RESUMEN
Notch-1 intervenes in the reparative processes of mucosa by controlling cell proliferation, differentiation and stem cell maintenance. Cigarette smoke alters airway epithelial homeostasis. The present study explored whether: Smokers showed altered Notch-1 expression; and whether in bronchial epithelial cells (16HBE): a) cigarette smoke extracts (CSE) altered the expression of Notch-1, of its ligand Jagged-1 (Jag-1) and the nuclear translocation of Notch-1; b) Notch-1 signaling activation as well as CSE modified Ki67, PCNA, p21, IL-33 expression, cell proliferation and repair processes. Notch-1 expression was assessed in the epithelium from large airway surgical samples from non-smoker and smoker subjects by immunohistochemistry.16HBE were cultured with/without CSE and Jag-1. A Notch-1 inhibitor (DAPT) was used as control. The expression of Notch-1, Jag-1, Ki67, PCNA, p21, IL-33 and cell proliferation (by CFSE) were all assessed by flow cytometry. Notch-1 nuclear expression was evaluated by immunofluorescence and western blot analysis. Repair processes were assessed by wound assay. Smokers had cytoplasmic but not nuclear Notch-1 expression. Although CSE increased Notch-1 expression, it counteracted Notch-1 signaling activation since it reduced Jag-1 expression and Notch-1 nuclear translocation. Notch-1 signaling activation by Jag-1 increased Ki67, PCNA and repair processes but reduced intracellular IL-33 and p21 expression without affecting cell proliferation. DAPT counteracted the effects of Notch-1 activation on PCNA and IL-33. CSE increased Ki67, PCNA, p21 and IL-33 expression but reduced cell proliferation and repair processes. In conclusion, cigarette smoke exposure, limiting Notch-1 signaling activation and hindering repair processes, amplifies injury processes in bronchial epithelial cells.
Asunto(s)
Apoptosis/efectos de los fármacos , Bronquios/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Receptores Notch/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Contaminación por Humo de Tabaco/efectos adversos , HumanosRESUMEN
BACKGROUND: Cigarette smoke is considered a risk factor for lung and colorectal cancer. A convincing link between epithelial-to-mesenchymal transition (EMT) with colorectal cancer progression and therapeutic resistance has emerged. Deregulated expression of E-Cadherin and Claudin-1 and increased miR-21 expression and invasiveness represent hallmarks of EMT. The effects of cigarette smoke exposure on EMT in colorectal adenocarcinoma cells are largely unknown. AIM: The aim of the study is to evaluate the effect of cigarette smoke extract (CSE) on miR-21, Claudin-1 and E-Cadherin, molecules associated to EMT in colorectal cancer cells. METHODS: A human colorectal adenocarcinoma cell line (Caco-2) was treated with CSE at different concentration (5% and 10%) and for different time points (3â¯h and 24â¯h). Metabolic activity (by MTS assay), cell necrosis/cell apoptosis (evaluating Propidium Iodide/Annexin V expression by flow cytometry), miR-21, Claudin-1 and E-Cadherin gene expression were evaluated by Real time PCR. Cell permeability, actin polymerization and cancer cell migration was assessed by Trans-Epitelial Electrical Resistance (TEER), Phalloidin expression and matrigel system, respectively. RESULTS: CSE at all the tested concentrations and at all time points reduced cell necrosis. CSE at 10% increased miR-21 and reduced the metabolic activity, cell necrosis, Claudin-1 and E-cadherin mRNA at 3â¯h. Cell permeability, actin polymerization and cancer cell migration were all increased upon CSE exposure. CONCLUSION: These results showed that CSE increasing miR-21, Claudin-1 and E-Cadherin and enhancing the aggressiveness of cancer cells, may concur to colorectal cancer progression.
