kamio ◽  
Rie Onishi ◽  
...  
Keyword(s):  
T Cell ◽  
Interleukin 2 ◽  
Leukemic Cells ◽  
T Cell Leukemia ◽  
Cell Leukemia ◽  
Adult T Cell Leukemia ◽  
Interleukin 2 Receptor ◽  
Β Chain

Analysis of In Vitro Effects of Bortezomib (Velcade) on T Cell Prolymphocytic Leukemia (T-PLL).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4426-4426
Author(s):  
Fulya Ozpuyan ◽  
Paul N. Meyer ◽  
Hytham Al-Masri ◽  
Hongyu Ni ◽  
Serhan Alkan
Keyword(s):  
T Cell ◽  
Lymphoproliferative Disorder ◽  
Flow Cytometric Analysis ◽  
Leukemic Cells ◽  
Cell Phenotype ◽  
Base Line ◽  
Prolymphocytic Leukemia ◽  
Bortezomib Treatment ◽  
Flow Cytometric

Abstract T-cell prolymphocytic leukemia (T-PLL) is an aggressive lymphoproliferative disorder with postthymic T cell phenotype and prolymphocytic morphology. In the majority of patients, the leukemic process progresses rapidly and patients die shortly after diagnosis (median survival of 7 months). Bortezomib, the first proteasome inhibitor to be approved for use in haematological malignancies such as multiple myeloma, is beginning to be utilized as an effective anti-neoplastic agent in other hematopoietic and non-hematopoietic neoplastic disorders. We report here the in vitro apoptotic effects of bortezomib on leukemic cells isolated from three T-PLL patients. Interestingly, one of the patient’s leukemia developed in the setting of immunosupression due to transplant therapy (post-transplant lymphoproliferative disorder). Flow cytometric analysis of leukemic cells of the three patients showed CD8, double CD4+CD8+ and double CD4−CD8− immunophenotypic features. All cases showed monoclonal band pattern by T-cell receptor (TCR) gene rearrangement as analyzed by the PCR amplification of the TCR gamma heavy chain gene. Freshly isolated leukemic cells with the CD8 phenotype T-PLL analyzed for apoptosis after ficoll hypaque separation and cultured in the presence of various concentration of Bortezomib (0.001, 0.01, 0.1, 1, and 10 uM) for dose curve analysis. Apoptosis of the leukemic cells was determined by Annexin-V and 7-AAD staining and flow cytometric analysis after incubation at 24 and 72 hours, respectively. Samples treated for 72 hours showed higher rate of apoptosis compared to 24 hours: 10 uM (62% increase above the base line of control cells), 1 uM (58%), 0.1 uM (55%), 0.01 uM (40%) and 0.001 uM (0%) concentrations while samples treated for 24 hours with 10 uM showed (42% increase above the base line of control cells) and 1 uM (33% increase above the base line). Light microscopic analysis of leukemic cells treated with Bortezomib confirmed that the majority of cells undergo apoptosis with Bortezomib treatment as it revealed nuclear fragmentation and apoptotic bodies. Leukemic cells recovered from cryopreservation from the second and third T-PLL patient samples analyzed also showed significant increase in early and late apoptosis at 24 hours with Bortezomib treatment (10nm). These results suggest that Bortezomib may provide an alternate therapy in the treatment of T-PLL. Future collaborative efforts investigating efficacy with Bortezomib as a single agent or in combination with other therapeutic agents will be crucial to improving survival for patients with this disease.

Prevalence of Immunophenotypes in Neoplastic Hematology At Laboratorios Fatima De Michoacan

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4829-4829
Author(s):  
Gregorio Campos-Cabrera ◽  
Salvador Campos-Cabrera ◽  
Virgina Campos-Cabrera ◽  
Maria Mora-Torres ◽  
Miguel-Angel Gomez-Guijosa ◽  
...  
Keyword(s):  
T Cell ◽  
Lymphoblastic Leukemia ◽  
Lymphoproliferative Disorders ◽  
Lymphocytic Leukemia ◽  
Cell Leukemia ◽  
Cell Precursor ◽  
Aberrant Expression ◽  
Flow Cytometric ◽  
Monoclonal Gammapathies ◽  
Low Prevalence

