Evaluation of Genotoxic Potential of 2-(Bromoacetamido) Phenylboronic Acid on Balb/C Mice Peripheral Blood Cells Using in Vivo Micronucleus Assay = تقييم قدرة حمض برومو أسيتا أميدو فينل بورونك على إحداث سمية جينية باستخدام اختبار تكوين النوى الصغيرة في خلايا دم الفئران البيض

2017 ◽  
Vol 10 (2) ◽  
pp. 103-112
Author(s):  
Ahmad M. Khalil ◽  
Wala’ K. Salman ◽  
Khaled M. Al-Qaoud
Keyword(s):  
Peripheral Blood ◽  
Blood Cells ◽  
Phenylboronic Acid ◽  
Micronucleus Assay ◽  
Peripheral Blood Cells ◽  
Genotoxic Potential

Allogeneic Donor NK Cell Infusion and IL-2 for Patients with Refractory B-Cell Non-Hodgkin Lymphoma.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3036-3036
Author(s):  
Veronika Bachanova ◽  
Linda J. Burns ◽  
David H. McKenna ◽  
Julie Curtsinger ◽  
Sarah Cooley ◽  
...  
Keyword(s):  
B Cell ◽  
Nk Cells ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Cell Expansion ◽  
Nk Cell ◽  
Peripheral Blood Cells ◽  
Non Hodgkin Lymphoma ◽  
Pre Treatment

Abstract Abstract 3036 Poster Board II-1012 The potential role of allogeneic natural killer (NK) cells for therapy of refractory lymphoma is supported by the curative potential of allogeneic transplantation for lymphoid malignancies. Haploidentical donor derived NK cells may overcome Class I MHC Ag mediated inhibition and deliver an NK versus lymphoma effect. In a Phase II study we evaluated allogeneic NK cell infusions with Rituximab and IL-2 in a non-transplant setting to determine the expansion of NK cells in vivo and the clinical response in patients with refractory B-cell non-Hodgkin lymphoma (NHL). Six patients with advanced NHL received conditioning with Rituximab 375mg/m2 days -8,-1,+6,+15; Cyclophosphamide 60 mg/kg IV day -5; Fludarabine 25 mg/m2 IV days -6 through -2 as immunosupression to permit homeostatic expansion of allogeneic donor NK cells. Peripheral blood cells were obtained by lymphapheresis from unmobilized, HLA-haploidentical donors and selected for “killer immunoglobulin receptor” (KIR) ligand mismatch when available (3 out of 6 patients). Donor peripheral blood cells were enriched for NK cells with the Miltenyi CliniMACS device by depletion of T (CD3+) cells. The donor NK cells were then activated by overnight incubation with IL-2 (1,000 U/mL) and infused at a median nucleated cell dose of 2.27 ±0.4 × 107/kg. Subcutaneous IL-2 10×106 units (qod x 6 doses) was given to facilitate NK cell survival and expansion. All patients were evaluable for toxicity and efficacy. Patients tolerated the NK infusion well with only transient grade 1-2 toxicity and 5 received all 6 scheduled doses of IL-2. IL-2 activated donor NK cell products showed > 55% cytotoxicity against K562 targets. After IL-2 therapy, we observed a median absolute lymphocyte count of 980 ±440/μL. All cells were of recipient origin with no detectable donor NK cells. Importantly, in all patients the median number of host regulatory T cells (T regs phenotype CD4+Foxp3+CD127−) post treatment was significantly increased compared to pre-treatment (day 14 T regs: 134 ±141 cells/μL versus pre-treatment T regs: 24 ±12 cells/μL; P=0.06). To investigate the possibility of NK trafficking to affected lymph nodes, we performed fine needle aspiration of palpable tumor in 1 patient and demonstrated a low level of donor DNA by RFLP testing (2.5% donor chimerism). Simultaneous absence of NK cells in peripheral blood in the same patient suggested NK cell tissue homing to lymphoma-bearing nodes. Three patients achieved a partial remission (PR), one of whom proceeded to non-myeloablative cord blood allograft 2 month after NK cell infusion; two remain in partial remission after 1 and 4 months of follow-up. The trial failed to achieve prospective statistical parameters established to detect circulating NK cell expansion rate and will be modified. Conclusions This “proof of principle” study demonstrated lack of in vivo expansion of haploidentical NK cells in peripheral blood of patients with lymphoma. However, we identified host factors that interfered with NK cell expansion, including T reg proliferation and possibly inadequate immunosupression, and additionally, the finding of donor DNA in sites of tumor suggested donor NK cell localization to extravascular or tumor sites. Novel approaches to adoptive NK cell therapy trials should incorporate strategies to eliminate or prevent T reg expansion using alternate lymphodepleting regimens. Disclosures No relevant conflicts of interest to declare.

scholarly journals Evaluation of the genotoxic potential of Austroplenckia populnea (Reiss) Lundell chloroform fraction from barkwood extract in rodent cells in vivo

