scholarly journals DECREASED CD10 EXPRESSION IN THE BONE MARROW NEUTROPHILS OF HIV POSITIVE PATIENTS

2010 ◽  
Vol 2 (3) ◽  
pp. e2010032 ◽  
Author(s):  
Annemarie Van de Vyver ◽  
Adele Visser
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Marked Reduction ◽  
Bone Marrow Aspirate ◽  
Hiv Positive ◽  
Morphological Abnormality ◽  
Cd10 Expression ◽  
Haemopoietic Cells ◽  
Hiv 1 ◽  
Qualitative Changes

Background: HIV-1 infection is associated with various quantitative and qualitative changes in haemopoietic cells. Clear distinction between primary myelodysplastic syndrome (MDS) and secondary dysplasia may not always be possible. Adjunctive analyses used in the diagnosis of MDS include cytogenetics and flow cytometry (FCM). Much focus has been placed on establishing FCM guidelines aiding in the diagnosis of MDS, and to distinguish this condition from secondary dysplastic changes. One of the parameters often cited is the CD10 expression on the granulocyte population, as this marker denotes granulocytic maturation. Aims: To determine the expression level of CD10 on granulocytes in HIV positive patients. Methods: In total, 117 HIV-1 positive and 29 HIV-1 negative patients were included in this study. Bone marrow aspirate samples were evaluated in terms of morphological abnormality as well as CD10 expression on the granulocytic population. Results: The average CD10 expression among the HIV-1 positive patients were markedly reduced, at 18.4%, and 113 patients (96.6%) of these patients had expression levels below 50%. Discussion: Disease conditions causing secondary dysplasia, especially HIV-1 infection, is associated with a marked reduction in CD10 expression on the granulocyte population independent from the presence of myelodysplastic features. This marker is therefore of doubtful significance as a diagnostic tool in distinguishing between primary and secondary dysplasia.  

Therapy Related Leukemia in HIV Patients after Treatment for Non-Hodgkin Lymphoma

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5284-5284
Author(s):  
Jed Katzel ◽  
Sanford Kempin ◽  
David Vesole ◽  
Portia Lagmay-Fuentes ◽  
William Cook ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Radiation Therapy ◽  
Antiretroviral Therapy ◽  
Burkitt’S Lymphoma ◽  
Burkitt's Lymphoma ◽  
Hiv Positive ◽  
Secondary Malignancies ◽  
Hiv Patients ◽  
Hiv Negative

Abstract Therapy related secondary malignancies after NHL are well characterized in the HIV negative population. The increased risk of secondary leukemia is most commonly associated with alkylating agents, topoisomerase II inhibitors and radiation therapy. We describe 2 patients with HIV-associated Burkitt’s lymphoma who subsequently developed acute leukemia. Case Report 1: An HIV positive 60 yr old male was diagnosed with Burkitt’s lymphoma five years after beginning antiretroviral therapy. Lymph node flow cytometry demonstrated: CD10+, CD19+, CD20+, KAPPA+. He achieved a complete remission after completing the Vanderbilt regimen (cyclophosphamide, methotrexate, bleomycin, vincristine, and doxorubicin) (McMaster M, et al. J Clin Oncol. 1991:9:941–946). Eight years later, he presented with acute myelomonocytic leukemia (M4) after a myelodysplastic prodrome. Flow cytometry demonstrated CD11c+, CD13+, CD33+, CD34+, CD 64+ and cytogenetics showed 5q(−) and 20q(−). He received induction chemotherapy with arsenic trioxide and low dose cytarabine. He did not achieve a remission, and died 2 months later. Case Report 2: A 45 yr old male presented with severe abdominal pain, and fever. During laparotomy, he was found to have a cecal mass consistent with Burkitt’s lymphoma. A bone marrow biopsy also showed Burkitt’s lymphoma: CD10+, CD19+, CD20+, CD22+, CD 38+, CD45+, CD71+. He was subsequently diagnosed with HIV with a CD4 count of 60/uL. He was treated with CODOX-M (cyclophosphamide, doxorubicin, vincristine, methotrexate, IT cytarabine, IT methotrexate) and IVAC (Ifosfamide, etoposide, cytarabine, IT methotrexate) (Magrath I, et al. J Clin Oncol1996;14:925–934) achieving a CR. He remained on antiretroviral therapy throughout his course. Two years later, he presented with thrombocytopenia. A bone marrow aspirate was consistent with precursor B-cell ALL CD19+, CD34+, CD79a+ and TdT+ distinct from the previous Burkitt’s lymphoma. He was treated with the L20 (Clarkson B, et al. Haematol Blood Transfus1990;33:397–408) protocol achieving a durable CR. He continued his retroviral therapy during his treatment. Conclusions: HIV positive patients have an increased propensity to develop malignancy independent of prior chemotherapy or radiotherapy exposure. In the era of HAART, the survival of HIV positive patients has markedly improved. Although the role of chemotherapy and radiation therapy are well documented as causative agents of neoplasia, the risk of HAART therapy with toxicity of nuclear, cytoplasmic and cell membrane effects potentially inducing malignancies is less well defined. Many of these agents are considered oncogenic in animal models but have not been proven to cause tumors in humans. However, it is conceivable, given the cellular toxicities of antineoplastic and antiretroviral therapy, that in combination they could cause myelodysplasia or further lymphodysplasia. It is too early to know if HIV patients are at a greater risk for development of secondary malignancies as a long-term complication of chemotherapy. However, because recent studies have demonstrated that HIV+ patients on highly active antiretroviral therapy (HAART) have comparable responses to chemotherapy compared to HIV negative patients, we recommend that patients continue HAART while receiving treatment for malignancy. Close surveillance for the appearance of secondary leukemias is warranted.

