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1.
Invest New Drugs ; 37(4): 702-710, 2019 08.
Artículo en Inglés | MEDLINE | ID: mdl-30569244

RESUMEN

Purpose Vorinostat is a potent HDAC inhibitor that sensitizes head and neck squamous cell carcinoma (HNSCC) to cytotoxic therapy while sparing normal epithelium. The primary objective of this Phase I study was to determine the maximally tolerated dose (MTD) and safety of Vorinostat in combination with standard chemoradiation therapy treatment in HNSCC. Patients and Methods Eligible patients had pathologically confirmed Stage III, IVa, IVb HNSCC, that was unresectable or borderline resectable involving the larynx, hypopharynx, nasopharynx, and oropharynx. Vorinostat was administered at the assigned dosage level (100-400 mg, three times weekly) in a standard 3 + 3 dose escalation design. Vorinostat therapy began 1 week prior to initiation of standard, concurrent chemoradiation therapy and continued during the entire course of therapy. Results Twenty six patients met eligibility criteria and completed the entire protocol. The primary tumor sites included tonsil (12), base of tongue (9), posterior pharyngeal wall (1), larynx (4) and hypopharynx (3). Of the 26 patients, 17 were HPV-positive and 9 were HPV-negative. The MTD of Vorinostat was 300 mg administered every other day. Anemia (n = 23/26) and leukopenia (n = 20/26) were the most commonly identified toxicities. The most common Grade3/4 events included leukopenia (n = 11) and lymphopenia (n = 17). No patient had Grade IV mucositis, dermatitis or xerostomia. The median follow time was 33.8 months (range 1.6-82.9 months). Twenty four of 26 (96.2%) patients had a complete response to therapy. Conclusion Vorinostat in combination with concurrent chemoradiation therapy is a safe and highly effective treatment regimen in HNSCC. There was a high rate of complete response to therapy with toxicity rates comparable, if not favorable to existing therapies. Further investigation in Phase II and III trials is strongly recommended.


Asunto(s)
Antineoplásicos/administración & dosificación , Quimioradioterapia , Neoplasias de Cabeza y Cuello/terapia , Carcinoma de Células Escamosas de Cabeza y Cuello/terapia , Vorinostat/administración & dosificación , Anemia/inducido químicamente , Antineoplásicos/efectos adversos , Quimioradioterapia/efectos adversos , Cisplatino/administración & dosificación , Erupciones por Medicamentos , Femenino , Humanos , Leucopenia/inducido químicamente , Masculino , Dosis Máxima Tolerada , Persona de Mediana Edad , Mucositis/inducido químicamente , Análisis de Supervivencia , Resultado del Tratamiento , Vorinostat/efectos adversos , Pérdida de Peso
2.
Oncogene ; 36(2): 263-274, 2017 01 12.
Artículo en Inglés | MEDLINE | ID: mdl-27292259

RESUMEN

Glioblastoma (GBM) represents the most common and aggressive histologic subtype among malignant astrocytoma and is associated with poor outcomes because of heterogeneous tumour cell population including mature non-stem-like cell and immature stem-like cells within the tumour. Thus, it is critical to find new target-specific therapeutic modalities. Protein arginine methyltransferase enzyme 5 (PRMT5) regulates many cellular processes through its methylation activity and its overexpression in GBM is associated with more aggressive disease. Previously, we have shown that silencing of PRMT5 expression in differentiated GBM cell lines results in apoptosis and reduced tumour growth in mice. Here, we report the critical role of PRMT5 in GBM differentiated cells (GBMDC) grown in serum and GBM neurospheres (GBMNS) grown as neurospheres in vitro. Our results uncover a very significant role for PRMT5 in GBMNS self-renewal capacity and proliferation. PRMT5 knockdown in GBMDC led to apoptosis, knockdown in GBMNS led to G1 cell cycle arrest through upregulation of p27 and hypophoshorylation of retinoblastoma protein, leading to senescence. Comparison of impact of PRMT5 on cellular signalling by the Human Phospho-Kinase Array and chromatin immunoprecipitation-PCR revealed that unlike GBMDC, PRMT5 regulates PTEN expression and controls Akt and ERk activity in GBMNS. In vivo transient depletion of PRMT5 decreased intracranial tumour size and growth rate in mice implanted with both primary tumour-derived GBMNS and GBMDC. This is the first study to identify PTEN as a potential downstream target of PRMT5 and PRMT5 is vital to support both mature and immature GBM tumour cell populations.


Asunto(s)
Neoplasias Encefálicas/patología , Glioblastoma/patología , Fosfohidrolasa PTEN/metabolismo , Proteína-Arginina N-Metiltransferasas/metabolismo , Esferoides Celulares/citología , Animales , Neoplasias Encefálicas/metabolismo , Ciclo Celular , Autorrenovación de las Células , Senescencia Celular , Glioblastoma/metabolismo , Humanos , Ratones , Trasplante de Neoplasias , Transducción de Señal , Esferoides Celulares/metabolismo , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/metabolismo
3.
Leukemia ; 30(4): 789-99, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26536822

