RESUMEN
The incidence of non-Hodgkin's lymphoma in individuals infected with HIV is approximately 60- to 100-fold increased over the general population. The majority of patients with AIDS-related lymphoma (ARL) present with stage III-IV disease and with B-symptoms. They often have multiple extranodal localisations, with a high incidence of central nervous system involvement. Histologically, most tumours are either diffuse large cell lymphomas or Burkitt lymphomas. Several factors, such as disrupted immune surveillance, Epstein-Barr virus infection, chronic antigenic stimulation, cytokine dysregulation and the acquisition of genetic lesions, are thought to contribute to the pathogenesis. Patients with ARL have a poor prognosis: overall survival ranges from 1.5 to 18 months. The most important adverse prognostic factors are poor performance status, a low CD4+ cell count and a history of opportunistic infections. Results of treatment with polychemotherapy compare unfavourably to results in patients without HIV infection. Since the advent of highly active antiretroviral therapy (HAART), there appears to be a decrease in the incidence of ARL. In addition, the use of HAART in combination with chemotherapy and the use of new treatment modalities may improve the outcome of this disease.
Asunto(s)
Linfoma Relacionado con SIDA/etiología , Linfoma Relacionado con SIDA/terapia , Linfoma no Hodgkin/etiología , Linfoma no Hodgkin/terapia , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Terapia Antirretroviral Altamente Activa , Factores de Crecimiento de Célula Hematopoyética/uso terapéutico , Humanos , Linfoma Relacionado con SIDA/epidemiología , Linfoma no Hodgkin/epidemiología , PronósticoRESUMEN
Oral mucositis is a frequent side effect of myeloablative chemo- and radiotherapy preceding stem cell transplantation. It causes pain, poor food intake, and is a port of entry for infection. We studied whether GM-CSF applied topically in the oral cavity can prevent or ameliorate this mucositis. In 36 consecutive patients undergoing a stem cell transplantation, we performed a double-blind placebo-controlled study of 300 micrograms GM-CSF in a 2% methylcellulose gel daily versus a 2% methylcellulose gel alone. Both were locally applied in the oral cavity. The primary end-point was mucositis as measured by the WHO toxicity scale for mucositis, oral assessment scale, and a subjective pain scale, all scored daily. The secondary end-points were need to give parenteral nutrition and morphine, incidence of fever and infections, and duration of neutropenia and hospitalization. No differences were found in the median subjective pain scores, WHO scores, and oral assessment scores between the placebo and the GM-CSF groups. In both groups, nine patients required morphine for pain control. Ten patients in the placebo group and 11 in the GM-CSF group received parenteral nutrition. Documented infections, use of broad-spectrum antibiotics, and number of days with fever were similar in the placebo and the GM-CSF groups. The duration of neutropenia below 0.5 x 10(9)/l (median 14.5 days in the placebo group versus 17 days in the GM-CSF group) and the duration of hospitalization (28.5 versus 29 days) was also not significantly different. We found no beneficial effect of 300 micrograms GM-CSF dissolved in a 2% methylcellulose gel applied locally for chemo- and radiotherapy-induced mucositis in patients undergoing a stem cell transplantation.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Trasplante de Células Madre Hematopoyéticas , Estomatitis/prevención & control , Administración Tópica , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Método Doble Ciego , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/uso terapéutico , Humanos , Masculino , Persona de Mediana Edad , Mucosa Bucal , Agonistas Mieloablativos/efectos adversos , Estudios Prospectivos , Estomatitis/inducido químicamenteRESUMEN
Functional studies revealed that two groups of B chronic lymphocytic leukemia (B-CLL) can be distinguished based on their capacity to mount a proliferative response following B-cell antigen receptor (BCR) cross-linking. The molecular basis for the functional distinction between these B-CLL groups most probably resides within or proximal to the BCR since non-responsive B-CLL, in marked contrast to responsive B-CLL, do not respond to BCR ligation with tyrosine phosphorylation of cellular substrates and increases in the free intracellular [Ca++]. Detailed biochemical analysis showed overall structural identity between responsive and non-responsive B-CLL with respect to both transmembrane and intracellular associates of the BCR complex. However expression levels of the protein tyrosine kinase syk, which is a key enzyme for the early signalling through the BCR, were found to be markedly lower in non-proliferating B-CLL. Here we will review current functional and biochemical data on responding and non-responding B-CLL and discuss the relevance of these findings for disease progression and our insight into the immunobiology of B-CLL.