Flow Cytometry Primary 10 Colours Panel for Chronic Lympho Proliferative Diseases Diagnosis

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5190-5190
Author(s):  
Jonathan Brauner ◽  
Ingrid Beukinga ◽  
Zoulikha Amraoui ◽  
Zaina Kassengera ◽  
Michel Toungouz ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
B Cells ◽  
B Lymphocytes ◽  
Plasma Cells ◽  
Cell Lymphoma ◽  
Cd10 Expression ◽  
Complete Characterisation

Abstract Abstract 5190 Objectives: Definition of the primary antibodies panel for 10 colours flow cytometry able to describe normal and clonal T, B lymphocytes and plamocytes in blood and bone marrow. Once clonalities are detected, the complete characterisation of Chronic Lymphoproliferative Diseases (CLPD) is supported by secondary panels chosen based on the results of CD5/CD10 expression for clonal B lymphocytes, CD27/CD38 for plasmatocytes and CD3/CD27 for clonal T cells. Materials and Methods: Blood and bone marrow of patients (N=50) with CLPD (mainly B-CLL). Samples are enumerated by haematology analyzer DxH 800 then 106 cells are washed three times, stained with the antibodies combination and red blood cells lysed with Versalyse (TM. Beckman Coulter). The samples were analysed on a 10 colours Navios flow cytometer (Beckman Coulter Fullerton, CA). The staining panel consists of 14 antibodies (CD45, CD8, CD4, CD5, CD3, CD19, CD38, λ, κ, CD23, CD5, CD10, CD14, CD27) conjugated with 10 different fluorochromes. The fixed gating strategy allows linking Navios analysis software to the middleware Remisol which drives the choice of the secondary panel. In some cases a third tube is performed for Ki67 or Zap-70 intra-cytoplasmic staining. Results: Monocytes are removed on the basis of their CD14/CD4 expression. B lymphocytes are CD19 positive. Normal naïve/memory B cells, hematogones and plasma cells are defined by their CD27, CD10 and CD38 expression. Eventual monoclonality is sought by analysis of the distribution of Kappa and Lambda light chains. A first classification of B cell lymphoma is achieved with the CD5 and CD10 expression of the clone (CD5+/CD10−: B-CLL MCL and few MZL, CD5−/CD10−: MZL and related, CD5−/CD10+ DLBCL and FL). Analysis of CD27, CD20 and CD23 expression allows discriminating between CD5+/CD10- lymphomas. All the 50 samples were correctly detected as CLPD and the automated Remisol choice of the second panel fit to the final diagnosis of all the cases of this small series. T lymphocytes are defined by their CD3 and CD5 expression. The analysis of CD4/CD8 balance and CD27/CD5 distribution are first line test when T cell clonality is suspected. There is a special gating to detect CD3-CD4+ T cell lymphoma and double negativity of CD4 and CD8 is a surrogate marker for gamma/delta T cells. NK cells are mentioned as not-T not-B lymphocytes, without specific staining. Conclusion/Discussion:This 10 colours 14 antibodies panel allows describing in one tube normal T and B cells, hematogones, memory and naives B cells plasma cells and detects T and B clonalities. This panel follows a similar logic than the Euroflow LST tube but with 10 colours and with Beckman Coulter's technology and antibodies. Moreover, this combination helps discriminating rapidly the CD5+/CD10- lymphomas while the complete characterisation of CD5 negative lymphomas only require less than 6 antibodies second tube. This is a paperless (all the process is driven and controlled by Remisol), fast and inexpensive diagnostic approach (always less than 20 antibodies required). Disclosures: Pradier: Beckman Coulter: Consultancy, Membership on an entity's Board of Directors or advisory committees.

