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1.
Mol Psychiatry ; 26(9): 5320-5333, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-32576965

RESUMO

Recent theories of cortical function construe the brain as performing hierarchical Bayesian inference. According to these theories, the precision of prediction errors plays a key role in learning and decision-making, is controlled by dopamine and contributes to the pathogenesis of psychosis. To test these hypotheses, we studied learning with variable outcome-precision in healthy individuals after dopaminergic modulation with a placebo, a dopamine receptor agonist bromocriptine or a dopamine receptor antagonist sulpiride (dopamine study n = 59) and in patients with early psychosis (psychosis study n = 74: 20 participants with first-episode psychosis, 30 healthy controls and 24 participants with at-risk mental state attenuated psychotic symptoms). Behavioural computational modelling indicated that precision weighting of prediction errors benefits learning in health and is impaired in psychosis. FMRI revealed coding of unsigned prediction errors, which signal surprise, relative to their precision in superior frontal cortex (replicated across studies, combined n = 133), which was perturbed by dopaminergic modulation, impaired in psychosis and associated with task performance and schizotypy (schizotypy correlation in 86 healthy volunteers). In contrast to our previous work, we did not observe significant precision-weighting of signed prediction errors, which signal valence, in the midbrain and ventral striatum in the healthy controls (or patients) in the psychosis study. We conclude that healthy people, but not patients with first-episode psychosis, take into account the precision of the environment when updating beliefs. Precision weighting of cortical prediction error signals is a key mechanism through which dopamine modulates inference and contributes to the pathogenesis of psychosis.


Assuntos
Dopamina , Transtornos Psicóticos , Teorema de Bayes , Encéfalo , Humanos , Aprendizagem , Imageamento por Ressonância Magnética , Recompensa
2.
Br J Cancer ; 121(3): 282, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31217480

RESUMO

A correction to this paper has been published and can be accessed via a link at the top of the paper.

4.
Sci Rep ; 14(1): 17614, 2024 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-39080434

RESUMO

We used a probabilistic reversal learning task to examine prediction error-driven belief updating in three clinical groups with psychosis or psychosis-like symptoms. Study 1 compared people with at-risk mental state and first episode psychosis (FEP) to matched controls. Study 2 compared people diagnosed with treatment-resistant schizophrenia (TRS) to matched controls. The design replicated our previous work showing ketamine-related perturbations in how meta-level confidence maintained behavioural policy. We applied the same computational modelling analysis here, in order to compare the pharmacological model to three groups at different stages of psychosis. Accuracy was reduced in FEP, reflecting increased tendencies to shift strategy following probabilistic errors. The TRS group also showed a greater tendency to shift choice strategies though accuracy levels were not significantly reduced. Applying the previously-used computational modelling approach, we observed that only the TRS group showed altered confidence-based modulation of responding, previously observed under ketamine administration. Overall, our behavioural findings demonstrated resemblance between clinical groups (FEP and TRS) and ketamine in terms of a reduction in stabilisation of responding in a noisy environment. The computational analysis suggested that TRS, but not FEP, replicates ketamine effects but we consider the computational findings preliminary given limitations in performance of the model.


Assuntos
Ketamina , Transtornos Psicóticos , Reversão de Aprendizagem , Esquizofrenia , Humanos , Esquizofrenia/fisiopatologia , Esquizofrenia/tratamento farmacológico , Transtornos Psicóticos/tratamento farmacológico , Transtornos Psicóticos/fisiopatologia , Transtornos Psicóticos/psicologia , Masculino , Feminino , Adulto , Reversão de Aprendizagem/fisiologia , Adulto Jovem , Adolescente , Psicologia do Esquizofrênico , Estudos de Casos e Controles
5.
Nat Genet ; 26(4): 484-9, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11101851

RESUMO

Notch receptors are involved in cell-fate determination in organisms as diverse as flies, frogs and humans. In Drosophila melanogaster , loss-of-function mutations of Notch produce a 'neurogenic' phenotype in which cells destined to become epidermis switch fate and differentiate to neural cells. Upon ligand activation, the intracellular domain of Notch (ICN) translocates to the nucleus, and interacts directly with the DNA-binding protein Suppressor of hairless (Su(H)) in flies, or recombination signal binding protein Jkappa (RBP-Jkappa) in mammals, to activate gene transcription. But the precise mechanisms of Notch-induced gene expression are not completely understood. The gene mastermind has been identified in multiple genetic screens for modifiers of Notch mutations in Drosophila. Here we clone MAML1, a human homologue of the Drosophila gene Mastermind, and show that it encodes a protein of 130 kD localizing to nuclear bodies. MAML1 binds to the ankyrin repeat domain of all four mammalian NOTCH receptors, forms a DNA-binding complex with ICN and RBP-Jkappa, and amplifies NOTCH-induced transcription of HES1. These studies provide a molecular mechanism to explain the genetic links between mastermind and Notch in Drosophila and indicate that MAML1 functions as a transcriptional co-activator for NOTCH signalling.


