RESUMEN
Several studies have suggested a link between human microbiome and rheumatoid arthritis (RA) development. Porphyromonas gingivalis seems involved in RA initiation and progression, as supported by the high occurrence of periodontitis. In this case-control study, we analysed tongue P. gingivalis presence and quantification in a large healthy and RA cohort. We enrolled 143 RA patients [male/female (M/F) 32/111, mean ± standard deviation (s.d.), age 57·5 ± 19·8 years, mean ± s.d. disease duration 155·9 ± 114·7 months); 36 periodontitis patients (M/F 11/25, mean ± s.d., age 56 ± 9·9 years, mean ± s.d. disease duration 25·5 ± 20·9 months); and 57 patients (M/F 12/45, mean ± s.d., age 61·4 ± 10·9 years, mean ± s.d. disease duration 62·3 ± 66·9 months) with knee osteoarthritis or fibromyalgia. All subjects underwent a standard cytological swab to identify the rate of P. gingivalis/total bacteria by using quantitative real-time polymerase chain reaction. The prevalence of P. gingivalis resulted similarly in RA and periodontitis patients (48·9 versus 52·7%, P = not significant). Moreover, the prevalence of this pathogen was significantly higher in RA and periodontitis patients in comparison with control subjects (P = 0·01 and P = 0·003, respectively). We found a significant correlation between P. gingivalis rate in total bacteria genomes and disease activity score in 28 joints (DAS28) (erythrocyte sedimentation rate) (r = 0·4, P = 0·01). RA patients in remission showed a significantly lower prevalence of P. gingivalis in comparison with non-remission (P = 0·02). We demonstrated a significant association between the percentage of P. gingivalis on the total tongue biofilm and RA disease activity (DAS28), suggesting that the oral cavity microbiological status could play a role in the pathogenic mechanisms of inflammation, leading to more active disease.
Asunto(s)
Artritis Reumatoide/inmunología , Infecciones por Bacteroidaceae/inmunología , Microbiota/inmunología , Periodontitis/inmunología , Porphyromonas gingivalis/fisiología , Lengua/patología , Adulto , Anciano , Artritis Reumatoide/epidemiología , Infecciones por Bacteroidaceae/epidemiología , Biopelículas , Estudios de Casos y Controles , Estudios de Cohortes , Progresión de la Enfermedad , Femenino , Humanos , Italia/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Lengua/microbiologíaRESUMEN
This review presents a possible new approach against cancer, as represented by inhibition of proton pumps, a mechanism used by tumour cells to avoid intracellular accumulation of toxic substances. Proton pump inhibitors (PPIs) belong to a family of pro-drugs that are currently used in the treatment of peptic diseases needing acidity to be activated. PPIs target the acidic tumour mass, where they are metabolized, thus blocking proton traffic. Proton pump inhibition triggers a rapid cell death as a result of intracellular acidification, caspase activation and early accumulation of reactive oxygen species into tumour cells. As a whole, the devastating effect of PPIs on tumour cells suggest the triggering of a fatal cell toxification. Many human tumours, including melanoma, osteosarcoma, lymphomas and various adenocarcinomas are responsive to PPIs. This appears highly conceivable, in as much as almost all human tumours are acidic and express high levels of proton pumps. Paradoxically, metastatic tumours appear to be more responsive to PPIs being more acidic than the majority of primary tumours. However, two clinical trials test the effectiveness of PPIs in chemosensitizing melanoma and osteosarcoma patients. Indeed, tumour acidity represents a very potent mechanism of chemoresistance. A majority of cytotoxic agents, being weak bases, are quickly protonated outside and do not enter the cells, thus preventing drugs to reach specific cellular targets. Clinical data will provide the proof of concept on the use of PPIs as a new class of antitumour agent with a very low level of systemic toxicity as compared with standard chemotherapeutic agents.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Inhibidores de la Bomba de Protones/farmacología , Antineoplásicos/uso terapéutico , Apoptosis/fisiología , Espacio Extracelular/química , Humanos , Concentración de Iones de Hidrógeno , Espacio Intracelular/química , Metástasis de la Neoplasia , Inhibidores de la Bomba de Protones/uso terapéutico , Bombas de Protones/químicaRESUMEN
