scholarly journals Studies of T-lymphocytes in preleukemic disorders and acute nonlymphocytic leukemia: in vitro radiosensitivity, mitogenic responsiveness, colony formation, and enumeration of lymphocytic subpopulations

Blood ◽  
1983 ◽  
Vol 61 (3) ◽  
pp. 449-455 ◽  
Author(s):  
SJ Knox ◽  
BR Greenberg ◽  
RW Anderson ◽  
LS Rosenblatt
Keyword(s):  
Whole Blood ◽  
Colony Formation ◽  
Tritiated Thymidine ◽  
Lymphocyte Stimulation Test ◽  
Acute Nonlymphocytic Leukemia ◽  
Clinically Normal ◽  
Normal Individuals ◽  
Increased Risk ◽  
X Irradiation

Abstract Tritiated thymidine incorporation in a whole blood lymphocyte stimulation test (LST) and lymphocyte colony formation (CFU-L) from whole blood were measured following in vitro x-irradiation. Lymphocytes from patients with myelodysplastic disorders, acute nonlymphocytic leukemia, and patients at increased risk for leukemia because of their primary disease and/or cytotoxic therapy were found to be significantly more sensitive to in vitro x-irradiation than lymphocytes from clinically normal individuals. Cloning efficiencies and mitogenic responsiveness of patient lymphocytes were significantly depressed as compared to normal values. Using monoclonal antibodies to specific surface markers, quantitative abnormalities in lymphocytic subpopulations from myelodysplastic patients also were observed. These findings are suggestive of a defect at the T-cell level that may directly or indirectly affect hematopoiesis.

scholarly journals Studies of T-lymphocytes in preleukemic disorders and acute nonlymphocytic leukemia: in vitro radiosensitivity, mitogenic responsiveness, colony formation, and enumeration of lymphocytic subpopulations

Blood ◽  
1983 ◽  
Vol 61 (3) ◽  
pp. 449-455 ◽  
Author(s):  
SJ Knox ◽  
BR Greenberg ◽  
RW Anderson ◽  
LS Rosenblatt
Keyword(s):  
Whole Blood ◽  
Colony Formation ◽  
Tritiated Thymidine ◽  
Lymphocyte Stimulation Test ◽  
Acute Nonlymphocytic Leukemia ◽  
Clinically Normal ◽  
Normal Individuals ◽  
Increased Risk ◽  
X Irradiation

Tritiated thymidine incorporation in a whole blood lymphocyte stimulation test (LST) and lymphocyte colony formation (CFU-L) from whole blood were measured following in vitro x-irradiation. Lymphocytes from patients with myelodysplastic disorders, acute nonlymphocytic leukemia, and patients at increased risk for leukemia because of their primary disease and/or cytotoxic therapy were found to be significantly more sensitive to in vitro x-irradiation than lymphocytes from clinically normal individuals. Cloning efficiencies and mitogenic responsiveness of patient lymphocytes were significantly depressed as compared to normal values. Using monoclonal antibodies to specific surface markers, quantitative abnormalities in lymphocytic subpopulations from myelodysplastic patients also were observed. These findings are suggestive of a defect at the T-cell level that may directly or indirectly affect hematopoiesis.

scholarly journals Increased radiosensitivity of a subpopulation ot T-lymphocyte progenitors from patients with Fanconi's anemia

Blood ◽  
1981 ◽  
Vol 57 (6) ◽  
pp. 1043-1048
Author(s):  
SJ Knox ◽  
FD Wilson ◽  
BR Greenberg ◽  
M Shifrine ◽  
LS Rosenblatt ◽  
...  
Keyword(s):  
T Lymphocyte ◽  
Whole Blood ◽  
Colony Formation ◽  
Lymphocyte Stimulation Test ◽  
Con A ◽  
Clinically Normal ◽  
Fanconi's Anemia ◽  
Fanconi’S Anemia ◽  
Significant Difference