Assuntos
Proteínas de Ligação a DNA , Proteínas de Drosophila , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Transativadores/genética , Transativadores/metabolismo , Células 3T3 , Animais , Células COS , Drosophila/genética , Genes de Insetos , Humanos , Camundongos , Dados de Sequência Molecular , Mutação , Receptores Notch , Especificidade da Espécie , Fatores de Transcrição , Transfecção
6.
J Exp Med ; 183(3): 811-20, 1996 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-8642285

RESUMO

Bcr/Abl is a chimeric oncogene that can cause both acute and chronic human leukemias. Bcr/Abl-encoded proteins exhibit elevated kinase activity compared to c-Abl, but the mechanisms of transformation are largely unknown. Some of the biological effects of Bcr/Abl overlap with those of hematopoietic cytokines, particularly interleukin 3 (IL-3). Such effects include mitogenesis, enhanced survival, and enhanced basophilic differentiation. Therefore, it has been suggested that p210Bcr/Abl and the IL-3 receptor may activate some common signal transduction pathways. An important pathway for IL-3 signaling involves activation of the Janus family kinases (JAKs) and subsequent tyrosyl phosphorylation of STAT proteins (signal transducers and activators of transcription). This pathway directly links growth factor receptors to gene transcription. We analyzed JAK activation, STAT protein phosphorylation, and the formation of specific DNA-binding complexes containing STAT proteins, in a series of leukemia cell lines transformed by Bcr/Abl or other oncogenes. We also examined these events in cell lines transformed by a temperature sensitive (ts) mutant of Bcr/Abl, where the kinase activity of Abl could be regulated. STAT1 and STAT5 were found to be constitutively phosphorylated in 32D, Ba/F3, and TF-1 cells transformed by Bcr/Abl, but not in the untransformed parental cell lines in the absence of IL-3. Phosphorylation of STAT1 and STAT5 was also observed in the human leukemia cell lines K562 and BV173, which express the Bcr/Abl oncogene, but not in several Bcr/Abl-negative leukemia cell lines. Phosphorylation of STAT1 and STAT5 was directly due to the tyrosine kinase activity of Bcr/Abl since it could be activated or deactivated by temperature shifting of cells expressing the Bcr/Abl ts mutant. DNA-STAT complexes were detected in all Bcr/Abl-transformed cell lines and they were supershifted by antibodies against STAT1 and STAT5. DNA-STAT complexes in 32Dp210Bcr/Abl cells were similar, but not identical, to those formed after IL-3 stimulation. It is interesting to note that JAK kinases (JAK1, JAK2, JAK3, and Tyk2) were not consistently activated in Bcr/Abl-positive cells. These data suggest that STATs can be activated directly by Bcr/Abl, possibly bypassing JAK family kinase activation. Overall, our results suggest a novel mechanism that could contribute to some of the major biological effects of Bcr/Abl transformation.


Assuntos
Transformação Celular Neoplásica , Proteínas de Ligação a DNA/metabolismo , Proteínas de Fusão bcr-abl/biossíntese , Proteínas do Leite , Fosfotirosina/metabolismo , Proteínas Tirosina Quinases/metabolismo , Sequência de Bases , Sítios de Ligação , Linhagem Celular , Linhagem Celular Transformada , Núcleo Celular/metabolismo , Citocinas/farmacologia , Ativação Enzimática , Proteínas de Fusão bcr-abl/metabolismo , Células HL-60 , Células-Tronco Hematopoéticas , Humanos , Dados de Sequência Molecular , Oligodesoxirribonucleotídeos , Fosforilação , Fosfotirosina/análise , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Fator de Transcrição STAT1 , Fator de Transcrição STAT5 , Transdução de Sinais , Transativadores/metabolismo , Transfecção , Células Tumorais Cultivadas , Tirosina/metabolismo
7.
J Exp Med ; 162(3): 1111-6, 1985 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-3928803

RESUMO

Gamma interferon induced surface expression of interleukin 2 (IL-2) receptors on normal human monocytes and the monocytoid cell lines U937 and HL60. These receptors were detected by anti-IL-2 receptor monoclonal antibodies, and U937 IL-2 receptors were indistinguishable from T lymphocyte IL-2 receptors by immunoprecipitation. Also, U937 IL-2 receptors bound biologically active IL-2. These results suggest a role for monocyte IL-2 receptors in T cell/monocyte interaction during an immune response.


