scholarly journals Laboratory characteristics of patients with variant Phiadelphia chromosome positive leukemia

2020 ◽  
Author(s):  
Qiling Song ◽  
Yangliu Guo ◽  
Dongsheng Wang ◽  
Qingsong Liu
Keyword(s):  
Flow Cytometry ◽  
Effective Treatment ◽  
Myeloid Leukemia ◽  
Bone Marrow Cells ◽  
Karyotype Analysis ◽  
Chromosome 9 ◽  
Flow Cytometry Analysis ◽  
Bone Morphology ◽  
Leukemia Patient ◽  
The Difference

Abstract Background Variant Philadelphia chromosomes are characterized by the involvement of another chromosome in addition to chromosome 9 or 22. To detect the difference between variant Philadelphia chromosomes positive leukemia and classic Philadelphia chromosomes positive leukemia. And to help diagnose and treat variant Philadelphia chromosomes positive leukemia. Methods In this study, Peripheral blood and bone morrow cell morphology test was used to analysis bone morphology of variant Ph positive patients. Karyotype analysis was used to find out variant Ph chromosomes. Flow cytometry analysis was used for immunology analysis. BCR/ABL was detected by PCR to monitor change of molecular genetics in variant Ph positive patients. Results From 48 patients with Ph positive leukemia, we found out 3 variant Ph positive leukemia. Compared with the classic Ph positive leukemia patients, the hemogram in the variant Ph positive leukemia patients was more variant for presenting hypomyelodysplasia or hyperactive. Compared with classic chronic myeloid leukemia which neutrophilic myelocyte, metamyelocyte and stab granulocyte is increasing in, both the morphological testing of bone marrow cells smear and flow cytometry analysis were indicated that proportion of myeloblast and promyelocyte increased in variant Philadelphia chromosomes leukemia. What’s more, the variant Ph with breakpoint 4q31 was the first report in leukemia patient. Same as the classic Ph positive leukemia patient, formal and effective treatment could prolong the survival of patients with variant Ph positive leukemia. Conclusions Compared with classic Ph positive leukemia, the hemogram was more variant in variant Ph. The myeloid morphology in the patients with variant Ph was more immature than that of in the patients with classic Ph. Reporting new cases of complex variant translocations, which can refer to new breakpoints that can eventually be recurrent and important for the understanding of this leukemia. Formal and effective treatment are necessary for variant Ph positve leukemia.

The Growth Inhibition and Apoptosis Induction of Acute Myeloid Leukemia Blast Population by the Blocking IGF-IR Pathway.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4515-4515
Author(s):  
Si Sun ◽  
Yanli He ◽  
Xingbing Wang ◽  
Wei Liu ◽  
Jun Liu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Bone Marrow Cells ◽  
Pi3k Pathway ◽  
Leukemia Cell Line ◽  
Free System ◽  
Acute Myeloid ◽  
Marrow Cells

Abstract The insulin-like growth factor-1receptor (IGF-1R) is overexpressed in a variety of tumors and has been associated with cancer development. Here, we analysis the IGF-IR expression on the bone marrow cells from 45 newly diagnosed patients with acute myeloid leukemia (AML) by flow cytometry. IGF-1R universally expressed on AML blasts and the leukemia cell line HL-60, did not show significant correlation with FAB subtypes. However, the bone marrow cells from AML patients with high myeloblast counts (>80%) generally showed brighter IGF-IR expressions, which indicated the IGF-IR pathway might play an important role for AML blast proliferation and survival. Indeed, blocking the IGF-1R pathway by neutralizing monoclonal antibodies could reduce the proliferation of HL-60 cells by 38.28% at 48 hr. This inhibitory effect on blast growth was observed in 4 of 5 AML samples. In the same IGF-1R blocking treatment, the apoptosis of HL-60 cells was significantly induced, resulting in apoptosis of 57% of the cell population with the measurement of Annexin V vs PI staining by flow cytometry. The control contained only 20% apoptotic cells. We also demonstrated that the blockade of the IGF-1R pathway inhibited the phophorylation of the PI3K pathway component Akt in HL-60 cells when cultured in a serum free system with a supplement of 50ng/ml exogenous IGF. Since PI3K pathway activation greatly contributes to the proliferation, survival and drug resistance of AML, it is of interest to study whether blockading IGF-IR could also inhibit the PI3K pathway in primary AML blasts and synergize other anti-leukemia agents to improve the therapeutic effectiveness. Conclusions: IGF-IR may play an important role in the proliferation and survival of the AML blast population; Blocking the IGF-IR pathway could significantly inhibit the growth of AML blasts and considerably induce the apoptosis of AML blasts; IGF-IR could become a critical molecular target in anti-leukemia drug discovery.