RESUMEN

Changes in the enzymatic activity of protein arginine methyltransferase (PRMT) 5 have been associated with cancer; however, the protein's role in acute myeloid leukemia (AML) has not been fully evaluated. Here, we show that increased PRMT5 activity enhanced AML growth in vitro and in vivo while PRMT5 downregulation reduced it. In AML cells, PRMT5 interacted with Sp1 in a transcription repressor complex and silenced miR-29b preferentially via dimethylation of histone 4 arginine residue H4R3. As Sp1 is also a bona fide target of miR-29b, the miR silencing resulted in increased Sp1. This event in turn led to transcription activation of FLT3, a gene that encodes a receptor tyrosine kinase. Inhibition of PRMT5 via sh/siRNA or a first-in-class small-molecule inhibitor (HLCL-61) resulted in significantly increased expression of miR-29b and consequent suppression of Sp1 and FLT3 in AML cells. As a result, significant antileukemic activity was achieved. Collectively, our data support a novel leukemogenic mechanism in AML where PRMT5 mediates both silencing and transcription of genes that participate in a 'yin-yang' functional network supporting leukemia growth. As FLT3 is often mutated in AML and pharmacologic inhibition of PRMT5 appears feasible, the PRMT5-miR-29b-FLT3 network should be further explored as a novel therapeutic target for AML.


Asunto(s)
Arginina/química , Metilación de ADN , Epigénesis Genética/genética , Epigenómica , Histonas/química , Leucemia Mieloide Aguda/genética , MicroARNs/genética , Proteína-Arginina N-Metiltransferasas/genética , Animales , Apoptosis , Western Blotting , Proliferación Celular , Inmunoprecipitación de Cromatina , Regulación hacia Abajo , Citometría de Flujo , Regulación Leucémica de la Expresión Génica , Humanos , Técnicas para Inmunoenzimas , Leucemia Mieloide Aguda/patología , Ratones , Ratones Endogámicos NOD , Ratones SCID , Proteína-Arginina N-Metiltransferasas/antagonistas & inhibidores , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Activación Transcripcional , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de Xenoinjerto
4.
Curr Hematol Malig Rep ; 10(4): 456-67, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26449716

RESUMEN

While Epstein-Barr virus (EBV) was initially discovered and characterized as an oncogenic virus in B cell neoplasms, it also plays a complex and multifaceted role in T/NK cell lymphomas. In B cell lymphomas, EBV-encoded proteins have been shown to directly promote immortalization and proliferation through stimulation of the NF-κB pathway and increased expression of anti-apoptotic genes. In the context of mature T/NK lymphomas (MTNKL), with the possible exception on extranodal NK/T cell lymphoma (ENKTL), the virus likely plays a more diverse and nuanced role. EBV has been shown to shape the tumor microenvironment by promoting Th2-skewed T cell responses and by increasing the expression of the immune checkpoint ligand PD-L1. The type of cell infected, the amount of plasma EBV DNA, and the degree of viral lytic replication have all been proposed to have prognostic value in T/NK cell lymphomas. Latency patterns of EBV infection have been defined using EBV-infected B cell models and have not been definitively established in T/NK cell lymphomas. Identifying the expression profile of EBV lytic proteins could allow for individualized therapy with the use of antiviral medications. More work needs to be done to determine whether EBV-associated MTNKL have distinct biological and clinical features, which can be leveraged for risk stratification, disease monitoring, and therapeutic purposes.


Asunto(s)
Infecciones por Virus de Epstein-Barr/virología , Células Asesinas Naturales/virología , Linfoma de Células T/virología , Humanos , Pronóstico
5.
Bone Marrow Transplant ; 48(9): 1212-7, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23584442

RESUMEN

Positron emission tomography/computed tomography (PET/CT)-positive findings before autologous SCT (auto-SCT) are associated with inferior PFS and OS in patients with relapsed Hodgkin's and diffuse large B-cell lymphoma. We classified pre-transplant PET/CT performed before auto-SCT as positive or negative to evaluate the impact of pre-transplant PET/CT in mantle cell lymphoma (MCL). In 29 patients, 17 were PET/CT(-) and 12 were PET/CT(+). PET/CT(+) patients were younger (P=0.04), had lower MCL International Prognostic Index (MIPI, P=0.04) scores, but increased bulky adenopathy >5 cm (45% vs 13%, P=0.09). With a median follow-up of 27 months (range: 5-55 months), 7 patients relapsed (4 in the PET/CT(-) group and 3 in the PET/CT(+) group) with 2 deaths in the PET/CT(+) group without a documented relapse. The estimated 2-year PFS was 64% (95% confidence interval (CI): 0.30-0.85) vs 87% (95% CI: 0.57-0.97) in PET/CT(+) and PET/CT(-) patients, respectively (P=0.054). OS was significantly decreased in PET/CT(+) patients (P=0.007), with 2-year estimates of 60% (95% CI: 0.23-0.84) vs 100% in PET/CT(-) patients. A positive pre-transplant PET/CT is associated with a poor prognosis in patients with MCL. Additional factors may impact the prognostic value of PET/CT, as several PET/CT(+) patients remain in remission.