Assuntos
Interferon gama/farmacologia , Interleucina-2/metabolismo , Leucemia Mieloide/imunologia , Monócitos/metabolismo , Receptores Imunológicos/metabolismo , Anticorpos Monoclonais/imunologia , Diferenciação Celular , Divisão Celular , Linhagem Celular , Humanos , Leucemia Mieloide/patologia , Monócitos/efeitos dos fármacos , Receptores Imunológicos/imunologia , Receptores de Interleucina-2 , Linfócitos T/metabolismo
8.
J Exp Med ; 162(6): 2035-52, 1985 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-3934319

RESUMO

Restriction on cytolytic T lymphocyte (CTL)-target cell-interactions are studied in the primate S. oedipus, a naturally occurring A + B----A bone marrow-chimeric species. We show that the T cell, B cell, and myelomonocytic progenitor cell populations are chimeric in this species. We selected animals for study that are populated by fully major histocompatibility complex (MHC)-disparate hematopoietic cell populations, using a functional assay system. We then developed an in vitro system for analyzing at the clonal level the genetic restrictions on the trinitrophenyl-specific CTL-target cell interactions of this species. In this system, we have shown that tolerance to foreign MHC determinants was not, of itself, sufficient to facilitate the generation of CTL specific for target cells expressing those foreign MHC determinants. Rather, a marked preference for the expansion of CTL clones with a restriction for target cells bearing the host animals' MHC determinants was seen. Hematopoietically derived cells did not affect the repertoire of these T lymphocytes. These studies represent the first demonstration of the phenomenon of an environment dictating interactional restrictions on CTL in a naturally occurring bone marrow-chimeric animal. This is also the first demonstration of the profound influence of the environment on the repertoire of the T lymphocyte in a primate species.


Assuntos
Medula Óssea/imunologia , Callitrichinae/imunologia , Comunicação Celular , Quimera , Citotoxicidade Imunológica , Saguinus/imunologia , Linfócitos T Citotóxicos/imunologia , Animais , Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/fisiologia , Medula Óssea/fisiologia , Células da Medula Óssea , Células Clonais/imunologia , Células Clonais/fisiologia , Feminino , Haptenos/imunologia , Antígenos de Histocompatibilidade/genética , Antígenos de Histocompatibilidade/imunologia , Teste de Histocompatibilidade , Humanos , Ativação Linfocitária , Masculino , Linfócitos T Citotóxicos/fisiologia , Gêmeos
9.
J Exp Med ; 180(5): 1665-73, 1994 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-7525835

RESUMO

Induction and maintenance of a state of T cell unresponsiveness to specific alloantigen would have significant implications for human organ transplantation. Using human histocompatibility leukocyte antigen DR7-specific helper T cell clones, we demonstrate that blockade of the B7 family of costimulatory molecules is sufficient to induce alloantigen-specific T cell clonal anergy. Anergized cells do not respond to alloantigen and a variety of costimulatory molecules, including B7-1, B7-2, intercellular adhesion molecule-1 (ICAM-1), and lymphocyte function-associated molecule (LFA)-3. However, after culture in exogenous interleukin (IL)-2 for at least 7 d, anergized cells can respond to alloantigen in the presence of LFA-3. LFA-3 costimulation subsequently restores responsiveness to alloantigen in the presence of previously insufficient costimulatory signals. Expression of CD2R epitope is downregulated on anergic cells and is restored after 7 d of IL-2 culture. The loss of the CD2R is temporally associated with the inability of anergized cells to respond to LFA-3. These results suggest that in addition to blockade of B7 family members, inhibition of CD2 and, potentially, other costimulatory pathways that might reverse anergy will be necessary to maintain prolonged alloantigen-specific tolerance.


