Immature Blood Cells Research Articles

Abstract B35: Establishment of patient-derived xenograft models of acute leukemias.

Abstract Acute leukemia, a hematological malignancy, is characterised by a rapid increase of functionally immature blood cells. Acute Myeloid Leukemia (AML) and Acute Lymphoblastic Leukemia (ALL) are distinguished by their cells of origin. If untreated, AML and ALL are life-threatening and patients show different responses to current standards of care (SoC) therapy. The majority of acute leukemias are genetically heterogeneous and therefore a challenge for the development of new targeted therapies. There is a need to develop robust animal models to investigate leukemia in vivo. Patient-derived xenograft (PDX) mouse models recapitulate many clinical features of cancer. They have proven to be useful tools for the evaluation of novel therapeutic strategies and biomarkers, and for the study of cancer biology. The aim of our work was to establish PDX models of AML and ALL and to characterize their sensitivity to different therapeutic agents. Bone marrow aspirates were obtained from primary and relapsed AML/ALL patients and were intravenously (i.v.) and/or subcutaneously (s.c.) inoculated into immunodeficient mice. Mice were sacrificed when signs of disease were obvious and single cell suspensions of spleens (i.v.) or tumor fragments (s.c.) were transferred to new recipient mice. Chemosensitivity to SoC treatments was tested. In our study several AML-PDX and ALL-PDX were established. These models show different sensitivity to SoC treatment. The models will be used for further investigations like gene expression or mutation analysis or biomarker identification. The established PDX models of AML and ALL are suitable tools for preclinical drug development. In order to cover the complex heterogeneity of acute leukemias, the establishment of further PDX models is currently in progress. Citation Format: Antje Siegert, Bernadette Brzezicha, Annika Wulf-Goldenberg, Iduna Fichtner, Jens Hoffmann. Establishment of patient-derived xenograft models of acute leukemias. [abstract]. In: Proceedings of the AACR Special Conference on Hematologic Malignancies: Translating Discoveries to Novel Therapies; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(17 Suppl):Abstract nr B35.

Induction of autophagy is a key component of all-trans-retinoic acid-induced differentiation in leukemia cells and a potential target for pharmacologic modulation

Acute myeloid leukemia (AML) is characterized by the accumulation of immature blood cell precursors in the bone marrow. Pharmacologically overcoming the differentiation block in this condition is an attractive therapeutic avenue, which has achieved success only in a subtype of AML, acute promyelocytic leukemia (APL). Attempts to emulate this success in other AML subtypes have thus far been unsuccessful. Autophagy is a conserved protein degradation pathway with important roles in mammalian cell differentiation, particularly within the hematopoietic system. In the study described here, we investigated the functional importance of autophagy in APL cell differentiation. We found that autophagy is increased during all-trans-retinoic acid (ATRA)-induced granulocytic differentiation of the APL cell line NB4 and that this is associated with increased expression of LC3II and GATE-16 proteins involved in autophagosome formation. Autophagy inhibition, using either drugs (chloroquine/3-methyladenine) or short-hairpin RNA targeting the essential autophagy gene ATG7, attenuates myeloid differentiation. Importantly, we found that enhancing autophagy promotes ATRA-induced granulocytic differentiation of an ATRA-resistant derivative of the non-APL AML HL60 cell line (HL60-Diff-R). These data support the development of strategies to stimulate autophagy as a novel approach to promote differentiation in AML.

Molecular mechanisms of angioimmunoblastic T-cell lymphoma

Angioimmunoblastic T-cell lymphoma (AITL) is a distinct subtype of peripheral T-cell lymphoma. The origins of AITL tumor cells were thought to be follicular helper T (TFH) cells based on the common features of these two cell types. Recent findings suggest a multistep model for the development of AITL. The immature blood cells evolve into premalignant (prelymphoma) cells by acquisition of premalignant mutations including TET2 and/or DNMT3A mutations. The premalignant cells finally develop into full-blown tumor cells by accumulation of tumor-specific RHOA mutations. Combinations of premalignant and tumor-specific mutations may induce development of AITL.

