scholarly journals A new solid-phase immunoassay for terminal deoxynucleotidyl transferase: analysis of TdT antigen in cells, plasma, and serum

Blood ◽  
1985 ◽  
Vol 65 (1) ◽  
pp. 41-45 ◽  
Author(s):  
MS Coleman ◽  
ML Cibull ◽  
GL Manderino
Keyword(s):  
Bone Marrow ◽  
Enzyme Activity ◽  
Horseradish Peroxidase ◽  
Peripheral Blood ◽  
Solid Phase ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Polystyrene Bead ◽  
Secondary Antibody ◽  
Terminal Transferase ◽  
Solid Phase Immunoassay

Abstract A solid-phase immunoassay for terminal deoxynucleotidyl transferase has been developed using a primary antibody-coated polystyrene bead and secondary antibody conjugated with horseradish peroxidase. The immunoassay was compared with assays for enzyme activity and detection of antigen with immunofluorescence using cells from peripheral blood and bone marrow from patients with leukemia or lymphoma. In each instance, the solid-phase immunoassay correlated correctly with cellular samples judged to be positive by other tests. However, the level of detection of terminal transferase antigen in plasma or serum of patients with leukemia did not reflect accurately the level of terminal transferase in neoplastic cells. The solid-phase immunoassay was greater than 100-fold more sensitive than conventional assays for enzyme activity, rendering it potentially useful for quantitatively monitoring terminal transferase in patients with leukemia.

scholarly journals A new solid-phase immunoassay for terminal deoxynucleotidyl transferase: analysis of TdT antigen in cells, plasma, and serum

Blood ◽  
1985 ◽  
Vol 65 (1) ◽  
pp. 41-45
Author(s):  
MS Coleman ◽  
ML Cibull ◽  
GL Manderino
Keyword(s):  
Bone Marrow ◽  
Enzyme Activity ◽  
Horseradish Peroxidase ◽  
Peripheral Blood ◽  
Solid Phase ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Polystyrene Bead ◽  
Secondary Antibody ◽  
Terminal Transferase ◽  
Solid Phase Immunoassay

A solid-phase immunoassay for terminal deoxynucleotidyl transferase has been developed using a primary antibody-coated polystyrene bead and secondary antibody conjugated with horseradish peroxidase. The immunoassay was compared with assays for enzyme activity and detection of antigen with immunofluorescence using cells from peripheral blood and bone marrow from patients with leukemia or lymphoma. In each instance, the solid-phase immunoassay correlated correctly with cellular samples judged to be positive by other tests. However, the level of detection of terminal transferase antigen in plasma or serum of patients with leukemia did not reflect accurately the level of terminal transferase in neoplastic cells. The solid-phase immunoassay was greater than 100-fold more sensitive than conventional assays for enzyme activity, rendering it potentially useful for quantitatively monitoring terminal transferase in patients with leukemia.

Solid-phase enzyme immunoassay of terminal deoxynucleotidyl transferase evaluated.

1987 ◽  
Vol 33 (2) ◽  
pp. 293-296 ◽  
Author(s):  
M Fleisher ◽  
R Stankievic ◽  
D Schwartz ◽  
M K Schwartz
Keyword(s):  
Bone Marrow ◽  
Whole Blood ◽  
Mononuclear Cells ◽  
Solid Phase ◽  
Linear Regression Analysis ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Human Leukemia ◽  
Solid Phase Immunoassay ◽  
Analysis Of Results ◽  
Good Concordance

Abstract We evaluated a newly developed solid-phase immunoassay (EIA) of terminal deoxynucleotidyl transferase (TdT, EC 2.7.7.3) and compared it with the enzymatic assay of TdT involving DNA polymerase. We assessed the precision, performance characteristics, and clinical efficacy of the EIA procedure, using 249 specimens of peripheral blood and bone marrow and 118 specimens of whole blood. On linear regression analysis of results for these 249 samples as measured by the two procedures, the correlation coefficient was 0.87. Distribution of TdT in mononuclear cells isolated from whole blood and bone marrow of subjects in several disease categories indicated good concordance between the two assay procedures. The EIA procedure is precise, can be performed on whole blood without first isolating mononuclear cells, is nonisotopic, and shows potential as a quantitative indicator for the differential diagnosis and monitoring of human leukemia.