Assuntos
Antígenos CD2/fisiologia , Anergia Clonal , Imunoconjugados , Isoantígenos/imunologia , Abatacepte , Antígenos CD/fisiologia , Antígenos de Diferenciação/fisiologia , Antígeno B7-1/fisiologia , Antígenos CD58 , Antígeno CTLA-4 , Células Clonais , Epitopos , Antígeno HLA-DR7/fisiologia , Humanos , Interleucina-2/farmacologia , Glicoproteínas de Membrana/fisiologia
10.
J Exp Med ; 157(1): 114-29, 1983 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-6571733

RESUMO

Fetal hematopoietic cells that express the common acute lymphoblastic leukemia antigen (CALLA) were purified from both fetal liver and fetal bone marrow by immune rosetting with sheep erythrocytes coated with rabbit anti-mouse immunoglobulin and by fluorescence-activated cell sorting. Dual fluorescence techniques disclosed that these cells were heterogenous with respect to the expression of a series of differentiation and activation antigens defined by monoclonal antibodies. Thus, whereas all CALLA+ cells were Ia+ and expressed two activation antigens, J2 and T10, only 30-50% expressed B1 antigen. Furthermore, using methanol-fixed cells, it could be shown that approximately 20% contained intracytoplasmic mu chains (cyto-mu) and that approximately 15% were positive for the terminal transferase enzyme (TdT) marker. The CALLA+ fetal cells thus closely resemble the childhood acute lymphoblastic leukemia cell with respect to surface marker phenotype. A population of CALLA- cells devoid of mature erythroid and myeloid surface markers was found to contain higher numbers of TdT+ cells but lower numbers of cyto-mu, B1, and Ia+ cells than the CALLA+ subset. In vitro analysis of normal, purified CALLA+ cells demonstrated that incubation at 37 degrees C with J5 monoclonal antibody specific for CALLA resulted in the specific modulation of surface antigen. Similar results have previously been obtained with CALLA+ tumor cells. Although phenotypic analysis of CALLA+ cells suggests that these cells are relatively immature lymphoid cells, CALLA+ cells do not appear to contain either myeloid precursor cells (CFU-G/M) or the earliest lymphoid stem cells.


Assuntos
Antígenos de Neoplasias/análise , Células-Tronco Hematopoéticas/imunologia , Leucemia Linfoide/imunologia , Anticorpos Monoclonais/imunologia , Antígenos de Superfície/análise , Células da Medula Óssea , Feto/imunologia , Humanos , Fígado/citologia
11.
Br J Cancer ; 102(2): 351-60, 2010 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-20010940

RESUMO

BACKGROUND: Epithelial-to-mesenchymal transition (EMT) is associated with decreased adhesion and acquisition of metastatic potential of breast cancer cells. Epithelial-to-mesenchymal transition is mediated, in part, by two transcription repressors, Snail and Slug, that are known to be targets of the Notch signaling pathway, and JAGGED1-induced Notch activation increases EMT. However, the events that lead to increased Notch activity during EMT of breast cancer cells are unknown. METHODS: The accumulation of hypoxia inducible factors (HIFs) under hypoxia was detected by western blot analysis, and their effects on Notch signaling were measured by an in vitro Notch reporter assay. The expression of Notch target genes under hypoxia was tested by real-time PCR. The knockdown of HIF-1alpha was mediated by retroviral delivery of shRNA. The expression of Slug and Snail under hypoxia was measured by real-time PCR. Breast cancer cell migration and invasion under hypoxia were tested with cell migration and invasion kits. RESULTS: Hypoxia increased the expression of Notch target genes such as HES1 and HEY1 in breast cancer cells, as was expression of Notch receptors and ligands. The mechanism is likely to involve the accumulation of HIF-1alpha and HIF-2alpha in these cells by hypoxia, which synergised with the Notch co-activator MAML1 in potentiating Notch activity. Hypoxia inducible factor-1alpha was found to bind to HES1 promoter under hypoxia. Knockdown of HIF-1alpha with shRNA inhibited both HES1 and HEY1 expression under hypoxia. Hypoxia increased the expression of Slug and Snail, and decreased the expression of E-cadherin, hallmarks of EMT. Notch pathway inhibition abrogated the hypoxia-mediated increase in Slug and Snail expression, as well as decreased breast cancer cell migration and invasion. CONCLUSION: Hypoxia-mediated Notch signaling may have an important role in the initiation of EMT and subsequent potential for breast cancer metastasis.


