RESUMEN
Acute promyelocytic leukemia (subtype M3) is characterized by malignant promyelocytes exhibiting an abundance of abnormally large or aberrant primary granules. Myeloperoxidase (MPO) activity of these azurophilic granules, as assessed by cytochemical staining, is unusually intense. In addition, M3 is universally associated with a chromosomal translocation, t(15;17)(q22;q11.2). In this report, the MPO gene was localized to human chromosome 17 (q12-q21), the region of the breakpoint on chromosome 17 in the t(15;17), by somatic cell hybrid analysis and in situ chromosomal hybridization. By means of MPO complementary DNA clones for in situ hybridization and Southern blot analysis, the effect of this specific translocation on the MPO gene was examined. In all cases of M3 examined, MPO is translocated to chromosome 15. Genomic blot analyses indicate rearrangement of MPO in leukemia cells of two of four cases examined. These findings suggest that MPO may be pivotal in the pathogenesis of acute promyelocytic leukemia.
Asunto(s)
Leucemia Mieloide Aguda/enzimología , Peroxidasa/genética , Translocación Genética , Médula Ósea/análisis , Mapeo Cromosómico , Cromosomas Humanos Par 15 , Cromosomas Humanos Par 17 , ADN/genética , Enzimas de Restricción del ADN , ADN Recombinante , Humanos , Leucemia Mieloide Aguda/genética , Hibridación de Ácido Nucleico , Plásmidos , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
Erythroleukemia (EL) is a rare form of myelogenous leukemia the classification and definition of which has evolved over the course of its 80-year descriptive history. In 1976 the French American British (FAB) Cooperative Group included EL within the classification system of acute myelogenous leukemias as AML-M6, and agreed on a quantitative standard to be used in the diagnosis of this disorder. The standards were revised in 1985 to the form in use today. We selected a series of 15 cases from our records which specifically fit the FAB criteria for AML-M6. Extensive direct comparison between our series and the old literature is not practical because of the changes which have occurred in classification and definition of the disease. Overall we found a rough correlation between the clinical and laboratory data shown in the old literature on EL and data from our cases. These cases underscore characteristic laboratory features which correspond to what is now defined as AML-M6: these patients present with pancytopenia, frequent peripheral blood nRBCs and no, or few, peripheral blood blasts. In addition, we note the presence of a hybrid myeloid-erythroid blast in the bone marrow in this disease and suggest that this may be characteristic of this type of AML. Old literature on EL has generally shown it to be a disease of the elderly, yet we found a subset of younger patients whose clinical outcome was significantly better than that of the older patients. Finally, EL has historically been viewed as a disease in which patients progress from a prodrome through erythroleukemia to other acute myeloid leukemia (AML) subtypes. Consistent with this idea, half of our 15 patients had been previously diagnosed with myelodysplastic syndrome or received chemotherapy. On the other hand only one of the 15 patients converted to another type of AML during his course.
Asunto(s)
Leucemia Eritroblástica Aguda/epidemiología , Adolescente , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Anemia/etiología , Biomarcadores de Tumor , Médula Ósea/patología , Aberraciones Cromosómicas , Células Precursoras Eritroides/patología , Femenino , Humanos , Incidencia , Leucemia Eritroblástica Aguda/clasificación , Leucemia Eritroblástica Aguda/complicaciones , Leucemia Eritroblástica Aguda/patología , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/diagnóstico , Síndromes Mielodisplásicos/patología , Estudios RetrospectivosRESUMEN
An unusual myelodysplastic syndrome with similar features in two patients is described. The entity is characterized by a maturation arrest at the myelocyte stage, strikingly clumpy chromatin, and a clinical course marked primarily by difficulties caused by anemia and thrombocytopenia. Electron microscopic description of the characteristic abnormal clumpy chromatin cells is included. While the disorder is unquestionably a myelodysplastic syndrome, it is clearly distinct from chronic myelogenous leukemia, chronic myelomonocytic leukemia, and the spectrum of refractory anemias with excess blasts.
Asunto(s)
Anomalía de Pelger-Huët/patología , Preleucemia/patología , Anciano , Cromatina/ultraestructura , Humanos , Microscopía ElectrónicaRESUMEN
Morphologic observation of the peripheral blood smear from a patient with chronic granulocytic leukemia suggested the presence of eosinophilic and basophilic granules in the same individual granulocyte. To unambiguously identify eosinophilic and basophilic granules simultaneously in the same preparation, the authors developed a cytochemical staining procedure using basophil-specific toluidine blue and eosinophil-specific cyanide-resistant peroxidase. Using this new dual stain, they demonstrated that ten out of ten chronic granulocytic leukemia patients they examined had cells that contained both eosinophilic and basophilic granules. The identity of the granules was corroborated by electron microscopic studies. These observations suggest that lineage confusion is common in chronic granulocytic leukemia.