scholarly journals Prognostic significance of terminal transferase activity in childhood acute lymphoblastic leukemia: a prospective analysis of 164 patients

Blood ◽  
1982 ◽  
Vol 60 (6) ◽  
pp. 1267-1276 ◽  
Author(s):  
JJ Hutton ◽  
MS Coleman ◽  
S Moffitt ◽  
MF Greenwood ◽  
P Holland ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukemia ◽  
Complete Remission ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Lymphoblastic Leukemia ◽  
Prognostic Significance ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Central Nervous System Relapse ◽  
Transferase Activity

Abstract Whether the level of terminal deoxynucleotidyl transferase (TdT) activity in mononuclear cells from bone marrow and peripheral blood has prognostic significance has been analyzed prospectively in 164 children with T and non-T, non-B marked acute lymphoblastic leukemia (ALL). TdT was measured at diagnosis to assess its value as a predictor of duration of remission and length of survival. It was measured repeatedly during remission to assess whether it could predict relapse. Ninety-seven percent of the children achieved a complete remission of their disease, and 40% relapsed during the study. The level of TdT activity in blasts at diagnosis varied 1000-fold from patient to patient. There was no statistically significant relationship between TdT activity in cells at diagnosis and the achievement of complete remission, the duration of remission, or length of survival. TdT activity was significantly increased in the bone marrow of 65% of patients at the time of marrow morphological relapse, but was rarely increased in marrow from patients with isolated testicular or central nervous system relapse. Wide fluctuations in TdT activity were characteristically seen in mononuclear cells from the marrow and peripheral blood of patients with ALL at all stages of their disease. An isolated high value of TdT activity in the bone marrow or peripheral blood cannot be taken as evidence of impending relapse. Quantitative measurements of TdT activity alone on mononuclear cells from bone marrow and peripheral blood are helpful in differential diagnosis, but cannot guide therapy of children with ALL.

scholarly journals Terminal deoxynucleotidyl transferase is found in prothymocytes

1976 ◽  
Vol 144 (2) ◽  
pp. 543-548 ◽  
Author(s):  
AE Silverstone ◽  
H Cantor ◽  
G Goldstein ◽  
D Baltimore
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Bone Marrow Cells ◽  
Marrow Cell ◽  
T Cell Differentiation ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Murine Bone Marrow ◽  
Terminal Transferase ◽  
Separate Pathway ◽  
Marrow Cells

Terminal deoxynucleotidyl transferase is an enzyme which has the unique property of polymerizing polydeoxynucleotides onto a primer in the absence of a template (1,2). This enzyme is found both in the thymus and the bone marrow of birds, rodents, and humans (3-7). Whether the marrow cells that contain terminal transferase are related to thymocytes, or are on a separate pathway of differentiation, is not yet known (7,8). To determine the lineage of the murine bone marrow cells that have terminal transferase, we have investigated whether these cells have the antigen Thy-1 induced on the cells by treatment with thymopoietin (9). Thymopoietin is known to induce a set of characteristic T-cell markers including the Thy-1 alloantigen on the surface of a subpopulation of bone marrow cells committed to T-cell differentiation (prothymocytes) (10). Destruction of Thy- 1-positive cells after exposure to thymopoietin allows elimination of a substantial fraction of those bone marrow cells that can repopulate an irradiated thymus (11). We find that such an elimination after induction with the thymic polypeptide removes a substantial amount of terminal transferase from the bone marrow cell population, suggesting that at least one-half of the marrow cells bearing this enzyme are related to those found in the thymus.

scholarly journals Some Factors Involved in the Effect of X-Irradiation on the Phosphatase Activity of Hematopoietic Tissues

