RESUMEN
Adult T cell leukemia (ATL) is associated with human T cell leukemia virus type 1 (HTLV-1) infection, and almost all ATL patients have the complication of hypercalcemia. To understand the mechanism of the high incidence of hypercalcemia in ATL, we studied the expression of a parathyroid hormone-related protein (PTHrP) gene that has been proposed as a causative factor of hypercalcemia in some solid tumors. The polymerase chain reaction coupled with reverse transcription of mRNA was applied to RNA from peripheral blood mononuclear cells. Cells from all 13 ATL patients examined showed abundant expression of the PTHrP gene, while cells from uninfected normal subjects did not. Significant expression of PTHrP gene was also detected in HTLV-1 carriers without any symptoms and in patients with HTLV-1-associated myelopathy or tropical spastic paraparesis. PTHrP mRNA levels correlated with the number of infected cells that were estimated by the integrated HTLV-1 DNA. These results suggest that HTLV-1-infected cells are expressing the PTHrP gene. This concept was further supported by the finding that the HTLV-1 trans-activator, the tax gene product, caused trans-activation of the PTHrP gene promoter linked to the CAT gene. These observations might explain the general expression of the PTHrP gene in ATL patients and the high incidence of hypercalcemia in ATL.
Asunto(s)
Regulación Viral de la Expresión Génica , Genes Virales , Infecciones por HTLV-I/genética , Virus Linfotrópico T Tipo 1 Humano/genética , Leucemia-Linfoma de Células T del Adulto/genética , Hormona Paratiroidea/genética , Proteínas/genética , Transactivadores/genética , Activación Transcripcional , Secuencia de Bases , Portador Sano , Línea Celular , ADN Viral/genética , Humanos , Datos de Secuencia Molecular , Sondas de Oligonucleótidos , Proteína Relacionada con la Hormona Paratiroidea , Plásmidos , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , Mapeo Restrictivo , Transcripción Genética , TransfecciónRESUMEN
A significant increase in CD25 antigen-positive cells by IL-1 was observed in cells of a patient with M7 acute myelogenous leukemia. Basal proliferation and expression of CD25 antigen by the M7 leukemic cells were inhibited by addition of anti-IL-1 beta antibody in a dose-dependent manner, but not by rabbit anti-IL-1 alpha antibody. Culture supernatants of these leukemic cells contained IL-1 activity, which was specifically inhibited by addition of anti-IL-1 beta antibody, and Northern blot analysis detected intracellular IL-1 beta mRNA. These results indicated that autocrine secretion of IL-1 beta was involved in proliferation of some myelogenous leukemic cells.
Asunto(s)
Interleucina-1/fisiología , Leucemia Mieloide Aguda/patología , Antígenos de Superficie/análisis , Antígenos de Superficie/genética , División Celular , Regulación de la Expresión Génica , Humanos , Técnicas Inmunológicas , Interleucina-1/genética , Leucemia Mieloide Aguda/fisiopatología , Hibridación de Ácido Nucleico , ARN Mensajero/genética , Transcripción Genética , Células Tumorales CultivadasRESUMEN
Using sheep erythrocytes and liposomes, an inhibitory effect of gangliosides has been shown on the activation of the alternative pathway of complement. However, in studies using human erythrocytes, we found that gangliosides had hemolytic activity that was possibly mediated through activation of the alternative pathway. Pretreatment of human erythrocytes obtained from healthy volunteers or paroxysmal nocturnal hemoglobinuria (PNH) patients with a ganglioside mixture purified from human erythrocytes enhanced their susceptibility to homologous human complement, and resulted in dose-dependent hemolysis. The enhancement was more marked in PNH erythrocytes than control cells. Protease treatment of the ganglioside mixture did not change its hemolytic activity, but sialidase treatment abolished the activity. Among the major erythrocyte gangliosides, II3NeuAc-LacCer (GM3) was the most potent hemolytic agent. Gangliosides purified from bovine brain were also active, while neither nonsialylated glycosphingolipids, the ceramide moiety, or sialic acid alone were active. Sialic acid residues in the ganglioside molecules were essential to this activity, but the amount of the residue or the source of the gangliosides seemed not to be important. Several treatments inhibiting the alternative but not classical complement pathway markedly reduced the ganglioside hemolytic activity. This novel bioactivity of gangliosides was thus suggested to be mediated partly by activation of the alternative pathway.