OBJECTIVE: To explore whether the climate has played a role in the COVID-19 outbreak, we compared virus lethality in countries closer to the Equator with others. Lethality in European territories and in territories of some nations with a non-temperate climate was also compared. MATERIALS AND METHODS: Lethality was calculated as the rate of deaths in a determinate moment from the outbreak of the pandemic out of the total of identified positives for COVID-19 in a given area/nation, based on the COVID-John Hopkins University website. Lethality of countries located within the 5th parallels North/South on 6 April and 6 May 2020, was compared with that of all the other countries. Lethality in the European areas of The Netherlands, France and the United Kingdom was also compared to the territories of the same nations in areas with a non-temperate climate. RESULTS: A lower lethality rate of COVID-19 was found in Equatorial countries both on April 6 (OR=0.72 CI 95% 0.66-0.80) and on May 6 (OR=0.48, CI 95% 0.47-0.51), with a strengthening over time of the protective effect. A trend of higher risk in European vs. non-temperate areas was found on April 6, but a clear difference was evident one month later: France (OR=0.13, CI 95% 0.10-0.18), The Netherlands (OR=0.5, CI 95% 0.3-0.9) and the UK (OR=0.2, CI 95% 0.01-0.51). This result does not seem to be totally related to the differences in age distribution of different sites. CONCLUSIONS: The study does not seem to exclude that the lethality of COVID-19 may be climate sensitive. Future studies will have to confirm these clues, due to potential confounding factors, such as pollution, population age, and exposure to malaria.
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Clima , Infecciones por Coronavirus/mortalidad , Neumonía Viral/mortalidad , Estaciones del Año , Tiempo (Meteorología) , Betacoronavirus , Brunei/epidemiología , Burundi/epidemiología , COVID-19 , Congo/epidemiología , Infecciones por Coronavirus/epidemiología , Ecuador/epidemiología , Guinea Ecuatorial/epidemiología , Europa (Continente) , Francia/epidemiología , Gabón/epidemiología , Humanos , Islas del Oceano Índico/epidemiología , Indonesia/epidemiología , Kenia/epidemiología , Malasia/epidemiología , Melanesia/epidemiología , Micronesia/epidemiología , Países Bajos/epidemiología , Pandemias , Papúa Nueva Guinea/epidemiología , Neumonía Viral/epidemiología , Rwanda/epidemiología , SARS-CoV-2 , Samoa/epidemiología , Santo Tomé y Príncipe/epidemiología , Seychelles/epidemiología , Singapur/epidemiología , Somalia/epidemiología , Timor Oriental/epidemiología , Clima Tropical , Uganda/epidemiología , Reino Unido/epidemiologíaRESUMEN
Tumour cells release vesicular structures, defined as microvesicles or exosomes, carrying a large array of proteins from their originating cell. The expression of antigenic molecules recognized by T cells has originally suggested a role for these organelles as a cell-free antigen source for anticancer vaccines. However, recent evidence shows that tumour exosomes may also exert a broad array of detrimental effects on the immune system, ranging from apoptosis in activated antitumour T cells to impairment of monocyte differentiation into dendritic cells and induction of myeloid suppressive cells. Immunosuppressive exosomes of tumour origin can be found in neoplastic lesions and sera from cancer patients, implying a potential role of this pathway in in vivo tumour progression. Through the expression of molecules involved in angiogenesis promotion, stromal remodelling, delivery of signalling pathways through growth factor/receptor transfer, chemoresistance and genetic intercellular exchange, tumour exosomes could represent a versatile tool for moulding host environment. Hence, their secretion by neoplastic cells may in the future become a novel pathway to target for therapeutic intervention in cancer patients.
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Vesículas Citoplasmáticas/fisiología , Neoplasias/inmunología , Transducción de Señal , Antígenos de Neoplasias/inmunología , Citocinas/inmunología , Citocinas/metabolismo , Vesículas Citoplasmáticas/inmunología , Células Dendríticas/inmunología , Humanos , Inmunoterapia , Neoplasias/patología , Neoplasias/fisiopatología , Neoplasias/terapia , Linfocitos T Citotóxicos/inmunologíaRESUMEN
As it is well-known, the characterization plan of an old landfill site is the first stage of the project for the treatment and reclamation of contaminated lands. It is a preliminary in-situ study, with collection of data related to pollution phenomena, and is aimed at defining the physical properties and the geometry of fill materials as well as the possible migration paths of pollutants to the surrounding environmental targets (subsoil and groundwater). To properly evaluate the extent and potential for subsoil contamination, waste volume and possible leachate emissions from the landfill have to be assessed. In such perspective, the integrated use of geophysical methods is an important tool as it allows a detailed 3D representation of the whole system, i.e. waste body and hosting environment (surrounding rocks). This paper presents a very accurate physical and structural characterization of an old landfill and encasing rocks obtained by an integrated analysis of data coming from a multi-methodological geophysical exploration. Moreover, drillings were carried out for waste sampling and characterization of the landfill body, as well as for calibration of the geophysical modeling.