Asunto(s)
Trasplante de Células Madre Hematopoyéticas/métodos , Linfoma de Células del Manto/diagnóstico , Linfoma de Células del Manto/cirugía , Imagen Multimodal/métodos , Tomografía de Emisión de Positrones/métodos , Tomografía Computarizada por Rayos X/métodos , Adulto , Anciano , Supervivencia sin Enfermedad , Femenino , Humanos , Linfoma de Células del Manto/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Retrospectivos , Acondicionamiento Pretrasplante/métodos , Trasplante Homólogo , Resultado del Tratamiento
6.
Oncogene ; 26(25): 3644-53, 2007 May 28.
Artículo en Inglés | MEDLINE | ID: mdl-17530018

RESUMEN

Alemtuzumab (Campath-1H) is a humanized IgG1 monoclonal antibody that targets the human CD52 antigen. CD52 is expressed by a variety of lymphoid neoplasms and most human mononuclear cell subsets. In 2001, alemtuzumab was approved for marketing in the United States and Europe for use in patients with fludarabine-refractory chronic lymphocytic leukemia (CLL). In heavily pretreated patients with CLL, the overall response rate (ORR) is approximately 35%, and in previously untreated patients the ORR is greater than 80%, with a recent randomized study suggesting it is superior to alkylator-based therapy. Importantly, alemtuzumab is effective in patients with high-risk del(17p13.1) and del(11q22.3) CLL. Alemtuzumab combination studies with fludarabine and/or monoclonal antibodies such as rituximab have demonstrated promising results. Alemtuzumab is also being studied in CLL patients as consolidation therapy for treatment of minimal residual disease, in preparation for stem cell transplantation and to prevent acute and chronic graft versus host disease. Alemtuzumab is frequently associated with acute 'first-dose' reactions when administered intravenously, but is much better tolerated when administered subcutaneously without loss of therapeutic efficacy. Additional potential adverse events associated with alemtuzumab administration include myelosuppression as well as profound cellular immune dysfunction with the associated risk of viral reactivation and other opportunistic infections. Additional studies detailing the mechanism of action of alemtuzumab as well as new strategies for prevention of opportunistic infections will aid in the future therapeutic development of this agent.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/uso terapéutico , Anticuerpos Antineoplásicos/inmunología , Anticuerpos Antineoplásicos/uso terapéutico , Inmunoterapia , Leucemia Linfocítica Crónica de Células B/inmunología , Leucemia Linfocítica Crónica de Células B/terapia , Alemtuzumab , Anticuerpos Monoclonales/administración & dosificación , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales Humanizados , Anticuerpos Antineoplásicos/administración & dosificación , Anticuerpos Antineoplásicos/efectos adversos , Antígenos CD/inmunología , Antígenos de Neoplasias/inmunología , Antígeno CD52 , Ciclofosfamida/uso terapéutico , Glicoproteínas/inmunología , Humanos , Leucemia Linfocítica Crónica de Células B/patología , Vidarabina/análogos & derivados , Vidarabina/uso terapéutico
7.
Gynecol Endocrinol ; 19(5): 253-8, 2004 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-15726913

RESUMEN

Our objective was to investigate the role of previous abdominal-pelvic surgery in the asymmetric distribution of pelvic endometriosic lesions. This was a retrospective study carried out at the Clinic of Obstetrics and Gynecology, University of Ancona, Italy, and included 238 patients with histological confirmation of endometriosis. The interventions were surgical treatment, at laparoscopy or laparotomy, for pelvic pain and endometriosis. The main outcome measure(s) were endometriotic lesions and adhesions in the pelvis found during surgery and the dinical records of the patients. We found unilateral lesions in 149 patients (62.6%): the right side of the pelvis affected in 55 patients (36.9%) and the left side in 94 patients (63.1%) (p < 0.01). In the group of patients who had undergone previous abdominal surgery, we found lesions on the right side in 26 cases (32.5%), and on the left in 54 cases (67.5%) (p < 0.01). We found that the patients who had undergone previous abdominal surgery had significantly more adhesions than those with no previous surgery (80/116 vs. 73/122, p = 0. 002). As a new finding, we have demonstrated that the left side of asymmetric distribution of intrapelvic macroscopic lesions is preserved and more evident in patients with previous abdominal surgery, including previous appendectomy. These data seem to be in agreement with our previous supposition of a possible interaction between previous abdominal surgery and the mechanisms of endometriosis development.


Asunto(s)
Endometriosis/patología , Endometriosis/cirugía , Complicaciones Posoperatorias , Abdomen/cirugía , Adulto , Endometriosis/etiología , Femenino , Humanos , Laparoscopía , Laparotomía , Dolor Pélvico , Estudios Retrospectivos , Adherencias Tisulares/diagnóstico , Adherencias Tisulares/epidemiología , Adherencias Tisulares/patología
8.
J Clin Invest ; 108(6): 887-94, 2001 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-11560958

RESUMEN

Epstein-Barr virus-associated lymphoproliferative disease (EBV-LPD) is a potentially life-threatening complication in immune-deficient patients. We have used the severe combined immune deficient (SCID) mouse engrafted with human leukocytes (hu-PBL-SCID) to evaluate the use of human cytokines in the prevention of EBV-LPD in vivo. Daily low-dose IL-2 therapy can prevent EBV-LPD in the hu-PBL-SCID mouse, but protection is lost if murine natural killer (NK) cells are depleted. Here we demonstrate that combined therapy with human GM-CSF and low-dose IL-2 is capable of preventing EBV-LPD in the hu-PBL-SCID mouse in the absence of murine NK cells. Lymphocyte depletion experiments showed that human NK cells, CD8(+) T cells, and monocytes were each required for the protective effects of GM-CSF and IL-2 combination therapy. This treatment resulted in a marked expansion of human CD3(+)CD8(+) lymphocytes in vivo. Using HLA tetramers complexed with EBV immunodominant peptides, a subset of these lymphocytes was found to be EBV-specific. These data establish that combined GM-CSF and low-dose IL-2 therapy can prevent the immune deficiencies that lead to fatal EBV-LPD in the hu-PBL-SCID mouse depleted of murine NK cells, and they point to a critical role for several human cellular subsets in mediating this protective effect.