Asunto(s)
Leucemia Linfocítica Crónica de Células B/inmunología , Receptores de Antígenos de Linfocitos B/sangre , División Celular/inmunología , Diagnóstico Diferencial , Progresión de la Enfermedad , Humanos , Leucemia Linfocítica Crónica de Células B/diagnóstico , Fosforilación , Proteínas Tirosina Quinasas/metabolismoRESUMEN
Granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) are frequently used in the clinical management of neutropenia. These cytokines not only enhance the proliferation of myeloid precursor cells but also influence the function of mature leukocytes. In a previous study, we found that the in vivo effects of G-CSF on neutrophils differed from those in vitro. In the present study, we investigated the effects of a single dose of recombinant GM-CSF (7.5 microg/kg, subcutaneously) on neutrophils, eosinophils, and monocytes in healthy volunteers. We analyzed leukocyte kinetics, phenotypical changes, neutrophil degranulation, and systemic cytokine production. After GM-CSF injection, phenotypical changes included upregulation of CD11b on all three cell types and a decreased expression of L-selectin and Fc(gamma)RIII on neutrophils. Neutrophil degranulation was evident from the increased plasma concentrations of lactoferrin and elastase. GM-CSF induced the release of interleukin-8 (IL-8), but not of IL-6 or tumor necrosis factor alpha. In comparison to the results from our previous study with G-CSF in healthy volunteers, GM-CSF induced a stronger activation of mature neutrophils but had a much less pronounced effect on the production and maturation of neutrophil precursors. These data may help to guide the choice between the two cytokines in different clinical situations.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Interleucina-8/metabolismo , Neutrófilos/efectos de los fármacos , Adulto , Gránulos Citoplasmáticos/metabolismo , Eosinófilos/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Humanos , Inmunofenotipificación , Inyecciones Subcutáneas , Interleucina-6/metabolismo , Cinética , Selectina L/biosíntesis , Selectina L/genética , Lactoferrina/metabolismo , Recuento de Leucocitos/efectos de los fármacos , Elastasa de Leucocito , Antígeno de Macrófago-1/biosíntesis , Antígeno de Macrófago-1/genética , Masculino , Monocitos/efectos de los fármacos , Elastasa Pancreática/metabolismo , Receptores de IgG/biosíntesis , Receptores de IgG/genética , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/farmacología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
In this study we investigated the applicability of 99mTc-labeled CD19 monoclonal antibody (mAb) for tumor imaging in patients with B cell non-Hodgkin's lymphoma. A 1-mg sample of murine CD19 mAb was labeled with approximately 550 MBq [99mTc]pertechnetate. The labeled mAb was administered i.v. to seven patients, four without and three with pretreatment with 10 mg unlabeled CD19 mAb. The number of circulating B cells was decreased by 44 +/- 5% 1 h after injection of the radiolabeled mAb. Peripheral B cells were coated with CD19, resulting in partial modulation of CD19, most pronounced in the three pretreated patients. Whole-body images were obtained with a gamma camera and compared with results obtained by conventional imaging techniques. Initially, blood-pool activity dominated, whereas 24 h after injection the radioactivity was mainly located in the spleen, kidneys and liver. In two patients, a lesion in the spleen appeared as an unlabeled spot. In one patient, a lesion in the femur, which was detected by computed tomography (CT) and gallium-67 scans, was also seen on the CD19 scan from 1 h after administration of the radioimmunoconjugate onwards. Good imaging of bone marrow infiltration was observed in one of three patients. Lymph node involvement was not observed in any of the patients in whom affected lymph nodes were detected by CT or gallium-67 scan. In conclusion, in the present study radioimmunodetection with 99mTc-labeled CD19 mAb was found to be inferior to CT and gallium-67 scanning in the diagnosis of patients with B cell non-Hodgkin's lymphoma.
Asunto(s)
Anticuerpos Monoclonales , Antígenos CD19/análisis , Linfoma de Células B/diagnóstico por imagen , Radioinmunodetección , Tecnecio , Adulto , Anciano , Animales , Antígenos CD19/inmunología , Femenino , Humanos , Linfoma de Células B/inmunología , Masculino , Ratones , Proyectos PilotoRESUMEN
Diagnosis of meningeal localization of lymphoid malignancies by means of cytologic examination of the cerebrospinal fluid (CSF) can be difficult. Thus far no reliable CSF tumor markers have been identified. CD27 is a transmembrane disulfide-linked 55-kD homodimer present on most peripheral blood T cells and on a subset of B cells. CD27 is also expressed on human malignant B cells and high levels of soluble CD27 can be present in the serum of patients with B-cell malignancies. The aim of this study is to determine prospectively the diagnostic value of CSF sCD27 as a tumor marker in patients with meningeal localization of lymphoid malignancies. CSF sCD27 levels were determined by sandwich enzyme-linked immunosorbent assay. The optimal cut-off value using receiver operator characteristics curves was found to be 10 U/mL. sCD27 levels were normal in all 50 control patients (lumbar disc protrusion) and in 39 of 40 samples obtained from patients with either solid tumors or acute myeloid leukemia. Of 104 CSF samples from 70 children with acute lymphoblastic leukemia (ALL) or non-Hodgkin's lymphoma (NHL) undergoing routine central nervous system (CNS) staging, sCD27 was false positive and false negative in only one sample each. In 70 samples from 45 patients suspected of meningeal localization of ALL or NHL, the sCD27 test had an excellent sensitivity (100%) and specificity (82%). In 7 patients with positive CSF studied longitudinally, sCD27 levels correlated very well with remission and relapse. sCD27 levels were not nonspecifically increased by the administration of cytostatic drugs. Finally, sCD27 was also elevated in the 4 patients studied with primary central nervous system lymphoma (PCNSL). CSF sCD27 is a promising tumor marker in patients with either meningeal localization of lymphoid malignancies or PCNSL, and can be useful in the differential diagnosis of CNS involvement by either lymphoid malignancies or solid tumors.