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Monitoreo del Ambiente , Contaminación Ambiental/análisis , Eliminación de Residuos , Agua Subterránea , Instalaciones de Eliminación de Residuos , Contaminantes Químicos del AguaRESUMEN
Activating BRAF or NRAS mutations have been found in 80% of human sporadic melanomas, but only one of these genetic alterations could be detected in each tumour. This suggests that BRAF and NRAS 'double mutants' may not provide advantage for tumour growth, or may even be selected against during tumorigenesis. However, by applying mutant-allele-specific-amplification-PCR method to short-term melanoma lines, one out of 14 tumours was found to harbour both BRAFV600E and the activating NRASQ61R mutations. On the other hand, analysis of 21 melanoma clones isolated by growth in soft agar from this tumour indicated that 16/21 clones harboured a BRAFV600E, but were wild-type for NRAS, whereas the remaining had the opposite genotype (NRASQ61R/wild-type BRAF). When compared to BRAFV600E clones, NRASQ61R clones displayed reduced growth in soft agar, but higher proliferative ability in vitro in liquid medium and even in vivo after grafting into SCID/SCID mice. These data suggest that NRAS and BRAF activating mutations can coexist in the same melanoma, but are mutually exclusive at the single-cell level. Moreover, the presence of NRASQ61R or BRAFV600E is associated with distinct in vitro and in vivo growth properties of neoplastic cells.
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Genes ras , Melanoma/genética , Melanoma/metabolismo , Mutación , Proteínas Proto-Oncogénicas B-raf/genética , Animales , Secuencia de Bases , Línea Celular Tumoral , Femenino , Genotipo , Humanos , Ratones , Ratones SCID , Datos de Secuencia Molecular , Trasplante de NeoplasiasRESUMEN
Macropinocytosis, a ruffling-driven process that allows the capture of large material, is an essential aspect of normal cell function. It can be either constitutive, as in professional phagocytes where it ends with the digestion of captured material, or induced, as in epithelial cells stimulated by growth factors. In this case, the internalized material recycles back to the cell surface. We herein show that activation of Rho GTPases by a bacterial protein toxin, the Escherichia coli cytotoxic necrotizing factor 1 (CNF1), allowed epithelial cells to engulf and digest apoptotic cells in a manner similar to that of professional phagocytes. In particular, we have demonstrated that 1) the activation of all Rho, Rac, and Cdc42 by CNF1 was essential for the capture and internalization of apoptotic cells; and 2) such activation allowed the discharge of macropinosomal content into Rab7 and lysosomal associated membrane protein-1 acidic lysosomal vesicles where the ingested particles underwent degradation. Taken together, these findings indicate that CNF1-induced "switching on" of Rho GTPases may induce in epithelial cells a scavenging activity, comparable to that exerted by professional phagocytes. The activation of such activity in epithelial cells may be relevant, in mucosal tissues, in supporting or integrating the scavenging activity of resident macrophages.
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Apoptosis , Toxinas Bacterianas/farmacología , Citotoxinas/farmacología , Células Epiteliales/efectos de los fármacos , Proteínas de Escherichia coli , Pinocitosis/fisiología , Proteínas de Unión al GTP rho/metabolismo , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Compartimento Celular , Células Cultivadas , Citotoxinas/genética , Citotoxinas/metabolismo , Endosomas , Activación Enzimática , Células Epiteliales/fisiología , Humanos , Lisosomas/metabolismo , Macrófagos/citología , Macrófagos/fisiología , Células Tumorales Cultivadas , Células U937 , Proteína de Unión al GTP cdc42/metabolismo , Proteínas de Unión al GTP rab/metabolismo , Proteínas de Unión a GTP rab7 , Proteínas de Unión al GTP rac/metabolismoRESUMEN
Tumor microenvironment is one of the major obstacles to the efficacy of chemotherapy in cancer patients. The abnormal blood flow within the tumor results in uneven drug distribution. Electrochemotherapy (ECT) is a tumor treatment that adopts the systemic or local delivery of anticancer drugs with the application of permeabilizing electric pulses having appropriate amplitude and waveforms. This allows the use of lipophobic drugs that frequently have a narrow therapeutic index maintaining at the same time a reduced patient morbidity and preserving appropriate anticancer efficacy. Its use in humans is addressed to the treatment of cutaneous neoplasms or the palliation of skin tumor metastases, and a standard operating procedure has been devised. On the other hand, in veterinary oncology this approach is gaining popularity, thus becoming a first line treatment for different cancer histotypes, in a variety of clinical conditions due to its high efficacy and low toxicity. This review summarizes the state of the art in veterinary oncology as a preclinical model and reports the new protocols in terms of drugs and therapy combination that have been developed.