Asunto(s)
Infecciones por Virus de Epstein-Barr/prevención & control , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Interleucina-2/farmacología , Trastornos Linfoproliferativos/prevención & control , Animales , Modelos Animales de Enfermedad , Infecciones por Virus de Epstein-Barr/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Humanos , Inmunidad Celular/efectos de los fármacos , Interleucina-2/administración & dosificación , Células Asesinas Naturales/inmunología , Transfusión de Leucocitos , Trastornos Linfoproliferativos/inmunología , Ratones , Ratones SCID , Subgrupos de Linfocitos T/efectos de los fármacos , Subgrupos de Linfocitos T/inmunología , Trasplante Heterólogo
9.
Cancer J Sci Am ; 6 Suppl 1: S45-51, 2000 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-10685658

RESUMEN

PURPOSE: The drastic increase in the incidence of non-Hodgkin's lymphoma in patients infected with HIV-1 is testimony to the fact that our immune system is critical for the prevention of certain malignancies. Preclinical and clinical studies were conducted to (1) gain further insight into defects in immunity that can lead to malignant transformation and (2) determine if certain immune deficiencies could be corrected by cytokines delivered at doses that result in near-physiologic concentrations in vivo. METHODS: We have used the severe combined immune deficient mouse engrafted with human peripheral blood leukocytes from healthy individuals who are seropositive for the Epstein-Barr virus to study the spontaneous development of malignant Epstein-Barr virus-positive human B-cell lymphoproliferative disorder. RESULTS: We have demonstrated in this model that, in the absence of CD4+ T cells, cytokine replacement with low-dose interleukin (IL)-2 therapy can prevent Epstein-Barr virus-positive human B-cell lymphoproliferative disorder by interacting with mouse natural killer and human CD8+ T cells. We review our clinical experience with administration of low-dose IL-2 therapy in patients with HIV-1-related cancer, noting minimal toxicity and significant immune modulation. We provide evidence that this therapy can favorably alter the type 1 cytokine profile in vivo in these patients, and improve the cellular response to infectious insults in vitro. CONCLUSION: Early clinical studies with low-dose IL-2 therapy in patients with HIV-1-related lymphoma suggest that this therapy may have a role in the prevention and treatment of this disease.


Asunto(s)
VIH-1 , Interleucina-2/uso terapéutico , Linfoma Relacionado con SIDA/terapia , Animales , Humanos , Interferón gamma/biosíntesis , Interleucina-2/efectos adversos , Células Asesinas Naturales/inmunología , Linfoma Relacionado con SIDA/inmunología , Ratones , Ratones SCID
10.
Curr Opin Oncol ; 11(6): 516-21, 1999 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-10550017

RESUMEN

AIDS-lymphoma is a heterogeneous disease that most likely results from the complex interaction of several contributing factors, including chronic antigenic activation of B lymphocytes, dysregulated cytokine and co-stimulatory networks, infection with potentially oncogenic viruses (human herpesvirus-8 [HHV-8], Epstein-Barr virus), and accumulation of secondary genetic mutations. Cytokines are believed to play an important role in the immunologic decline that favors opportunistic infection and malignancy in advanced HIV infection. Recent work has provided some evidence that cytokine therapy can partially reverse the immune dysregulation seen in AIDS. This suggests that cytokines are likely to have an important role in both the pathogenesis and treatment or prevention of AIDS-lymphoma.


Asunto(s)
Citocinas/inmunología , Citocinas/uso terapéutico , Linfoma Relacionado con SIDA/inmunología , Linfoma Relacionado con SIDA/terapia , Humanos
11.
J Immunol ; 163(1): 500-6, 1999 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-10384154

RESUMEN

There are experimental data which suggest that the primary immune effector cell responsible for maintaining immune surveillance against the outgrowth of EBV-transformed B cells in humans is the CTL, but in vivo proof of this is lacking. In this study we perform a series of cellular and molecular assays to characterize an autologous, endogenous immune response against a transplantation-associated, monoclonal, EBV+ posttransplant lymphoproliferative disorder (PTLD). Following allogeneic bone marrow transplantation, a patient developed a monoclonal PTLD of donor B cell origin. With a decrease in immune suppression, we document the emergence of endogenous, donor-derived CD3+CD8+ CTLs, followed by regression of the PTLD. The TCR Vbeta repertoire went from a polyclonal pattern prior to the development of PTLD to a restricted TCR Vbeta pattern during the outgrowth and regression of PTLD. Donor-derived CD3+CD8+ T lymphocytes displayed MHC class I-restricted cytolytic activity against the autologous EBV+ B cells ex vivo without additional in vitro sensitization. The striking temporal relationship between the endogenous expansion of a TCR Vbeta-restricted, CD3+CD8+ population of MHC class I-restricted CTL, and the regression of an autologous monoclonal PTLD, provides direct evidence in humans that endogenous CD3+CD8+ CTLs can be responsible for effective immune surveillance against malignant transformation of EBV+ B cells.