In vitro radiation survival of peripheral blood T lymphocytes was studied in 15 clinically normal adults and 4 patients with Fanconi's anemia. Tritiated thymidine incorporation in a whole blood lymphocyte stimulation test (LST) and a newly developed whole blood T-lymphocyte colony assay were used to measure lymphocyte blastogenesis and colony formation in response to phytohemagglutinin (PHA) or concanavalin-A (Con-A) stimulation. Lymphocyte colony formation was found to be consistently more sensitive than the LST for detection of low-level radiation effects using both normal cells and lymphocytes from Fanconi's anemia patients. Lymphocytes from patients with Fanconi's anemia were significantly more sensitive to in vitro x-irradiation than lymphocytes from clinically normal individuals as measured by their ability to divide when stimulated by PHA in the LST (patients, D37 = 198 R; normals, D37 = 309 R, p = 0.057) and colony formation assay (patients, D37 = 53 R; normals, D37 = 109 R, p = 0.016). No significant difference in the radiosensitivity of the Con-A response was observed between the two groups. The PHA-responsive T-lymphocyte subpopulation in Fanconi's anemia patients appears to be intrinsically defective. The nature of this defect, significance in the disease process, and relevancy of these findings to the establishment of radiation protection standards are discussed.

scholarly journals Increased radiosensitivity of a subpopulation ot T-lymphocyte progenitors from patients with Fanconi's anemia

Blood ◽  
1981 ◽  
Vol 57 (6) ◽  
pp. 1043-1048 ◽  
Author(s):  
SJ Knox ◽  
FD Wilson ◽  
BR Greenberg ◽  
M Shifrine ◽  
LS Rosenblatt ◽  
...  
Keyword(s):  
T Lymphocyte ◽  
Whole Blood ◽  
Colony Formation ◽  
Lymphocyte Stimulation Test ◽  
Con A ◽  
Clinically Normal ◽  
Fanconi's Anemia ◽  
Fanconi’S Anemia ◽  
Significant Difference

Abstract In vitro radiation survival of peripheral blood T lymphocytes was studied in 15 clinically normal adults and 4 patients with Fanconi's anemia. Tritiated thymidine incorporation in a whole blood lymphocyte stimulation test (LST) and a newly developed whole blood T-lymphocyte colony assay were used to measure lymphocyte blastogenesis and colony formation in response to phytohemagglutinin (PHA) or concanavalin-A (Con-A) stimulation. Lymphocyte colony formation was found to be consistently more sensitive than the LST for detection of low-level radiation effects using both normal cells and lymphocytes from Fanconi's anemia patients. Lymphocytes from patients with Fanconi's anemia were significantly more sensitive to in vitro x-irradiation than lymphocytes from clinically normal individuals as measured by their ability to divide when stimulated by PHA in the LST (patients, D37 = 198 R; normals, D37 = 309 R, p = 0.057) and colony formation assay (patients, D37 = 53 R; normals, D37 = 109 R, p = 0.016). No significant difference in the radiosensitivity of the Con-A response was observed between the two groups. The PHA-responsive T-lymphocyte subpopulation in Fanconi's anemia patients appears to be intrinsically defective. The nature of this defect, significance in the disease process, and relevancy of these findings to the establishment of radiation protection standards are discussed.

scholarly journals Differences in cell cycle characteristics among patients with acute nonlymphocytic leukemia

Blood ◽  
1987 ◽  
Vol 69 (6) ◽  
pp. 1647-1653 ◽  
Author(s):  
A Raza ◽  
Y Maheshwari ◽  
HD Preisler
Keyword(s):  
Cell Cycle ◽  
Tritiated Thymidine ◽  
S Phase ◽  
Total Cell ◽  
Acute Nonlymphocytic Leukemia ◽  
Cell Cycle Time ◽  
Myeloid Leukemias ◽  
History Of