Understanding multiple myeloma

Understanding multiple myeloma

Nonmarrow hematopoiesis occurs in a hyaluronic-acid-rich node and duct system in mice.

A hyaluronic-acid-rich node and duct system (HAR-NDS) was found on the surface of internal organs of mice, and inside their blood and lymph vessels. The nodes (HAR-Ns) were filled with immune cells of the innate system and were especially enriched with mast cells and histiocytes. They also contained hematopoietic progenitor cells (HPCs), such as granulocyte-macrophage, erythroid, multipotential progenitors, and mast cell progenitors (MCPs). MCPs were the most abundant among the HPCs in HAR-Ns. Their frequency was fivefold higher than that of the MCPs in bone marrow. In addition, the system contained pluripotent stem cells (PSCs) capable of producing CD45(-)Flk1(+) hemangioblast-like cells, which subsequently generated various types of HPCs and differentiated blood cells. Although HAR-Ns did not appear to harbor enough number of cells capable of long-term reconstitution or short-term radioprotection of lethally irradiated recipients, bone marrow cells were able to engraft in the HAR-NDS and reconstitute hematopoietic potentials of the system. PSCs and HPCs were consistently found in intravenous, intralymphatic, and intestinal HAR-ND. We infer that PSCs and HPCs reside in the HAR-ND and that this novel system may serve as an alternative means to traffic immature and mature blood cells throughout the body.

Chemotherapy Drug Scheduling for the Induction Treatment of Patients With Acute Myeloid Leukemia

Leukemia is an immediately life-threatening cancer wherein immature blood cells are overproduced, accumulate in the bone marrow (BM) and blood and causes immune and blood system failure. Treatment with chemotherapy can be intensive or nonintensive and can also be life-threatening since only relatively few patient-specific and leukemia-specific factors are considered in current protocols. We have already presented a mathematical model for one intensive chemotherapy cycle with intravenous (i.v.) daunorubicin (DNR), and cytarabine (Ara-C). This model is now extended to nonintensive subcutaneous (SC) Ara-C and for a standard intensive chemotherapy course (four cycles), consistent with clinical practice. Model parameters mainly consist of physiological patient data, indicators of tumor burden and characteristics of cell cycle kinetics. A sensitivity analysis problem is solved and cell cycle parameters are identified to control treatment outcome. Simulation results using published cell cycle data from two acute myeloid leukemia patients are presented for a course of standard treatment using intensive and nonintensive protocols. The aim of remission-induction therapy is to debulk the tumor and achieve normal BM function; by treatment completion, the total leukemic population should be reduced to at most 10(9) cells, at which point BM hypoplasia is achieved. The normal cell number should be higher than that of the leukemic, and a 3-log reduction is the maximum permissible level of population reduction. This optimization problem is formulated and solved for the two patient case studies. The results clearly present the benefits from the use of optimization as an advisory tool for treatment design.

Systemic Juvenile Idiopathic Arthritis–like Syndrome in Mice Following Stimulation of the Immune System With Freund's Complete Adjuvant: Regulation by Interferon‐γ