Asunto(s)
Gangliósidos/fisiología , Hemólisis/fisiología , Cromatografía en Capa Delgada , Activación de Complemento , Citometría de Flujo , Gangliósidos/análisis , Gangliósidos/farmacología , Hemoglobinuria Paroxística/sangre , Hemólisis/efectos de los fármacos , Humanos , Relación Estructura-ActividadRESUMEN
To determine the origin of circulating parathyroid hormone fragments, hormonal peptides released from bovine parathyroid tissue in a physiologically responsive in vitro "perifusion" system were analyzed by gel exclusion chromatography and region-specific radioimmunoassays. When exposed to low Ca++, the tissue released large quantities of intact hormone (parathyroid hormone 1--84) as well as amino- and carboxyl-terminal fragments. Fragments of the hormone were also released when the tissue was exposed to high Ca++, but the carboxyl fragments comprised a much greater proportion of the hormonal peptides released. Control experiments indicated that fragmentation of the hormone occurred within the gland and not after it was secreted. These experiments provide direct evidence, therefore, that release of fragments from the parathyroid gland may contribute to the immunologic heterogeneity of the hormone in the circulation.
Asunto(s)
Glándulas Paratiroides/metabolismo , Hormona Paratiroidea/metabolismo , Animales , Calcio/farmacología , Bovinos , Cromatografía en Gel , Técnicas In Vitro , Glándulas Paratiroides/efectos de los fármacos , Hormona Paratiroidea/inmunología , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/metabolismo , RadioinmunoensayoRESUMEN
The gene coding for the protein LD78 was isolated from stimulated human tonsillar lymphocytes by differential hybridization. The gene product consisted of 92 amino acids with characteristics of cytokines. LD78 gene transcripts were detected in eight of eight fresh samples of cells from patients with acute nonlymphocytic leukemia (ANLL) by Northern blot analysis. ANLL cells with monocytic features gave the strongest bands. RNA transcripts were found in two of three samples of cells from patients with adult T cell leukemia (ATL), eight of nine samples from patients with acute lymphocytic leukemia (ALL) of B cell lineage, and one of the three samples from patients with T cell ALL. KG-1, HL-60, HUT 102, MT-2, and MJ cell lines expressed the LD78 gene constitutively. The LD78 protein was detected in culture supernatants and cell lysates of HUT 102, MT-2, MJ, and fresh ATL cells by Western blot analysis. This protein was not found in culture supernatants or cell lysates of monocytic leukemia cells and HL-60 cells, although LD78 transcripts were found in those cells. The discrepancy between gene and protein expression might be explained by the stability of the mRNA. Thus, the protein may be involved in the neoplastic transformation of hematopoietic cells.
Asunto(s)
Factores Biológicos/biosíntesis , Expresión Génica , Leucemia/metabolismo , Proteínas de Neoplasias/biosíntesis , Factores Biológicos/análisis , Factores Biológicos/genética , Western Blotting , Quimiocina CCL4 , Citocinas , ADN/genética , ADN/aislamiento & purificación , Humanos , Cinética , Leucemia Mieloide Aguda/metabolismo , Leucemia de Células T/metabolismo , Linfocitos/química , Proteínas Inflamatorias de Macrófagos , Proteínas de Neoplasias/análisis , Proteínas de Neoplasias/genética , Hibridación de Ácido Nucleico , Tonsila Palatina/citología , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , ARN Mensajero/análisis , ARN Mensajero/genética , Transcripción Genética , Células Tumorales CultivadasRESUMEN
The adult T cell leukemia (ATL) is a T cell neoplasm etiologically associated with human T lymphotropic virus type I (HTLV-I) infection. ATL cells often abnormally express interleukin 2 (IL-2) receptors, and ATL patients may show clinical evidence of hypercalcemia, osteolytic bone lesions, or increased bone turnover. Whereas interleukin 1 (IL-1) is not generally recognized as a product of T cells, this cytokine is capable of both altering IL-2 receptor expression and activating osteoclasts. Thus, we investigated the possibility that primary ATL leukemic T cells and HTLV-I-infected long-term ATL cell lines produce IL-1. S1 nuclease protection assays demonstrated that primary leukemic ATL cells from five out of six patients, as well as one patient with T4+ chronic lymphocytic leukemia, contained considerable quantities of IL-1 beta messenger RNA (mRNA) and small amounts of IL-1 alpha mRNA. These primary leukemic T cells also released biologically active IL-1 protein as evaluated in the murine thymocyte comitogenesis bioassay. In contrast to primary tumor cells, four out of six long-term ATL cell lines produced variable amounts of IL-1 alpha mRNA in the absence of detectable IL-1 beta mRNA as measured by S1 nuclease protection. These data demonstrate that IL-1 gene (especially IL-1 beta) expression occurs in many primary HTLV-I-infected leukemic T cells raising the possibility that this mediator may play a role in the pathological changes associated with this leukemia. Also, these studies show that the pattern of IL-1 alpha and IL-1 beta gene expression differs between primary ATL tumor cells and long-term cultured ATL cell lines, indicating an interesting biological difference in these two HTLV-I-infected cell populations.