Assuntos
Neoplasias da Mama/metabolismo , Caderinas/biossíntese , Movimento Celular/fisiologia , Hipóxia/fisiopatologia , Receptores Notch/biossíntese , Fatores de Transcrição Hélice-Alça-Hélice Básicos/biossíntese , Neoplasias da Mama/fisiopatologia , Proteínas de Ciclo Celular/biossíntese , Feminino , Proteínas de Homeodomínio/biossíntese , Humanos , Invasividade Neoplásica/fisiopatologia , Transdução de Sinais , Fatores de Transcrição HES-1
12.
Science ; 250(4982): 805-8, 1990 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-2237430

RESUMO

The product of the cdc2 gene, designated p34cdc2, is a serine-threonine protein kinase that controls entry of eukaryotic cells into mitosis. Freshly isolated human T lymphocytes (G0 phase) were found to have very low amounts of p34cdc2 and cdc2 messenger RNA. Expression of cdc2 increased 18 to 24 hours after exposure of T cells to phytohemagglutinin, coincident with the G1 to S transition. Antisense oligodeoxynucleotides could reduce the increase in cdc2 expression and inhibited DNA synthesis, but had no effect on several early and mid-G1 events, including blastogenesis and expression of interleukin-2 receptors, transferrin receptors, c-myb, and c-myc. Induction of cdc2 required prior induction of c-myb and c-myc. These results suggest that cdc2 induction is part of an orderly sequence of events that occurs at the G1 to S transition in T cells.


Assuntos
Proteína Quinase CDC2/genética , Fase G1 , Regulação da Expressão Gênica , Fase S , Linfócitos T/citologia , Sequência de Bases , Northern Blotting , Proteína Quinase CDC2/biossíntese , Células Cultivadas , DNA/biossíntese , DNA/genética , Citometria de Fluxo , Genes do Retinoblastoma , Genes myc , Humanos , Ativação Linfocitária , Dados de Sequência Molecular , Fosforilação , Reação em Cadeia da Polimerase , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-myb , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Linfócitos T/metabolismo
13.
Leukemia ; 21(3): 462-71, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17252014

RESUMO

Activating mutations in the NOTCH1 gene have been found in about 60% of patients with T-cell acute lymphoblastic leukemia (T-ALL). In order to study the molecular mechanisms by which altered Notch signaling induces leukemia, a zebrafish model of human NOTCH1-induced T-cell leukemia was generated. Seven of sixteen mosaic fish developed a T-cell lymphoproliferative disease at about 5 months. These neoplastic cells extensively invaded tissues throughout the fish and caused an aggressive and lethal leukemia when transplanted into irradiated recipient fish. However, stable transgenic fish exhibited a longer latency for leukemia onset. When the stable transgenic line was crossed with another line overexpressing the zebrafish bcl2 gene, the leukemia onset was dramatically accelerated, indicating synergy between the Notch pathway and the bcl2-mediated antiapoptotic pathway. Reverse transcription-polymerase chain reaction analysis showed that Notch target genes such as her6 and her9 were highly expressed in NOTCH1-induced leukemias. The ability of this model to detect a strong interaction between NOTCH1 and bcl2 suggests that genetic modifier screens have a high likelihood of revealing other genes that can cooperate with NOTCH1 to induce T-ALL.


Assuntos
Transformação Celular Neoplásica/genética , Leucemia-Linfoma de Células T do Adulto/etiologia , Proteínas Proto-Oncogênicas c-bcl-2/fisiologia , Receptor Notch1/fisiologia , Animais , Animais Geneticamente Modificados , Apoptose , Fatores de Transcrição Hélice-Alça-Hélice Básicos/fisiologia , Feminino , Raios gama , Perfilação da Expressão Gênica , Regulação Leucêmica da Expressão Gênica , Rearranjo Gênico da Cadeia alfa dos Receptores de Antígenos dos Linfócitos T , Genes bcl-2 , Humanos , Leucemia-Linfoma de Células T do Adulto/genética , Masculino , Mosaicismo , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/fisiologia , Transplante de Neoplasias , Oncogenes , Quimera por Radiação , Tolerância a Radiação , Receptor Notch1/genética , Proteínas Recombinantes de Fusão/fisiologia , Transdução de Sinais , Fatores de Tempo , Peixe-Zebra , Proteínas de Peixe-Zebra/fisiologia
14.
J Clin Invest ; 79(1): 100-6, 1987 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-3491836

RESUMO

Expression of the granulocyte-macrophage colony-stimulating factor (GM-CSF) gene was studied by Northern blot analysis in normal human hematopoietic cells and a series of leukemias. GM-CSF messenger (m)RNA was detected in activated T cells, but not in normal bone marrow cells, monocytes, or nonactivated T cells. In contrast, leukemic cells from 11 of 22 cases of acute myeloblastic leukemia expressed GM-CSF transcripts. Biologically active CSF was detected in supernatant conditioned by 6 of these 11 leukemias. Expression of the GM-CSF gene was not detected in "common" (pre-B cell) acute lymphoblastic leukemia (11 cases tested) or chronic myeloid leukemia (4 cases tested). These results show that the GM-CSF gene is constitutively expressed in a subset of patients with AML, and further suggest that expression of this gene could contribute to the abnormal growth properties characteristic of AML.