The proliferative characteristics of myeloid leukemias were defined in vivo after intravenous infusions of bromodeoxyuridine (BrdU) in 40 patients. The percentage of S-phase cells obtained from the biopsies (mean, 20%) were significantly higher (P = .00003) than those determined from the bone marrow (BM) aspirates (mean, 9%). The post- BrdU infusion BM aspirates from 40 patients were incubated with tritiated thymidine in vitro. These double-labeled slides were utilized to determine the duration of S-phase (Ts) in myeloblasts and their total cell cycle time (Tc). The Ts varied from four to 49 hours (mean, 19 hours; median, 17 hours). Similarly, there were wide variations in Tc of individual patients ranging from 16 to 292 hours (mean, 93 hours; median, 76 hours). There was no relationship between Tc and the percentage of S-phase cells, but there was a good correlation between Tc and Ts (r = .8). Patients with relapsed acute nonlymphocytic leukemia (ANLL) appeared to have a longer Ts and Tc than those studied at initial diagnosis. A subgroup of patients at either extreme of Tc were identified who demonstrated clinically documented resistance in response to multiple courses of chemotherapy. We conclude that Ts and Tc provide additional biologic information that may be valuable in understanding the variations observed in the natural history of ANLL.

Effect of Progesterone and Synthetic Progestins on Whole Blood Clot Formation and Erythrocyte Structure

2017 ◽  
Vol 23 (3) ◽  
pp. 607-617 ◽  
Author(s):  
Albe C. Swanepoel ◽  
Odette Emmerson ◽  
Etheresia Pretorius
Keyword(s):  
Whole Blood ◽  
Blood Clot ◽  
Thrombotic Event ◽  
Clot Formation ◽  
Erythrocyte Aggregation ◽  
Erythrocyte Morphology ◽  
Increased Risk ◽  
Blood Clots ◽  
Membrane Ultrastructure

AbstractCombined oral contraceptive (COC) use is a risk factor for venous thrombosis (VT) and related to the specific type of progestin used. VT is accompanied by inflammation and pathophysiological clot formation, that includes aberrant erythrocytes and fibrin(ogen) interactions. In this paper, we aim to determine the influence of progesterone and different synthetic progestins found in COCs on the viscoelasticity of whole blood clots, as well as erythrocyte morphology and membrane ultrastructure, in an in vitro laboratory study. Thromboelastography (TEG), light microscopy, and scanning electron microscopy were our chosen methods. Our results point out that progestins influence the rate of whole blood clot formation. Alterations to erythrocyte morphology and membrane ultrastructure suggest the presence of eryptosis. We also note increased rouleaux formation, erythrocyte aggregation, and spontaneous fibrin formation in whole blood which may explain the increased risk of VT associated with COC use. Although not all COC users will experience a thrombotic event, individuals with a thrombotic predisposition, due to inflammatory or hematological illness, should be closely monitored to prevent pathological thrombosis.

scholarly journals The effect of chemotherapy on the kinetics and proliferative capacity of normal and tumorous tissues in vivo

Blood ◽  
1975 ◽  
Vol 45 (1) ◽  
pp. 107-118 ◽  
Author(s):  
SH Rosenoff ◽  
JM Bull ◽  
RC Young
Keyword(s):  
Bone Marrow ◽  
Dna Synthesis ◽  
Colony Formation ◽  
Antitumor Agents ◽  
Tritiated Thymidine ◽  
Chemotherapeutic Agents ◽  
Proliferative Capacity

Abstract The proliferative state of a given tissue is a major determinant of its sensitivity to both phase-specific and cycle-specific chemotherapeutic agents. To study the extent of injury induced by antitumor agents to normal and tumorous tissues, a technique for following DNA synthesis as reflected in the incorporation of tritiated thymidine (3H-TdR) into DNA was compared to the conventional radioautographic technique of the labeling index (LI) and to the functional kinetic technique of granulocyte colony formation in vitro. Alterations in DNA synthesis induced by a single dose of cyclophosphamide in normal and tumorous tissues in vivo paralleled in many respects the changes seen when the more time-consuming techniques of the LI or granulocyte colony formation were employed. However, the recovery of granulocyte colony formation after cyclophosphamide therapy laged behind the recovery of DNA synthesis in the bone marrow, obscuring a kinetic event of potential therapeutic significance. The determination of DNA synthesis simultaneously in normal and tumorous tissues in vivo was easy to perform and supplied therapeutically pertinent results comparatively quickly.

scholarly journals Molecular Basis and Clinical Application of Growth-Factor-Independent In Vitro Myeloid Colony Formation in Chronic Myelomonocytic Leukemia