Treatment with 8F4, An Anti-PR1/HLA-A2 T Cell Receptor-Like Antibody, Reduces Established Acute Myeloid Leukemia and Eliminates Leukemia Stem Cells in Mice

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 766-766
Author(s):  
Anna Sergeeva ◽  
Hong He ◽  
Kathryn Ruisaard ◽  
Karen Clise-Dwyer ◽  
Lisa S St. John ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Myeloid Leukemia ◽  
Bone Marrow Cells ◽  
Cell Receptor ◽  
Leukemia Stem Cells ◽  
Nsg Mice ◽  
Secondary Transfer

Abstract Abstract 766 PR1 (VLQELNVTV) is an HLA-A2-restricted leukemia-associated peptide from proteinase 3 and neutrophil elastase that is recognized by PR1-specific cytotoxic T lymphocytes that contribute to cytogenetic remission of myeloid leukemia. We developed a high affinity T cell receptor (TCR)-like mouse monoclonal antibody (8F4) that binds to a conformational epitope of the PR1/HLA-A2 complex. Flow cytometry and confocal microscopy of 8F4-labeled cells showed significantly higher PR1/HLA-A2 expression on AML blasts compared with normal leukocytes. Moreover, 8F4 mediated complement dependent cytolysis of AML blasts and Lin−CD34+CD38− leukemia stem cells (LSC), but not normal leukocytes. To investigate in vivo biological effects 8F4 on established leukemia, we established xenografts of primary human HLA-A2-positive AML in sublethally irradiated NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice. Leukemia engraftment was monitored in peripheral blood by flow cytometry. Mice with established PR1/HLA-A2-expressing leukemia were treated with twice-weekly intravenous injections of 200 μg 8F4 or isotype control antibody. Flow cytometry and histology analysis of tissues was used to assess leukemia burden and level of engraftment. After 5 weeks of treatment AML was reduced 300-fold in bone marrow of 8F4-treated mice compared to isotype-treated control animals (0.07 ± 0.06% hCD45+cells versus 20.4 ± 4.1%, n=5 mice per group). Moreover, leukemia stem cells (LSC, CD34+CD38−Lin-) were no longer detected in bone marrow of 8F4-treated mice, compared to 0.88 ± 0.24% in isotype-treated mice. Equally, AML was evident in the liver and spleen of isotype-treated mice (1.1 ± 0.16% and 0.32 ± 0.17%, respectively), but was undetectable in 8F4-treated mice (p<0.001). Similar results were obtained with AML from two additional patients, one with secondary AML (CMML) and one with AML-M7. Bone marrow contained 6.2 ± 3.0% (n=3) AML versus 41 ± 15% (n=2 mice; p=0.06) in the first case and 0.16 (n=1) versus 7.0 ± 4.1 (n=2) in the second case after 2–3 weeks of twice-weekly injection. To confirm 8F4-mediated elimination of LSC, we performed secondary transfer experiment with 1×106 bone marrow cells from 8F4- and isotype-treated mice, transplanted into recipient NSG mice, irradiated with 250 cGy. AML was undetectable in mice that received bone marrow from 8F4-treated animals versus 4.1 ± 2.4% (n=4) in bone marrow of mice that received cells from isotype- treated mice, determined at 16 weeks after secondary transfer. Because PR1/HLA-A2 expression on normal hematopoietic cells (HSC) is similar to LSC in AML patients, we sought to determine whether 8F4 treatment of NSG mice xenografted with CD34-selected umbilical cord blood resulted in elimination of xenograft. Fourteen weeks after transplant stable chimerism (4.1 - 7.7% hCD45+ cells) was established, mice were treated with 50 μg 8F4 intravenously and peripheral blood was monitored weekly for chimerism. Human CD45+ cells decreased to 0.35 – 0.95% by week 1, but increased to 1.9 – 2.1 % hCD45+ cells at week 3. Bone marrow at week three contained myeloid (CD13+CD33+) and lymphoid (CD19+) cells showing that while 8F4 has off- target effects against normal hematopoietic cells, HSC are preserved. This is consistent with our previous studies that showed no 8F4-mediated effect on colony formation of normal bone marrow cells. In conclusion, these results show that anti-PR1/HLA-A2 monoclonal antibody 8F4 is biologically active in vivo and selectively eliminates LSC, but not normal HSC. This justifies continued study of 8F4 as a novel therapy for AML. Disclosures: No relevant conflicts of interest to declare.