Systemic juvenile idiopathic arthritis (JIA) is unique among the rheumatic diseases of childhood, given its distinctive systemic inflammatory character. Inappropriate control of innate immune responses following an initially harmless trigger is thought to account for the excessive inflammatory reaction. The aim of this study was to generate a similar systemic inflammatory syndrome in mice by injecting a relatively innocuous, yet persistent, immune system trigger: Freund's complete adjuvant (CFA), containing heat-killed mycobacteria. Given the central role of interferon-γ (IFNγ) in immune regulation, we challenged wild-type (WT) and IFNγ-knockout (KO) BALB/c mice with CFA, and analyzed their clinical symptoms and biologic characteristics. The production of cytokines and the effects of anticytokine antibodies were investigated. In WT mice, CFA injection resulted in splenomegaly, lymphadenopathy, neutrophilia, thrombocytosis, and increased cytokine expression. In the absence of IFNγ, these symptoms were more pronounced and were accompanied by weight loss, arthritis, anemia, hemophagocytosis, abundance of immature blood cells, and increased levels of interleukin-6 (IL-6), all of which are reminiscent of the symptoms of systemic JIA. CFA-challenged IFNγ-KO mice showed increased expression of IL-17 by CD4+ T cells and by innate γ/δ T cells. Inflammatory and hematologic changes were prevented by treatment with anti-IL-12/IL-23p40 and anti-IL-17 antibodies. Immune stimulation of IFNγ-KO mice with CFA produces a systemic inflammatory syndrome reflecting the clinical, biologic, and histopathologic picture of systemic JIA. The protective function of IFNγ in preventing anemia and overall systemic inflammation is a striking observation. The finding that both adaptive and innate T cells are important sources of IL-17 may be of relevance in the pathogenesis of systemic JIA.

NonHodgkin's Lymphoma with Peritoneal Localization

The gastrointestinal tract is the most common extranodal site involved with lymphoma accounting for 5–20% of all cases. Lymphoma can occur at any site of the body, but diffuse and extensive involvement of the peritoneal cavity is unusual and rare. We report a case of diffuse large B-cell lymphoma in a 57-year-old female infiltrating the peritoneum and omentum and presenting with ascites and pleural effusion. The performed examinations did not discover any pathological findings affecting the digestive tract or parenchymal organs, except for diffuse thickening of the peritoneum and omentum. Peripheral, mediastinal, or retroperitoneal lymphadenopathy was not registered. The blood count revealed only elevated leukocytes and on examination there were no immature blood cells in the peripheral blood. The cytology from the ascites and pleural effusion did not detect any malignant cells. Due to the rapid disease progression the patient died after twenty-two days of admission. The diagnosis was discovered postmortem with the histological examination and immunohistochemical study of the material taken during the surgical laparoscopy performed four days before the lethal outcome. Although cytology is diagnostic in most cases, laparoscopy with peritoneal biopsy is the only procedure which can establish the definitive diagnosis of peritoneal lymphomatosis.

The spectrum of paraneoplastic cutaneous vasculitis in a defined population: incidence and clinical features.

Cutaneous vasculitis may be associated with malignancies, and may behave as a paraneoplastic syndrome. This association has been reported in a variable proportion of patients depending on population selection. We conducted the current study to assess the frequency, clinical features, treatment, and outcome of paraneoplastic vasculitis in a large unselected series of 766 patients with cutaneous vasculitis diagnosed at a single university hospital.Sixteen patients (10 men and 6 women; mean age ± standard deviation, 67.94 ± 14.20 yr; range, 40–85 yr) presenting with cutaneous vasculitis were ultimately diagnosed as having an underlying malignancy. They constituted 3.80% of the 421 adult patients. There were 9 hematologic and 7 solid underlying malignancies. Skin lesions were the initial clinical presentation in all of them, and the median interval from the onset of cutaneous vasculitis to the diagnosis of the malignancy was 17 days (range, 8–50 d). The most frequent skin lesions were palpable purpura (15 patients). Other clinical manifestations included constitutional syndrome (10 patients) and arthralgia and/or arthritis (4 cases). Hematologic cytopenias (11 cases) as well as immature peripheral blood cells (6 cases) were frequently observed in the full blood cell count, especially in those with vasculitis associated with hematologic malignancies.Specific treatment for vasculitis was prescribed in 10 patients; nonsteroidal antiinflammatory drugs (4 patients), corticosteroids (3 patients), chloroquine (1 patient), antihistamines (1 patient), and cyclophosphamide (1 patient). Ten patients died due to the malignancy and 6 patients recovered following malignancy therapy. Patients with paraneoplastic vasculitis were older, more frequently had constitutional syndrome, and less frequently had organ damage due to the vasculitis than the remaining patients with cutaneous vasculitis.In summary, cutaneous paraneoplastic vasculitis is an entity not uncommonly encountered by clinicians. The most common underlying malignancy is generally hematologic. In these cases the presence of cytopenias and immature cells may be red flags for the diagnosis of cancer. In patients with paraneoplastic cutaneous vasculitis, the prognosis depends on the underlying neoplasia.