2020 ◽  
Vol 21 (17) ◽  
pp. 6057
Author(s):  
Klaus Geissler ◽  
Eva Jäger ◽  
Agnes Barna ◽  
Michael Gurbisz ◽  
Temeida Graf ◽  
...  
Keyword(s):  
Growth Factor ◽  
Colony Formation ◽  
Molecular Basis ◽  
Strong Predictor ◽  
Chronic Myelomonocytic Leukemia ◽  
Molecular Profile ◽  
Molecular Features ◽  
Increased Risk ◽  
Myelomonocytic Leukemia

We have originally reported that colony-forming units granulocyte/macrophage (CFU-GM) formation is an in vitro feature of chronic myelomonocytic leukemia (CMML) and a strong predictor for short survival. Elucidation of the molecular basis underlying this in vitro phenomenon could be helpful to define molecular features that predict inferior outcome in patients. We studied the correlation between the mutational landscape and spontaneous colony formation in 164 samples from 125 CMML patients. As compared to wildtype samples, spontaneous in vitro CFU-GM formation was significantly increased in samples containing mutations in NRAS, CBL and EZH2 that were confirmed as independent stimulatory factors by multiple regression analysis. Inducible expression of mutated RAS but not JAK2 was able to induce growth factor independence of Ba/F3 cells. Whereas high colony CFU-GM growth was a strong unfavorable parameter for survival (p < 0.00001) and time to transformation (p = 0.01390), no single mutated gene had the power to significantly predict for both outcome parameters. A composite molecular parameter including NRAS/CBL/EZH2, however, was predictive for inferior survival (p = 0.00059) as well as for increased risk of transformation (p = 0.01429). In conclusion, we show that the composite molecular profile NRAS/CBL/EZH2 derived from its impact on spontaneous in vitro myeloid colony formation improves the predictive power over single molecular parameters in patients with CMML.

scholarly journals Blood manufacturing methods affect red blood cell product characteristics and immunomodulatory activity

2018 ◽  
Vol 2 (18) ◽  
pp. 2296-2306 ◽  
Author(s):  
Ruqayyah J. Almizraq ◽  
Philip J. Norris ◽  
Heather Inglis ◽  
Somaang Menocha ◽  
Mathijs R. Wirtz ◽  
...  
Keyword(s):  
Whole Blood ◽  
Cytokine Production ◽  
White Blood Cells ◽  
Adverse Outcomes ◽  
Immunomodulatory Activity ◽  
Red Cell ◽  
Cell Of Origin ◽  
Increased Risk ◽  
Manufacturing Methods

AbstractTransfusion of red cell concentrates (RCCs) is associated with increased risk of adverse outcomes that may be affected by different blood manufacturing methods and the presence of extracellular vesicles (EVs). We investigated the effect of different manufacturing methods on hemolysis, residual cells, cell-derived EVs, and immunomodulatory effects on monocyte activity. Thirty-two RCC units produced using whole blood filtration (WBF), red cell filtration (RCF), apheresis-derived (AD), and whole blood–derived (WBD) methods were examined (n = 8 per method). Residual platelet and white blood cells (WBCs) and the concentration, cell of origin, and characterization of EVs in RCC supernatants were assessed in fresh and stored supernatants. Immunomodulatory activity of RCC supernatants was assessed by quantifying monocyte cytokine production capacity in an in vitro transfusion model. RCF units yielded the lowest number of platelet and WBC-derived EVs, whereas the highest number of platelet EVs was in AD (day 5) and in WBD (day 42). The number of small EVs (&lt;200 nm) was greater than large EVs (≥200 nm) in all tested supernatants, and the highest level of small EVs were in AD units. Immunomodulatory activity was mixed, with evidence of both inflammatory and immunosuppressive effects. Monocytes produced more inflammatory interleukin-8 after exposure to fresh WBF or expired WBD supernatants. Exposure to supernatants from AD and WBD RCC suppressed monocyte lipopolysaccharide-induced cytokine production. Manufacturing methods significantly affect RCC unit EV characteristics and are associated with an immunomodulatory effect of RCC supernatants, which may affect the quality and safety of RCCs.