Asunto(s)
Adenocarcinoma/metabolismo , Antígenos CD/metabolismo , Cadherinas/metabolismo , Movimiento Celular , Fumar Cigarrillos/efectos adversos , Claudina-1/metabolismo , Neoplasias Colorrectales/metabolismo , MicroARNs/metabolismo , Humo/efectos adversos , Adenocarcinoma/genética , Adenocarcinoma/patología , Antígenos CD/genética , Células CACO-2 , Cadherinas/genética , Claudina-1/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Progresión de la Enfermedad , Transición Epitelial-Mesenquimal , Regulación Neoplásica de la Expresión Génica , Humanos , MicroARNs/genética , Invasividad Neoplásica , Transducción de SeñalRESUMEN
BACKGROUND: Alterations in the nasal epithelial barrier homeostasis and increased interleukin 33 (IL-33) expression contribute to the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP). AIMS: As Notch-1 signaling is crucial in repair processes of mucosa, the current study assessed Notch-1/Jagged-1 signaling and IL-33 in the epithelium of nasal polyps biopsies from allergic (A-CRSwNP; n = 9) and not allergic (NA-CRSwNP; n = 9) subjects by immunohistochemistry. We also assessed, in a model of nasal epithelial cells, the effects of stimulation of Notch-1 with Jagged-1 on the expression of IL-33 (by flow cytometry, immunofluorescence, and immunocytochemistry), Jagged-1 (by flow cytometry), and p-CREB transcription factor (by western blot analysis). RESULTS: Ex vivo (a) in normal epithelium, the expression of Notch-1 and IL-33 were higher in NA-CRSwNP than in A-CRSwNP; (b) in metaplastic epithelium, the expression of Notch-1, Jagged-1, and IL-33 were higher in NA-CRSwNP than in A-CRSwNP; (c) in hyperplastic epithelium, the expression of Notch-1, Jagged-1, and IL-33 were higher in A-CRSwNP than in NA-CRSwNP; and (d) in basal epithelial cells, no differences were observed in the expression of Jagged-1, IL-33, and Notch-1. The expression of Notch-1 significantly correlated with the expression of IL-33. In vitro, stimulation of Notch-1 with Jagged-1 induced the expression of (a) Jagged-1; (b) IL-33; and (c) p-CREB transcription factor. The inhibitor of Notch-1, DAPT, reduced all the effects of Jagged-1 on nasal epithelial cells. CONCLUSIONS: The data herein provided support, for the first time, a putative role of Notch-1/Jagged-1 signaling in the overexpression of IL-33 in the epithelium of nasal polyps from patients with CRSwNP.
Asunto(s)
Células Epiteliales/metabolismo , Interleucina-33/metabolismo , Mucosa Nasal/metabolismo , Pólipos Nasales/metabolismo , Receptor Notch1/metabolismo , Rinitis Alérgica/metabolismo , Sinusitis/metabolismo , Adulto , Línea Celular , Enfermedad Crónica , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Células Epiteliales/inmunología , Células Epiteliales/patología , Femenino , Humanos , Proteína Jagged-1/metabolismo , Masculino , Persona de Mediana Edad , Mucosa Nasal/inmunología , Mucosa Nasal/patología , Pólipos Nasales/inmunología , Pólipos Nasales/patología , Fosforilación , Rinitis Alérgica/inmunología , Rinitis Alérgica/patología , Transducción de Señal , Sinusitis/inmunología , Sinusitis/patología , Regulación hacia Arriba , Adulto JovenRESUMEN
TGF-beta-targeting structural and inflammatory cells has been implicated in the mechanisms leading to the inflammatory and restructuring processes in asthma, suggesting an impact of TGF-beta1 signaling on the development and persistency of this disease. We investigated the potential early involvement of TGF-beta1 activity in the immunological and molecular mechanisms underlying progression of inflammation in childhood asthma. We evaluated the levels of TGF-beta1 in induced sputum supernatants (ISSs) and the expression of small mother cell against decapentaplegic (Smad) 2 and Smad7 proteins in induced sputum cells (ISCs) from children with intermittent asthma (IA), moderate asthma (MA) and control subjects (C). Furthermore, we investigated the regulatory role of TGF-beta1 activity on eosinophil and neutrophil adhesion to epithelial cells using adhesion assay, and on the granulocyte expression of adhesion molecule CD11b/CD18 Macrophage-1 antigen (MAC-1), by flow cytometry. We found that the levels of TGF-beta1 are increased in ISSs of IA and MA in comparison to C, concomitantly to the activation of intracellular signaling TGFbeta/Smads pathway in ISCs. In MA, TGF-beta1 levels correlated with the number of sputum eosinophils and neutrophils. Furthermore, we showed the ability of sputum TGF-beta1 to promote eosinophil and neutrophil adhesion to epithelial cells, and to increase the expression of MAC-1 on the granulocyte surface. This study shows the activation of TGFbeta/Smad signaling pathway in the airways of children with IA and, despite the regular ICS treatment, in children with MA, and provides evidence for the contribution of TGF-beta1 in the regulation of granulocyte activation and trafficking.