Assuntos
Interleucina-3/genética , Leucemia Mieloide Aguda/genética , Reações Antígeno-Anticorpo , Bioensaio , Células Cultivadas , Regulação da Expressão Gênica , Humanos , Interleucina-3/imunologia , RNA Mensageiro/genética
15.
J Clin Invest ; 77(1): 13-20, 1986 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-3455931

RESUMO

The expression of HLA-DR antigens by normal myeloid progenitor cells (CFU-GM) has been linked to inhibition of colony growth by prostaglandin E (PGE), while resistance to the inhibitory effects of PGE in chronic myeloid leukemia (CML) has been attributed to a lower fraction of HLA-DR+ CFU-GM in this disease. However, we have previously shown that virtually all CFU-GM in normal bone marrow (NBM) as well as CML peripheral blood express HLA-DR antigens, which raises the possibility that these surface molecules may not be the sole determinants of a progenitor cell's sensitivity to PGE. In order to evaluate the relationship between HLA-DR expression and prostaglandin inhibition, we partially purified NBM progenitor cells using fluorescence-activated cell sorting to prepare cell fractions with high and low HLA-DR antigen density. Normal progenitor cells with high DR density tended to form monocyte colonies in agar culture, whereas the low DR density fraction was enriched for granulocyte colony-forming cells. Inhibition by PGE was greatest in the high DR+ fraction and was largely restricted to monocyte progenitor cells. Inhibition of CFU-GM by PGE was less in CML than in NBM, but this decreased inhibition correlated with a significantly lower number of monocyte-CFU in CML. These data suggest that high HLA-DR antigen density may select for normal progenitor cells that are committed to monocyte differentiation and are, therefore, more likely to be inhibited by PGE. The relative deficit of monocyte progenitor cells in CML may partially explain the phenomenon of PGE resistance in this disease.


Assuntos
Alprostadil/farmacologia , Inibidores do Crescimento/farmacologia , Células-Tronco Hematopoéticas/imunologia , Antígenos de Histocompatibilidade Classe II/análise , Leucemia Mieloide/imunologia , Células da Medula Óssea , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Granulócitos/classificação , Granulócitos/citologia , Granulócitos/imunologia , Antígenos HLA-DR , Hematopoese/efeitos dos fármacos , Células-Tronco Hematopoéticas/patologia , Humanos , Leucemia Mieloide/patologia , Macrófagos/classificação , Macrófagos/citologia , Macrófagos/imunologia , Suspensões , Ensaio Tumoral de Célula-Tronco
16.
J Clin Invest ; 100(7): 1708-15, 1997 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-9312168

RESUMO

The BCR/ABL oncogene encodes an activated tyrosine kinase that causes human chronic myelogenous leukemia. The mechanism of transformation, however, is complex and not well understood. One of the important contributions of BCR to transformation is believed to be dimerization or oligomerization of ABL, thereby activating ABL tyrosine kinase activity. We reasoned that if ABL was dimerized through other mechanisms, activation of the tyrosine kinase activity should also result, and the activated kinase may also be transforming. Erythropoietin is known to activate its receptor by causing dimerization, and therefore a synthetic oncogene was created by linking the extracytoplasmic and transmembrane domains of the EPO receptor with c-ABL. This chimeric receptor was stably expressed in Ba/F3 cells and, in the absence of EPO, had no detectable biological effect on the cells. EPO, however, induced a rapid, dose-dependent activation of ABL tyrosine kinase activity and phosphorylation of several cellular proteins. The major target proteins have been identified, and are very similar to the known substrates of BCR/ABL, including Shc, CBL, CRKL, and several proteins in the cytoskeleton. EPO treatment also resulted in biological effects that were remarkably similar to those of BCR/ABL, including improved viability, altered integrin function, and a weak mitogenic signal. The biological effects were in part dose-dependent, in that low EPO concentrations enhanced viability but did not cause proliferation. At high EPO doses, kinase activation was maximal, and a mitogenic effect was also revealed. In nude mice, Ba/F3 cells expressing this chimeric receptor did not cause detectable disease without administration of pharmacologic doses of EPO. If EPO was given intraperitoneally 5 days a week, however, a dose-dependent lethal leukemia resulted. This ligand-regulatable oncogene mimics some of the biological effects of BCR/ABL, and analysis of ABL mutants in this system will be useful to dissect the signaling pathways that cause CML.