Asunto(s)
Basófilos/patología , Gránulos Citoplasmáticos/patología , Eosinófilos/patología , Leucemia Mieloide/patología , Histocitoquímica , Humanos , Recuento de Leucocitos , Microscopía Electrónica , Coloración y EtiquetadoRESUMEN
Macrocytosis is most commonly associated with vitamin B12 and folate deficiencies, followed by alcoholism, liver disease, and malignant neoplasms. Many laboratories have observed that in recent years macrocytosis increasingly has been associated with zidovudine treatment of acquired immune deficiency syndrome. One hundred consecutive inpatients in a large metropolitan urban hospital with mean corpuscular volumes greater than 110 fL were studied; 44% were patients with acquired immune deficiency syndrome being treated with zidovudine, 19% were alcoholics, and 12% had malignant neoplasms. Only 3% were folate deficient and just 4% were vitamin B12 deficient. This study suggests that zidovudine has become the most common cause of macrocytosis in the hospitalized urban patient population and that vitamin B12 and folate deficiencies have decreased in proportion.
Asunto(s)
Eritrocitos Anormales , Zidovudina/efectos adversos , Síndrome de Inmunodeficiencia Adquirida/sangre , Síndrome de Inmunodeficiencia Adquirida/tratamiento farmacológico , Alcoholismo/sangre , Índices de Eritrocitos , Enfermedades Hematológicas/inducido químicamente , Enfermedades Hematológicas/etiología , Humanos , Neoplasias/sangre , Deficiencia de Vitamina B/sangreRESUMEN
After the introduction of Coulter S-Cal, a bias became apparent between platelet counts obtained from instruments calibrated with this material and those obtained from a Clay Adams Ultra-Flo. Statistical methods were used to compare platelet counts obtained from the Coulter S-Plus IV, Ortho ELT-800, Clay Adams Ultra-Flo 100, and phase microscopy. At a P value of 0.01, paired t analysis revealed statistically significant biases between the Ultra-Flo and each of the other methods. Significant biases were also found between phase microscopy and each of the other methods, although these were of a smaller magnitude. The results indicate the necessity for users of multiple platelet counting methods to conduct comprehensive interinstrument evaluations, particularly when altering methods of calibration.
Asunto(s)
Recuento de Plaquetas/métodos , Humanos , Microscopía de Contraste de Fase , Valores de Referencia , Estadística como AsuntoRESUMEN
Dark pigmented inclusions were found in the monocytes and polymorphonuclear leukocytes of a patient with widely metastatic malignant melanoma. Special stains demonstrated the granules were composed of melanin. The authors are not aware of any previous case report of patients with metastatic malignant melanoma that had leukocytic melanin inclusions. Prospective search for this phenomenon in malignant melanoma patients may allow such a finding to be used as a diagnostic tool.
Asunto(s)
Leucocitos/análisis , Melaninas/sangre , Melanoma/sangre , Neoplasias Cutáneas/sangre , Humanos , Masculino , Persona de Mediana Edad , Monocitos/análisis , Metástasis de la Neoplasia , Neutrófilos/análisisRESUMEN
Polymerase chain reaction-based methods for the detection of translocation-induced gene rearrangements are now widely used for diagnostics and patient monitoring. This article concentrates on two of the best studied chromosome translocations resulting in specific gene rearrangements and oncogene activation: the Philadelphia translocation of chronic myelogenous leukemia and acute leukemias, and the t(14;18) translocation of follicular lymphomas.
Asunto(s)
Aberraciones Cromosómicas , Leucemia/genética , Linfoma/genética , Reacción en Cadena de la Polimerasa , Proteínas de Fusión bcr-abl/genética , Reordenamiento Génico , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva , Cromosoma Filadelfia , Translocación GenéticaRESUMEN
In the last decade our understanding of acute promyelocytic leukemia (APL) has advanced tremendously. The recognition of all-trans retinoic acid (ATRA) as a powerful therapeutic agent paralleled the cloning of the t(15;17) breakpoint. RtPCR for the PML-RARA hybrid mRNA has become the hallmark of molecular diagnosis and molecular monitoring in APL. Current techniques are useful in predicting complete remission and a possible cure in many patients who repeatedly test negative by PCR. Standardizing techniques and improving the sensitivity of the assay are important. Doing this in a way so that clinically relevant minimal residual disease can be distinguished from "indolent disease" remains among the future challenges in APL.