2009 ◽  
Vol 69 (4) ◽  
pp. 1141-1147 ◽  
Author(s):  
JC. Ribeiro ◽  
SF. Andrade ◽  
JK. Bastos ◽  
EL. Maistro
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Chromosome Aberrations ◽  
Blood Cells ◽  
Bone Marrow Cells ◽  
Chloroform Fraction ◽  
Peripheral Blood Cells ◽  
Swiss Mice ◽  
Marrow Cells

The genotoxic effect of the Austroplenckia populnea chloroform fraction from barkwood extract was tested in vivo on peripheral blood cells of Swiss mice with the comet assay (SCGE), and the clastogenic effect was investigated on peripheral blood cells of Swiss mice and bone marrow cells of Wistar rats, with the micronucleus and chromosome aberrations tests. The animals were treated by gavage with 3 concentrations of the extract: 300, 600 and 900 mg.kg-1. Peripheral blood cells of Swiss mice were collected 4 and 24 hours after the treatment to the SCGE assay and 48 and 72 hours to the micronucleus test. Bone marrow cells of Wistar rats were collected 24 hours after the treatment to the micronucleus and chromosome aberration tests. The results showed that the A. populnea chloroform fraction induced an increase in the average number of DNA damage in peripheral blood cells at the three concentrations tested, but this increase was not statistically significant. In the micronucleus and chromosome aberrations test, no significant increase was observed in the mean number of micronucleated polychromatic erythrocytes (MNPCE) of Swiss mice or MNPCE or chromosome aberrations for the rat bone marrow cells, for any of the tested doses. Our findings enable us to conclude that by the comet assay, A. populnea chloroform fraction from barkwood extract showed no genotoxic effects, and by the micronucleus and chromosome aberration tests, the extract fraction showed no clastogenic/aneugenic effects on the rodent cells tested.

PERIPHERAL BLOOD PLASMACYTOSIS FOLLOWING SYSTEMIC EXPOSURE TO PHYTOLACCA AMERICANA (POKEWEED)

PEDIATRICS ◽  
1966 ◽  
Vol 38 (3) ◽  
pp. 490-493
Author(s):  
B. E. Barker ◽  
P. Farnes ◽  
P. H. LaMarche
Keyword(s):  
Peripheral Blood ◽  
Blood Cells ◽  
Plasma Cells ◽  
Human Peripheral Blood ◽  
Systemic Exposure ◽  
Peripheral Blood Cells ◽  
Phytolacca Americana ◽  
Oral Ingestion

MITOGENIC properties of extracts from Phytolacca americana (pokeweed, scoke, inkberry) for human peripheral blood cells in vitro have been reported from this laboratory. Subsequently, the appearance of leukocytes typical of early and late members of the plasmacytic series was described in the peripheral bloods of two of the authors (P.F. and B.E.B.), who received accidental systemic exposure to the mitogen. Effects of phytomitogens on human cells in vivo are not established, although a number of investigators have administered phytohemagglutinin from Phaseolus vulgaris to patients with aplastic anemia, hoping that the "transformed" lymphocytes might possess hematopoietic potentialities and repopulate the marrow with useful cells. Interpretations of such studies have been complicated by the small numbers of patients studied, the natural course of this group of anemias, simultaneous use of other therapy, and other variables. More information has been accumulated about the effects of phytomitogens on peripheral blood cells in vitro. The process of "transformation" or blastogenesis of lymphocytes, whether induced by plant extracts or specific antigenic stimuli, appears to involve an immune mechanism, although morphologic differentiation toward the plasmacytic series is not seen in the in vitro environment. Recently, we have had the opportunity to study serial blood films from children who received systemic exposure to pokeberry, either through proven oral ingestion (berries recovered from vomitus), or by exposure of fresh cuts and abrasions to pokeberry juice in the course of handling the berries. In each of these instances large cells morphologically typical of plasmablasts and proplasmacytes, and mature plasma cells were found in the peripheral blood films for periods up to 2 weeks following exposure. Mitotic cells were present in the peripheral blood during the first 10 days after exposure (Fig. 1 and 2).

The Morphology and Cellular Characterization of Thrombocytes in Adult Xenopus laevis.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3947-3947
Author(s):  
Takako Ishida ◽  
Miyako Obuchi-Shimoji ◽  
Takeshi Kuribara ◽  
Nami Nogawa ◽  
Tomoyuki Tahara ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Xenopus Laevis ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Structural Characteristics ◽  
Flow Cytometric Analysis ◽  
Peripheral Blood Cells ◽  
Morphological Aspect ◽  
Cell Counts