Blood ◽  
1959 ◽  
Vol 14 (10) ◽  
pp. 1128-1136 ◽  
Author(s):  
EDWIN M. UYEKI ◽  
PAUL R. SALERNO
Keyword(s):  
Bone Marrow ◽  
Enzyme Activity ◽  
Peripheral Blood ◽  
Adenosine Triphosphatase ◽  
Lymphoid Tissues ◽  
X Ray ◽  
Adenosine Triphosphatase Activity ◽  
Assay Medium ◽  
X Irradiation ◽  
Total Body

Abstract Factors which modify lymphoid distribution of tissues were found to modify the adenosine triphosphatase activity of these tissues. Starvation or cortisone injection, which produces destructive changes in lymphoid tissues, was found to increase the enzyme activity of spleen and thymus tissues. The greater increment of enzyme activity of the thymus as compared to that of the spleen was correlated with its normally higher content of lymphoid tissue. The increase in adenosine triphosphatase activity of hematopoietic tissues appears to be associated with the type of cells present in the assay medium. With respect to peripheral blood leukocytes of the rat, the cell type is confined largely to lymphocytes and granulocytes. The increase in adenosine triphosphatase activity of the leukocytes after total-body x-ray was seen to parallel the increase in granulocytes present in the assay medium. The ratio of granulocytes to lymphocytes is not appreciably altered in dog peripheral blood after exposure to total-body x-ray; the adenosine triphosphatase activity similarly was not significantly altered. After total-body x-ray (390 r and 780 r), cells isolated from the rat bone marrow displayed a fivefold increase in adenosine triphosphatase activity. This increase was seen to correspond with an increase in the ratio of segmented leukocytes and reticuloendothelial cells and a decrease in the immature forms of the erythroid and myeloid cells. The heterogeneous cell mixtures used for our assay procedures permit the observation that total-body x-irradiation results in an increased enzyme activity of the isolated cells of the peripheral blood, bone marrow and spleen tissue of the rat. The increased enzyme activity was associated with the increased ratio of cells with high enzyme activity present in the assay medium.

scholarly journals Importance of Flow Cytometry in the Differential Diagnosis of Hairy Cell Leukemia in the State of Rio Grande Do Norte, Brazil

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 14-15
Author(s):  
Aldair Sousa Paiva ◽  
Alessandra Suelen Jardim Silva ◽  
Victor lima Soares ◽  
Gustavo Henrique de Medeiros Oliveira ◽  
Lenilton Silva DA Silva Júnior ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Differential Diagnosis ◽  
Peripheral Blood ◽  
Complete Blood Count ◽  
Conflicts Of Interest ◽  
Specific Treatment ◽  
Leukemic Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Cell Leukemia