Assuntos
Eritropoetina/farmacologia , Regulação Neoplásica da Expressão Gênica , Leucemia Experimental/induzido quimicamente , Proteínas Tirosina Quinases/biossíntese , Proteínas Proto-Oncogênicas c-abl/biossíntese , Receptores da Eritropoetina/biossíntese , Sequência de Aminoácidos , Animais , Linhagem Celular , Relação Dose-Resposta a Droga , Ativação Enzimática , Injeções Intraperitoneais , Leucemia Experimental/genética , Ligantes , Camundongos , Camundongos Nus , Dados de Sequência Molecular , Fosforilação , Proteínas Tirosina Quinases/genética , Proteínas Proto-Oncogênicas c-abl/genética , Receptores da Eritropoetina/genética , Proteínas Recombinantes de Fusão/biossíntese
17.
J Clin Invest ; 98(2): 521-8, 1996 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-8755665

RESUMO

Cell adhesion to the extracellular matrix is largely mediated by adhesion molecules of the integrin family and is often diminished upon oncogenic transformation. However, we show here that the chronic myelogenous leukemia oncogene Bcr/Abl has positive effects on VLA-4 and VLA-5 integrin function. The presence of Bcr/Abl in the GM-CSF- or IL-3-dependent hematopoietic cell lines MO7e, 32D, and BaF/3 enhanced cell binding to both soluble and immobilized fibronectin. The effect was due to enhanced function of the VLA-5 integrin fibronectin receptor and not to increased surface expression. In parallel, Bcr/Abl stimulated cell adhesion to the VLA-4 integrin ligand VCAM-1. Stimulation of VLA-5 function directly correlated with induction of Bcr/Abl tyrosine kinase activity in a temperature-sensitive kinase mutant. Thus, Bcr/Abl stimulates integrin-dependent cell adhesion, by a mechanism involving increased ligand binding, with the tyrosine kinase activity of Bcr/Abl likely playing a key role. Consistent with these results, hematopoietic precursor cells from chronic myelogenous leukemia patients also showed increased adhesion to fibronectin.


Assuntos
Adesão Celular , Proteínas de Fusão bcr-abl/biossíntese , Células-Tronco Hematopoéticas/fisiologia , Integrinas/fisiologia , Animais , Linhagem Celular , Fibronectinas , Proteínas de Fusão bcr-abl/fisiologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Immunoblotting , Integrina alfa4beta1 , Interleucina-3/farmacologia , Cinética , Leucemia Experimental , Leucemia Megacarioblástica Aguda , Camundongos , Receptores de Fibronectina/fisiologia , Receptores de Retorno de Linfócitos/fisiologia , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Fatores de Tempo , Transfecção , Células Tumorais Cultivadas , Molécula 1 de Adesão de Célula Vascular/fisiologia
18.
J Clin Invest ; 68(4): 932-41, 1981 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-6945311

RESUMO

A series of monoclonal antibodies have been characterized that define four surface antigens (MY3, MY4, MY7, and MY8) of human myeloid cells. They were derived from a fusion of the NS-1 plasmacytoma cell line with splenocytes from a mouse immunized with human acute myelomonocytic leukemia cells. MY3 and MY4 are expressed by normal monocytes and by greater than 90% of patients with acute monocytic leukemia or acute myelomonocytic leukemia, but are detected much less often on other types of myeloid leukemia. MY7 is expressed by granulocytes, monocytes, and 5% of normal bone marrow cells. 79% of all acute myeloblastic leukemia (AML) patients tested (72 patients) express MY7 without preferential expression by any AML subtype. MY8 is expressed by normal monocytes, granulocytes, all peroxidase-positive bone marrow cells, and 50% of AML patients. MY3, MY4, and MY8 define myeloid differentiation antigens in that they are not detected on myeloid precursor cells and appear at discrete stages of differentiation. These antigens are not expressed by lymphocytes, erythrocytes, platelets, or lymphoid malignancies. The monoclonal antisera defining these antigens have been used to study differentiation of normal myeloid cells and malignant cell lines.