Asunto(s)
Leucemia Promielocítica Aguda/diagnóstico , Trasplante de Médula Ósea , Humanos , Interferones/farmacología , Leucemia Promielocítica Aguda/genética , Leucemia Promielocítica Aguda/terapia , Neoplasia Residual , Reacción en Cadena de la Polimerasa , Translocación Genética , Tretinoina/uso terapéuticoRESUMEN
The granule proteins are among the most abundant and characteristic proteins of myeloid cells. They are essential for the antimicrobial activity of these cells and they provide important markers for the differentiation stage of the myeloid series and for the diagnosis of myeloid leukemias. In acute promyelocytic leukemia (APL) there is high production of myeloperoxidase, and its cytochemical detection as well as the t(15;17) chromosomal translocation are important markers in the diagnosis of this acute myelogenous disease. The expression of other granule protein genes in APL has not been systematically determined. We have used the reverse transcriptase-polymerase chain reaction (RT-PCR) method to determine the pattern of expression of granule protein genes at the mRNA level in APL cells. We have examined the expression of the primary granule proteins defensin, myeloperoxidase, elastase, and cathepsin G; the secondary granule proteins lactoferrin, collagenase, and transcobalamin; as well as lysozyme, a protein reportedly found in both primary and secondary granules. mRNAs for all of these granule proteins were present in normal bone marrow mononuclear cells. We found that APL cells from three patients contain, in addition to myeloperoxidase mRNA, mRNAs for elastase, cathepsin G, and lysozyme. One patient had faint but detectable lactoferrin mRNA signal, but collagenase and transcobalamin mRNAs were not detectable in this patient. Defensin mRNA was found in one of the three APL patients, and all the primary granule protein mRNAs measured were found to be expressed in the APL cell line NB4. None of the secondary granule protein mRNAs measured were detectable in NB4 cells. After treatment with retinoic acid (RA), which induces neutrophil maturation of these cells, weak induction of lactoferrin and collagenase but not transcobalamin was observed. However, in view of the weak transcobalamin signal observed in normal bone marrow, the absence of transcobalamin in RA-induced NB4 cells must be interpreted with caution. Interestingly, elastase and cathepsin G mRNA disappeared after RA induction, whereas defensin and myeloperoxidase mRNAs remained present. These findings indicate that granule protein mRNAs are regulated separately and differently, and that only minimal expression of secondary granule protein genes can occur in APL cells.
Asunto(s)
Leucemia Promielocítica Aguda/metabolismo , Biosíntesis de Proteínas , ARN Mensajero/análisis , Actinas/biosíntesis , Secuencia de Bases , Proteínas Sanguíneas/biosíntesis , Catepsinas/biosíntesis , Colagenasas/biosíntesis , ADN Complementario , Defensinas , Humanos , Lactoferrina/biosíntesis , Datos de Secuencia Molecular , Muramidasa/biosíntesis , Elastasa Pancreática/biosíntesis , Peroxidasa/biosíntesis , Reacción en Cadena de la Polimerasa , Transcobalaminas/biosíntesis , Células Tumorales CultivadasRESUMEN
The acute promyelocytic leukemia cell line, NB4, can be induced to differentiate to mature granulocytes by retinoic acid treatment. A novel retinoic acid-inducible cDNA clone, designated RI58, was isolated from a cDNA library constructed from retinoic acid-treated NB4 cells by differential hybridization. RI58 cDNA encodes a protein of 58kDa which has a similarity in its amino acids sequence to interferon (IFN)-inducible proteins. In addition, RI58 was induced by recombinant human IFN-alpha (rhIFN-alpha) in NB4 cells. RI58 was detectable within 4 hours post-stimulation with rhIFN-alpha, while it took as long as 1day after retinoic acid stimulation. Culture supernatant from retinoic acid-treated NB4 cells also induced RI58 expression similarly as rhIFN-alpha. This activity in culture supernatant was inhibited by anti-leukocyte IFN antiserum which showed specific reactivity to rhIFN-alpha. These results indicate that RI58 is induced by retinoic acid stimulation through autocrinally secreted IFN-alpha from NB4 cells. In the retinoic acid-treated NB4 cells, the expression of RI58 was increased along the process of differentiation. On the other hand, it was expressed constitutively in untreated non-hematopoietic cell lines and mature hematopoietic cell lines.