Asunto(s)
Antígenos de Neoplasias/líquido cefalorraquídeo , Biomarcadores de Tumor/líquido cefalorraquídeo , Proteínas del Líquido Cefalorraquídeo/análisis , Infiltración Leucémica/líquido cefalorraquídeo , Linfoma no Hodgkin/líquido cefalorraquídeo , Linfoma no Hodgkin/patología , Meninges/patología , Invasividad Neoplásica/diagnóstico , Neoplasias/líquido cefalorraquídeo , Neoplasias/patología , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/líquido cefalorraquídeo , Adulto , Neoplasias Encefálicas/líquido cefalorraquídeo , Neoplasias Encefálicas/patología , Niño , Diagnóstico Diferencial , Ensayo de Inmunoadsorción Enzimática , Reacciones Falso Positivas , Humanos , Recurrencia Local de Neoplasia , Estudios Prospectivos , Curva ROC , Inducción de Remisión , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Microglobulina beta-2/líquido cefalorraquídeoRESUMEN
Twenty-one previously untreated multiple myeloma (MM) patients and 10 previously treated patients with refractory or relapsed disease received two or three cycles of intermediate-dose melphalan (70 mg/m2) (IDM), administered intravenously every 6 weeks. Seven previously untreated patients received three and all other patients received two courses of IDM. The objective of the study was to reduce the toxicity of high-dose melphalan (140 mg/m2) (HDM) while maintaining its cytotoxic efficacy and secondly to ensure the possibility of collecting sufficient numbers of peripheral blood stem cells (PBSC) for transplantation. 18 (85%) previously untreated patients responded, of whom four achieved CR (18%). In addition five out of 10 previously treated patients with refractory or relapsed disease responded although bone marrow toxicity in this category was a major drawback. Toxicity was moderate, consisting of alopecia and moderate bone marrow suppression: the granulocyte count dropped below 0.5 x 10(9)/l and platelets below 25 x 10(9)/l for a median of 8 and 6 d, respectively. No serious infections occurred and the majority of patients attended the out-patient clinic. In 12/14 previously untreated patients sufficient peripheral blood CD34+ cells for harvest were present in the repopulation phase after the first IDM. In nine patients peripheral blood stem cells were collected and eight patients have undergone successful transplantation. Repeated IDM followed by filgrastim is highly effective in untreated MM and may be safely administered to reduce tumour load prior to PBSC collection. Autologous stem cells harvested after repeated IDM have a full long-term repopulating capacity.