The Alkaloids and Bioactivities of Ethanol Extract from a Traditional Remedy of Vietnam

2020 ◽  
Vol 10 (1) ◽  
pp. 20-25
Author(s):  
Nguyen H. Sa ◽  
Trinh T. Thuy ◽  
Tran D. Quan ◽  
Dao D. Thien ◽  
Nguyen T. Tam ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Chemical Constituents ◽  
Clinical Manifestations ◽  
Herbal Remedies ◽  
Flow Cytometry Analysis ◽  
Oral Toxicity ◽  
Experimental Conditions ◽  
Acute Myeloid

Background and Objective: In Vietnam, “Dai trang hoan ba Giang” (BG), which belongs to herbalist “Ba Giang”, has been one of the famous herbal remedies for ulcerative colitis and diarrhea for over 100 years. However, up to the present, the main BG’s chemical constituents have not been investigated. Therefore, this study aims to investigate the phytochemistry and antiproliferative activity of isolated compounds from BG on Acute Myeloid Leukemia (OCI-AML) cells as well as evaluating its safety by the acute and subchronic toxicological tests. Methods: Compounds from herbmaterial were isolated by using column chromatography. Their structures were determined by combining spectral analysis and comparison with reported data. The concentrations of components were were determined by HPLC/DAD analysis. Anti-proliferative activity in OCI-AML cell line of isolated compounds was carried out by flow cytometry analysis. The acute and subchronic oral toxicity in micewere appraised by observingdaily for clinical manifestations of toxicity while the experiment lasted. Results: In this study, driedpowders of materials were extracted by 70% EtOH to afford an extract (DTBG). Two compounds, including palmatine (1) and berberine (2) were isolated from DTBG. The DTBG inhibited the growth of OCI-AML by a significant increase of caspase-8-independent apoptosis as measured by propidium iodide flow cytometry analysis and caspase-3 activation. In the acute study, DTBG is well tolerated, non-toxic and non-lethal to mice under the present experimental conditions. In thesubchronic study in mice, DTBG did not impact on weight gain, blood, liver and kidney function. Conclusion: Overall, the main chemical constituents of DTBG and its effect on proliferation of acute myeloid leukemia cells were reported for the first time. Moreover, the results suggest that, DTBG did not express any significant toxic effect in mice. Hence, the extract can be utilized for pharmaceutical formulations.

A Novel t(9;22;11) Translocation Variant Involving 11q23 in a Patient with Chronic Myeloid Leukemia

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 7-7
Author(s):  
Lin Yuehui ◽  
Qinglong Zheng ◽  
Tong Wu ◽  
DAN LIU
Keyword(s):  
Flow Cytometry ◽  
In Situ Hybridization ◽  
Chronic Myeloid Leukemia ◽  
Fluorescence In Situ Hybridization ◽  
Myeloid Leukemia ◽  
Conflicts Of Interest ◽  
Fusion Gene ◽  
Philadelphia Chromosome ◽  
Flow Cytometry Analysis