Asunto(s)
Asma/metabolismo , Pulmón/metabolismo , Transducción de Señal , Factor de Crecimiento Transformador beta1/metabolismo , Administración por Inhalación , Adolescente , Corticoesteroides/administración & dosificación , Factores de Edad , Asma/diagnóstico , Asma/tratamiento farmacológico , Asma/inmunología , Asma/fisiopatología , Broncodilatadores/administración & dosificación , Estudios de Casos y Controles , Adhesión Celular , Línea Celular , Niño , Eosinófilos/inmunología , Eosinófilos/metabolismo , Células Epiteliales/inmunología , Células Epiteliales/metabolismo , Femenino , Granulocitos/inmunología , Granulocitos/metabolismo , Humanos , Pulmón/efectos de los fármacos , Pulmón/inmunología , Pulmón/fisiopatología , Antígeno de Macrófago-1/metabolismo , Masculino , Neutrófilos/inmunología , Neutrófilos/metabolismo , Fosforilación , Mucosa Respiratoria/inmunología , Mucosa Respiratoria/metabolismo , Índice de Severidad de la Enfermedad , Transducción de Señal/efectos de los fármacos , Proteína Smad2/metabolismo , Proteína smad7/metabolismo , Esputo/metabolismoRESUMEN
In this study we wanted to analyse the pattern of the immune response to the Parietaria major allergen Par j 1 in freshly purified peripheral blood mononuclear cell (PBMC) from healthy subjects. We observed that Par j 1 was capable of inducing IFN-γ production by CD3â» and CD16âº/CD56⺠cells exclusively in healthy individuals. Furthermore, a multiparametric analysis allowed us a better definition of two IFN-γ-Par j 1 specific populations (IFN-γ(dim) and IFN-γ(high)) characterized by the presence of different proportions of NKT and NK cells. We also identified the concomitant presence of a subset of IL-10⺠NK cells. Moreover, CFSE staining showed that the Par j 1 preferentially induced the proliferation of CD3â»/CD56âº/CD335⺠cells. Finally, a subset of CD4âº/CD25âº/FoxP3âº/IL-10â» T cells was identified. The result of this pilot study suggest that during a tolerogenic response, the major allergen of the Parietaria pollen works as an activator of both the innate and the adaptive human immune system.
Asunto(s)
Inmunidad Adaptativa , Alérgenos/farmacología , Inmunidad Innata , Células Asesinas Naturales/efectos de los fármacos , Células T Asesinas Naturales/efectos de los fármacos , Parietaria/inmunología , Proteínas de Plantas/farmacología , Adulto , Alérgenos/biosíntesis , Alérgenos/genética , Antígenos CD/genética , Antígenos CD/inmunología , Células Cultivadas , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/inmunología , Expresión Génica/efectos de los fármacos , Humanos , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-10/genética , Interleucina-10/inmunología , Células Asesinas Naturales/citología , Células Asesinas Naturales/inmunología , Células T Asesinas Naturales/citología , Células T Asesinas Naturales/inmunología , Proteínas de Plantas/biosíntesis , Proteínas de Plantas/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologíaRESUMEN
SETTING: Dendritic cells (DC) could regulate between the protective and pathogenic immune responses following tuberculous infection. In this paper we investigated if their early infection in the lungs represents a plausible alternative to cross-priming with mycobacterial antigens acquired from infected macrophages. OBJECTIVE: To determine the extent and time course of infection of lung DCs following intranasal inoculation of BALB/c mice with green fluorescent protein (GFP) tagged Bacillus Calmette-Guerin (BCG). RESULTS: A fraction of GFP-BCG infected lung cells were classified as monocytic DCs with the CD11c+IA+33D1+CD8a- phenotype. These cells represented 5-18% of the total GFP+ cells, the bulk of which were macrophages. The infected DCs could be separated by cell size into two fractions with similar cell surface staining properties during the 2-72 h period after infection. An unexpected difference was observed for the time course of infection between DCs and macrophages: DC infection peaked at 48 h followed by decline at 72 h, while the proportion of infected macrophages remained steady during the same period. CONCLUSION: The presented results are direct evidence that monocytic DCs are recruited to the lungs and take up live bacilli within 48 h of intranasal infection with GFP-BCG. This finding is pertinent for the regulation of pulmonary and systemic immune responses and possibly for the dissemination of mycobacterial infection by DCs.