Asunto(s)
Antineoplásicos Alquilantes/uso terapéutico , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Trasplante de Células Madre Hematopoyéticas , Melfalán/uso terapéutico , Mieloma Múltiple/tratamiento farmacológico , Adulto , Anciano , Antineoplásicos Alquilantes/efectos adversos , Supervivencia sin Enfermedad , Esquema de Medicación , Femenino , Filgrastim , Estudios de Seguimiento , Humanos , Masculino , Melfalán/efectos adversos , Persona de Mediana Edad , Mieloma Múltiple/terapia , Proteínas Recombinantes/uso terapéuticoRESUMEN
BACKGROUND: Chemotherapy intensification may lead to new forms of toxicity such as hemolytic uremic syndrome. METHODS: Three patients are described who developed this complication 4 to 6 months after high dose chemotherapy followed by autologous stem cell support. The literature on this subject is reviewed. RESULTS: One patient was conditioned with BEAC (carmustine, etoposide, cytosine arabinoside, and cyclophosphamide) and received autologous bone marrow. The other two underwent triple peripheral stem cell transplantation after conditioning with CTC (carboplatin, cyclophosphamide, and thiotepa). Symptoms were hypertension, microangiopathic hemolytic anemia, thrombocytopenia, and renal insufficiency. One patient had a retinal vein thrombosis. One patient died of a cardiac arrest shortly after the diagnosis was made. The remaining two achieved a partial remission: one with fresh frozen plasma without plasmapheresis and fresh frozen plasma, but improved on high dose intravenous immunoglobulin and vincristine. CONCLUSIONS: Hemolytic uremic syndrome is a serious complication of the more intensive chemotherapy made possible by stem cell support. Because of the rapidly growing indications for this approach, an increase in this type of vascular complication is expected.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Síndrome Hemolítico-Urémico/etiología , Adulto , Antimetabolitos Antineoplásicos/administración & dosificación , Antimetabolitos Antineoplásicos/efectos adversos , Antineoplásicos/administración & dosificación , Antineoplásicos/efectos adversos , Antineoplásicos Alquilantes/administración & dosificación , Antineoplásicos Alquilantes/efectos adversos , Antineoplásicos Fitogénicos/administración & dosificación , Antineoplásicos Fitogénicos/efectos adversos , Trasplante de Médula Ósea/efectos adversos , Carboplatino/administración & dosificación , Carboplatino/efectos adversos , Carmustina/administración & dosificación , Carmustina/efectos adversos , Ciclofosfamida/administración & dosificación , Ciclofosfamida/efectos adversos , Citarabina/administración & dosificación , Citarabina/efectos adversos , Etopósido/administración & dosificación , Etopósido/efectos adversos , Resultado Fatal , Femenino , Humanos , Hipertensión/etiología , Inmunoglobulinas Intravenosas/uso terapéutico , Masculino , Persona de Mediana Edad , Plasma , Inducción de Remisión , Insuficiencia Renal/etiología , Oclusión de la Vena Retiniana/etiología , Tiotepa/administración & dosificación , Tiotepa/efectos adversos , Trombocitopenia/etiología , Trasplante Autólogo , Vincristina/uso terapéuticoRESUMEN
Members of the Tumour Necrosis Factor-Receptor (TNFR) family play an essential role in the control of lymphoid cell growth and differentiation. The ligand of one of its lymphoid-specific members, CD27, was recently characterized as CD70, a type II transmembrane molecule with homology to TNF that is expressed on activated T and B cells. Ligation of CD27 by its natural ligand generates a potent costimulatory signal for cytokine production and proliferation of activated T cells. In contrast to normal B cells, where CD27 expression is confined to germinal centre cells and to a small subset of circulating B lymphocytes, CD27 expression is found on a large array of distinct B-cell neoplasia. Here, we review recent data on the expression and function of TNFR family members on normal and malignant lymphocytes and propose a role for CD27-CD70 interaction in B-cell development.
Asunto(s)
Antígenos CD , Linfocitos B/citología , Leucemia de Células B/patología , Linfoma de Células B/patología , Proteínas de la Membrana/fisiología , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/fisiología , Linfocitos B/patología , Ligando CD27 , División Celular/fisiología , HumanosRESUMEN
BACKGROUND: High-dose chemoradiotherapy combined with autologous bone marrow transplantation can cure patients with disseminated, aggressive non-Hodgkin's lymphoma in whom first-line chemotherapy has failed. In contrast, cure is rare with second-line chemotherapy. It has been suggested that patients with slow responses to the initial phase of first-line chemotherapy are at high risk for relapse. Therefore, such patients are potential candidates for early bone marrow transplantation. METHODS: To investigate whether patients with slow responses, defined as only a partial response after three courses of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), would benefit from early transplantation, we conducted a prospective, randomized trial. The early application of high-dose chemoradiotherapy and autologous bone marrow transplantation was compared with the continuation of CHOP therapy for another five courses. Patients with complete responses after three courses of CHOP (fast responses) and patients who responded partially but still had tumor-positive marrow continued with another five courses of CHOP. The study end points were the response rate, overall survival, disease-free survival, and event-free survival. RESULTS: Of 286 patients who could be evaluated for the rapidity of their response after three courses of CHOP, 38 percent had fast