Asunto(s)
Regulación de la Expresión Génica , Interleucina-1/genética , Leucemia/genética , Adulto , Línea Celular , Infecciones por Deltaretrovirus/genética , Humanos , Leucemia/metabolismo , Leucemia/patología , Activación de Linfocitos , ARN Mensajero/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismoRESUMEN
Paroxysmal nocturnal hemoglobinuria (PNH) erythrocytes lack complement regulatory membrane proteins and are susceptible to complement. Although the critical role of complement in intravascular hemolysis in PNH is accepted, the precise mechanism of complement activation in vivo is unknown. Accordingly, in a PNH patient who was suffering from a hemolytic precipitation soon after a common cold-like upper respiratory infection, we analyzed the erythrocytes with lectins and by flow cytometry to detect membrane alteration that lead to complement activation. The lectin reactivity of erythrocytes showed the expression of cryptantigen Th. The patient serum at the time of the hemolysis induced the expression of Th on erythrocytes from PNH patients and from healthy volunteers in vitro, whereas neither the patient serum after recovery from the hemolysis nor blood type-matched control serum from healthy donor showed this activity. Moreover, autologous serum selectively hemolyzed Th+ PNH erythrocytes, but not Th- PNH erythrocytes, or Th+ control erythrocytes. Hemolysis was not observed either in complement-inactivated serum or in blood type-matched cord blood serum, which lacks natural antibodies to cryptantigens. These findings indicate that the immunoreaction of infection-induced Th with natural antibody on PNH erythrocytes is a trigger of the complement activation, leading to intravascular hemolysis.
Asunto(s)
Antígenos/análisis , Eritrocitos/inmunología , Hemoglobinuria Paroxística/sangre , Hemólisis , Adolescente , Antígenos CD/análisis , Antígenos CD59 , Activación de Complemento , Membrana Eritrocítica/inmunología , Hemoglobinuria Paroxística/inmunología , Humanos , Masculino , Glicoproteínas de Membrana/análisisRESUMEN
In paroxysmal nocturnal hemoglobinuria (PNH), impaired glycosyl-phosphatidylinositol (PI)-anchoring of membrane proteins such as decay-accelerating factor has been known to lead to increased susceptibility to complement. Moreover, abnormal expression of non-PI-anchoring glycoproteins such as C3b/C4b receptor (CR1) or glycophorin-alpha also has been shown in PNH. Therefore, we biochemically analyzed glycosphingolipids (GSL) as one of the membrane glycoconjugates of PNH erythrocytes. Erythrocytes of all seven PNH patients showed altered expression of sialosyl GSL (gangliosides) as compared with the control erythrocytes of healthy donors. Both a sialosylparagloboside (IV6NeuAc-nLc4Cer) among four major gangliosides and some minor gangliosides in normal erythrocytes variably disappeared in erythrocytes from the peripheral blood of PNH patients. As one of the possible mechanisms of altered expression of gangliosides in PNH erythrocytes, structural analysis suggested impaired sialylation of GSL. These results suggest not only the altered metabolism of gangliosides in PNH erythrocytes, but also a metabolic disorder of membrane glycoconjugates as a new feature of PNH.