FRI0228 Cutaneous vasculitis as a presenting manifestation of neoplasia

BackgroundCutaneous Leukocytoclastic Vasculitis (CLV) may be associated with malignancies, and sometimes it behaves as a paraneoplastic syndrome. This association has been reported in a variable proportion of CLV patients (from...

Clinical analysis of orbital chloroma

To analyze and summarize the clinical aspects, diagnostic methods and treatment principle of the orbit chloroma. It was a retrospective case series study. The clinical aspects of 17 orbital chloroma cases were analyzed, which included symptoms, signs, imageology, diagnosis, treatment and prognosis. Orbit chloroma was more prevalent in males, and usually occurred in children and individuals under 14 years old. The main symptoms were exophthalmos and orbit masses. B ultrasonic examination was performed in 16 cases, which found that the mass was located in the temporal side of the orbit, with irregular shape and clear edge. Color doppler imaging was performed in 5 cases, which found that the sufficient blood supply signal was the main manifestation. CT scan was performed in 11 cases, which revealed that the mass was located in temporal side of the orbit, with irregular shape, regular density, a clear edge and seldom destruction of the orbital bone. Blood examination was performed in 17 cases, which revealed the increase of white blood cells and the presence of numerous immature blood cells. Sixteen cases were referred to the department of hematology for treatment. In 4 following cases, 2 cases were died within 3 months. Orbital chloroma can be easily misdiagnosed as other types of malignant orbital tumor in children. The diagnosis of orbital chloroma can be established by radiology and hematological examinations. This tumor can be treated by chemotherapy and radiotherapy.

Predictive Value of Immature Reticulocyte and Platelet Fractions in Hematopoietic Recovery of Allograft Patients

Hematopoietic progenitor cell transplantation is the treatment of choice for patients with malignant hematologic diseases. Neutrophil (NEUT) and platelet (PLT) counts are used to evaluate hematologic engraftment of transplanted patients. Recent-generation hematology analyzers offer an alternative way to evaluate immature peripheral blood (PB) cell fractions, which may also give an indication of hematopoietic recovery. The immature reticulocyte fraction (IRF) and immature platelet fraction (IPF) in PB samples may provide early indicators of transplant success. We evaluated the predictive value of IRF and IPF for the hematologic recovery of 46 adult patients undergoing allogeneic PB progenitor cell transplantation. We observed that IRF recovery anticipated by 4 days compared with NEUT recovery (11 vs 15 d) and IPF by 2 days compared with PLT (10 vs 12 d). The recovery was different for patients undergoing a nonmyeloablative regimen (NMA); we observed an early IRF recovery by 5 days compared with NEUT (10 vs 15 d) and a IPF compared with PLT recovery by 2 days (9 vs 11 d). We also observed significant correlations between NEUT and PLT recovery with recoveries of the new parameters IRF and IPF. We concluded that IRF and IPF predicted hematopoietic recovery. For allografted patients after NMA regimens, prediction was even more clinically relevant. These immature fractions open new perspectives for monitoring patient transfusion support through the posttransplantation recovery.

Synergistically Regulated Genes and Pathways in Leukemias Induced by Co-Expression of BCR-ABL and Nup98-HoxA9.