responses, 47 percent had slow responses, and 15 percent had no response. Among 106 patients with slow responses who had lymphoma-negative marrow, 69 patients (65 percent) were randomized. Seventy-four percent of the CHOP group and 68 percent of the transplantation group had complete remissions (P = 0.54). At four years the rates of overall, disease-free, and event-free survival were 85, 72, and 53 percent, respectively, in the CHOP group and 56, 60, and 41 percent in the transplantation group (P > 0.10). The disease-free survival in both groups did not differ significantly from that of nonrandomized patients with fast responses (54 percent at four years). CONCLUSIONS: The early application of high-dose, marrow-ablative chemoradiotherapy with autologous bone marrow transplantation does not improve the outcome in patients with aggressive non-Hodgkin's lymphoma that responds slowly to first-line CHOP chemotherapy.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Trasplante de Médula Ósea , Linfoma no Hodgkin/terapia , Adolescente , Adulto , Terapia Combinada , Ciclofosfamida/administración & dosificación , Ciclofosfamida/uso terapéutico , Supervivencia sin Enfermedad , Doxorrubicina/administración & dosificación , Doxorrubicina/uso terapéutico , Femenino , Humanos , Linfoma no Hodgkin/tratamiento farmacológico , Linfoma no Hodgkin/radioterapia , Masculino , Persona de Mediana Edad , Prednisona/administración & dosificación , Prednisona/uso terapéutico , Pronóstico , Estudios Prospectivos , Inducción de Remisión , Resultado del Tratamiento , Vincristina/administración & dosificación , Vincristina/uso terapéuticoRESUMEN
We describe 3 patients with Corynebacterium jeikeium sepsis in neutropenic phase during treatment for acute myeloid leukaemia. Fever was the first symptom. All had a central venous catheter which was removed. Two patients developed subcutaneous nodules containing pus when the neutrophil count recovered; 1 had intracutaneous and pulmonary lesions. They were treated with vancomycin and recovered when the neutrophil count started to rise. A review of 80 neutropenic patients with C. jeikeium sepsis reported in the literature, together with our 3 cases indicates that risk factors for infection are the presence of a central venous catheter, being an adult male or postmenopausal female, profound and prolonged neutropenia and exposure to multiple antibiotics. Skin lesions are reported in 48% and pulmonary lesions in 36% of the patients. The overall mortality is 34% but in patients with recovery of the bone marrow only 5%. Therefore haematopoietic growth factors should be considered in neutropenic patients with C. jeikeium infection.
Asunto(s)
Bacteriemia/etiología , Infecciones por Corynebacterium/etiología , Corynebacterium/clasificación , Huésped Inmunocomprometido , Leucemia Mieloide Aguda/complicaciones , Adulto , Anciano , Antibacterianos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Bacteriemia/tratamiento farmacológico , Bacteriemia/fisiopatología , Corynebacterium/aislamiento & purificación , Infecciones por Corynebacterium/tratamiento farmacológico , Infecciones por Corynebacterium/fisiopatología , Quimioterapia Combinada/uso terapéutico , Femenino , Humanos , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/inmunología , Masculino , Persona de Mediana Edad , Neutropenia/complicaciones , PronósticoRESUMEN
In four healthy volunteers, we analyzed in detail the immediate in vivo effects on circulating neutrophils of subcutaneous administration of 300 micrograms of granulocyte colony-stimulating factor (G-CSF). Neutrophil activation was assessed by measurement of degranulation. Mobilization of secretory vesicles was shown by a decrease in leukocyte alkaline phosphatase content of the circulating neutrophils. Furthermore, shortly postinjection, Fc gamma RIII was found to be upregulated from an intracellular pool that we identified by immunoelectron microscopy as secretory vesicles. Intravascular release of specific granules was shown by increased plasma levels of lactoferrin and by upregulation of the expression of CD66b and CD11b on circulating neutrophils. Moreover, measurement of fourfold elevated plasma levels of elastase, bound to its physiologic inhibitor alpha 1-antitrypsin, indicated mobilization of azurophil granules. However, no expression of CD63, a marker of azurophil granules, was observed on circulating neutrophils. G-CSF--induced mobilization of secretory vesicles and specific granules could be mimicked in whole blood cultures in vitro, in contrast to release of azurophil granules. Therefore, we postulate that the most activated neutrophils leave the circulation, as observed shortly postinjection, and undergo subsequent stimulation in the endothelial microenvironment, resulting in mobilization of azurophil granules. Our data demonstrate that G-CSF should be regarded as a potent immediate activator of neutrophils in vivo.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos/farmacología , Activación Neutrófila/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Adulto , Fosfatasa Alcalina/biosíntesis , Antígenos CD/biosíntesis , Antígenos de Diferenciación/biosíntesis , Moléculas de Adhesión Celular , Gránulos Citoplasmáticos/metabolismo , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Inyecciones Subcutáneas , Antígeno de Macrófago-1/biosíntesis , Masculino , Microscopía Inmunoelectrónica , Neutrófilos/metabolismo , Elastasa Pancreática/biosíntesis , Receptores de IgG/biosíntesisRESUMEN