Asunto(s)
Eritrocitos/análisis , Gangliósidos/sangre , Hemoglobinuria Paroxística/sangre , Adulto , Anciano , Cromatografía en Capa Delgada , Membrana Eritrocítica/análisis , Femenino , Técnica del Anticuerpo Fluorescente , Gangliósidos/aislamiento & purificación , Hemólisis , Humanos , Masculino , Lípidos de la Membrana/sangre , Lípidos de la Membrana/aislamiento & purificación , Proteínas de la Membrana/sangre , Proteínas de la Membrana/aislamiento & purificación , Persona de Mediana Edad , Valores de ReferenciaRESUMEN
Cytokine LD78 is a human counterpart of the mouse macrophage inflammatory protein 1 alpha/hematopoietic stem cell inhibitor. Promoters of the LD78 alpha and LD78 beta genes showed similar inducible activities in two leukemic cell lines, K562 and Jurkat, but the induction mechanisms differed between the two cell lines. Further characterization of the LD78 alpha promoter indicated that multiple positive and negative regulatory elements are present, some of which are differentially required for induction and repression of the promoter activity in different cells. One of the negative regulatory elements, ICK-1, functioned in both cell lines in the absence and presence of stimulation and was shown to be a recognition site for positive and negative transcriptional factors. This ICK-1 element contained a direct repeat, and similar repeats were also found in the negative regulatory elements of hematopoietic growth factor interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) gene promoters. Nuclear extracts from K562 and Jurkat cells formed several protein-DNA complexes with the LD78 alpha ICK-1 element, one of which was also observed with the IL-3 and GM-CSF ICK-1 elements. Results from in vivo and in vitro analyses suggested that the protein forming this complex functions as a negative factor. The binding affinity of this protein, ICK-1A, to the LD78 alpha ICK-1 element was low and was significantly affected by the incubation temperature and the salt concentration in the binding buffer. ICK-1B, another protein bound specifically by the LD78 alpha ICK-1 element, was shown to be a positive factor important for induction of the promoter. These results suggested that ICK-1A plays an important role in balanced expression of LD78, IL-3, and GM-CSF during hematopoietic cell growth and differentiation.
Asunto(s)
Citocinas/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Interleucina-3/genética , Monocinas/genética , Proteínas de Neoplasias/genética , Regiones Promotoras Genéticas , Secuencias Reguladoras de Ácidos Nucleicos , Factores de Transcripción/metabolismo , Animales , Secuencia de Bases , Núcleo Celular/metabolismo , Quimiocina CCL3 , Quimiocina CCL4 , Cloranfenicol O-Acetiltransferasa/genética , Cloranfenicol O-Acetiltransferasa/metabolismo , Sondas de ADN , Humanos , Leucemia Eritroblástica Aguda , Leucemia de Células T , Luciferasas/genética , Luciferasas/metabolismo , Proteínas Inflamatorias de Macrófagos , Metilación , Ratones , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Oligodesoxirribonucleótidos , Regiones Promotoras Genéticas/efectos de los fármacos , Mapeo Restrictivo , Eliminación de Secuencia , Homología de Secuencia de Ácido Nucleico , Acetato de Tetradecanoilforbol/farmacología , Transfección , Células Tumorales CultivadasRESUMEN
Adult T-cell leukemia-associated antigen (ATLA) and adult T-cell leukemia virus (ATLV) antigens were localized in the MT-2 cell system by the immunocolloidal gold method using 71 human sera having various anti-ATLA titers and rabbit anti-ATLV antisera. In the thin-section method with anti-ATLA-positive human sera and rabbit antisera, protein A-gold particles were preferentially observed on and around sectioned adult T-cell leukemia (ATL) virions located in pericellular aggregates and within the cytoplasmic vacuoles but nowhere else in a significant number. The number of gold particles per ATL virion was statistically well correlated with the anti-ATLA titers of human sera applied (p less than 0.005). The preembedding method showed that pericellular ATL virions were specifically tagged with protein A-gold, but the antigens in question were not expressed on the plasma membrane of MT-2 cells. The absence of ATLA and ATLV antigens on the plasma membrane constitutes a unique pathobiological feature of ATL and ATLV as compared with other retrovirus systems.
Asunto(s)
Antígenos de Neoplasias/análisis , Leucemia/inmunología , Glicoproteínas de Membrana , Linfocitos T/inmunología , Adulto , Línea Celular , Membrana Celular/inmunología , Coloides , Oro , Humanos , Sueros Inmunes , Inmunodifusión , Indicadores y ReactivosRESUMEN
The antigen defined by MCS-2 monoclonal antibody (mAb) was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of surface and internally labeled cells. Surface iodination revealed that MCS-2 antigen on the surfaces of acute myelogenous leukemia cells, HL-60 cells, and polymorphonuclear leukocytes (PMN) had the same molecular weight (Mr 150,000) and that their autoradiographic bands were also the same. Internal labeling of HL-60 cells with [35S]methionine followed by immunoprecipitation revealed two bands whose molecular weights were 150,000 and 130,000. HL-60 cells gave stronger bands than did PMN. The intensity of the Mr 130,000 band became weaker, when internally pulse-labeled cells were cultured in the absence of labeled methionine, suggesting that Mr 130,000 glycoprotein was a precursor protein of Mr 150,000 glycoprotein. MCS-2 mAb precipitated two bands from tunicamycin-treated HL-60 cells whose apparent molecular weights were 100,000 and 110,000. When cells were cultured with MCS-2 mAb, expression of the antigen decreased rapidly (within 10 min). The degree of suppression was more prominent in PMN than in acute myelogenous leukemia and in myelomonocytic cell lines. Reexpression of MCS-2 antigen by PMN after removal of the mAb from the culture medium was not observed, but it occurred rapidly in myelomonocytic cell lines, although it was blocked by pretreatment of the cells with cycloheximide. These findings suggested that the less-marked suppression of MCS-2 antigen expression by cell lines was due to its greater synthesis by these cells. These findings suggested that MCS-2 mAb reacted with identical molecules, which were recognized by other mAbs belonging to CD13. Furthermore, modulation of MCS-2 antigen was observed by MCS-2 mAb itself.