Abstract Abstract 4829 Introduction: Medical indication of flow cytometric immunophenotyping includes diagnosis, classification, prognosis and disease monitoring. This is an important tool that each time is more used in hematology. Third world countries could not stay away from this technology, effort should be made to get access to it and not only make diagnosis in morphology. Objective: To determinate the prevalence of immunophenotypes in neoplastic hematology in our region (Central-West Mexico). Material and Methods: Bone marrow samples referred to Laboratorios Fatima de Michoacan for flow cytometry immunophenotyping for neoplastic hematological disease. The protocol is based on the Report on the Second Latin American Consensus Conference for Flow Cytometric Immunophenotyping of Hematological Malignancies (Cytometry Part B (Clinical Cytometry) 2005;70B:39–44), and Immunophenotyping of acute leukemia and lymphoproliferative disorders: a consensus proposal of the European LeukemiaNet Work Package 10 (Leukemia 2011;25:567–574). Results: One hundred and seventy two cases were diagnosed. Fourteen myelodisplastic syndromes were detected. Forty five acute myeloid leukemia; M0-M1 twenty two cases, 5 with aberrant expression of CD7, one with CD19 and one CD20; M2 four cases, 1 with aberrant expression of CD 2 and 1 with CD7; M3 four cases; M4 one case; M5 twelve cases, 3 aberrant expression of CD7; M6 and M7 one case each. Sixty five B cell precursor acute lymphoblastic leukemia; BI twenty three cases, 2 with aberrant expression of CD7, and 2 aberrant expression of CD13 and one with aberrant expression of CD 33; BII twenty cases, 1 aberrant expression of CD2, 1 aberrant expression of CD5, and 4 aberrant expression of CD33; BIII twenty one cases, 1 aberrant expression of CD3; BIV one case. Five T cell precursor acute lymphoblastic leukemia. Thirty four chronic lymphoproliferative disorders; eighteen B cell chronic lymphocytic leukemia, two T cell chronic lymphocytic leukemia, seven follicular lymphoma, three mantle cell lymphoma, three splenic lymphoma with villous lymphocytes and one hairy cell leukemia One adult T cell leukemia/lymphoma. One natural killer cell leukemia (CD94+, perforin +, granzyme +) Seven monoclonal gammapathies. Conclusions: It is important to create the experience with new diagnostic tools based on the regional protocols. Low prevalence of AML M2 presumably because of classic morphologic features. Low prevalence of monoclonal gammapathies because for recent incorporation to diagnosis, treatment and response criteria; it is expected that in future this prevalence will arise. This data is complemented, whenever it is possible, with chromosomal analysis to determinate the risk and treatment. Disclosures: No relevant conflicts of interest to declare.

Hyperdiploid Acute Lymphoblastic Leukemia With 51 to 65 Chromosomes: A Distinct Biological Entity With a Marked Propensity to Undergo Apoptosis

Blood ◽  
1999 ◽  
Vol 93 (1) ◽  
pp. 315-320 ◽  
Author(s):  
Chikako Ito ◽  
Masa-aki Kumagai ◽  
Atsushi Manabe ◽  
Elaine Coustan-Smith ◽  
Susana C. Raimondi ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Lymphoblastic Leukemia ◽  
Interleukin 1 ◽  
Flow Cytometric Analysis ◽  
Growth Potential ◽  
Leukemic Cells ◽  
Biological Entity ◽  
Allogeneic Bone ◽  
Cell Recovery ◽  
Flow Cytometric

To determine the cellular basis for the excellent clinical outcome of hyperdiploid acute lymphoblastic leukemia (ALL), defined by a modal chromosome number of 51 to 65, we assessed the growth potential of leukemic cells from 129 children with newly diagnosed ALL. Flow cytometric analysis was used to compare leukemic cell recoveries at the beginning and at the end of 7-day cultures on allogeneic bone marrow–derived stromal layers. The median percentage of cell recovery after culture was 91% (range, <1% to 550%). Among the 25 hyperdiploid cases, only two had cell recoveries above the median value, compared with 63 of 104 cases with different ploidies (P< .001); 21 had recoveries within the first quartile, in contrast to only 12 of the 104 other cases. Cell recoveries in the 16 cases with duplications of chromosomes 4 and 10, a feature previously associated with a superior outcome, were all within the first quartile. Flow cytometric studies indicated that rapid induction of apoptosis was the underlying cause of low cell recoveries in cases with hyperdiploidy. The demise of hyperdiploid cells on stroma was not due to failure to adhere with stromal elements (as shown by electron microscopy) or to deficiencies of interleukin-1 (IL-1), IL-2, IL-3, IL-4, IL-6, IL-7, IL-11, stem-cell factor, interferon- (IFN-), tumor necrosis factor- (TNF-), or to combinations of these cytokines. Inactivation of IL-4, IFN- and TNF-, which if secreted by stromal layers could be toxic to ALL cells, failed to improve the survival of hyperdiploid blasts. We conclude that leukemic cells bearing 51 to 65 chromosomes have a marked propensity to undergo apoptosis. The stringent survival requirements of these cells, together with their potentially higher sensitivity to antileukemic drugs, may well account for the high cure rates achieved in patients with this form of ALL.

scholarly journals Expression of two alpha chains on the surface of T cells in T cell receptor transgenic mice.