Abstract 3473Poster Board III-410Leukemia arises when normal hematopoiesis is altered causing an over-production of immature blood cells. In most cases, mutations appear to occur early in hematopoietic development affecting the more primitive stem/progenitor cell compartment, generating a leukemia stem cell (LSC). The BCR-ABL translocation is a common abnormality linked to chronic myeloid leukemia (CML), predisposing patients to the acute form of the disease, blast crisis (bcCML). The genetic interplay between BCR-ABL and the secondary mutations required for acute leukemia is poorly understood. Therefore, to better define the transformation process, we adapted a strategy previously described by McMurray et al in which genes synergistically regulated by two cooperating oncogenes are identified. This approach was shown to identify key targets of cancer cell survival and potential targets for therapeutic intervention. To implement this strategy for bcCML we employed a mouse model in which primitive (lineage depleted) mouse bone marrow cells are engineered through retroviral transduction to express BCR-ABL alone or in combination with another oncogene associated with human leukemia; Nup98-HoxA9. We generated leukemic mice and isolated bone marrow cells using fluorescence activated cell sorting (FACS). Each oncogene was tagged with a distinct fluorescent marker (green fluorescent protein for BCR-ABL and yellow fluorescent protein for Nup98-HoxA9) allowing for isolation of purified cell populations. From six independent cohorts, 4 distinct primitive (lineage negative) cell populations were purified: 1) BCR-ABL+, 2) Nup98-HoxA9+, BCR-ABL+/Nup98/HoxA9+, and BCR-ABL−/Nup98HoxA9− (i.e endogenous normal cells). Probes from each cell population were analyzed using mouse 430 Plus microarray chips from Affymetrix. Genome wide microarray analysis was performed by first comparing gene expression between non-malignant (endogenous normal) and bcCML (both oncogenes) cells providing a list of genes differentially expressed in the disease state. This gene-set was further refined by comparison to cells expressing each oncogene independently, identifying 78 genes that are synergistically regulated by both BCR-ABL and Nup98-HoxA9 in primitive leukemic cells. We term this gene set, CRGs, for cooperation response genes, the nomenclature previously established by McMurray et al. The aberrant expression of 8 specific CRGs (serpinB2, AREG, EREG, pla2g7, csf2, selp, irf8, mycn) was verified using quantitative real-time PCR, and found to be in close agreement with the gene array data. Next, pathway analysis of the CRG gene set was performed using the Database for Annotation, Visualization and Integrated Discovery (DAVID). Pathways identified by this analysis include activation of JAK-STAT, Hedgehog, and Erb2 signaling, as well as increased xenobiotic metabolism. Interestingly, our data set suggests that activation of Erb2 signaling by AREG and EREG acts through STAT5, thereby influencing cell survival. In addition, drug metabolism through cytochrome P450 is observed to be up-regulated in primitive leukemia cells, indicating an enhanced ability to tolerate therapeutic agents. Taken together, the analyses performed thus far have identified gene targets implicated as central to the growth of primitive leukemia cells. Going forward, we will evaluate how the 78 synergistically-regulated genes are modulated in response to various forms of therapy, and determine whether perturbation of specific genes/pathways can selectively inhibit/eradicate LSC. Disclosures:No relevant conflicts of interest to declare.

Function of retinoid acid receptor α and p21 in all-trans-retinoic acid-induced acute T-lymphoblastic leukemia apoptosis

All-trans-retinoic acid (ATRA) is a morphogenetic signalling molecule derived from vitamin A and is used clinically to target acute promyelocytic leukemia by inducing differentiation of immature blood cells. Retinoid signals are mediated by retinoic acid (RA) receptors (RARs) and retinoid X receptors (RXRs). Retinoic acid receptors consist of RARα, RARβ and RARγ isotypes. Among these components, RARα is preferentially bound to ATRA, which is used to treat acute T-lymphoblastic leukemia, yet the conditions and mechanisms remain unknown. In this study, we have demonstrated that, in human acute T-lymphoblastic leukemia Molt3 cells, inhibition of RA-induced proliferation results from massive cell death characterised by apoptosis. The effect of ATRA:RARα binding on apoptosis in Molt3 cells has been investigated. Consequently, it has been shown that, in RA-treated Molt3 cells, upregulation of p21 due to RA accompanies caspase 3/PARP activation which precedes the occurrence of apoptosis.