Asunto(s)
Antígenos de Diferenciación/análisis , Leucemia Mieloide Aguda/inmunología , Neutrófilos/inmunología , Anticuerpos Monoclonales/inmunología , Antígenos de Superficie/análisis , Diferenciación Celular , Humanos , Peso Molecular , Células Tumorales Cultivadas/inmunologíaRESUMEN
Chromosomes of six patients with plasma cell leukemia and one patient with leukemic macroglobulinemia were examined from peripheral blood, bone marrow, and/or pleural fluid. All the patients had a clonal chromosomal abnormality. The modal chromosome number was near tetraploid in two, pseudodiploid in two, and hypodiploid in three patients. Rearrangements of chromosome 1 were found in all the patients. The most consistent abnormality was a large marker involving the long arm of No. 1, found in six patients, including the patient with macroglobulinemia. Each patient had one to four large markers which resulted in partial trisomy to hexasomy for the long arm of No. 1. The translocations occurred with No. 9 in two, with No. 16 in two, and Nos. 8, 17, and 18 each in one patient. The survival time from the diagnosis was less than 1 year in five of them and over 2 years in one. The only patient whose cells lacked an extra 1q lived for over 3 years. Five 14q+ marker chromosomes were detected in three patients. The donor chromosome was No. 11 in one of these and was undetermined in the others; the size of each 14q+ marker seemed quite different which suggested different donor chromosomes. Loss of a sex chromosome was found in five patients. Loss of No. 13 and gain of No. 7 or 7q were each found in two patients. Rearrangement or deletion of the short arm of No. 8 was found in five patients. A rearrangement of 9p was found in three patients. The myeloma cells had a different morphology in the peripheral blood, bone marrow, and/or pleural fluid before and after the leukemic phase of one patient; however, chromosome analysis revealed the same clone despite the altered morphology.
Asunto(s)
Aberraciones Cromosómicas/genética , Leucemia de Células Plasmáticas/genética , Leucemia/genética , Macroglobulinemia de Waldenström/genética , Adulto , Anciano , Bandeo Cromosómico , Trastornos de los Cromosomas , Femenino , Humanos , Cariotipificación , Leucemia/complicaciones , Masculino , Persona de Mediana Edad , Macroglobulinemia de Waldenström/complicacionesRESUMEN
A neutrophil chemotactic factor (human interleukin 8, human granulocyte-macrophage colony-stimulating factor)-producing cell line, named KHM-5M, was established from a patient with an undifferentiated thyroid carcinoma, neutrophilia, and malignant pleurisy with many neutrophils and a few malignant cells. The cell line was transplanted into nude rats, and the infiltration of neutrophils was observed in and around the transplanted tumor tissue. Neutrophil chemotactic activity was predicted from the clinical features and pathological findings in this case. The extreme chemotactic activity of the neutrophils was demonstrated in conditioned medium from KHM-5M cells using the modified Boyden chamber technique. With sodium dodecyl sulfate-polyacrylamide gel electrophoresis, at least two neutrophil chemotactic activities in conditioned medium from the cell line were observed. The levels of these activities derived from KHM-5M cells were screened by measuring conditioned medium from the COS cells, which expressed a complementary DNA library from the KHM-5M cells. Chemotactic activities (human interleukin 8, human granulocyte-macrophage colony-stimulating factor) were identified by DNA cloning. These results show that the KHM-5M cells derived from an undifferentiated thyroid carcinoma produce multicytokines and suggest that those cytokines modified some pathological features in this case.