scholarly journals A rare morphological and immunophenotypic presentation of adult B-cell acute lymphoblastic leukemia with blasts in the monocytic zone on CD45 side scatter

2019 ◽  
Vol 5 (3) ◽  
pp. 82
Author(s):  
Shuaeb Bhat ◽  
Saleem Hussain
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Acute Lymphoblastic Leukemia ◽  
B Cell ◽  
Bone Marrow Aspirate ◽  
Lymphoblastic Leukemia ◽  
Prognostic Significance ◽  
Flow Cytometric Analysis ◽  
Blood Film ◽  
High Side

<p class="abstract">We present a case of B-acute lymphoblastic leukemia in an elderly patient who presented with severe weakness and pancytopenia. The patient was a 75 year old Female whose blasts had an unusual morphology in form of coarse azurophilic granules and cytoplasmic blebs and on flow cytometry the blasts were present in the bright CD45 zone with a high side scatter. Bone marrow aspirate sample was subjected to multicolour flow cytometry using Beckman Coulter Navios® which is an 8 colour flow cytometer.  Flow cytometric analysis of the bone marrow aspirate showed blasts in the monocytic zone with a precursor B cell immunophenotype. Complete blood counts showed pancytopenia with peripheral blood film not showing any blasts. Bone marrow aspirate smears showed 20% blasts with coarse azurophilic granules and cytoplasmic blebs. The position of the blasts in this case which were in monocytic zone giving them a bright expression of CD45 and a high side scatter on the CD45 side scatter. This is not the usual position for blasts in B- acute lymphoblastic leukemia as these blasts are less complex. A bright expression of CD45 by blasts in B- acute lymphoblastic leukemia is known to be associated with a poor prognosis but the clinical significance of blasts being bright CD45 with a high side scatter is a very rare occurrence and more number of cases with a similar presentation are required to determine a prognostic significance.</p>

scholarly journals Development of Somatic NRAS Mutation Associated with Rapid Transition from Germline GATA2 Mutation Associated Myelodysplastic Syndrome to Acute Myeloid Leukemia

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 3616-3616 ◽  
Author(s):  
Yanqin Yang ◽  
Yubo Zhang ◽  
Jun Zhu ◽  
Catherine E. Lai ◽  
Jingrong Tang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Acute Myeloid Leukemia ◽  
Myelodysplastic Syndrome ◽  
Myeloid Leukemia ◽  
Research Funding ◽  
Bone Marrow Aspirate ◽  
Nras Mutation ◽  
Trisomy 8 ◽  
Acute Myeloid