Background:The progression of Philadelphia chromosome positive chronic myeloid leukemia (CML) is frequently accompanied by cytogenetic evolution, commonly unbalanced chromosomal changes. but balanced chromosomal translocations are very rare in CML, especially translocations involving the 11q23. The few reported cases with blast phase (BP) of CML carrying a 11q23 rearrangement results in insufficient responses to tyrosine kinase inhibitors (TKIs) and possess a poor prognosis. Methods:Cytogenetic analysis , fluorescence in situ hybridization (FISH), RNA sequencing (RNA-seq), targeted genomic sequencing and multi-parametric flow cytometry analysis were performed to identify the chromosome translocations and pathogenic gene alterations in a 36-year-old female with myeloid BP of CML. Results: In BP, the bone marrow (BM) aspiration showed 61% myeloid blasts; Multi-parametric flow cytometry analysis revealed the abnormal myeloid blasts expression of the following antigens: CD117, CD13, CD33, CD38, partially expressed CD15, CD64. Chromosome analysis revealed a t(11;22)(q23;q11) translocation in addition to the t(9;22)(q34;q11). Fluorescence in situ hybridization (FISH) test confirmed that the t(11;22)(q23;q11) involved the mixed lineage leukemia(MLL) gene on 11q23 and RNA sequence revealed MLL-SEPT5 and BCR/ABL1(p210) fusion transcripts positive. Mutations on 339 commonly mutated genes in hematologic malignancies were analyzed by targeted next-generation sequencing showed ASXL1 p.G949Vfs*2 mutation. The patient failed to respond to both imatinib and dasatinib despite the absence of resistance-associated mutations in the BCR/ABL1 gene and she had a myeloid blast crisis at 19 months after initiation of first- and second-generation TKI treatment. After BP, she received ponatinb, a third-generation TKI with chemotherapy. Regretly she didn't achieve a complete remission(CR) and was in the process of salvage transplantation at present. Conclusions:The presence of 11q23 rearrangements in BC of CML is rare and most likely accounts for the adverse clinical outcome. We first report a patient who diagnosed CML with t(11;22)(q23;q11) and MLL-SEPT5 fusion gene positive in BP of CML. The clinical course was aggressive, and therapy was poorly tolerated. Disclosures No relevant conflicts of interest to declare.

scholarly journals The translocation (6;9) (p23;q34) shows consistent rearrangement of two genes and defines a myeloproliferative disorder with specific clinical features

Blood ◽  
1992 ◽  
Vol 79 (11) ◽  
pp. 2990-2997 ◽  
Author(s):  
D Soekarman ◽  
M von Lindern ◽  
S Daenen ◽  
B de Jong ◽  
C Fonatsch ◽  
...  
Keyword(s):  
Myeloid Leukemia ◽  
Bone Marrow Cells ◽  
Molecular Level ◽  
Myeloproliferative Disorders ◽  
Chromosome 9 ◽  
Refractory Anemia ◽  
Karyotypic Analysis ◽  
Cytogenetic Aberration ◽  
Dna And Rna ◽  
Cdna Preparation

Abstract Translocation (6;9)(p23;q34) is a cytogenetic aberration that can be found in specific subtypes of both acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). This translocation is associated with an unfavourable prognosis. Recently, the genes involved in the t(6;9) were isolated and characterized. Breakpoints in both the dek gene on chromosome 6 and the can gene on chromosome 9 appear to occur in defined regions, which allows us to diagnose this type of leukemia at the molecular level. Moreover, because of the translocation a chimeric dek-can mRNA is formed which, as we show here, is an additional target for diagnosis via cDNA-preparation and the polymerase chain reaction (PCR). We studied 17 patients whose blood cells and/or bone marrow cells showed a t(6;9) with karyotypic analysis. Fourteen patients suffered from AML, one patient had a refractory anemia with excess of blasts in transformation (RAEBt), one patient had an acute myelofibrosis (AMF), and one patient a chronic myeloid leukemia (CML). In nine cases studies at the DNA and RNA levels were possible while in seven cases only the DNA could be analyzed. In one case only RNA was available. Conventional Southern blot analysis showed the presence of rearrangements of both the dek gene and the can gene. In both genes, breakpoints cluster in one intron in the patients investigated. The presence of a consistent chimeric dek-can product after cDNA preparation followed by the PCR was demonstrated. We conclude from our data that the t(6;9) is found in myeloproliferative disorders with typical clinical characteristics. This translocation results in highly consistent abnormalities at the molecular level.