Asunto(s)
Factores Quimiotácticos/metabolismo , Neutrófilos/fisiología , Neoplasias de la Tiroides/metabolismo , Anciano , Animales , Secuencia de Bases , Factores Quimiotácticos/genética , Quimiotaxis de Leucocito , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/fisiología , Humanos , Interleucina-8/genética , Interleucina-8/fisiología , Masculino , Datos de Secuencia Molecular , Trasplante de Neoplasias , Oligodesoxirribonucleótidos/química , Reacción en Cadena de la Polimerasa , Ratas , Ratas Desnudas , Neoplasias de la Tiroides/patología , Células Tumorales CultivadasRESUMEN
The annual age- and sex-specific human T-cell leukemia virus type I carrier rate of blood donors in Kumamoto, Kyushu, Japan from 1986 to 1990 revealed that the carrier rates of all the age groups below 50 years declined linearly in both sexes (P less than 0.005). Furthermore, the annual declining rates relative to the carrier rates of 16-19-year-old and 20-29-year-old males were higher than those of all of the older males (P less than 0.02), and all female age groups below 50 years had higher relative declining rates than 50-64-year-old females (P less than 0.05). Although several factors, such as a notification program at obstetric clinics, methodological and technical improvement of the assays, wider knowledge of human T-cell leukemia virus type I infection in the latter years, and immigration of individuals from a nonendemic area, might cause an absolute decline of the carrier rate of the blood donors, these factors could not explain the acceleration of the relative declining rate among younger donors. Therefore, this acceleration represents the tendency of the general population.
Asunto(s)
Donantes de Sangre , Portador Sano/epidemiología , Infecciones por HTLV-I/epidemiología , Adolescente , Adulto , Factores de Edad , Femenino , Predicción , Humanos , Japón/epidemiología , Funciones de Verosimilitud , Masculino , Persona de Mediana EdadRESUMEN
Cell surface expression of the T-cell receptor (TCR)/CD3 complex on the cells from 11 acute type adult T-cell leukemia (ATL) and 4 lymphoma type ATL patients was examined by flow cytometry. Cells from 10 of 11 acute ATL patients were TCR alpha beta+ and CD3+, and their mean fluorescence intensities were low (TCR alpha beta, 25.3-84.6; CD3, 22.8-87.8). Cells from two of four lymphoma type ATL did not express this complex, and the other two were CD3+, TCR alpha beta-. In contrast, the mean fluorescence intensity of the TCR/CD3 complex in cells from a patient with T4 chronic lymphocytic leukemia was not low (TCR alpha beta, 129.9; CD3, 117.1). mRNA expressions of the TCR alpha, beta, and CD3 gamma, delta, epsilon, and zeta chains were examined by Northern blots. ATL cells from two acute and two lymphoma types expressed amounts of this complex equal to or greater than those expressed by T4 chronic lymphocytic leukemia. CD3 delta and TCR beta mRNA in ATL and T4 chronic lymphocytic leukemia cells were equally stable to actinomycin D treatment. The synthesis of CD3 zeta protein by ATL cells was detected by Western blotting assay. On the basis of these findings, we discuss the possible involvement of the TCR/CD3 complex in activation of ATL cells.
Asunto(s)
Antígenos de Diferenciación de Linfocitos T/análisis , Leucemia de Células T/inmunología , Receptores de Antígenos de Linfocitos T/análisis , Adulto , Anciano , Anciano de 80 o más Años , Antígenos de Diferenciación de Linfocitos T/genética , Complejo CD3 , Dactinomicina/farmacología , Femenino , Humanos , Leucemia de Células T/genética , Masculino , Persona de Mediana Edad , ARN Mensajero/análisis , Receptores de Antígenos de Linfocitos T/genética , Células Tumorales CultivadasRESUMEN
The use of antigen-presenting dendritic cells (DCs) is currently proposed for tumor immunotherapy through generation of CTLs to tumor antigens in cancer patients. In this study, DCs were differentiated using granulocyte-macrophage colony-stimulating factor and tumor necrosis factor-alpha from CD34+ hematopoietic progenitor cells that had been mobilized into the peripheral blood. To use the phagocytic activity of DCs for processing and presentation of tumor antigens, we established DC clusters containing immature DCs by preserving proliferating cell clusters without mechanical disruption. After an 11-day culture, the developed clusters contained not only typical mature DCs but also immature DCs that showed active phagocytosis of latex particles, suggesting that the clusters consisted of DCs of different maturational stages. These heterogeneous clusters could present an exogenous protein antigen, keyhold limpet hemocyanin, to both CD4+ and CD8+ T lymphocytes. Furthermore, in three acute myelogeneous leukemia patients, clusters pulsed with autologous irradiated leukemic cells could also induce antileukemic CTLs. The mechanical disruption of clusters abrogated the induction of CTLs to leukemic cells as well as to hemocyanin. This observation gives an important information for the use of heterogeneous DC clusters derived from autologous peripheral blood CD34+ cells in the case of immunotherapy for leukemia.