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Antineoplásicos/uso terapéutico , Protocolos Clínicos , Electroquimioterapia , Neoplasias/veterinaria , Medicina Veterinaria , Animales , Gatos , Perros , Humanos , Neoplasias/terapiaRESUMEN
CD95(APO-1/Fas)-mediated apoptosis of bystander uninfected T cells exerts a major role in the HIV-1-mediated CD4+ T-cell depletion. HIV-1 gp120 has a key role in the induction of sensitivity of human lymphocytes to CD95-mediated apoptosis through its interaction with the CD4 receptor. Recently, we have shown the importance of CD95/ezrin/actin association in CD95-mediated apoptosis. In this study, we explored the hypothesis that the gp120-mediated CD4 engagement could be involved in the induction of susceptibility of primary human T lymphocytes to CD95-mediated apoptosis through ezrin phosphorylation and ezrin-to-CD95 association. Here, we show that gp120/IL-2 combined stimuli, as well as the direct CD4 triggering, on human primary CD4(+)T lymphocytes induced an early and stable ezrin activation through phosphorylation, consistent with the induction of ezrin/CD95 association and susceptibility to CD95-mediated apoptosis. Our results provide a new mechanism through which HIV-1-gp120 may predispose resting CD4(+)T cell to bystander CD95-mediated apoptosis and support the key role of ezrin/CD95 linkage in regulating susceptibility to CD95-mediated apoptosis.
Asunto(s)
Apoptosis/fisiología , Proteína gp120 de Envoltorio del VIH/toxicidad , Fosfoproteínas/metabolismo , Linfocitos T/metabolismo , Receptor fas/metabolismo , Apoptosis/efectos de los fármacos , Antígenos CD4/metabolismo , Proteínas del Citoesqueleto , Humanos , Interleucina-2/metabolismo , Activación de Linfocitos/fisiología , FosforilaciónRESUMEN
We have recently shown that a human CD4+ T cell line (CEM-SS) acquires the permissiveness to M-tropic strains and primary isolates of HIV-1 after transplantation into SCID mice. This permissiveness was associated with the acquisition of a memory (CD45RO+) phenotype as well as of a functional CCR5 coreceptor. In this study, we have used this model for invest-igating in vivo the relationships between HIV-1 infection, apoptosis and T cell differentiation. When an in vivo HIV-1 infection was performed, the CEM cell tumors grew to a lower extent than the uninfected controls. CEM cells explanted from uninfected SCID mice (ex vivo CEM) underwent a significant level of spontaneous apoptosis and proved to be CD45RO+, Fas+ and Fas-L+, while Bcl-2 expression was significantly reduced as compared to the parental cells. Acute HIV-1 infection markedly increased apoptosis of uninfected ex vivo CEM cells, through a Fas/Fas-L-mediated autocrine suicide/fratricide, while parental cells did not undergo apoptosis following viral infection. The susceptibility to apoptosis of ex vivo CEM cells infected with the NSI strain of HIV-1, was progressively lost during culture, in parallel with the loss of Fas-L and marked changes in the Bcl-2 cellular distribution. On the whole, these results are strongly reminiscent of a series of events possibly occurring during HIV-1 infection. After an initial depletion of bystander CD4+ memory T cells during acute infection, latently or chronically infected CD4+ T lymphocytes are progressively selected and are protected against spontaneous apoptosis through the development of an efficient survival program. Studies with human cells passaged into SCID mice may offer new opportunities for an in vivo investigation of the mechanisms involved in HIV-1 infection and CD4+ T cell depletion.