scholarly journals In Vitro Growth of Granulocytic and Mononuclear Cell Colonies From Blood of Normal Individuals

Blood ◽  
1971 ◽  
Vol 37 (2) ◽  
pp. 131-135 ◽  
Author(s):  
PAUL A. CHERVENICK ◽  
DANE R. BOGGS
Keyword(s):  
Tritiated Thymidine ◽  
Maximum Size ◽  
Growth Characteristics ◽  
Cell Type ◽  
Equal Frequency ◽  
Mixed Cell ◽  
Normal Individuals ◽  
Monocytes And Macrophages ◽  
Peripheral Leukocytes

Abstract Colonies of eosinophils, neutrophils, monocytes, and macrophages were grown in vitro from peripheral leukocytes of normal individuals. The concentration of circulating nucleated cells capable of giving rise to such colonies is considerably less than in marrow. In blood obtained from six healthy adults, between 4.5 and 21.6 colonies were observed per 106 nucleated cells plated. Growth characteristics of these colonies were similar to those arising from marrow cells. Colony formation was observed after 6-10 days of incubation and increased to a maximum size of 200-1000 cells after 18-20 days and then began to undergo degeneration. Colonies of eosinophils were the most frequent cell type observed. Neutrophil and monocyte colonies were the next most common and occurred with about equal frequency; colonies of mixed cell type were less frequent than the above three types. These colonies were mainly observed at about 3 weeks of incubation. Colonies examined after 4 weeks of incubation consisted primarily of large histiocytes and macrophages. Studies with tritiated thymidine (3H-TDR) revealed that all colonies examined contained labeled cells, indicating that these colonies arose from cell proliferation.

scholarly journals Comparison of three common whole blood platelet function tests for in vitro P2Y12 induced platelet inhibition

2019 ◽  
Vol 50 (1) ◽  
pp. 135-143 ◽  
Author(s):  
Joao D. Dias ◽  
Torben Pottgiesser ◽  
Jan Hartmann ◽  
Daniel Duerschmied ◽  
Christoph Bode ◽  
...  
Keyword(s):  
Platelet Function ◽  
Whole Blood ◽  
Linear Models ◽  
Point Of Care ◽  
Platelet Function Tests ◽  
Blood Samples ◽  
P2y12 Inhibitors ◽  
Function Tests ◽  
Increased Risk

Abstract In the context of interventional cardiology, platelet function testing may identify patients treated with P2Y12-inhibitors at an increased risk of mortality, thrombosis and bleeding. Several whole blood point-of-care platelet function analyzers are available; however, inter-device differences have not been examined systematically. To compare three platelet function tests under standardized in vitro conditions. Healthy volunteer (n = 10) blood samples were spiked with increasing concentrations of ticagrelor (0–7500 ng/mL) and/or ASA (0–3280 ng/mL), measured on three platelet function analyzers (TEG®6s, Multiplate®, and VerifyNow®) and respective Effective Concentration (EC) levels EC10, EC50 and EC90 were calculated. Repeatability was assessed in a separate group of pooled blood samples (n = 10) spiked with ticagrelor at EC10, EC50 and EC90. ASA had no impact on ADP-activated channels for all three devices. TEG®6s was able to distinguish (p ≤ 0.05) between all ticagrelor EC zones; VerifyNow® and Multiplate® were able to distinguish between three and two zones, respectively. Multiplate® showed the largest window between EC10 and EC90 (19–9153 ng/mL), followed by TEG®6s (144–2589 ng/mL), and VerifyNow® (191–1100 ng/mL). Drug effect models distribution of disagreements were identified for TEG®6s (5.0%), VerifyNow® (8.3%), and Multiplate® (13.3%). TEG®6s showed the smallest average coefficient of variation between EC conditions (5.1%), followed by Multiplate® (14.1%), and VerifyNow® (17.7%). Linear models could be generated between TEG®6s and Multiplate®, but not VerifyNow®. Significant differences were found between whole blood point-of-care platelet function analyzers and the clinical impact of these differences needs to be further investigated.

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