Asunto(s)
Células Dendríticas/inmunología , Pulmón/microbiología , Infecciones por Mycobacterium/inmunología , Administración Intranasal , Animales , Antígenos Bacterianos/inmunología , Antígenos CD/inmunología , Antígenos de Superficie/inmunología , Vacuna BCG/administración & dosificación , Tamaño de la Célula , Proteínas Fluorescentes Verdes , Sustancias Luminiscentes , Pulmón/patología , Macrófagos/inmunología , Ratones , Ratones Endogámicos BALB C , Monocitos/inmunología , Infecciones por Mycobacterium/patología , Factores de Tiempo , Tuberculosis Pulmonar/inmunología , Tuberculosis Pulmonar/patologíaRESUMEN
Much evidence now indicates that human leucocyte antigen (HLA) class I and class II transgenic (Tg) mice can be of value in analysing HLA-restricted presentation of T-cell epitopes relevant to experimental models of autoimmune diseases. One area where this has been applied is the characterization of myelin epitopes presented by HLA class II molecules in experimental model of multiple sclerosis (experimental allergic encephalomyelitis (EAE)). As a first step towards humanized disease models in HLA Tg mice, we have analysed immune response of lymph node cells of HLA-DR1 Tg mice immunized with the human myelin basic protein (MBP) peptides 13-33, 87-106 and 139-154 bound by HLA-DR1. We report here that HLA-DR1 Tg mice display a hierarchy of response in vivo and in vitro to MBP epitopes depending on the binding affinity to DRB*0101 molecule. In fact, the 13-33 epitope induced a strong T helper 1 (Th1) response accompanied by high T-cell precursor frequency and caused mild EAE, while the two other epitopes gave poor (139-154) or no disease (87-106), and these data correlate with in vitro Th1 response. These data could prove a useful tool in understanding the role played by different MBP epitopes in EAE.
Asunto(s)
Encefalomielitis Autoinmune Experimental/genética , Encefalomielitis Autoinmune Experimental/inmunología , Epítopos de Linfocito T/inmunología , Antígeno HLA-DR1/inmunología , Esclerosis Múltiple/inmunología , Proteína Básica de Mielina/inmunología , Secuencia de Aminoácidos , Animales , Modelos Animales de Enfermedad , Femenino , Predisposición Genética a la Enfermedad , Antígeno HLA-DR1/genética , Humanos , Ganglios Linfáticos/inmunología , Masculino , Ratones , Ratones Transgénicos , Datos de Secuencia Molecular , Esclerosis Múltiple/genética , Proteína Básica de Mielina/genética , Fragmentos de Péptidos/inmunologíaRESUMEN
The identification of immunodominant and universal mycobacterial peptides could be applied to vaccine design and have an employment as diagnostic reagents. In this paper we have investigated the fine specificity, clonal composition and HLA class II restriction of CD4+ T cell clones specific for an immunodominant epitope spanning amino acids 91-110 of the 16-kDa protein of Mycobacterium tuberculosis. Twenty-one of the tested 28 clones had a Th1 profile, while seven clones had a Th0 profile. None of the clones had a Th2 profile. While the TCR AV gene usage of the clones was heterogeneous, a dominant TCR BV2 gene family was used by 18 of the 28 clones. The CDR3 regions of BV2+ T cell clones showed variation in lengths, but a putative common motif R-L/V-G/S-Y/W-E/D was detected in 13 of the 18 clones. Moreover, the last two to three residues of the putative CDR3 loops, encoded by conserved BJ sequences, could also play a role in peptide recognition. Antibody blockade and fine restriction analysis using HLA-DR homozygous antigen-presenting cells established that 16 of 18 BV2+ peptide-specific clones were DR restricted and two clones were DR-DQ and DR-DP restricted. Additionally, five of the 18 TCRBV2+ clones recognized peptide 91-110 in association with both parental and diverse HLA-DR molecules, indicating their promiscuous recognition pattern. The ability of peptide 91-110 to bind a wide range of HLA-DR molecules, and to stimulate a Th1-type interferon (IFN)-gamma response more readily, encourage the use of this peptide as a subunit vaccine component.