Asunto(s)
Trasplante de Médula Ósea/inmunología , Herpesvirus Humano 4/inmunología , Trastornos Linfoproliferativos/inmunología , Complicaciones Posoperatorias/inmunología , Linfocitos T Citotóxicos/inmunología , Adulto , Linfocitos B/inmunología , Linfocitos B/virología , Trasplante de Médula Ósea/efectos adversos , Trasplante de Médula Ósea/patología , División Celular/inmunología , Células Clonales , Pruebas Inmunológicas de Citotoxicidad , Reordenamiento Génico de la Cadena beta de los Receptores de Antígenos de los Linfocitos T , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/inmunología , Leucemia Mielógena Crónica BCR-ABL Positiva/terapia , Activación de Linfocitos , Trastornos Linfoproliferativos/patología , Trastornos Linfoproliferativos/virología , Masculino , Complicaciones Posoperatorias/patología , Receptores de Antígenos de Linfocitos T alfa-beta/biosíntesis , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Linfocitos T Citotóxicos/patología , Linfocitos T Citotóxicos/virología
12.
J Clin Invest ; 101(6): 1373-8, 1998 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-9502779

RESUMEN

This study was undertaken to determine if prolonged daily subcutaneous administration of ultra low dose IL-2 could influence the constitutive endogenous production of a type 1 (IFN-gamma) cytokine in patients with AIDS or AIDS-associated malignancies. Using a quantitative reverse transcription PCR assay, we demonstrate that daily administration of one type 1 cytokine, IL-2, for 3 mo increases significantly the constitutive endogenous gene expression of another type 1 cytokine, IFN-gamma, in vivo. The predominant source of IFN-gamma appears to be IL-2-expanded natural killer cells and CD8(+) T cells. Moreover, PBMC obtained from these patients during IL-2 therapy showed normalization of a profound deficit in IFN-gamma protein production after stimulation with extracts from infectious agents in vitro. Our data suggest that prolonged exogenous administration of a type 1 cytokine in a nontoxic fashion to patients with AIDS and AIDS-associated malignancies can enhance significantly the endogenous type 1 cytokine profile in vivo. Consequently, ultra low dose IL-2 therapy has the potential to improve the immunodeficient hosts' immune response to infectious pathogens that require IFN-gamma for clearance.


Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/terapia , Interferón gamma/metabolismo , Interleucina-2/administración & dosificación , Interleucina-2/uso terapéutico , Linfoma Relacionado con SIDA/terapia , Sarcoma de Kaposi/terapia , Síndrome de Inmunodeficiencia Adquirida/inmunología , Síndrome de Inmunodeficiencia Adquirida/metabolismo , Recuento de Linfocito CD4 , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Citometría de Flujo , Expresión Génica , Humanos , Inmunidad Innata , Huésped Inmunocomprometido/efectos de los fármacos , Huésped Inmunocomprometido/inmunología , Inmunoterapia/métodos , Interferón gamma/genética , Interleucina-10/genética , Interleucina-10/metabolismo , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Leucocitos Mononucleares/metabolismo , Lipopolisacáridos/inmunología , Recuento de Linfocitos , Linfoma Relacionado con SIDA/inmunología , Linfoma Relacionado con SIDA/metabolismo , Reacción en Cadena de la Polimerasa , Sarcoma de Kaposi/inmunología , Sarcoma de Kaposi/metabolismo , Subgrupos de Linfocitos T/inmunología
13.
Blood ; 90(5): 1737-46, 1997 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-9292506

RESUMEN

The poor prognosis associated with patients afflicted with the acquired immunodeficiency syndrome and primary central nervous system lymphoma (AIDS-PCNSL) is due in part to the intrinsic resistance of this Epstein-Barr virus (EBV)-associated tumor to conventional antineoplastic therapy. Fas (CD95) is a transmembrane protein receptor that transmits an intracellular signal leading to rapid programmed cell death following ligation with its natural ligand or anti-Fas antibodies. Fas expression and function were assessed in AIDS-PCNSL biopsy samples and in EBV+ human B-cell tumors that spontaneously developed in severe combined immune deficient (SCID) mice engrafted with human lymphocytes (hu-PBL-SCID mice). All tumors samples showed high-density surface expression of Fas by flow cytometry or immunohistochemical staining. Cells from two AIDS-PCNSL biopsy samples that did not express pan B-cell markers did not express Fas antigen. All tumors examined were susceptible to Fas-mediated apoptosis, as measured by standard assays for endonucleolytic cleavage of DNA. The response to Fas-mediated apoptosis was dependent on log-fold increases in the concentration of immobilized anti-Fas antibody, but could also be induced with a mobilized anti-Fas antibody. No evidence for intrinsic resistance to Fas-mediated apoptosis (ie, secreted or truncated forms of Fas) could be shown. Radiation-induced apoptosis of neoplastic EBV+ B cells was enhanced by activation of Fas, and prolonged exposure to interleukin-2 increased both Fas expression and Fas-induced apoptosis. As the normal brain parenchyma appears to have either low-density or absent expression of Fas, and antineoplastic therapy can be selectively delivered to the CNS with little systemic toxicity, local delivery of Fas-activating molecules could prove to be a useful component in the multimodal treatment of AIDS-PCNSL.