We performed a detailed kinetic study on the in vivo effect of a single subcutaneous dose of granulocyte colony-stimulating factor (G-CSF; 300 micrograms) in four healthy individuals on the expression and function of neutrophil Fc gamma receptors (Fc gamma R). G-CSF did not induce Fc gamma RI (CD64) on circulating neutrophils. However, neutrophils newly formed in response to G-CSF were Fc gamma RI positive and were able to perform antibody-dependent cellular cytotoxicity in an Fc gamma RI-dependent way. Fc gamma RII (CD32) expression was not changed significantly. Fc gamma RIII (CD16, phosphatidylinositol-linked) expression, slightly increased immediately (30 minutes) postinjection, was found to be strongly decreased on the newly formed population. For comparison, we studied the expression of the PI-linked proteins leukocyte alkaline phosphatase (LAP) and CD14. Intracellular levels of LAP mirrored the biphasic expression pattern as membrane-bound Fc gamma RIII. In contrast, CD14 expression on neutrophils was initially constant, followed by high levels on the newly formed neutrophils. Soluble CD14 levels were found to be elevated transiently, whereas peak levels of soluble Fc gamma III were observed as late as 6 days postinjection. In conclusion, we have shown that G-CSF results in an immunophenotypically and functionally altered neutrophil population for an important part as a result of its effect on myeloid precursor cells.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos/farmacología , Células Madre Hematopoyéticas/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Adulto , Fosfatasa Alcalina/sangre , Citotoxicidad Celular Dependiente de Anticuerpos/efectos de los fármacos , Antígenos CD/análisis , Antígenos de Diferenciación Mielomonocítica/análisis , Northern Blotting , Femenino , Humanos , Inmunofenotipificación , Recuento de Leucocitos , Receptores de Lipopolisacáridos , Masculino , Neutrófilos/inmunología , Neutrófilos/fisiología , Receptores de IgG/análisis , Receptores de IgG/genética , Proteínas Recombinantes/farmacologíaRESUMEN
In this study, we investigated the effect of recombinant human interleukin-6 (IL-6) on colony-forming cells for granulocytes and macrophages (CFU-GM) cultured in suspension. IL-6 when used alone did not induce proliferation of highly purified CD34+ human hematopoietic progenitors. Moreover, no influence of IL-6 was observed on the proliferation induced by granulocyte-macrophage colony-stimulating factor (GM-CSF) or granulocyte (G)-CSF. However, a marked survival enhancement (GM-CSF 228 +/- 42%, p < 0.01, and G-CSF 137 +/- 9%, p < 0.05) was observed when CD34+ cells were preincubated with IL-6 for 6 days. This survival effect became even more pronounced under serum-poor conditions (GM-CSF 380 +/- 80%, p < 0.01, and G-CSF 180 +/- 20%, p < 0.01) and could also be demonstrated at the single cell level in a colony-forming assay. By analysis of subpopulations of CD34+ bone marrow (BM) cells selected on the basis of CD45RO expression, the observed IL-6-mediated survival effect was found to be restricted to the CFU-GM containing CD45RO- subset. Our data show that IL-6 is a survival factor for CFU-GM.
Asunto(s)
Células de la Médula Ósea , Interleucina-6/fisiología , Células Madre/citología , Antígenos CD/análisis , Antígenos CD34 , Médula Ósea/inmunología , Médula Ósea/fisiología , División Celular/efectos de los fármacos , División Celular/fisiología , Separación Celular , Supervivencia Celular/fisiología , Células Cultivadas , Citometría de Flujo , Factor Estimulante de Colonias de Granulocitos/farmacología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Factores de Crecimiento de Célula Hematopoyética/fisiología , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/inmunología , Humanos , Interleucina-6/farmacología , Antígenos Comunes de Leucocito/análisis , Proteínas Recombinantes/farmacología , Células Madre/inmunología , Células Madre/fisiología , Factores de TiempoRESUMEN
In this study we have examined hFc gamma RI expression during myelopoiesis. Normal bone marrow (BM) cells were found to express hFc gamma RI up to the metamyelocyte stage. A different Fc gamma RI expression pattern was observed in an in vitro model of myelopoiesis. Purified CD34-positive BM cells, cultured for 12 to 14 days with granulocyte colony-stimulating factor (G-CSF), differentiate into a population of mature granulocytic cells. In these cultures, in which hFc gamma RI was virtually absent on the initial CD34-positive BM cells, hFc gamma RI was strongly induced by G-CSF after only 5 days. During final maturation the cells remained hFc gamma RI positive. This expression was confirmed functionally by antibody-sensitized erythrocytes (EA)-rosette assays. Moreover, the mature myeloid cells were found to express mRNA encoding for hFc gamma RI, whereas reverse-transcriptase polymerase chain reaction analysis showed that both hFc gamma RIA and hFc gamma RIB genes were expressed. In contrast, on peripheral blood (PB) polymorphonuclear neutrophil leukocytes (PMN) the in vitro effect of G-CSF as to hFc gamma RI induction was limited. Therefore, we conclude that, with respect to hFc gamma RI expression on PMN, G-CSF acts on myeloid precursor cells rather than on mature cells. This conclusion could be strengthened by in vivo administration of a single dose of G-CSF to a healthy volunteer. After a 12-hour lag time, hFc gamma RI expressing PMNs were detected in the peripheral blood. This study shows that hFc gamma RI is an early myeloid differentiation marker that is lost during normal final maturation. However, committed myeloid progenitor cells can be strongly induced by G-CSF to express hFc gamma RI, ultimately resulting in mature granulocytic cells expressing the high-affinity receptor for IgG. This expression may have important consequences for the functional capacity of these cells.