Granulocytic sarcoma of the skin

An 89-year-old woman with refractory anemia presented with a painful neck mass that measured 5 cm × 3 cm ([Figure 1][1]A). Her peripheral leukocyte count was (118 × 109/L), with 98% immature blood cells. A biopsy of the mass showed massive infiltration of the subcutaneous connective tissue by

Murine Hematopoietic Stem Cells and Multipotent Progenitors Express Truncated Intracellular Form of c-kit Receptor

The c-kit receptor plays a vital role in self-renewal and differentiation of hematopoietic stem cells (HSCs) and multipotent progenitors (MPPs). We have discovered that besides c-kit, the murine multipotent HSC/MPP-like cell line EML expresses the transcript and protein for a truncated intracellular form of c-kit receptor, called tr-kit. Notably, the tr-kit transcript and protein levels were down-regulated during cytokine-induced differentiation of the HSC/MPP-like cell line EML into myeloerythroid lineages. These findings prompted us to analyze tr-kit expression in purified murine fetal liver and bone marrow cell populations containing long-term repopulating (LTR) HSCs, short-term repopulating (STR) HSCs, MPPs, lineage-committed progenitors, and immature blood cells. Remarkably, these studies have revealed that in contrast to more widespread expression of c-kit, tr-kit is transcribed solely in cell populations enriched for LTR-HSCs, STR-HSCs, and MPPs. On the other hand, cell populations in which HSCs and MPPs are either present at a much lower frequency or are absent altogether, cells representing more advanced stages of differentiation into lymphoid and myeloid lineages do not express tr-kit. The observation that tr-kit is co-expressed with c-kit only in more primitive HSC- and MPP-enriched cell populations raises an exciting possibility that tr-kit functions either as a new component of the stem cell factor (SCF)/c-kit pathway or is involved in a novel signaling pathway, present exclusively in HSC and MPPs. Taken together, these findings necessitate functional characterization of tr-kit and analysis of its potential role in the self-renewal, proliferation, and/or differentiation of HSC and multipotent progenitors.

Establishing a Bayesian predictive survival model adjusting for random effects

Myelodysplastic syndrome (MDS), sometimes referred to as pre-leukemia or smoldering leukemia, is a group of diseases usually characterized by failure of the bone marrow to produce enough normal blood cells. In about one-third of patients, the disease transforms into acute leukemia. In high-risk MDS, the bone marrow contains too many immature blood cells known as blasts. Patients with high-risk MDS survive for an average of 6–12 months. We have taken data from a large clinical trial and re-examined it considering the pre-leukemia as a random event in a Weibull distribution model. The issue becomes that of examining the affect of the variable on the predictive survival. We have taken the intercept, stress effect and shape parameter of the distribution to be random effects as well with realistic prior distributions based on previous shapes of the survival experience of subjects with this disease. We demonstrate how the model performs under relevant clinical conditions. The conditions are all tested using a Bayesian statistical approach allowing for the robust testing of the model parameters under various stress conditions which we introduce into the model. The convergence of the parameters to stable values are seen in trace plots which follow the convergence patterns This allows for precise estimation for determining clinical conditions under which the survival pattern will change. We give a numerical example of our results. The major platform for the theoretical development follows the Bayesian methodology and the multiple parameter Weibull model with random effects having carefully chosen hyper parameters. We have implemented the basic infrastructure for the analysis using the commercially available WinBugs software employing the Markov Chain Monte Carlo (MCMC) methodology.