Abstract In primates and rodents, platelets originate from the bone marrow megakaryocytes through a unique differentiation process with nuclear polyploidization, cytoplasmic maturation and proplatelet formation. In contrast, circulating thrombocytes of most non-mammalian vertebrates are particularly distinctive; the cells are large and nucleated. Adult Xenopus laevis may be an useful non-mammalian model for analyzing dynamic hematopoiesis because they are individually tolerable for time lapse analysis in vivo with sequential blood sampling, whereas classification of cell types has not been established yet. Microstructures of Xenopus thrombocytes observed with electron microscope exhibited structural characteristics largely resembling zebrafish thrombocytes with nucleated spindle cellular features (Thattaliyath et al., Blood 2005), and they had lobulated nuclear chromatin, granules, microparticles and open canalicular system-like-structures as in mammalian megakaryocytes. Since thrombocyte identification based on the morphological aspect was not sufficient, chemical staining with acetylecholinesterase and thiazole orange were performed. Additionally, mice were immunized by Xenopus peripheral blood cells to generate monoclonal antibodies, and two hybridomas producing IgG, respectively T12 and T5, were screened. T12+ (T12 positive) cells were morphologically typical thrombocytes. Flow cytometric analysis revealed that T12+ cells were also positive to anti-human GpIIb/IIIa polyclonal antibodies, and approximately 2-3% of whole peripheral blood cells were T12+/GpIIb/IIIa+ that distributed in FSClow/SSClow fraction. When T12 was injected into Xenopus to deplete T12+ cells in vivo, the detectable level of T12 in the circulation lasted for more than several weeks. Peripheral thrombocyte counts predominantly began to decrease immediately and reached their nadir at day 3, but white blood cell counts were not changed. RNA-rich blood cells considered as younger cells were then increasingly appeared, and finally the cell counts recovered to normal levels at day 10–15, indicating that in vivo depletion of T12+ cells induced thrombopoiesis and/or release of mature thrombocytes from the pool. T5 recognizing cells were classified into two populations by immunostaining and flow cytometry; T5+/GpIIb/IIIa+ cells were morphologically thrombocytic as the cells recognized by T12, while T5+/GpIIb/IIIa− cells were spherical and similar appearance to lymphocytic cells. These observations raised some possibilities e.g.; antigen of T5 was a membrane protein common to both lymphocytes and thrombocytes, or T5+/GpIIb/IIIa− cells were thrombocyte progenitors at earlier development stage than T12+/GpIIb/IIIa+ cells. Nevertheless only a few percent of T12+ and T5+ cells resided in peripheral blood, immunostaining revealed that the proportions of T12+/T5+ and T5+ cells in spleen were 10% and 70%, and T12+/T5+ and T5+ cells in liver were 5% and 20%, respectively. These suggest that spleen is predominantly involved in thrombopoiesis and/or thrombocyte storage in adult Xenopus. As T12 and T5 can be used successfully in flow cytometry and magnetic cell sorting, they should contribute us directly to elucidate the origin of circulating Xenopus thrombocytes and their cellular development process.

scholarly journals In Vivo Effects of Myeloablative Alkylator Therapy on Survival and Differentiation of MGMTP140K-Transduced Human G-CSF-Mobilized Peripheral Blood Cells

2006 ◽  
Vol 13 (5) ◽  
pp. 1016-1026 ◽  
Author(s):  
Shanbao Cai ◽  
Jennifer R. Hartwell ◽  
Ryan J. Cooper ◽  
Beth E. Juliar ◽  
Emi Kreklau ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Blood Cells ◽  
Peripheral Blood Cells ◽  
In Vivo Effects ◽  
Mobilized Peripheral Blood

scholarly journals Tyrosine phosphorylation of CRKL in Philadelphia+ leukemia

Blood ◽  
1994 ◽  
Vol 84 (6) ◽  
pp. 1731-1736 ◽  
Author(s):  
J ten Hoeve ◽  
RB Arlinghaus ◽  
JQ Guo ◽  
N Heisterkamp ◽  
J Groffen
Keyword(s):  
Tyrosine Phosphorylation ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Catalytic Domain ◽  
Lymphoblastic Leukemia ◽  
Adaptor Protein ◽  
Myelogenous Leukemia ◽  
Peripheral Blood Cells

Abstract The chimeric BCR/ABL protein is characteristic of Philadelphia (Ph)+ leukemia because it is the direct product of the Ph translocation and it has been shown to play a causal role in the genesis of leukemia. The BCR/ABL protein exhibits a deregulated tyrosine-kinase activity capable of phosphorylating different cellular substrates in vivo and in vitro. CRKL, an adaptor protein consisting of SH2 and SH3 domains in the absence of a catalytic domain, is one potential in vivo substrate of BCR/ABL. Previous experiments have shown that CRKL is phosphorylated on tyrosine in the chronic myelogenous leukemia (CML) cell line K562 and that CRKL is a substrate for ABL and for BCR/ABL in COS-1 cells. In the current study, we show that in peripheral blood cells a direct correlation exists between the presence of BCR/ABL and the phosphorylation status of CRKL. In Ph- peripheral blood cells, CRKL is present only in the nonphosphorylated form. In contrast, all BCR/ABL+ CML and acute lymphoblastic leukemia patient samples examined showed clear tyrosine-phosphorylation of CRKL. This result strongly suggests that CRKL is a biologically significant substrate for BCR/ABL and is likely to play a major role in the development of Ph+ leukemia.

Sign in / Sign up

Export Citation Format

Share Document