Introduction:Hairy Cell Leukemia (HCL) is a B-cell non-Hodgkin's Lymphoma (B-NHL) representing about 2% of chronic leukemias, is manifested in adults with an average age of 55 years old or more and the ratio of male: female is 5:1, being more common among white people. It is characterized by the presence of neoplastic lymphocytes with cytoplasmic projections (villous cells), a characteristic commonly observed in other DLPCs such as variant HCL (HCL-v) and splenic villous cell lymphoma (SVCL), being the immunophenotyping by flow cytometry determinant in the differential diagnosis of these neoplasms. HCL is characterized by splenomegaly, hepatomegaly, pancytopenia in peripheral blood (PB) with leukopenia, anemia, neutropenia, monocytopenia, and thrombocytopenia. It has a low number of circulating tumor cells, spleen, liver, and bone marrow (BM) infiltration.Objective:To investigate, by flow cytometry, patients with lymphocytosis and presence of villous lymphocytes in the characterization of HCL and HCV-v and SMZL.Methodology:Were investigated samples of peripheral blood (SP) and bone marrow (MO) from 27 patients previously diagnosed with DLPC and presence of villous lymphocytes which were by flow cytometry with a panel of monoclonal antibodies (MoAb) conjugated to fluorochromes and targeted to T-lymphocytes: CD1a, CD2, CD3, CD5, CD7, subpopulation T-helper (CD3/CD4) and T-cytotoxic (CD3/CD8), in addition to TCR a/b and TCR g/d; Natural Killer cells: CD16-56; B-lymphocytes: CD19, CD20, CD21, CD22, CD23, CD79b, CD200, IgM, IgG, IgD, anti-kappa and anti-lambda, in addition to CD10, TdT (Terminal deoxynucleotidyl Transferase), CD103, CD123, CD11c, CD25, CD38, CD138, CD45 and CD14. At the same time, a complete blood count with differential white blood cell count and investigation of clinical and demographic data such as age, sex and ethnicity / race were also performed.Results:The distribution of patients according to ethnicity and gender, there was a predominance of white individuals and males. The age group most affected was in patients older than 60 years. All patients expressed pan-B antigens on leukemic cells with expression of CD19, CD22 / CD20 (Forte), sIgH, associated with clonal restriction for immunoglobulin light chain (kappa= 20 and lambda= 7), associated with FMC7 expression, HLADR, CD38 and CD45 strong and negativity to CD10, CD138, CD200, CD23, CD5, TdT and related T antigens. Sixteen cases were categorized as HCL, six HCLv and five SVCL. The immunophenotyping of HCL cases was positive for CD103, CD25, CD123 and CD11c. HCLv was negative for CD103 in three cases and CD25 and SVCL negative for CD103, CD123 and CD11c and CD25 in all cases.Conclusions:The precise diagnosis of HCL has fundamental importance because each NHL-B has a specific treatment, besides emphasizing the sensitivity and speed of the IFC regarding diagnosis and follow-up. Disclosures No relevant conflicts of interest to declare.

scholarly journals Prognostic significance of terminal transferase activity in childhood acute lymphoblastic leukemia: a prospective analysis of 164 patients

Blood ◽  
1982 ◽  
Vol 60 (6) ◽  
pp. 1267-1276
Author(s):  
JJ Hutton ◽  
MS Coleman ◽  
S Moffitt ◽  
MF Greenwood ◽  
P Holland ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukemia ◽  
Complete Remission ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Lymphoblastic Leukemia ◽  
Prognostic Significance ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Central Nervous System Relapse ◽  
Transferase Activity

Whether the level of terminal deoxynucleotidyl transferase (TdT) activity in mononuclear cells from bone marrow and peripheral blood has prognostic significance has been analyzed prospectively in 164 children with T and non-T, non-B marked acute lymphoblastic leukemia (ALL). TdT was measured at diagnosis to assess its value as a predictor of duration of remission and length of survival. It was measured repeatedly during remission to assess whether it could predict relapse. Ninety-seven percent of the children achieved a complete remission of their disease, and 40% relapsed during the study. The level of TdT activity in blasts at diagnosis varied 1000-fold from patient to patient. There was no statistically significant relationship between TdT activity in cells at diagnosis and the achievement of complete remission, the duration of remission, or length of survival. TdT activity was significantly increased in the bone marrow of 65% of patients at the time of marrow morphological relapse, but was rarely increased in marrow from patients with isolated testicular or central nervous system relapse. Wide fluctuations in TdT activity were characteristically seen in mononuclear cells from the marrow and peripheral blood of patients with ALL at all stages of their disease. An isolated high value of TdT activity in the bone marrow or peripheral blood cannot be taken as evidence of impending relapse. Quantitative measurements of TdT activity alone on mononuclear cells from bone marrow and peripheral blood are helpful in differential diagnosis, but cannot guide therapy of children with ALL.

scholarly journals Importance of Detection of Intracellular Myeloperoxidase, CD13, CD79a, CD22, CD3 and Terminal Deoxynucleotidyl Transferase By Flow Cytometry Diagnosis of Acute Leukemias

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5194-5194
Author(s):  
Aldair Sousa Paiva ◽  
Hugo Diogenes De Oliveira Paiva ◽  
Geraldo Barroso Cavalcanti ◽  
Frank Bahia ◽  
Rodrigo Villar Freitas ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Monoclonal Antibodies ◽  
Acute Leukemia ◽  
Lymphoblastic Leukemia ◽  
Lymphoid Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Acute Leukemias ◽  
Cell Markers ◽  
Terminal Transferase