1993 ◽  
Vol 178 (5) ◽  
pp. 1807-1811 ◽  
Author(s):  
W R Heath ◽  
J F Miller
Keyword(s):  
T Cells ◽  
T Cell ◽  
Transgenic Mice ◽  
T Cell Receptor ◽  
Cd8 T Cells ◽  
Flow Cytometric Analysis ◽  
Cell Receptor ◽  
Specific Monoclonal Antibody ◽  
Beta Chain ◽  
Flow Cytometric

CD8+ T cells taken directly from mice expressing a Kb-specific T cell receptor (TCR) transgene expressed the transgenic TCR in a bimodal profile as detected by flow cytometric analysis using a clonotype-specific monoclonal antibody. Those cells expressing the lower density of the transgenic TCR expressed the transgenic beta chain and two different alpha chains on their surface. One alpha chain was the product of the alpha transgene, whereas the other was derived by endogenous rearrangement. This report provides the first demonstration that T cells isolated directly from mice may express two different TCR clonotypes on their surface. The potential consequences of this finding for studies using TCR transgenic mice and for the induction of autoimmunity are discussed.

scholarly journals Establishment of a karyotypically normal cytotoxic leukemic T-cell line from a T-ALL sample engrafted in SCID mice

Blood ◽  
1993 ◽  
Vol 81 (10) ◽  
pp. 2714-2722
Author(s):  
A Cesano ◽  
R O'Connor ◽  
PC Nowell ◽  
B Lange ◽  
SC Clark ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Line ◽  
Chromosomal Abnormalities ◽  
Lymphoblastic Leukemia ◽  
Interleukin 2 ◽  
Cell Subset ◽  
Scid Mice ◽  
Leukemic Cells ◽  
Cytotoxic T Cell ◽  
Tumoricidal Activity

Bone marrow (BM) cells from a child with an immature (CD3-) acute T lymphoblastic leukemia (T-ALL) bearing no chromosomal abnormalities failed to grow in long-term culture in the presence or absence of recombinant human (rh) growth factors but could be engrafted in severe combined immunodeficient (SCID) mice and induced leukemia. The leukemic cells recovered from the animal tissues could be adapted to grow in vitro in the presence of rh interleukin-2 (IL-2) and give rise to a growth factor-dependent cell line designated TALL-107. This cell line expresses T-cell-specific mature markers (CD2, CD3/T-cell receptor [TCR] alpha beta, CD8, CD56), and its growth can be inhibited by IL-4 of all the cytokines tested. Similar to the original leukemic blasts, TALL-107 cells are clonal, have rearranged TCR-beta, gamma, and delta loci, and a normal 46 XY karyotype. However, unlike the patient's BM cells, the TALL-107 cell line displays potent tumoricidal activity that is not major histocompatibility complex restricted. The magnitude of mRNA expression of perforin and serine esterases and of lytic activity depends on the doses of IL-2 added. TALL-107 cells can also be triggered by CD3- and CD2-specific monoclonal antibodies (MoAbs) to mediate reverse tumor cell lysis. In addition, this cell line produces high levels of interferon gamma and tumor necrosis factor alpha on stimulation with anti-CD3 and/or anti-CD2 MoAb both in the presence or absence of IL-2. The overall data indicate that the SCID mouse is able to support the functional maturation and expansion of a cytotoxic T- cell subset from some types of T-ALL.

Genetic expression of adenosine deaminase in human lymphoid malignancies

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1376-1380 ◽  
Author(s):  
TE Gan ◽  
PE Dadonna ◽  
BS Mitchell
Keyword(s):  
T Cell ◽  
Adenosine Deaminase ◽  
Cell Lines ◽  
Specific Activity ◽  
Lymphoblastic Leukemia ◽  
High Specific Activity ◽  
Lymphocytic Leukemia ◽  
Leukemic Cells ◽  
Gene Copy ◽  
Immunoreactive Protein

Abstract Adenosine deaminase (ADA) is an enzyme in the purine catabolic pathway that has been used as an enzymatic marker of T cell lymphoblastic malignancies due to its high specific activity in thymocytes and immature T cells. We have investigated whether the level of ADA activity in lymphoid leukemic cells correlates with the amount of ADA- specific RNA and/or immunoreactive protein in these cells as an initial step toward characterizing the nature of the genetic regulation of ADA expression during differentiation. We have found a good correlation between the steady state levels of ADA-specific RNA and ADA- immunoreactive protein in T lymphoblastic leukemic cell lines, mature T cell lines, a B lymphoblast cell line, and leukemic cells directly isolated from four patients with acute lymphoblastic leukemia and three patients with chronic lymphocytic leukemia. Southern blot analysis of DNA from these cells shows no evidence for differences in ADA gene copy number or gene rearrangement to account for the variability in ADA expression. We conclude that levels of ADA in lymphoid leukemic cells are directly related to the amount of ADA-specific mRNA present. These findings imply that ADA expression in leukemic cells reflects either the transcriptional activity of the ADA gene or the stability of ADA mRNA in these cells.

Sign in / Sign up

Export Citation Format

Share Document