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Apoptosis , Linfocitos T CD4-Positivos/virología , Infecciones por VIH/patología , VIH-1/fisiología , Receptor fas/metabolismo , Animales , Linfocitos T CD4-Positivos/trasplante , Línea Celular , Humanos , Ratones , Ratones SCID , Neoplasias Experimentales/fisiopatología , FenotipoRESUMEN
Cancer cell cannibalism is currently defined as a phenomenon in which an ensemble of a larger cell containing a smaller one, often in a big cytoplasmic vacuole, is detected in either cultured tumor cells or a tumor sample. After almost one century of considering this phenomenon as a sort of neglected curiosity, some recent studies have first proposed tumor cell cannibalism as a sort of "aberrant phagocytosis", making malignant cells very similar to professional phagocytes. Later, further research has shown that, differently to macrophages, exclusively ingesting exogenous material, apoptotic bodies, or cell debris, tumor cells are able to engulf other cells, including lymphocytes and erythrocytes, either dead or alive, with the main purpose to feed on them. This phenomenon has been associated to the malignancy of tumors, mostly exclusive of metastatic cells, and often associated to poor prognosis. The cannibalistic behavior increased depending on the microenvironmental condition of tumor cells, such as low nutrient supply or low pH, suggesting its key survival option for malignant cancers. However, the evidence that malignant cells may cannibalize tumor-infiltrating lymphocytes that act as their killers, suggests that tumor cell cannibalism could be a very direct and efficient way to neutralize immune response, as well. Tumor cell cannibalism may represent a sign of regression to a simpler, ancestral or primeval life style, similar to that of unicellular microorganisms, such as amoebas, where the goal is to survive and propagate in an overcrowded and very hostile microenvironment. In fact, we discovered that metastatic melanoma cells share with amoebas a transmembrane protein TM9SF4, indeed related to the cannibal behavior of these cells. This review attempts to provide a comprehensive description of the current knowledge about the role of TM9SF4 in cancer, highlighting its role as a key player in the cannibal behavior of malignant cancer cells. Moreover, we discuss differences and similarities between tumor cannibalism, entosis, phagocytosis and emperipolesis.
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Citofagocitosis , Neoplasias/patología , Amoeba/metabolismo , Animales , Supervivencia Celular , Emperipolesis , Entosis , Humanos , Proteínas de la Membrana/metabolismo , Neoplasias/inmunología , Neoplasias/metabolismo , FagocitosisRESUMEN
An inverted pH gradient across the cell membranes is a typical feature of malignant cancer cells that are characterized by extracellular acidosis and cytosol alkalization. These dysregulations are able to create a unique milieu that favors tumor progression, metastasis and chemo/immune-resistance traits of solid tumors. A key event mediating tumor cell pH alterations is an aberrant activation of ion channels and proton pumps such as (H+)-vacuolar-ATPase (V-ATPase). TM9SF4 is a poorly characterized transmembrane protein that we have recently shown to be related to cannibal behavior of metastatic melanoma cells. Here, we demonstrate that TM9SF4 represents a novel V-ATPase-associated protein involved in V-ATPase activation. We have observed in HCT116 and SW480 colon cancer cell lines that TM9SF4 interacts with the ATP6V1H subunit of the V-ATPase V1 sector. Suppression of TM9SF4 with small interfering RNAs strongly reduces assembly of V-ATPase V0/V1 sectors, thus reversing tumor pH gradient with a decrease of cytosolic pH, alkalization of intracellular vesicles and a reduction of extracellular acidity. Such effects are associated with a significant inhibition of the invasive behavior of colon cancer cells and with an increased sensitivity to the cytotoxic effects of 5-fluorouracil. Our study shows for the first time the important role of TM9SF4 in the aberrant constitutive activation of the V-ATPase, and the development of a malignant phenotype, supporting the potential use of TM9SF4 as a target for future anticancer therapies.