Abstract Background: The detection of Intracellular (IC) antigens by flow cytometry (FC) such as myeloperoxidase (MPO), cCD13, cCD79a, cCD22, cCD3 and Terminal deoxynucleotidyl Transferase (TdT) has become the useful tool in the differential diagnosis between acute myeloid leukemias (AML) and acute lymphoid leukemias (ALL). Through detection of myeloid antigens (MPO and cCD13), B cells precursors (cCD79a and cCD22) and precocity T-cells (cCD3) it has been possible to confirm the diagnosis of these acute leukemias. The detection of intracellular cell markers by FC usually requires previous permeabilization of fresh cell suspensions. TdT, also known as DNA nucleotidylexotransferase (DNTT) or terminal transferase, is a specialized DNA polymerase expressed in immature, pre-B, pre-T lymphoid cells, and acute lymphoblastic leukemia/lymphoma cells. TdT adds N-nucleotides to the V, D, and J exons of the TCR and BCR genes during antibody gene recombination, enabling the phenomenon of junctional diversity. In humans, terminal transferase is encoded by the DNTT gene. This antigen is expressed mostly in the nucleus cells from primary lymphoid organs, like the thymus and bone marrow. The TdT detection has also been shown to be useful in confirming the acute forms of B and T-lineage lymphoproliferative diseases by FC. The aim of this study was to demonstrate the importance of this cell markers' detection by FC in the differential diagnostic of acute leukemias. Methods: Bone marrow and/or peripheral blood leukemic cells from 50 cases of acute leukemia: 16 ALL and 36 AML. The cells were fixed and permeabilized in briefly exposed to Becton & Dickinson Lyse Solution at concentration of 10%, and subsequently labeled with monoclonal antibodies anti-MPO, TdT, CD3, CD13, CD22 and CD79a. Results: The MPO expression was observed in 35/36(97,22%) and cCD13 in all cases of AML and in none ALL patients. Three cases of MPO-positive ALL (FAB-L2) could be reclassified as M0-AML. These cases were CD34+;HLADR+;CD33-;CD13-;CD7+ and cCD13+. The intensity of TdT expression was observed in 15/16 (93.8%) of ALL and 5/36 (13.9%) of AML. The cCD22 and cCD79a were positive in 15/16 (93.8%) and all of pre-B ALL respectively and cCD3 was expressed in one case of Pre-T ALL that initial phenotype was CD34+/HLADR+/TdT+/CD7+ and sCD3-). Conclusions: These results indicate that monoclonal antibodies anti-MPO, cCD13, cCD79a, cCD22, cCD3 and TdT were excellent cell markers for the diagnosis and classification of acute leukemias and can be reliably detected by FC. This rapid and specific technique should be a valuable addition to routine immunophenotyping of acute leukemia. Disclosures No relevant conflicts of interest to declare.

scholarly journals N-acetyl-beta-glucosaminidase activity in normal and malignant leukocytes.

1975 ◽  
Vol 23 (10) ◽  
pp. 752-757 ◽  
Author(s):  
C E Reed ◽  
J M Bennett
Keyword(s):  
Bone Marrow ◽  
Enzyme Activity ◽  
Solid Tumor ◽  
Peripheral Blood ◽  
Large Series ◽  
Acute Nonlymphocytic Leukemia ◽  
Significant Elevation ◽  
Cytochemical Method

An improved cytochemical method demonstrating N-acetyl-beta-glucosaminidase in peripheral blood and bone marrow leukocytes is described. A significant elevation in enzyme activity in circulating monocytes from patients with solid tumor malignancies was observed. In a large series of cases of acute nonlymphocytic leukemia, elevated levels were found in the vast majority of those leukemias that had a predominant monocytic component identified either morphologically or by standard cytochemical methods. This reaction would appear to be useful as a monocyte marker.

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