Abstract There is increasing recognition of the role of inherited germline predisposition for myeloid disorders such as myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). The additional somatic genetic events required for development of a malignant phenotype are however poorly understood. A 25 year old woman was referred to the NHLBI hematology branch in March 2014 for a seven year history of pancytopenia. Her medical history included recurrent pneumonias, oral ulcers, severe varicella infection and arthralgias. Prior bone marrow examinations at ages 21 and 23 at outside institutions reported normocellular marrow, tri-lineage hematopoiesis and mild dyspoiesis. Cytogenetics were remarkable for trisomy 8 in 80% (aged 21) or 90% (aged 23) of metaphases. Previously unrecognized lymphedema was noted on examination. Peripheral blood counts showed WBC 2.28 K/ul [normal range: 3.98-10.04], HGB 9.9 g/dL [11.2-15.7], PLT: 67 K/ul [173-369], ALC: 0.36 K/ul [1.18-3.74] and AMC: 0.06 [0.24-0.86]. Peripheral blood flow cytometry demonstrated decreased CD3+ CD4+ (T) cells, CD19+ (B) cells and NK cells. HLA-DR15 negative. Bone marrow examination showed trilineage hematopoiesis, 50-60% cellularity, mild erythroid predominance and mildly increased, mildly atypical megakaryocytes. Blasts less than 5%. Bone marrow flow cytometry revealed severely decreased B-cells and monocytes, absent B-cell precursors, absent dendritic cells, inverted CD4:CD8 ratio, and atypical myeloid maturation pattern. Cytogenetics demonstrated stable trisomy 8 in 90% of metaphases. On the basis of this assessment the diagnosis of MDS was confirmed. Sanger sequencing revealed a GATA2 L375S mutation in the second zinc finger of known pathogenic significance. Four months later she developed increased fatigue and easy bruising with worsening thrombocytopenia (PLT: 10K/ul). Bone marrow was dramatically changed; now markedly hypercellular (90-100%) with diffuse sheets of immature cells consistent with blasts having fine chromatin, distinct or prominent nucleoli, and visible cytoplasm. Blasts were positive for CD33, CD56, CD64, CD123, and CD163; and were negative for CD34, CD14, and myeloperoxidase. Cytogenetics showed a new trisomy 20 in 65% of metaphases, in addition to previously seen trisomy 8 in 100%. A diagnosis of acute monoblastic leukemia (M5a subtype) was made. At both clinic visits bone marrow aspirate was collected on an IRB approved research sample acquisition protocol. Whole exome sequencing of 1ug DNA was performed using Agilent SureSelect v5 Exome enrichment Kits on an Illumina HiSeq 2000 with 100-bp paired-end reads (Macrogen, Rockville, MD). Data was mapped to hg19 (BWA) and processed using an in-house pipeline (Samtools/Picard/GATK/VarScan/Annovar). Mean read depth of target regions was 157 and 149. There was high correlation between both samples with the exception of a NRAS:NM_002524:exon3:c.C181A:p.Q61K mutation (57 of 180 reads) seen only in the later sample. Confirmatory ultra-deep sequencing for NRAS was performed using Illumina TruSight Myeloid Sequencing Panel on an Illumina MiSeq. No evidence of the NRAS Q61K mutation was found in the earlier March MDS bone marrow sample even when sequenced to a depth greater than 1750 reads (see figure). The mutation was confirmed in the August AML sample at a variant allele frequency of 35%. If heterozygous this would reflect a clone size of 70%, consistent with data from both cytogenetics (new trisomy 20 in 65% of metaphases) and the 76% blasts documented by bone marrow aspirate smear differential. We report here the rapid progression to AML in a patient with germline GATA2 MDS associated with development of a new trisomy 20 karyotype and a NRAS Q61K mutation. The NRAS mutation was not detectable after the patient achieved a complete remission following induction chemotherapy further supporting this association. This NRAS mutation has been implicated in the pathogenesis of multiple cancers by constitutive activation of proliferative signaling. GATA2 associated MDS is a high-risk pre-leukemic condition with the potential for rapid evolution to AML. This is the first report of acquired somatic mutations in the RAS/RTK signaling pathway in the context of germline GATA2 insufficiency associated with acute leukemic transformation. Figure 1. Figure 1. Disclosures Townsley: Novartis: Research Funding; GSK: Research Funding.