Assuntos
Antígenos de Neoplasias/análise , Antígenos de Superfície/análise , Medula Óssea/imunologia , Leucemia/imunologia , Animais , Anticorpos Monoclonais , Células da Medula Óssea , Diferenciação Celular , Linhagem Celular , Hematopoese , Humanos , Leucemia Mieloide Aguda/imunologia , Camundongos
19.
J Clin Invest ; 75(2): 746-53, 1985 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-3855866

RESUMO

The expression of differentiation-associated surface antigens by the clonogenic leukemic cells from 20 patients with acute myeloblastic leukemia (AML) was studied with a panel of seven cytotoxic monoclonal antibodies (anti-Ia, -MY9, -PM-81, -AML-2-23, -Mol, -Mo2, and -MY3). The surface antigen phenotypes of the clonogenic cells were compared with the phenotypes of the whole leukemic cell population, and with the phenotypes of normal hematopoietic progenitor cells. In each case the clonogenic leukemic cells were found within a distinct subpopulation that was less "differentiated" than the total cell population. Clonogenic leukemic cells from different patients could be divided into three phenotype groups. In the first group (7 of 20 cases), the clonogenic cells expressed surface antigens characteristic of the normal multipotent colony-forming cell (Ia, MY9). These cases tended to have "undifferentiated" (FAB M1) morphology, and the total cell population generally lacked expression of "late" monocyte antigens such as MY3 and Mo2. A second group (seven cases) of clonogenic cells expressed surface antigens characteristic of an "early" (day 14) colony-forming unit granulocyte-monocyte (CFU-GM), and a third group (six cases) was characteristic of a "late" (day 7) CFU-GM. The cases in these latter two groups tended to have myelomonocytic (FAB M4) morphology and to express monocyte surface antigens. These results suggest that the clonogenic cells are a distinct subpopulation in all cases of AML, and may be derived from normal hematopoietic progenitor cells at multiple points in the differentiation pathway. The results further support the possibility that selected monoclonal antibodies have the potential to purge leukemic clonogenic cells from bone marrow in some AML patients without eliminating critical normal progenitor cells.


Assuntos
Leucemia Mieloide Aguda/patologia , Células-Tronco Neoplásicas/patologia , Células-Tronco/patologia , Adulto , Idoso , Antígenos de Neoplasias , Antígenos de Superfície , Diferenciação Celular , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Leucemia Mieloide Aguda/imunologia , Masculino , Pessoa de Meia-Idade , Células-Tronco Neoplásicas/imunologia
20.
J Clin Invest ; 75(5): 1690-5, 1985 May.
Artigo em Inglês | MEDLINE | ID: mdl-2987309

RESUMO

SM-1 is a murine monoclonal antibody strongly reactive with a cell membrane antigen of small cell carcinoma (SCC) of the lung but unreactive with the membrane of most other carcinomas and normal tissues including normal bone marrow. We have found that in the presence of human complement, SM-1 antibody is highly cytotoxic to SCC cells. Using three treatments with antibody and complement, more than 99% of SCC cells in culture were lysed, as determined by the chromium release and clonogenic assays. Similar efficiency of SCC cell lysis was observed when one SM-1 antibody treatment was followed by three treatments with human complement. In contrast, there was little antibody-dependent lysis of non-small cell lung cancer cells, other carcinomas, and leukemia cell lines. The amount of chromium released from normal bone marrow cells treated with SM-1 antibody and complement was minimal and was mainly due to the effect of complement alone. Clonogenic assays, including colony-forming unit-granulocytic/monocytic, erythroid burst-forming unit, and colony-forming unit-granulocytic/erythroid/monocytic/megakaryocytic, also showed no significant SM-1 antibody-dependent cytotoxicity on normal bone marrow precursors. Since SM-1 antibody is selectively cytotoxic to SCC cells in the presence of human complement, it is a potentially useful agent for the selective eradication of tumor cell contamination in marrows of patients with metastatic small cell lung cancer and possibly for in vivo serotherapy.


Assuntos
Anticorpos Monoclonais/uso terapêutico , Carcinoma de Células Pequenas/terapia , Proteínas do Sistema Complemento/uso terapêutico , Citotoxicidade Imunológica , Neoplasias Pulmonares/terapia , Anticorpos Monoclonais/fisiologia , Temperatura Corporal , Células da Medula Óssea , Carcinoma de Células Pequenas/imunologia , Carcinoma de Células Pequenas/patologia , Linhagem Celular , Ensaio de Unidades Formadoras de Colônias , Relação Dose-Resposta Imunológica , Humanos , Fatores de Tempo , Ensaio Tumoral de Célula-Tronco
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