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Células de la Médula Ósea , Factor Estimulante de Colonias de Granulocitos/farmacología , Células Madre Hematopoyéticas/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Receptores de IgG/análisis , Antígenos CD/análisis , Antígenos CD34 , Secuencia de Bases , Células Cultivadas , Hematopoyesis , Células Madre Hematopoyéticas/fisiología , Humanos , Datos de Secuencia Molecular , Neutrófilos/inmunologíaRESUMEN
To study the receptors involved in the interaction between extracellular matrix proteins and hematopoietic progenitor cells, we analyzed the expression of beta 1 integrins on CD34+ bone marrow cells by means of immunoflowcytometry. Alpha 4 beta 1 and alpha 5 beta 1 were expressed, whereas alpha 1 beta 1, alpha 2 beta 1, alpha 3 beta 1, alpha 6 beta 1, and alpha v beta 1 were virtually absent. Furthermore, we assessed the alpha 4 and alpha 5 expression on committed myeloid progenitor cells. These colony-forming cells were detected in the alpha 4 dull fraction and the alpha 5 dull fraction. During myeloid differentiation, both in vivo and in vitro, a differential expression of alpha 4 beta 1 and alpha 5 beta 1 was observed. alpha 5 beta 1 was found to be lost at the myelocytic-metamyelocytic stage, before the loss of alpha 4 beta 1, at the band stage. Functional studies showed no binding of erythroid progenitor-depleted, CD34+ bone marrow cells to fibronectin. However, protein kinase C activation strongly induced fibronectin binding (68% of the cells). Inhibition experiments with specific antibodies and peptides showed the binding to be mediated by both alpha 4 beta 1 and alpha 5 beta 1. Also, colony-forming cells of granulocytes and macrophages were demonstrated to adhere to fibronectin in an activation-dependent way. During granulocyte colony-stimulating factor-induced in vitro maturation, the activation-dependent fibronectin binding capacity is gradually lost. We conclude that: (1) CD34+ bone marrow cells express alpha 4 beta 1 and alpha 5 beta 1; (2) the expression of alpha 4 beta 1 and alpha 5 beta 1 is differentially expressed during myeloid differentiation; and (3) binding of CD34+ bone marrow cells to fibronectin is activation dependent.
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Antígenos CD/análisis , Antígenos CD/fisiología , Médula Ósea/fisiología , Adhesión Celular , Fibronectinas/metabolismo , Células Madre Hematopoyéticas/fisiología , Integrinas/fisiología , Anticuerpos Monoclonales , Antígenos CD34 , Células de la Médula Ósea , Adhesión Celular/efectos de los fármacos , Ensayo de Unidades Formadoras de Colonias , Antígenos HLA-DR/análisis , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/inmunología , Humanos , Integrinas/inmunología , Valores de Referencia , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
In this study we have made a detailed analysis of growth factor (granulocyte-macrophage colony-stimulating factor [GM-CSF], granulocyte colony-stimulating factor [G-CSF], and macrophage colony-stimulating factor [M-CSF])-induced proliferation and differentiation of highly purified CD34+ committed human myeloid progenitor cells in suspension cultures. The results were compared with colony formation in semisolid medium. Proliferation in suspension cultures was determined by means of incorporation of [3H]thymidine, differentiation by flow cytometric immunophenotyping using a panel of monoclonal antibodies against monomyeloid antigens, and by morphology. A good correlation was found between the number of granulocyte-macrophage colony-forming units (CFU-GM) in semisolid medium and [3H]thymidine incorporation in suspension (r = 0.82), both assessed at day 11. Moreover, the frequency of proliferating cells as determined in suspension cultures by limiting dilution analysis was similar to the frequencies of CFU-GM as measured in semisolid medium. Studies on GM-CSF- and G-CSF-induced cell-growth kinetics revealed distinct proliferation patterns. Immunophenotypically the subsequent induction of the mature granulocytic antigens CD15 and CD67 was observed to be accompanied by a gradual loss of the HLA-DR antigen, whereas little monocytic differentiation was observed. M-CSF, although inducing no colony formation of CD34+ cells and minimal proliferation in suspension, induced monocytic differentiation, demonstrated by the expression of HLA-DR, CD14, and CD36 in the absence of CD15 and CD67. The observed immunophenotypical profiles were confirmed by the results of cytological characterization. Thus, the combined measurement of growth factor-induced proliferation and differentiation of progenitor cells in suspension cultures can be a useful alternative for the CFU-GM assay. Moreover, because small numbers of cells are required, it allows for detailed studies on cell-growth kinetics and developmental stages within the granulocytic and monocytic lineages.