Asunto(s)
Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Interferones/farmacología , Leucemia Mieloide Aguda/genética , Proteínas de Neoplasias/genética , Secuencia de Aminoácidos , Comunicación Autocrina/genética , Secuencia de Bases , Diferenciación Celular/genética , Línea Celular , Clonación Molecular , ADN Complementario/aislamiento & purificación , Humanos , Leucemia Mieloide Aguda/metabolismo , Datos de Secuencia Molecular , Proteínas de Neoplasias/biosíntesis , Homología de Secuencia de Aminoácido , Tretinoina/farmacología , Células Tumorales CultivadasRESUMEN
Protein markers are often used to corroborate the morphological subtyping of hematopoietic malignancy. Most commonly, surface markers are used for the phenotyping of hematopoietic cells; however, internal proteins have also been used as markers. Glycophorin, hemoglobin A, hemoglobin F, and transferrin have all been used as markers for the erythroid phenotype. We have recently shown that carbonic anhydrase is constitutively and aberrantly expressed in two erythroleukemic cell lines. We here show that it is also present in high levels in primary erythroleukemic blasts and that it is a useful marker for the M6 phenotype when classifying acute nonlymphocytic leukemia.
Asunto(s)
Anhidrasas Carbónicas/análisis , Pruebas Enzimáticas Clínicas , Eritroblastos/enzimología , Leucemia/diagnóstico , Enfermedad Aguda , Línea Celular , Diagnóstico Diferencial , Humanos , Leucemia/patología , Leucemia Eritroblástica Aguda/diagnóstico , Leucemia Eritroblástica Aguda/patología , Leucemia Linfoide/diagnóstico , Leucemia Linfoide/patología , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/patologíaRESUMEN
There is a three- to four-fold decrease in the content/cell of tRNAs for ten different amino acids four days after the induction of erythroid differentiation in Friend leukemia cells, consistent with the decrease in cell volume that occurs. Surprisingly, there is an approximately two-fold increase in the cellular content of each of these tRNAs between day 4 and day 6 after induction, indicating the net synthesis of tRNA late in induction. The tRNA changes affect all species and do not result in tRNA specialization for hemoglobin synthesis, as occurs in normal erythroid development. The tRNA content of imidazole-treated cells, which do not synthesize hemoglobin although they undergo other changes of erythroid differentiation, decreases initially as described above, but shows no increase from day 4 to day 6.
Asunto(s)
Hemoglobinas/biosíntesis , Leucemia Experimental/metabolismo , Aminoacil-ARN de Transferencia/metabolismo , Animales , Diferenciación Celular , Línea Celular , Dimetilsulfóxido/farmacología , Eritrocitos , Virus de la Leucemia Murina de Friend , Imidazoles/farmacología , Factores de TiempoRESUMEN
A 68-year-old female patient is described in whom acute lymphoblastic leukemia followed a long course of polycythemia vera. Chromosomal analysis of a peripheral blood specimen at the time of blastic transformation revealed two distinct clones: one characterized by a chromosomal abnormality frequently noted in polycythemia vera and the other by a rearrangement characteristically observed in lymphoid malignancies. These findings suggest the existence of two independent hematologic diseases: this phenomenon would not support the speculation from previous reports that lymphoproliferative disorders in patients with polycythemia vera arise from clonal evolution.
Asunto(s)
Aberraciones Cromosómicas/complicaciones , Policitemia Vera/complicaciones , Leucemia-Linfoma Linfoblástico de Células Precursoras/complicaciones , Anciano , Antineoplásicos/uso terapéutico , Recuento de Células Sanguíneas , Trastornos de los Cromosomas , Femenino , Humanos , Cariotipificación , Policitemia Vera/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/genéticaRESUMEN
Pure red cell aplasia has been reported to be associated with chronic lymphocytic leukemia. It has been proposed that this complication may be a result of T-cell populations that suppress erythropoiesis. It has been postulated that antithymocyte globulin might reverse this abnormality by eliminating the population of suppressor T cells responsible for this inhibition. We treated a 74-year-old man who had B-cell chronic lymphocytic leukemia and pure red cell aplasia that was refractory to cytotoxic and corticosteroid therapy with equine antithymocyte globulin and methylprednisolone sodium succinate. This therapy resulted in a durable complete remission of both the chronic lymphocytic leukemia and the pure red cell aplasia and was associated with normalization of helper/suppressor T-cell ratios in the bone marrow. Antithymocyte globulin should be investigated further as a therapeutic modality for patients with pure red cell aplasia associated with chronic lymphocytic leukemia.