Asunto(s)
Presentación de Antígeno , Antígenos de Neoplasias/inmunología , Células Dendríticas/inmunología , Células Madre Hematopoyéticas/citología , Leucemia Mieloide/inmunología , Células Madre Neoplásicas/inmunología , Linfocitos T Citotóxicos/inmunología , Enfermedad Aguda , Antígenos CD34 , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Adhesión Celular , Células Cultivadas , Células Dendríticas/citología , Hemocianinas/inmunología , Humanos , Inmunoterapia , Leucemia Mieloide/patología , Leucemia Mieloide/terapia , Prueba de Cultivo Mixto de Linfocitos , Ovalbúmina/inmunología , Fagocitosis , Estrés MecánicoRESUMEN
The antigenic profile of basal cell carcinoma (BCC) cells was analyzed by immunoperoxidase staining of 27 surgically removed BCC lesions with antiganglioside, antiadhesion molecule, and antihistocompatibility locus antigen (HLA) monoclonal antibodies (MoAb). The large majority of BCC lesions were stained by antiganglioside MoAb; among the latter the anti-GD3 ganglioside MoAb R24 displayed the broadest reactivity. The GD3 ganglioside expression by BCC cells, which was corroborated by thin layer chromatography immunostaining with MoAb R24, appears to be a proliferation dependent phenomenon. Among the adhesion molecules tested vitronectin receptor and CDw44 were found in up to 70% of the lesions tested, while intercellular adhesion molecule 1 (ICAM-1) was detected in only a low percentage of BCC cells in one lesion. ICAM-1 was not induced on BCC cells in five and three lesions removed 24 and 48 h, respectively, following the intralesional injection of gamma-interferon. The latter enhanced HLA Class I antigen expression and induced ICAM-1 expression by the surrounding keratinocytes; furthermore gamma-interferon induced HLA Class II antigen expression by a small percentage of BCC cells in three lesions. These results suggest that malignant transformation of keratinocytes is associated with a selective loss of susceptibility to induction by cytokines of ICAM-1 expression. Besides confirming the low HLA Class I and Class II antigen expression by BCC cells, the present investigation has shown a differential expression of distinct monomorphic determinants of HLA Class I antigens and a lower expression of HLA-A antigens than of HLA-B antigens by BCC cells. Furthermore, the present study has shown that HLA Class II antigens can be induced on BCC cells by cytokines.
Asunto(s)
Antígenos CD/análisis , Carcinoma Basocelular/patología , Moléculas de Adhesión Celular/análisis , Gangliósidos/análisis , Antígenos HLA/análisis , Linfocitos Infiltrantes de Tumor/patología , Neoplasias Cutáneas/patología , Anciano , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Carcinoma Basocelular/inmunología , Carcinoma Basocelular/cirugía , Cromatografía en Capa Delgada , Femenino , Gangliósidos/aislamiento & purificación , Antígenos HLA-D/análisis , Antígenos de Histocompatibilidad Clase I/análisis , Humanos , Técnicas para Inmunoenzimas , Molécula 1 de Adhesión Intercelular , Interferón gamma/farmacología , Linfocitos Infiltrantes de Tumor/inmunología , Masculino , Neoplasias Cutáneas/inmunología , Neoplasias Cutáneas/cirugía , Linfocitos T Citotóxicos/inmunologíaRESUMEN
The inhibitory effect of a diphtheria toxin-related interleukin 2 fusion protein, IL-2-toxin, on protein synthesis in adult T-cell leukemia/lymphoma (ATL) cells was examined in vitro. Peripheral blood ATL cells from 12 patients (six acute type, four chronic type, and two smoldering type ATL) and the lymph node cells from three ATL patients (two acute type and one lymphoma type ATL) were examined. At a concentration of 10(-8) M, IL-2-toxin inhibited protein synthesis by 60 to 98% in lymph node ATL cells, whereas protein synthesis in peripheral blood ATL cells was inhibited from 20 to 57% in acute type, and from 3 to 13% in chronic type. In contrast, IL-2-toxin had no measurable effect on T-cells from either patients with smoldering type ATL or normal controls. The cytopathic effects of IL-2-toxin were blocked by the addition of anti-CD25 monoclonal antibody, suggesting that the inhibition of protein synthesis in target cells was mediated by the IL-2 receptor (IL-2R). The degree of inhibition of protein synthesis, however, was not closely correlated with expression of CD25 antigen (low-affinity Mr 55,000 glycoprotein, IL-2R, Tac antigen) on ATL cells. There was an apparent correlation between the degree of inhibition and the rate of protein synthesis in ATL cells. We demonstrate that ATL cells from patients with acute or lymphoma type disease were more sensitive to IL-2-toxin than cells from chronic or smoldering disease. These findings suggest that the high affinity IL-2R present on acute and lymphoma type ATL cells may serve as a target for therapy with this recombinant chimeric toxin.