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Antígenos CD/análisis , Células de la Médula Ósea , Hematopoyesis/fisiología , Anticuerpos Monoclonales , Antígenos CD34 , Médula Ósea/inmunología , Médula Ósea/metabolismo , Diferenciación Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Células Cultivadas , ADN/metabolismo , Citometría de Flujo , Factor Estimulante de Colonias de Granulocitos/farmacología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/inmunología , Células Madre Hematopoyéticas/metabolismo , Humanos , Inmunofenotipificación , Factor Estimulante de Colonias de Macrófagos/farmacología , Timidina/metabolismo , TritioRESUMEN
Twelve male patients, eight with the acquired immunodeficiency syndrome (AIDS) and four with AIDS related complex (ARC), who had zidovudine associated neutropenia (less than 1 x 10(9) neutrophils/l) were treated with recombinant human granulocyte colony-stimulating factor (G-CSF) in a phase I/II study. Treatment consisted of daily subcutaneous injections with G-CSF in a weekly increasing dose of 0.4, 2, 5 or 10 micrograms/kg body weight until a neutrophil count of more than 3 x 10(9) neutrophils/l was observed. This effective dose was continued for up to 4 weeks, followed by 4 weeks observation period without G-CSF treatment. Two patients (both with ARC) reached target neutrophil counts at the lowest G-CSF dose, whereas nine patients needed 2 micrograms/kg. One patient discontinued treatment before he reached target neutrophil counts. Mean (+/- SD) neutrophil counts before and after 1 and 4 weeks of effective dose treatment were 0.65(+/- 0.188) x 10(9), 6.016(+/- 2.595) x 10(9) and 5.54(+/- 4.237) x 10(9)/l respectively (P less than 0.01). The number of monocytes increased from 0.171(+/- 0.113) to 0.501(+/- 0.274) and 0.474(+/- 0.374) x 10(9)/l after 1 and 4 weeks of treatment (P less than 0.01). Other haematologic parameters did not change significantly. Two weeks post-treatment the numbers of neutrophils and monocytes had returned to pre-treatment values. Mild side effects consisting of bone, joint or muscle pain were observed in three patients. Two patients (both with AIDS) did not complete the study. One patient stopped treatment because of fever and malaise, attributable to a generalized cytomegalovirus (CMV) infection and one patient had to stop zidovudine treatment because of severe thrombocytopenia. We conclude that G-CSF increases the number of circulating neutrophilic granulocytes in zidovudine-treated patients at relatively low doses and with few side-effects.
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Complejo Relacionado con el SIDA/tratamiento farmacológico , Síndrome de Inmunodeficiencia Adquirida/tratamiento farmacológico , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Neutropenia/terapia , Zidovudina/efectos adversos , Evaluación de Medicamentos , Humanos , Masculino , Neutropenia/inducido químicamente , Proteínas Recombinantes/uso terapéutico , Zidovudina/uso terapéuticoRESUMEN
The incidence and outcome of streptococcal septicemia was analyzed in 76 consecutive patients with newly diagnosed and relapsed acute myeloid leukemia. They received 215 courses of remission induction or intensive consolidation treatment. There were 31 different episodes of streptococcal septicemia in 27 patients, making these microorganisms the most frequently encountered bacteria in blood cultures. This high incidence coincided with the introduction of selective intestinal decontamination. In 24 episodes (20 patients) there was a fast recovery, but 7 patients developed pulmonary symptoms resulting in death due to respiratory failure in 5 of them. The infections all occurred in the phase of maximum bone marrow suppression 1-3 weeks after the start of the chemotherapy. Streptococcal septicemia was not limited to patients treated with cytosine arabinoside but also occurred in patients treated with other regimens of intensive chemotherapy. In 28 episodes there were no focal signs of infection, but in half there were symptoms of treatment induced gastrointestinal toxicity. The streptococci probably invade through oral and gastrointestinal mucosa damaged by the chemotherapy. Selective decontamination may play a promoting role.