Asunto(s)
Suero Antilinfocítico/uso terapéutico , Leucemia Linfoide/terapia , Aplasia Pura de Células Rojas/terapia , Linfocitos T/inmunología , Anciano , Linfocitos B , Terapia Combinada , Humanos , Leucemia Linfoide/sangre , Masculino , Hemisuccinato de Metilprednisolona/uso terapéutico , Aplasia Pura de Células Rojas/etiologíaRESUMEN
The levels of the erythrocyte proteins carbonic anhydrase (CA) and hemoglobin (Hb) change coordinately during human ontogeny. To further probe the coordinate gene expression of these two proteins in vitro, we used an immunoblotting technique to measure their levels during erythroid differentiation in normal human and murine erythroid progenitors, in human and murine erythroleukemia cells, and in normal murine erythroid progenitors infected with Friend virus. Levels of CA and Hb seem to gradually increase in normal differentiating stem cells. In contrast, both human and murine erythroleukemia cells show high levels of CA, but not of Hb, prior to induction of differentiation. Friend virus infection of normal murine progenitors appears to stimulate CA synthesis as an initial and integral step in transformation. In addition, both the erythroleukemia cells and the erythroid progenitors transformed with Friend virus seem to contain much higher levels of CA than Hb during the early stages of differentiation. This relationship is in marked contrast to normal erythroid differentiation, in which Hb levels are always higher than CA levels. Thus, neoplastic transformation seems to be associated with aberrant production of CA that does not correspond to a maturation arrest of the normal differentiation sequence.
Asunto(s)
Anhidrasas Carbónicas/genética , Leucemia Eritroblástica Aguda/genética , Animales , Línea Celular , Virus de la Leucemia Murina de Friend , Regulación de la Expresión Génica , Hemoglobinas/genética , Humanos , RatonesRESUMEN
The expression of the c-fos protooncogene was investigated by in situ hybridization in normal murine bone marrow cells. A strong signal was found in murine marrow cells having the morphologic features of erythroblasts. This result was confirmed in human marrow cells using a double labeling technique (in situ hybridization and immunocytochemistry). A majority (70%) of the cells expressing c-fos mRNA were glycophorin A-positive. In contrast, granulocytic precursors (CD 15-positive) or monocytes and their precursors (CD 14-positive cells) did not significantly hybridize with the c-fos probe. In addition, c-fos mRNA (2.2Kb) was detected by Northern blotting in RNA extracted from homogeneous populations of erythroblasts obtained by immune panning from fetal liver and from adult blood BFU-E-derived colonies. Fos protein was also detected in erythroblasts by immunofluorescence. The high level of c-fos mRNA previously found in hematopoietic tissue should therefore be related to the transcription of the c-fos gene during terminal erythroid differentiation.
Asunto(s)
Médula Ósea/análisis , Eritroblastos/análisis , Proto-Oncogenes , Animales , Sondas de ADN , Feto , Globinas/análisis , Glicoforinas/análisis , Humanos , Hígado/análisis , Ratones , Hibridación de Ácido Nucleico , ARN Mensajero/análisisRESUMEN
During the course of differentiation of early human myeloid cells toward monocytes and granulocytes, cell surface expression of the cell adhesion molecule, CD11b/CD18 (Mo1) increases dramatically and expression of myeloperoxidase (MPO), a bacteriocidal enzyme, decreases markedly. Using the inducible promyelocytic cell line HL-60 as a model, we studied the mRNA expression of these genes. Differentiation of these cells along both a monocytic and a granulocytic pathway demonstrated that the mRNA levels of the two subunits of CD11b/CD18 increased in a pattern temporally and quantitatively similar to the increase in cell surface expression of this heterodimer. In contrast, the expression of MPO mRNA decreased in a temporal and quantitative pattern similar to the known decrease in MPO protein during differentiation, suggesting that regulation of these myeloid-specific proteins may occur at the level of mRNA expression. These findings have important implications with regard to the nature of the block in differentiation in acute nonlymphocytic leukemia and the regulation of myeloid gene expression.