Asunto(s)
Antígenos Bacterianos/inmunología , Linfocitos T CD4-Positivos/inmunología , Citocinas/biosíntesis , Epítopos Inmunodominantes/inmunología , Mycobacterium tuberculosis/inmunología , Células Cultivadas , Regiones Determinantes de Complementariedad/inmunología , Antígenos HLA-DR/inmunología , Humanos , Interferón gamma/biosíntesis , Fragmentos de Péptidos/inmunología , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/inmunología , Tuberculosis/inmunologíaRESUMEN
Human T cells expressing Vgamma9Ndelta2-encoded TCR recognize several nonpeptide phosphoantigens in the absence of major histocompatibility complex restriction. As these cells respond differentially to increasing concentrations of structurally related phosphoantigens, such ligands constitute agonists of different strengths. By analyzing early cellular events and late effector responses of gammadelta T cells, we compared their patterns of stimulation by weak, medium and strong phosphoantigen agonists. We found that, although the early metabolic activation as assessed by cytosensor microphysiometry directly reflects the intensity of subsequent effector response by gammadelta cells, TCR down-modulation is dissociated from the latter. Weak and mid-range phosphoantigen agonists induce a time- and dose-dependent down-modulation of the gammadelta TCR, whereas strong phosphoantigen agonists induce little or no TCR down-regulation. This indicates that gammadelta TCR down-modulation does not match the extent of TCR signaling as assessed by microphysiometry or conventional effector responses (TNF-alpha production and cytotoxicity). This differential pattern of gammadelta cell activation by phosphoantigens could explain the stronger potencies of some of these agonists.
Asunto(s)
Antígenos/química , Antígenos/inmunología , Activación de Linfocitos/inmunología , Receptores de Antígenos de Linfocitos T gamma-delta/inmunología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Ácidos/metabolismo , Células Cultivadas , Células Clonales/inmunología , Células Clonales/metabolismo , Regulación hacia Abajo , Citometría de Flujo , Humanos , Concentración de Iones de Hidrógeno , Estructura Molecular , Fosforilación , Receptores de Antígenos de Linfocitos T gamma-delta/metabolismo , Linfocitos T/citología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Lymph node cells from TNCB-immune BALB/c mice fail to produce IFN-gamma when exposed to antigen in vitro. Conversely, lymph node cells of irradiated (550 rads) BALB/c mice produce IFN-gamma. Transfer experiments show that normal BALB/c mice contain cells which suppress IFN-gamma production. These downregulatory cells are CD4(+)alpha beta(+)and rearrange the invariant V alpha 14-J alpha 281 T cell receptor alpha chain, thus belonging to the NK T cell subset. Downregulatory cells probably act by producing IL-4 as their effect is blocked by mAb to IL-4.
Asunto(s)
Interferón gamma/biosíntesis , Células Asesinas Naturales/efectos de la radiación , Receptores de Antígenos de Linfocitos T alfa-beta/biosíntesis , Animales , Anticuerpos Monoclonales/metabolismo , Células Presentadoras de Antígenos , Antígenos de Protozoos/metabolismo , Antígenos CD4/biosíntesis , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD4-Positivos/parasitología , Antígenos CD8/biosíntesis , Separación Celular , Regulación hacia Abajo , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Interleucina-4/biosíntesis , Interleucina-4/inmunología , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/parasitología , Leishmania major/metabolismo , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/metabolismo , Ganglios Linfáticos/parasitología , Ganglios Linfáticos/efectos de la radiación , Masculino , Ratones , Ratones Endogámicos BALB C , Fenotipo , Bazo/inmunología , Bazo/parasitología , Bazo/efectos de la radiaciónRESUMEN
T cells mediate protection against tuberculosis, but little is known about their role during chemotherapy of patients with active disease. Here we examined the cytokine profile of CD4 T cells before and after four months of chemotherapy in six initial skin test anergic cases. Purified protein derivative (PPD) and 16-kDa antigen-reactive CD4 T-cell clones prior to therapy resided mostly in disease-associated body fluids and were of the Th0 (interferon (IFN)-gamma + interleukin (IL)-4) secreting profile. In contrast, the majority of postchemotherapy CD4 T-cell clones originated from blood and were of the IFN-gamma secreting Th1 type. However, the recognition of several peptides derived from the 16-kDa antigen was not significantly different between the Th1 and Th0 clones. We conclude that chemotherapy shifts CD4 T cells from the affected body fluids to the blood circulation, accompanied by a change from Th0 to Th1 cytokine profile.