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Neoplasias del Colon/patología , Resistencia a Antineoplásicos/fisiología , Proteínas de la Membrana/metabolismo , Invasividad Neoplásica/patología , ATPasas de Translocación de Protón Vacuolares/metabolismo , Western Blotting , Línea Celular Tumoral , Neoplasias del Colon/metabolismo , Transferencia Resonante de Energía de Fluorescencia , Humanos , Concentración de Iones de Hidrógeno , Inmunoprecipitación , Microscopía Confocal , Reacción en Cadena de la Polimerasa , TransfecciónRESUMEN
OBJECTIVES: To explore the possibility that HIV-1 budding and cellular adhesion molecules co-polarize at cell-to-cell contact sites. To investigate the incorporation of host-cell-derived adhesion molecules into HIV-1. METHODS: The cellular sites involved in HIV-1 budding were examined by transmission electron microscopy. Single and double immunocytochemistry staining was used to evaluate the cellular distribution of the viral matrix protein and adhesion molecules. Quantitative flow cytometry was used to measure the cellular expression of adhesion molecules. An immunocapture technique was used to measure the presence of cell-derived proteins on HIV-1. The captured virus was measured by a p24 antigen assay. The infectivity of virus captured by monoclonal antibodies was tested by measuring the virus antigen yield in supernatants after the addition of sensitive cells. RESULTS: Released and budding HIV-1 was mainly localized at the cell-to-cell contact regions. This feature was consistent with a polarized staining for the virus matrix protein p18 at cell-to-cell contact regions. Intercellular adhesion molecules (ICAM)-1 in HIV-1-infected cells were polarized on both isolated cells and syncytia, co-localizing with HIV-1 matrix protein. HIV-1 incorporated all the adhesion molecules expressed by the host cells, although without quantitative correlation with their cellular expression. CONCLUSIONS: HIV-1 is released at cell-to-cell membrane contact sites. Both ICAM-1 and virus matrix protein co-polarized on isolated cells and syncytia at the sites involved in the recruitment of uninfected cells. The impressive concentration of ICAM at cell sites where most virions are released may account for the acquisition of these membrane proteins by the HIV-1 progeny, and may be important for the cell-mediated spread.
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Antígenos de Diferenciación , Moléculas de Adhesión Celular/metabolismo , VIH-1/crecimiento & desarrollo , VIH-1/fisiología , Proteínas de la Matriz Viral/metabolismo , Antígenos CD/metabolismo , Sitios de Unión , Linfocitos T CD4-Positivos/ultraestructura , Linfocitos T CD4-Positivos/virología , Adhesión Celular , Línea Celular , Polaridad Celular , Proteína p24 del Núcleo del VIH/metabolismo , VIH-1/inmunología , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Antígeno-1 Asociado a Función de Linfocito/metabolismo , Microscopía ElectrónicaRESUMEN
Efficiency of Fas-mediated apoptosis of lymphoid cells is regulated, among other means, by a mechanism involving its association with ezrin, a cytoskeletal protein belonging to the 4.1 family of proteins. In the present work, we provide evidence for a further molecule that associates to ezrin in Fas-triggered apoptosis, the disialoganglioside GD3. In fact, as an early event, GD3 redistributed in membrane-associated domains in uropods and co-localized with ezrin. Co-immunoprecipitation analyses confirmed this result, indicating a GD3-ezrin association. Altogether, these results are suggestive for a role of GD3 in Fas/ezrin-mediated apoptosis, supporting the view that uropods contain a multimolecular signaling complex involved in Fas-mediated apoptosis.
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Apoptosis/fisiología , Gangliósidos/fisiología , Fosfoproteínas/metabolismo , Receptor fas/fisiología , Línea Celular , Cromatografía en Capa Delgada , Proteínas del Citoesqueleto , Gangliósidos/metabolismo , Humanos , Microscopía Confocal , Microscopía Electrónica de Rastreo , Microscopía Fluorescente , Pruebas de PrecipitinaRESUMEN
The highly metastatic ESb tumor is totally resistant to murine interferon-alpha/beta (IFN-alpha/beta) therapy, regardless of the number of cells injected or the route of inoculation. In contrast, as we show herein, mouse IFN-alpha1-transduced ESb tumor cells were inhibited markedly when injected subcutaneously into immunocompetent mice. IFN-producing ESb tumor rejection was mediated by the immune system, because these tumor cells grew normally in immunosuppressed mice. Tumor regression was accompanied by extensive necrosis and cellular infiltrates in the tumor area. These results further support the use of IFN-alpha in cytokine gene therapy of cancer and suggest the advantage of using gene transfer rather than cytokine administration to enhance an antitumor immune response.