Asunto(s)
Neoplasias Encefálicas/inmunología , Linfoma Relacionado con SIDA/inmunología , Receptor fas/análisis , Animales , Neoplasias Encefálicas/patología , Citometría de Flujo , Humanos , Linfoma Relacionado con SIDA/patología , Ratones , Ratones SCID , Receptor fas/inmunología
14.
Cancer J Sci Am ; 3 Suppl 1: S129-36, 1997 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-9457408

RESUMEN

PURPOSE: Congenital, acquired, and some iatrogenically induced immune deficiencies are characterized by an increased incidence of viral-associated cancers. Preclinical and clinical studies were conducted to understand the pathogenesis of immune-deficiency-associated cancer and its response to low-dose recombinant interleukin-2 (rIL-2) therapy, with the ultimate goal of applying this or other immune therapy in the treatment or prevention of immune-deficiency-associated lymphoma. METHODS: We have used the severe combined immune-deficient (SCID) mouse engrafted with human peripheral blood lymphocytes (PBL) from healthy Epstein-Barr virus seropositive donors to study the pathogenesis of malignant B-cell lymphoproliferative disease that commonly occurs in some immune-deficient patients. In this chimeric human (hu)-PBL-SCID mouse model, administration of daily low-dose rIL-2 interacts with murine natural killer cells and human CD8+ T cells to prevent the outgrowth of human Epstein-Barr virus lymphoproliferative disease. We have utilized the information gained from this chimeric mouse model to perform a phase I study of daily, subcutaneous, low-dose rIL-2 therapy in patients with both acquired immune deficiency syndrome (AIDS) and cancer. RESULTS: Plasma concentrations of rIL-2 were achieved in vivo comparable to those seen in our hu-PBL-SCID model, in the absence of significant (grade 3) clinical toxicity or an increase in the plasma human immune deficiency virus (HIV) RNA level. Significant expansion in human cells, particularly the CD3-CD56bright natural killer cell subset, resulted after 6 weeks of therapy. Results of the hu-PBL-SCID mouse model and the phase I study have led to a national trial of low-dose rIL-2 therapy in AIDS-associated lymphoma. CONCLUSION: Daily low-dose rIL-2 therapy may be effective in treating or preventing AIDS-associated lymphoma without amplifying HIV replication.


Asunto(s)
Inmunoterapia/métodos , Interleucina-2/uso terapéutico , Linfoma Relacionado con SIDA/terapia , Trastornos Linfoproliferativos/fisiopatología , Síndrome de Inmunodeficiencia Adquirida/complicaciones , Animales , Citocinas/fisiología , Modelos Animales de Enfermedad , Herpesvirus Humano 4/efectos de los fármacos , Humanos , Interleucina-10/fisiología , Interleucina-2/administración & dosificación , Interleucina-2/sangre , Interleucina-2/fisiología , Interleucina-6/fisiología , Transfusión de Linfocitos , Trastornos Linfoproliferativos/mortalidad , Ratones , Ratones SCID , Tasa de Supervivencia
15.
J Clin Invest ; 96(6): 2578-82, 1995 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-8675621

RESUMEN

Natural killer (NK) cells are large granular lymphocytes that constitutively express functional IL-2 receptors. We have shown that recombinant human IL-15 uses the IL-2 receptor to activate human NK cells and can synergize with recombinant human IL-12 to stimulate NK cell production of IFN-gamma in vitro. IFN-gamma production by NK cells is critical in the prevention of overwhelming infection by obligate intracellular microbial pathogens in several experimental animal models. Herein, we demonstrate that human monocytes produce IL-15 protein within 5 h of activation with LPS. Using an IL-15-neutralizing antiserum in a coculture of LPS-activated monocytes and NK cells, we demonstrate that monocyte-derived IL-15 is critical for optimal NK cell production of IFN-gamma. Endogenous IL-15 activates NK cells through the IL-2 receptor, and with endogenous IL-12, regulates NK cell IFN-gamma after monocyte activation by LPS. These in vitro studies are the first to characterize a function for endogenous IL-15, and as such, suggest an important role for IL-15 during the innate immune response. IL-15 may be an important ligand for the NK cell IL-2 receptor in vivo.


Asunto(s)
Interferón gamma/biosíntesis , Interleucinas/biosíntesis , Interleucinas/farmacología , Células Asesinas Naturales/inmunología , Activación de Linfocitos , Monocitos/inmunología , Receptores de Interleucina-2/fisiología , Animales , Anticuerpos Monoclonales , Western Blotting , Células Cultivadas , Técnicas de Cocultivo , Sinergismo Farmacológico , Ensayo de Inmunoadsorción Enzimática , Escherichia coli , Humanos , Inmunohistoquímica , Interleucina-12/farmacología , Interleucina-15 , Interleucinas/análisis , Células Asesinas Naturales/efectos de los fármacos , Cinética , Lipopolisacáridos/farmacología , Monocitos/efectos de los fármacos , Proteínas Recombinantes/farmacología
16.
Blood ; 85(12): 3577-85, 1995 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-7540068