VPS4A : A Novel Candidate Gene for Congenital Dyserythropoietic Anemia

Blood ◽  
2017 ◽  
Vol 130 (Suppl_1) ◽  
pp. 923-923
Author(s):  
Katie Giger ◽  
Georgios E Christakopoulos ◽  
Lisa Trump ◽  
Clarissa E. Johnson ◽  
Haripriya Sakthivel ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Transferrin Receptor ◽  
Bone Marrow Aspirate ◽  
Critical Role ◽  
Congenital Dyserythropoietic Anemia ◽  
Spindle Poles ◽  
Cd34 Cells ◽  
Cytoplasmic Bridges

Abstract CDAR (ClinicalTrials.gov Identifier: NCT02964494), a registry for patients with Congenital Dyserythropoietic Anemia (CDA) in North America, was recently created with the goal to provide a longitudinal database and associated biorepository to facilitate natural history studies and research on the molecular pathways involved in the pathogenesis of CDAs. A 2 y.o. female patient with transfusion dependent anemia, pathologic diagnosis of Congenital Dyserythropoietic Anemia type I (CDA-I), and neurodevelopmental delay was enrolled in CDAR. Next Generation sequencing and deletion/duplication assay identified no mutations in the known CDA-associated genes, including CDAN1 and C15orf41, which are causative for CDA-I. Whole-exome sequencing for the patient and her parents (family-trio design) revealed a novel, de novo VPS4A missense variant located in the last codon of exon 8, potentially affecting splicing. VPS4A is an ATPase which, in association with the endosomal sorting complex required for transport (ESCRT), has been shown to play a critical role in cell division of HeLa cells in vitro, concentrating at the spindle poles during mitosis and at the midbody during cytokinesis. The aim of this work is to validate the pathogenetic role of the VPS4A variant for CDA and further investigate the role of VPS4A in erythroblast mitosis and cytokinesis. Central review of the patient's bone marrow aspirate smears revealed bi-nucleated erythroblasts in the range of 3-7%, a criterion compatible with CDA-I. However, cytoplasmic bridges were noted (arrows in Figure 1A) rather than the nuclear bridges typical of CDA-I. Immunofluorescence staining performed on erythroblasts generated ex vivo from normal CD34+ cells verified that VPS4A localizes to the spindle poles during mitosis and the midbody during cytokinesis in dividing human erythroid cells analyzed by Imaging Flow Cytometry (Figure 1B). The level of VPS4A mRNA expression in the patient's reticulocytes was evaluated by qPCR using three different sets of primers and found to be decreased by 55-70% compared to control reticulocytes and knock-down of VPS4A in normal CD34+ cells resulted in erythroid cultures enriched in binucleated cells. Induced pluripotent stem cells (iPSCs) were generated from the patient's peripheral blood mononuclear cells after the family's consent. Erythroblasts produced from these iPSCs exhibited decreased VPS4A localization at the spindle poles and midbody and fail to divide properly, frequently maintaining cytoplasmic bridges as seen in the patient's bone marrow. Additionally, flow cytometry analysis of the patient's peripheral blood cells stained with anti-CD71 for the transferrin receptor and Thiazol Orange (TO) for RNA revealed a unique cell population which is TO negative, yet CD71 positive implying that VPS4A is also involved in reticulocyte maturation, likely participating in vesicle formation and the normal exocytosis of the transferrin receptor. VPS4A appears to play a critical role in erythroblast mitosis and cytokinesis, as well as erythrocyte maturation, and is a novel candidate gene for congenital dyserythropoietic anemia. Figure 1. A) Binucleated erythroblasts and cytoplasmic bridges (arrows) were noted on the patient's bone marrow aspirate smears. B) VPS4A localizes at the spindle poles (upper image) and midbody (lower image) in normal human erythroblasts. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

A Useful Flow Cytometry Panel To Exclude Myelodysplastic Syndrome.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4848-4848
Author(s):  
Ling Zhang ◽  
Jean R. Lopatequi ◽  
Sharron E. Kelly ◽  
Tobi Neer ◽  
Stephen Lee
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Test Group ◽  
Bone Marrow Examination ◽  
Normal Group ◽  
P Value ◽  
Aberrant Expression ◽  
Cd10 Expression ◽  
Group A ◽  
Group B