Asunto(s)
Toxina Diftérica/farmacología , Inmunotoxinas/farmacología , Interleucina-2/farmacología , Leucemia de Células T/inmunología , Leucemia-Linfoma de Células T del Adulto/inmunología , Linfocitos/metabolismo , Biosíntesis de Proteínas , Adulto , Anciano , Células Cultivadas , Femenino , Humanos , Cinética , Linfocitos/efectos de los fármacos , Masculino , Persona de Mediana Edad , Proteínas Recombinantes de Fusión/farmacologíaRESUMEN
Clonal proliferation of human T-lymphotropic virus type I (HTLV-I)-infected cells has been detected by Southern blot analysis and inverse PCR in patients with adult T-cell leukemia, patients with HTLV-I-associated diseases, and even in asymptomatic carriers. Combining inverse PCR with long PCR, we amplified the genomic DNA regions flanking the integration sites of the HTLV-I provirus to detect clones of infected cells. Inverse long PCR revealed that increased virus load was associated with an increase of both the number of cells in each clone and the number of clones. Clonal proliferations were found in both CD4- and CD8-positive cells in a carrier and a patient with HTLV-I-associated neuropathy/tropical spastic paraparesis. These HTLV-I-infected clones persisted over several years in the same carriers, and, moreover, most of the persistent clones were CD4 positive in a HTLV-I carrier. These findings indicate that HTLV-I infection plays an important role in the clonal expansion of lymphocytes and the prolonged survival of CD4-positive cells in vivo. Surviving T-lymphocytes may be susceptible to genetic changes, leading to the onset of leukemia.
Asunto(s)
Linfocitos T CD4-Positivos/patología , Linfocitos T CD8-positivos/patología , Portador Sano/patología , Replicación del ADN , Infecciones por HTLV-I/patología , Virus Linfotrópico T Tipo 1 Humano/genética , Reacción en Cadena de la Polimerasa/métodos , Adulto , Southern Blotting , Linfocitos T CD4-Positivos/virología , Linfocitos T CD8-positivos/virología , Portador Sano/virología , División Celular/genética , Infecciones por HTLV-I/genética , Infecciones por HTLV-I/virología , Humanos , Leucemia-Linfoma de Células T del Adulto/patología , Leucemia-Linfoma de Células T del Adulto/virología , Paraparesia Espástica Tropical/patología , Paraparesia Espástica Tropical/virología , Provirus/genética , Integración ViralRESUMEN
Adult T-cell leukemia-lymphoma (ATL) is a unique T-cell cancer first described in Japan. We estimate that more than 200 patients a year have been detected in Kyushu. The surface phenotype of ATL cells characterized by monoclonal antibodies is T3+, T4+, T8-, T11+, and Tac+. In all cases the serum is positive for anti-human T-cell leukemia (lymphotropic) virus (HTLV-I) antibodies and the ATL cells contain the proviral DNA of HTLV-I. Variations in the clinical features of atypical cases suggest a division of the spectrum of ATL into five types: acute (prototypic), chronic, smoldering, crisis, and lymphoma. Screening of the sera from healthy adults for presence of the anti-HTLV-I antibodies revealed that 3.6% of healthy individuals in Kumamoto Prefecture, which is located in the middle of Kyushu, were HTLV-I carriers. The percentage of positivity increased with age and was higher in females than in males. It varied from town to town, ranging from 0 to 17.6%. Family studies showed that the routes of natural infection of HTLV-I are from mother to child and also from husband to wife. The third route is blood transfusion. The borderline between the healthy carrier state and smoldering ATL remains unclear. In the endemic areas smoldering ATL is frequently diagnosed in patients with fungus infection of the skin, chronic lymphadenopathy, interstitial pneumonitis, chronic renal failure, and strongyloidiasis. In addition our experiences with a concurrence of lymphoma-type ATL in three sisters and spontaneous remissions in a patient with chronic ATL are cited.