RESUMEN

Human natural killer (NK) cells are large granular lymphocytes that constitutively express functional forms of the interleukin-2 receptor (IL-2R) and lyse tumor and virally infected cells without prior sensitization. NK cells with high density expression of CD56 (CD56bright) express the high affinity IL-2R and proliferate in response to low (picomolar) concentrations of IL-2. CD56dim NK cells express the intermediate affinity IL-2R and demonstrate enhanced cytotoxic activity without proliferation in response to high (nanomolar) concentrations of IL-2. In the present study, we characterized IL-10R expression on human NK cells and the functional consequences of IL-10 binding directly to highly purified subsets of CD56bright and CD56dim NK cells. Binding studies using 125I-IL-10 indicated that resting human NK cells constitutively express the IL-10 receptor protein at a surface density of approximately 90 receptor sites per cell, with a kd of approximately 1 nmol/L. Alone, IL-10 did not induce proliferation of CD56bright or CD56dim NK cell subsets. However, at low concentrations (0.5 to 5 ng/mL), IL-10 significantly augmented IL-2-induced proliferation of the CD56bright NK cell subset mediated via the high-affinity IL-2R. In the absence of IL-2, IL-10 was able to induce significant NK cytotoxic activity against NK-resistant tumor cell targets in both subsets of NK cells in a dose-dependent fashion. Furthermore, the combination of IL-10 and IL-2 had an additive effect on NK cytotoxic activity, whereas that of IL-10 and IL-12 did not. Production of interferon-gamma, tumor necrosis factor-alpha, and granulocyte-macrophage colony-stimulating factor by IL-2-activated NK cells was also significantly enhanced by IL-10. Neither resting nor activated human NK cells appear to produce human IL-10 protein. In summary, NK cells constitutively express the IL-10R protein in low density, and the functional consequences of IL-10 binding directly to human NK cell subsets appear to be stimulatory and dose-dependent. In contrast to its direct effects on human T cells and monocytes/macrophages, IL-10 potentiates cytokine production by human NK cells.


Asunto(s)
Interleucina-10/metabolismo , Células Asesinas Naturales/metabolismo , Receptores de Interleucina/biosíntesis , Antígenos CD/biosíntesis , Antígenos de Diferenciación de Linfocitos T/biosíntesis , Antígeno CD56 , División Celular/efectos de los fármacos , Células Cultivadas , Humanos , Interleucina-10/farmacología , Interleucina-2/farmacología , Células Asesinas Naturales/inmunología , ARN Mensajero/análisis , Ensayo de Unión Radioligante , Receptores de Interleucina-10
17.
Blood ; 85(4): 1063-74, 1995 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-7849294

RESUMEN

Both human (hu) and viral (v) interleukin-10 (IL-10) appear to be important cofactors in the survival and growth of lymphoblastoid cell lines infected with Epstein-Barr virus (EBV). When mice with severe combined immune deficiency (SCID) are injected with human peripheral blood lymphocytes (PBL) from normal individuals who are seropositive for EBV, the majority of hu-PBL-SCID mice will develop an EBV-associated lymphoproliferative disease (EBV-LPD) of human B-cell origin, not unlike some cases of EBV-LPD that are seen in immunocompromised individuals. The role of huIL-10 or vIL-10 in this chimeric mouse model of EBV-LPD is unknown. In the present study, we show that hu-PBL-SCID mice that develop EBV-LPD have significant elevation of serum huIL-10 levels compared with mice that do not develop EBV-LPD (P = .005). vIL-10 was undetectable in all animals. The EBV+ tumor samples express transcript for huIL-10 and huIL-10 receptor, express huIL-10 protein by immunohistochemical staining, and show specific binding of recombinant (r) huIL-10. In vitro analysis of the functional consequences of rhuIL-10 binding to IL-10 receptors on fresh EBV+ tumor cells shows that rhuIL-10 can prevent programmed cell death as well as promote proliferation and can do so at concentrations of huIL-10 found in vivo. Thus, huIL-10 production by EBV+ tumor cells may contribute directly to their malignant outgrowth in the hu-PBL-SCID mouse by two autocrine mechanisms: prevention of programmed cell death and proliferation. The implications of such findings with regard to EBV-LPD in humans is discussed.


Asunto(s)
Interleucina-10/biosíntesis , Transfusión de Linfocitos , Linfoma/inmunología , Animales , Antígenos Virales/análisis , Western Blotting , División Celular/efectos de los fármacos , ADN de Neoplasias/análisis , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Herpesvirus Humano 4/aislamiento & purificación , Herpesvirus Humano 4/patogenicidad , Humanos , Técnicas para Inmunoenzimas , Inmunohistoquímica , Interleucina-10/análisis , Interleucina-10/farmacología , Activación de Linfocitos , Linfoma/patología , Ratones , Ratones SCID , Proteínas Oncogénicas Virales/análisis , Proteínas Tirosina Quinasas/análisis , Proteínas Proto-Oncogénicas/análisis , Proteínas Proto-Oncogénicas c-bcl-2 , Proteínas Recombinantes/farmacología , Trasplante Heterólogo , Células Tumorales Cultivadas , Proteínas de la Matriz Viral/análisis
18.
Proc Natl Acad Sci U S A ; 91(16): 7553-7, 1994 Aug 02.
Artículo en Inglés | MEDLINE | ID: mdl-7519782

RESUMEN

The bcl-2 protein plays a central role in the regulation of programmed cell death in a variety of tissues and is pivotal to the survival of lymphocytes in vivo. The growth factors responsible for survival of normal lymphocytes are unknown but are likely to maintain viability in part through the regulation of bcl-2 expression. A subset of human natural killer (NK) cells (CD3-CD56bright) are unique among lymphocytes in their constitutive expression of c-kit, a tyrosine kinase cell surface receptor that binds c-kit ligand (KL). Alone, KL does not promote proliferation or further differentiation of CD56bright NK cells. We now report that, in the absence of serum or additional growth factors, KL prevents apoptosis of cultured CD56bright NK cells, as assessed by DNA fragmentation studies, and maintains viability, as measured by biologic responses (i.e., proliferation and cytotoxicity) to the subsequent addition of other cytokines. Furthermore, we demonstrate that KL induces CD56bright NK cells to express the bcl-2 protein. In the presence of anti-c-kit antibody, the tyrosine kinase inhibitor genistein, or bcl-2 antisense oligonucleotide, the protective effect of KL on the survival of CD56bright NK cells is dramatically reduced. These data demonstrate that the binding of KL to its tyrosine kinase receptor results in the upregulation of bcl-2, thereby preventing apoptosis in this subset of normal human lymphocytes. As soluble KL is plentiful in normal human serum, this survival mechanism may be operative for CD56bright NK cells in vivo.