Asunto(s)
Interferón gamma/biosíntesis , Interleucina-4/biosíntesis , Células TH1/inmunología , Tuberculosis/inmunología , Linfocitos T CD4-Positivos/inmunología , Cristalinas/inmunología , Humanos , Activación de Linfocitos , Células Th2/inmunología , Tuberculina/inmunología , Tuberculosis/tratamiento farmacológicoRESUMEN
The alphabeta and gammadelta T cell responses were analyzed in the peripheral blood of children affected by active tuberculosis (TB) and in healthy children who tested positive (PPD+) or negative (PPD-) for purified protein derivative. PPD+ healthy and diseased children responded equally well to PPD in vitro. In contrast, only 18% of PPD+ TB patients responded to peptide p38G derived from the 38-kDa protein of Mycobacterium tuberculosis. Analysis of the whole gammadelta T cell population and of its Vgamma9/Vdelta2 subset showed similar frequencies in PPD+ children with TB and in healthy PPD+ and PPD- children. Vgamma9/Vdelta2 cells from children with TB responded to 5 different phosphoantigens similarly to those from healthy PPD+ children, but healthy PPD- children responded very poorly. Chemotherapy had contrasting effects on the tested lymphocyte population, represented by increase of alphabeta and decline of Vgamma9/Vdelta2 T cell responses. T cell responses in childhood TB may be similar to those in adult TB.
Asunto(s)
Receptores de Antígenos de Linfocitos T alfa-beta , Receptores de Antígenos de Linfocitos T gamma-delta , Subgrupos de Linfocitos T , Tuberculosis/inmunología , Adolescente , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Tuberculosis/tratamiento farmacológicoRESUMEN
The repertoire of CD4+ T-lymphocytes was investigated in six patients affected by tuberculosis, who had a negative PPD skin test at diagnosis. Polyclonal CD4+ T-cell lines from the peripheral blood failed to proliferate to PPD and to the 16- or 38-kDa proteins of Mycobacterium tuberculosis, while CD4+ T-cell lines from the site of disease responded to PPD, and to the 16- and 38-kDa proteins, and derived epitopes in vitro. The repertoire of CD4+ T-cells accumulating at the site of disease was found to be widely heterogeneous as demonstrated by the finding that at least seven different peptides from the 16- and 38-kDa proteins were recognized by every patient. These results indicate that CD4+ T-cells localized at the site of disease in tuberculosis recognize a vast array of M. tuberculosis epitopes.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Mycobacterium tuberculosis/inmunología , Tuberculosis/inmunología , Secuencia de Aminoácidos , Antígenos Bacterianos/inmunología , Epítopos de Linfocito T/inmunología , Humanos , Lipoproteínas/inmunología , Meningitis Bacterianas/sangre , Meningitis Bacterianas/inmunología , Meningitis Bacterianas/patología , Datos de Secuencia Molecular , Pericarditis Tuberculosa/sangre , Pericarditis Tuberculosa/inmunología , Pericarditis Tuberculosa/patología , Pleuresia/sangre , Pleuresia/inmunología , Pleuresia/patología , Tuberculosis/sangre , Tuberculosis/patologíaRESUMEN
BACKGROUND: The role of T lymphocytes in graft rejection in xenotransplantation is still unclear. The ability of the human HLA class II molecules DR and DQ to function as xenoantigens was investigated in a murine model of skin grafting, using HLA-DR1 and -DQ6-transgenic mice. METHODS: Skin from HLA-DR1- or -DQ6-transgenic mice was transplanted in control littermates. Spleen cells from donors or recipients were tested in mixed lymphocyte reaction and cytotoxic assay. RESULTS: Skin from HLA-DR1-transgenic mice was rejected and spleen cells from rejecting mice were able to proliferate to donor cells, although no rejection was observed when the skin of HLA-DQ6-transgenic mice was engrafted in control littermates. No cytotoxicity was observed in any models. CONCLUSIONS: Taken all together these results clearly suggest a hierarchy in the xenogeneic potency of human HLA class II molecules, with the HLA-DR1 molecule functioning as a potent xenoantigen when compared with the HLA-DQ6 molecule.