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Técnicas de Transferencia de Gen , Interferón-alfa/administración & dosificación , Interferón-alfa/genética , Animales , Resistencia a Múltiples Medicamentos , Interferón beta/administración & dosificación , Interferón beta/genética , Masculino , Ratones , Ratones Endogámicos DBA , Neoplasias Experimentales/inmunología , Retroviridae/genética , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
We have investigated the effect of somatostatin (SOM) on the mitogen-induced activation of lamina propria mononuclear cells isolated from the human intestinal mucosa (LPMNC) and of the autologous peripheral blood lymphocytes (PBMNC). The occurrence of specific SOM receptors and their biological characteristics were also investigated. The counts of interleukin-2 receptor (IL-2R)-positive cells after mitogen stimulation were significantly lower in the presence of SOM. This effect of SOM appeared to be dose dependent, with SOM concentrations ranging between 1 pM and 1 microM. The amount of SOM required for the 50% inhibition of this expression was 1000 times lower in the LPMNC population than in the PBMNC. Binding studies showed that human LPMNC bear specific receptors for SOM and demonstrated that the affinity of these receptors was 1000 times higher than that of the SOM receptors present on the PBMNC (Kd 2.1 +/- 0.34 nM vs. 910 +/- 46 nM, respectively). The inhibitory effect of SOM on the proliferative response appeared to be restricted to PBMNC, with a maximal inhibition at 1 nM SOM, while LPMNC proliferative response was poorly affected. SOM inhibited the in vitro immunoglobulin production of both PBMNC and LPMNC over a wide range of concentrations, with a maximal inhibition at 1 nM. At this concentration the effect of SOM on IgA was more pronounced in the PBMNC than in the LPMNC. Our results lend support to the concept that in humans SOM plays a role in the modulation of the immune response at the level of the intestinal mucosa where cell-to-cell interactions between SOM releasing nerve fibers and cells and the immune system occur.
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Mucosa Intestinal/inmunología , Activación de Linfocitos/efectos de los fármacos , Somatostatina/farmacología , ADN/biosíntesis , Humanos , Inmunoglobulinas/biosíntesis , Mucosa Intestinal/efectos de los fármacos , Receptores de Interleucina-2/análisis , Receptores de Neurotransmisores/análisis , Receptores de Somatostatina , Somatostatina/metabolismoRESUMEN
BACKGROUND: The residual resistance of severe combined immunodeficiency (SCID) mice to human graft is the main factor in conditioning both the extent of human cell reconstitution and the xenograft-to-xenograft variability. We have recently shown that an early and massive murine granulocyte recruitment is the main event in the SCID mouse reaction to the human graft. METHODS: Here, we evaluate the importance of mouse granulocytes in the restriction of human cell engraftment in SCID mice. We injected SCID mice with a monoclonal antibody to murine granulocytes. RESULTS: Injection of this antibody resulted in a marked depletion of polymorphonuclear cells in the hematopoietic organs of SCID mice. This depletion was associated with a significant increase in both the growth of human cell lines of different hematopoietic origin and the engraftment of human peripheral blood leukocytes. Moreover, the abolishment of the early granulocyte reaction markedly reduced the xenograft-to-xenograft variation, a major shortcoming of these xenochimeric models. CONCLUSIONS: These results provide new insights into the control of the natural immune response of SCID mice against human graft. Furthermore, treatments aimed at controlling the acute inflammatory reaction of SCID mouse-to-human cell transplantation can be considered useful experimental approaches for increasing the xenograft-to-xenograft reproducibility.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Citocinas/biosíntesis , Granulocitos/inmunología , Transfusión de Leucocitos , Neutrófilos/inmunología , Transcripción Genética , Trasplante Heterólogo/inmunología , Animales , División Celular/inmunología , Línea Celular , Femenino , Humanos , Ratones , Ratones SCID , Reacción en Cadena de la Polimerasa , ARN Mensajero/biosíntesis , Bazo/inmunologíaRESUMEN
In this study, we describe the kinetics of host immune reactions occurring in mice with severe combined immunodeficiency (SCID) at different times after the intraperitoneal injection of human peripheral blood mononuclear leukocytes (huPBL). At 24 hr, a massive neutrophil recruitment and an induced expression of a wide spectrum of murine cytokine mRNA (i.e., interleukin [IL]-1 beta, IL-4, IL-6, IL-10, IL-12, tumor necrosis factor [TNF]-alpha and interferon [IFN]-gamma) occurred in the huPBL-SCID mouse peritoneal cavity. By using ELISAs specific for mouse cytokines, large amounts of IL-1-alpha, TNF-alpha, IL-6, and IFN-gamma were detected in the peritoneal washings of huPBL-SCID mice 1 day after intraperitoneal injection. IL-6 and IFN-gamma production persisted for up to 2 weeks after PBL transplantation. Medullary and extramedullary expansion of the SCID mouse hematopoietic cells also occurred in the chimeras as early as 1 week after injection, together with a marked thymic differentiation (murine CD4+/CD8+ cells) at 10-12 weeks after transplantation. On the whole, these results indicate that, after huPBL injection, SCID mice mount a complex multistage immune response. These host reactions should be taken into consideration for any accurate interpretation of results obtained using the huPBL-SCID model. The control of responses (by means of specific antibodies to murine cytokines and to granulocytes or through the use of anti-inflammatory drugs) may be helpful in improving the engraftment of huPBL in SCID mice and in furthering our knowledge of the T and B cell-independent natural immune reactions.