Asunto(s)
Apoptosis/efectos de los fármacos , Factores de Crecimiento de Célula Hematopoyética/farmacología , Células Asesinas Naturales/efectos de los fármacos , Proteínas Proto-Oncogénicas/biosíntesis , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Tirosina Quinasas Receptoras/metabolismo , Receptores del Factor Estimulante de Colonias/metabolismo , Antígenos CD , Antígenos de Diferenciación de Linfocitos T , Secuencia de Bases , Antígeno CD56 , Supervivencia Celular , Células Cultivadas , Humanos , Datos de Secuencia Molecular , Proteínas Proto-Oncogénicas c-bcl-2 , Proteínas Proto-Oncogénicas c-kit , Factor de Células Madre , Regulación hacia Arriba
19.
Proc Natl Acad Sci U S A ; 91(12): 5577-81, 1994 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-7911241

RESUMEN

When severe combined immune deficient (SCID) mice undergo i.p. injection with peripheral blood lymphocytes from normal human donors seropositive for EBV, a majority of these mice (hu-PBL-SCID mouse model) subsequently develop a fatal EBV+ lymphoproliferative disease (EBV-LPD) of human B-cell origin. Because T cells normally are critical in the control of EBV infection, we hypothesized that human T-cell dysfunction accounts for EBV-LPD in the hu-PBL-SCID mouse and that systemic administration of T-cell-derived cytokines would reestablish protective immunity against EBV-LPD. We show that the daily s.c. administration of a very low dose (500 international units) of polyethylene glycol-modified recombinant human interleukin 2 (PEG-IL-2) to hu-PBL-SCID mice can prevent the development of fatal EBV-LPD and significantly improves survival (78%), compared with the survival of hu-PBL-SCID mice treated with placebo (20%, P = 0.0008). Additional lymphocyte-depletion experiments showed that mouse natural killer cells and human CD8+ T cells provided cellular immunity necessary for the PEG-IL-2-mediated protective effect, whereas i.p. injection of human peripheral blood lymphocytes depleted of CD4+ T cells had no adverse effect when combined with PEG-IL-2 therapy and may have been beneficial. These data establish that very low-dose PEG-IL-2 therapy can overcome the immune deficiencies that lead to EBV-LPD in the hu-PBL-SCID mouse and point to the usefulness of this model for evaluating cytokine therapies in EBV-LPD. The use of low-dose IL-2 as a preventative immune therapy has potential application in immunocompromised individuals at high risk for EBV-LPD.


Asunto(s)
Infecciones por Herpesviridae/prevención & control , Herpesvirus Humano 4/patogenicidad , Interleucina-2/administración & dosificación , Células Asesinas Naturales/inmunología , Trastornos Linfoproliferativos/microbiología , Subgrupos de Linfocitos T/inmunología , Infecciones Tumorales por Virus/prevención & control , Animales , Linfocitos T CD4-Positivos/inmunología , Trastornos Linfoproliferativos/inmunología , Trastornos Linfoproliferativos/prevención & control , Ratones , Ratones SCID , Proteínas Recombinantes
20.
Cell Immunol ; 155(2): 508-16, 1994 May.
Artículo en Inglés | MEDLINE | ID: mdl-7514106

RESUMEN

The origin, lineage derivation, and sites of human natural killer (NK) cell differentiation are presently unresolved. The vast majority of NK cells found in peripheral blood have surface membrane expression of CD2 and CD16. Both antigens trigger activation pathways which require the zeta protein, a signal-transducing subunit of the CD3-T cell receptor (CD3-TCR) complex which is found as an isolated homodimer (zeta-zeta) or heterodimer (zeta-Fc epsilon RI gamma) in human NK cells. Unlike NK cells found in adult peripheral blood, NK cells derived in vitro from human CD34+ hematopoietic progenitor cells lack CD2 and CD16, and those found in fetal liver constitutively express CD3 epsilon and delta proteins. However, NK effectors derived in vitro from immature human CD3- thymocytes show striking phenotypic and functional similarities to adult human NK cells. In this report, we characterize zeta protein expression in CD3- thymocytes following short-term culture in recombinant (r)IL-2. CD3-CD56+ thymocyte NK effectors express the zeta protein as a disulphide-linked homodimer of 32 kDa, yet lack other protein components of the CD3-TCR complex. Both CD16+ and CD16- populations were found to express zeta, and within the CD16+ fraction, zeta is physically associated with CD16. These data provide evidence of additional similarities between adult peripheral blood NK cells and CD3-CD56+ NK effectors derived from human thymocytes, and suggest that under these experimental conditions, human NK cells can arise from early thymic precursors.


Asunto(s)
Células Asesinas Naturales/inmunología , Proteínas de la Membrana/biosíntesis , Receptores de Antígenos de Linfocitos T/biosíntesis , Subgrupos de Linfocitos T/inmunología , Timo/inmunología , Antígenos CD , Antígenos de Diferenciación de Linfocitos T , Complejo CD3 , Antígeno CD56 , Células Cultivadas , Preescolar , Humanos , Fenotipo , Receptores de IgG , Timo/citología
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