scholarly journals Separation of myeloid and erythroid progenitors based on expression of CD34 and c-kit

Blood ◽  
1995 ◽  
Vol 86 (11) ◽  
pp. 4076-4085 ◽  
Author(s):  
MO De Jong ◽  
G Wagemaker ◽  
AW Wognum
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Bone Marrow Cells ◽  
Low Frequency ◽  
Erythroid Progenitors ◽  
Color Flow ◽  
Novel Approach ◽  
Colony Forming Units ◽  
The Difference

In this report, a novel approach is described to physically separate erythroid progenitors from monocyte and granulocyte progenitors, based on the expression of CD34 and Kit. Using biotin-labeled human Kit ligand (KL) and flow cytometry, Kit was detectable on 2% to 3% of the nucleated cells in rhesus monkey bone marrow. Combination of biotin-KL with CD34 monoclonal antibodies (MoAb) showed that Kit was expressed on subsets of CD34low and CD34pos cells. Our data clearly demonstrate that CD34pos cells are more heterogeneous with respect to Kit expression than observed in studies using Kit MoAb. A small cluster, approximately 7% of the CD34pos cells, expressed CD34 at submaximal levels and stained brightly with biotinylated KL. This CD34pos/kithi fraction contained predominantly erythroid progenitors (burst-forming units- erythroid; BFU-E). The majority of the granulocytic and monocytic progenitors (colony-forming units-granulocyte/macrophage; CFU-GM) were CD34pos/kitmed. Some BFU-E were also detected in the CD34pos/kitmed and CD34low/kitpos fractions at low frequency. In the latter subset, most erythroid colony-forming units (CFU-E) were recovered. Using three- color flow cytometry, we analyzed expression of Kit in relation to that of CD34 and the class II major histocompatibility antigen, RhLA-DR. The most immature bone marrow cells that can be identified in vitro, ie, CD34pos/RhLA-DRlow cells, were kitmed. The CD34pos/kithi and CD34pos/kitneg subsets predominantly contained the more mature RhLA- DRbright cells. Our results demonstrate that erythroid precursors express c-kit at much higher levels than monomyeloid precursors and pluripotent progenitors. The difference in expression levels of CD34 and c-kit can be exploited to isolate BFU-E populations that are virtually devoid of nonerythroid cells.

The translocation (6;9), associated with a specific subtype of acute myeloid leukemia, results in the fusion of two genes, dek and can, and the expression of a chimeric, leukemia-specific dek-can mRNA

1992 ◽  
Vol 12 (4) ◽  
pp. 1687-1697 ◽  
Author(s):  
M von Lindern ◽  
M Fornerod ◽  
S van Baal ◽  
M Jaegle ◽  
T de Wit ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Bone Marrow Cells ◽  
Fusion Gene ◽  
Open Reading Frames ◽  
Chromosome 9 ◽  
Chromosome 6 ◽  
Large Gene ◽  
Molecular Masses ◽  
Acute Myeloid

The translocation (6;9) is associated with a specific subtype of acute myeloid leukemia (AML). Previously, it was found that breakpoints on chromosome 9 are clustered in one of the introns of a large gene named Cain (can). cDNA probes derived from the 3' part of can detect an aberrant, leukemia-specific 5.5-kb transcript in bone marrow cells from t(6;9) AML patients. cDNA cloning of this mRNA revealed that it is a fusion of sequences encoded on chromosome 6 and 3' can. A novel gene on chromosome 6 which was named dek was isolated. In dek the t(6;9) breakpoints also occur in one intron. As a result the dek-can fusion gene, present in t(6;9) AML, encodes an invariable dek-can transcript. Sequence analysis of the dek-can cDNA showed that dek and can are merged without disruption of the original open reading frames and therefore the fusion mRNA encodes a chimeric DEK-CAN protein of 165 kDa. The predicted DEK and CAN proteins have molecular masses of 43 and 220 kDa, respectively. Sequence comparison with the EMBL data base failed to show consistent homology with any known protein sequences.

Sign in / Sign up

Export Citation Format

Share Document