Asunto(s)
Citocinas/fisiología , Hematopoyesis , Leucocitos/inmunología , Ratones SCID/inmunología , Cavidad Peritoneal/patología , Timo/citología , Animales , Diferenciación Celular , Expresión Génica , Humanos , Inflamación/patología , Inyecciones Intraperitoneales , Ratones , Neutrófilos/inmunología , ARN Mensajero/genéticaRESUMEN
Similarly to HIV-infected cells, recombinant HIV-1 glycoprotein 120 induces acid-labile interferon production in peripheral blood mononuclear cells from healthy donors. Acid lability of this interferon is due to the presence of both IFN-alpha and -gamma molecules. In fact, although not revealed by neutralization of antiviral activity with antibody to IFN-gamma, the presence of IFN-gamma was shown both immunoenzymatically and by detection of specific mRNA in gp120-stimulated cells. The source of IFN-gamma appears to be a T cell present in the CD4-enriched subpopulation. Cultures treated with monoclonal antibodies to the ICAM-1 and LFA-1 adhesion molecules showed an impaired release of both IFN types after gp120 stimulation, suggesting a crucial role of cell-to-cell interactions in the process leading to IFN production. Our data suggest that the HIV envelope glycoprotein could be responsible for the induction of endogenous IFN-alpha and -gamma observed in AIDS patients.
Asunto(s)
Regulación de la Expresión Génica/efectos de los fármacos , Proteína gp120 de Envoltorio del VIH/farmacología , Interferón-alfa/biosíntesis , Interferón gamma/biosíntesis , Subgrupos de Linfocitos T/efectos de los fármacos , Moléculas de Adhesión Celular/metabolismo , Células Cultivadas , Humanos , Molécula 1 de Adhesión Intercelular , Interferón-alfa/genética , Interferón gamma/genética , Antígeno-1 Asociado a Función de Linfocito/metabolismo , ARN Mensajero/análisis , Proteínas Recombinantes/farmacologíaRESUMEN
Cellular adhesion molecules, such as ICAM-1, -2, and -3; LFA-1; and HLA class I and II are incorporated into HIV-1 virions during budding from infected cells. These virion-associated molecules can be involved in the adsorption to susceptible cells displaying the corresponding counterligands. A number of cytokines have been shown to upregulate the cellular expression of adhesion molecules, such as ICAM-1 and HLA-DR. In this study we investigated the effects of IFN-gamma on the incorporation of ICAM-1, LFA-1, and HLA-DR into mature HIV-1 progeny from chronically infected cells. The ability of such virus progeny to infect either CD4-positive or -negative cells was also investigated. The results indicate that IFN-gamma stimulates the expression of ICAM-1 and of HLA-DR on HIV-1-infected cells, whereas LFA-1 expression is unaffected. The same modifications were also observed on virus progeny, because specific MAbs to ICAM-1 and HLA-DR captured infectious HIV-1 from IFN-treated cells with higher efficiency as compared to virus from control cells, whereas virus binding to anti LFA-1 MAb was unchanged. Moreover, the HIV-1 progeny released from IFN-treated cells showed an increased ability to bind to and to infect CD4-negative cells, whereas the infectivity was basically unchanged for CD4-positive cells. Our results suggest that cytokines, as well as other soluble factors, may expand the host cell range of HIV-1, possibly through modifications of the cell-derived surface molecules on the virions.(ABSTRACT TRUNCATED AT 250 WORDS)