SIC – a new hematologic parameter for early diagnosis of sepsis in preterm infants

Introduction: Early and accurate diagnosis of sepsis in preterm infants is vital. Despite intensive research in the field of early and reliable sepsis screening tests, no satisfactorily sensitive and specific parameter has been found so far. Aim: To evaluate a new hematologic parameter for the early diagnosis of nosocomial sepsis in preterm infants in comparison to interleukin-8 (IL-8). This new hematological parameter called „SIC“ (Single Infection and Control parameter) is calculated on the basis of the immature information (IMI) channel by the Sysmex XE-2100 blood analyzer which represents information regarding immature blood cells after selective lyses of leukocytes. The parameter is gained automatically and simultaneously by routine blood count measurement which requires neither additional blood sampling from the patient nor an increased volume of blood. Methods: Twenty-five preterm infants below 32 weeks' of gestation with culture-positive nosocomial sepsis were compared to 49 controls of same gestational age. A ROC plot was calculated with regard to SIC and IL-8. Results: Adopting a cut-off of 1.07at the time of clinical sepsis suspicion, the following values for SIC were calculated: sensitivity 76,9%, specificity 87.7%, PPV 55,6 and NPV 95. For IL-8 with a cut-off of 70pg/ml, the respective values were: 60%, 72,4%, 30, and 90,2. The area under the ROC plot was greater for SIC than for IL-8 (SIC: 0.834 and IL-8 0.693). Conclusion: Based on the overall shape of the two ROC plots, the SIC test appeared to be superior to IL-8 for diagnosis of nosocomial bacterial sepsis in preterm infants. A prospective study for evaluation of the new parameter in a higher number of patients is in progress.

Three‐dimensional distribution patterns of CD34 antigen on nonactivated cord blood cells

Circulating immature blood cells are inhomogeneous in most cytometric parameters, including CD34 expression. This surface antigen forms patches, and we studied their spatial distribution pattern by processing staggered and digitalized microscopical images; the number, fluorescence intensity, and volume were determined in randomly identified CD34-positive cord blood cells in suspension. Quantitative fluorescence analysis of individual CD34-labeled cells was performed after acquisition of microscopical images in high resolution by using a CCD camera and adjacent deconvolution. Two major (n = 42 and n = 41) and one minor (n = 5) group of haematopoietic progenitors with different CD34+ distribution patterns were detected. All CD34 antigen patches are localized on the spherical cell membrane, and differ most significantly in fluorescence intensity (P < 10(-4)), antigen volume (P < 0.004), and patch number per cell (P < 0.02). If all parameters are employed, 96.6% of CD34+ cells are correctly classified into one of the three groups as shown by discriminant analysis. If fluorescence intensity is eliminated, recognition efficiency decreases to 68.2%, indicating that this is the most powerful single parameter. While total fluorescence intensity is most characteristic for each group, total patch volume and number per cell, and heterogeneity of patch volumes and their spatial distribution also contribute significantly to reliable classification into their groups.

Prognostic factors and risk assessment in chronic myelomonocytic leukemia: Validation study of the M.D. Anderson Prognostic Scoring System

Chronic myelomonocytic leukemia continues to be a poorly understood disease, defined by clinical rather than biological features, with no consensus on optimal therapy. In the past, patients were often assessed for risk using scoring systems developed for other diseases, notably the International Prognostic Scoring System commonly used for myelodysplastic syndrome. The M.D. Anderson Prognostic Scoring System, using hemoglobin, absolute lymphocyte count, peripheral blood immature cells, and bone marrow blasts, was developed specifically for CMML; it was based on retrospective analysis of 213 patients. This report re-examines the validity of this scoring system based on follow-up of the initial cohort and prospectively examines its validity in 250 new patients. Both the original MDAPS system and a modified version derived from data of the initial cohort after extended follow-up (substituting lactate dehydrogenase for bone marrow blasts) effectively stratify both patient cohorts by survival and provide a useful risk assessment tool and additional guidance during treatment decisions.