Abstract Background: Myelodysplastic syndromes (MDS) are a heterogeneous group of hematopoietic disorders, which can be difficult to diagnose based only on morphologic bone marrow examination. Karyotyping can be useful in diagnosing borderline cases. However, only 40% of patients with MDS have an abnormal karyotype. Flow cytometry(FCM) approaches have been described but not clearly defined in MDS due to use of different criteria. We devised a 10-parameter FCM panel, mainly including myeloid and erythroid maturation markers, to differentiate MDS marrows from normal marrows. Design: Bone marrow from 91 patients with cytopenia(s) or anemia were included in the study(10/2005–7/2006). Cases were divided into 3 groups: normal, not morphologically suspicious for MDS, 46 cases, equivocal, morphologically suggestive but not diagnostic of MDS, 20 cases, morphologically diagnostic of MDS, 25 cases. 10 FCM paremeters were performed and scored: hypogranularity,aberrant expression of CD56,lack of CD10 expression,decreased CD64 expression,&lack of CD13 or CD33 expression,&abnormal CD13/CD16 or CD16/CD11b pattern,increase of CD 34 expression gating on all cells excluding erythrocytes,decreased expression of CD71/Glycophorin gating on erythroid precursors. Karyotypings were analyzed. Results: and (see Table 1 and 2). Karyotyping were anlayzed in 86 cases. Cytogenetic abnormalities were found in 2.2% (1/44) of normal group, 5.3% (1/19) of equivocal group and 34.8% (8/43) of MDS group. In MDS group 7 of 8 patients (87.5%) who had both morphologic and cytogenetic diagnosis of MDS were scored &gt;=3 of 8 FCM parameters. Conclusions: Study showed the 8-parameter FCM panel was more predictive of MDS than 10-parameters one. Both CD13/CD16 & CD16/CD11b patterns were considered to be non-specific. In 8-parameter panel, zero score tended to rule out MDS, while score &gt;=3 suggested MDS. The most specific FCM parameters were hypogranularity, CD34, aberrant expression of CD56 or lack of CD10 expression by mature granulocytes. CD71/Glycophorin A might be useful in identifying dysplasia in erythroid lineage. Table 1. Comparison of FCM Parameters in Patients with/without MDS Parameters/Group Normal Group(A)(n=46) Equivocal Group(B)(n=20) MDS Group(C)(n=25) CHITEST(P value)(GRoup A vs C) * These two parameters were deleted in the 8-parameter panel due to their high frequency seen in normal group. Hypogranularity 4 (8.7%) 6 (30%) 18 (72%) 0.0001 CD56 0 (0.0%) 1 (5%) 6 (24%) 0.0005 CD10 7 (15.2%) 7 (35%) 15 (60%) 0.0001 CD64 6 (13.1%) 2 (10%) 6 (24%) 0.2396 CD13 0 (0.0%) 0 (0.0%) 2 (8.0%) 0.0516 CD33 0 (0.0%) 2 (10 %) 0 (0%) 0 CD13/CD16* 23 (50.0%) 7 (35%) 21 (84%) 0.0047 CD34 8 (17.4%) 7 (35%) 13 (52%) 0.0026 CD71 2 (4.3%) 5 (25%) 12 (48%) 0.0001 CD16/CD11b* 27 (58.7%) 8 (40%) 22 (88%) 0.0107 Table 2. Scoring with 8 FCM Parameters in Patients with/without MDS Score/Group Normal Group (A)* (n=46) Equivocal Group (B)* (n=20) MDS Group (C)* (n=25) * Fish Exact T Test: Group A vs C: p&lt;0.0001, A vs B: p=0.0006, B vs C: p=&lt;0.0001 Score=0 21 (45.7%) 3 (15%) 0 (0.0%) Score=1–2 25 (543 %) 14 (70%) 8 (32 %) Score&gt;3 0 (0.0%) 3 (15%) 17 (68%) *Mean score +/− SD 0.65+/−1.06 1.55+/−1.54 2.8+/−1.65

scholarly journals Flow cytometry of DNA content in human bone marrow: a critical reappraisal

Blood ◽  
1980 ◽  
Vol 55 (5) ◽  
pp. 734-740
Author(s):  
GM Dosik ◽  
B Barlogie ◽  
W Gohde ◽  
D Johnston ◽  
JL Tekell ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Dna Content ◽  
Bone Marrow Aspirate ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Blood Contamination ◽  
Dna Distribution ◽  
M Phase ◽  
Normal Human

Because cytokinetic studies of the human bone marrow aspirate as a prognostic factor and as a monitor of drug perturbation are frequently inconsistent, we investigated reproducibility of DNA distribution measured by flow cytometry of DNA content in patients with morphologically normal bone marrow. In 15 patients, correlation was noted between DNA distributions simultaneously obtained on right and left iliac crest bone marrow aspirates (r = .588), although considerable variation in individuals was encountered. Much better reproducibility (r = .879) was achieved using bilateral core biopsy of bone marrow in these same patients. In 60 samples, comparison of DNA distribution between bone marrow aspirate and simultaneously obtained biopsy revealed higher relative proportions of S and G2 + M phase cells in biopsies (p less than 0.001), suggesting peripheral blood contamination of aspirate material. Brisk shaking of biopsy specimens in saline expelled a representative sample in the supernatant that could be subjected to simultaneous cytomorphological and cytokinetic analysis. To improve reproducibility of DNA content determinations in normal human bone marrow, bone marrow biopsy should be utilized.

scholarly journals Study of myelodysplastic features in patients with myelodysplastic syndromes by multicolor flow cytometry

2019 ◽  
Vol 13 (4) ◽  
pp. 75-88
Author(s):  
O. Yu. Davydova ◽  
I. V. Galtseva ◽  
E. N. Parovichnikova ◽  
A. V. Kokhno ◽  
N. M. Kapranov ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Myelodysplastic Syndromes ◽  
Bone Marrow Aspirate ◽  
Screening Method ◽  
Control Group ◽  
Allogeneic Bone ◽  
Multicolor Flow Cytometry ◽  
Color Flow ◽  
5Q Deletion

Background . Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal diseases of the hematopoiesis system characterized by dismyelopoiesis and cytopenia, the presence of cytogenetic aberrations and a high risk of transformation into acute myeloid leukemias. Diagnosis of MDS requires a comprehensive approach and mandatory performance of cytological, cytochemical and cytogenetic studies of bone marrow aspirate, as well as histological examination of trephine biopsy. However, in some cases it is necessary to undergo a diagnostic test that would allow verification of the MDS. The study of bone marrow aspirate by multicolor flow cytometry (MFC) can be considered as an additional diagnostic criterion in the diagnosis of MDS.The objective of the study was to estimate the incidence of myelodysplastic features in patients with various forms of MDS by the MFC method. Materials and methods . The study included 79 patients with MDS: 8 with MDS with 5q deletion, 33 with MDS without excess blast cells and 38 with excess of blasts. A bone marrow aspirate test was performed by 6-color flow cytometry. The control group included 35 donors of allogeneic bone marrow. The analysis resulted in a conclusion on the Ogata score scale, the Wells prognostic scale and the combined Ogata–Wells scale. When using the screening method, the presence of two or more cytometric signs of MDS was detected in 60 (75.9 %) of 79 MDS patients. Wells score was higher in MDS group with an excess of blast than in others. Using the combined Ogata–Wells scale, cytometric aberrations were found in 70 (88.6 %) of 79 MDS patients. In patients with MDS with an excess of blasts, the incidence of increased CD34+ and/or CD117+ myeloid cells was higher than in MDS patients without an excess of blasts and an MDS with a 5q deletion. The frequency of abnormal cytometric parameters (anomalous expression of CD34, CD117, CD56+ myeloblasts) in these groups did not differ. In patients with isolated 5q deletion and MDS without excess of blasts, an increased proportion of CD7+CD34+ cells was more often detected than in MDS with an excess of blasts.Conclusion . Thus, cytometric abnormalities in MDS are common, even in patients without excess of blasts. The MFC method can be used as an additional diagnostic method in the initial diagnosis of MDS.

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