Asunto(s)
Antígenos HLA-DQ/fisiología , Antígenos HLA-DR/fisiología , Trasplante de Piel/inmunología , Trasplante Heterólogo/inmunología , Animales , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Distribución TisularRESUMEN
BACKGROUND: We analyzed the gammadelta T cell composition and responses in the peripheral blood and cerebrospinal fluid (CSF) of children affected by tuberculous meningitis (TBM) and in control children. MATERIALS AND METHODS: Peripheral blood and CSF samples were stimulated with different phosphoantigens and IL-2, and expansion of Vgamma9/Vdelta2 T cells assessed by FACS analysis. Vgamma9/Vdelta2 lines were obtained by culturing CSF or peripheral blood mononuclear cells (PBMC) in vitro with phosphoantigens and IL-2 for 2 months, and tested for proliferation and cytokine production in response to phosphoantigens. Vdelta2(D)Jdelta junctional sequence length was assessed by PCR. RESULTS: The repertoire of gammadelta T cells from the CSF of TBM patients was characterized by the predominance of Vgamma9/Vdelta2 T lymphocytes, which accounted for >80% of gammadelta T cells. Vgamma9/Vdelta2 cells from the CSF of TBM children responded to different synthetic and natural (mycobacterial) phosphoantigens and produced discrete amounts of IFN-gamma and TNF-alpha. The in vitro expansion of Vgamma9/Vdelta2 T cells from CSF and peripheral blood of TBM patients prominently decreased following chemotherapy, and similarly, the proportion of ex vivo unstimulated Vgamma9/Vdelta2 T cells in CSF of TBM patients decreased to levels detected in the CSF of control subjects. Vdelta2 CDR3 TCR analysis showed that the remaining Vdelta2 cells in the CSF of TBM patients were still polyclonal. CONCLUSIONS: These findings are consistent with an involvement of Vgamma9/Vdelta2 T cells in TBM. http://link. springer-ny.com/link/service/journals/00020/bibs/5n5p301. html
Asunto(s)
Receptores de Antígenos de Linfocitos T gamma-delta/metabolismo , Linfocitos T/inmunología , Tuberculosis Meníngea/tratamiento farmacológico , Tuberculosis Meníngea/inmunología , Antígenos/metabolismo , Antígenos Bacterianos/farmacología , Líquido Cefalorraquídeo/citología , Líquido Cefalorraquídeo/inmunología , Niño , Preescolar , Citocinas/metabolismo , Femenino , Humanos , Lactante , Recuento de Linfocitos , Masculino , Valor Predictivo de las Pruebas , Valores de Referencia , Linfocitos T/efectos de los fármacos , Tuberculosis Meníngea/diagnósticoRESUMEN
The specificity of CD4 T lymphocytes was investigated in 6 patients affected by tuberculosis who had negative tuberculin purified protein derivative (PPD) skin tests at diagnosis. Polyclonal CD4 T cell lines from the peripheral blood failed to proliferate to PPD and to the 16- or 38-kDa proteins of Mycobacterium tuberculosis, while CD4 cell lines from the disease site responded to PPD and to the 16- and 38-kDa proteins and derived epitopes in vitro. Four months after chemotherapy, the patients became responsive to PPD. The proliferative response to PPD and to the 16- or 38-kDa proteins and their derived peptides decreased in CD4 T cell lines from the disease site and increased in lines from the peripheral blood. These results indicate that CD4 T cells recognizing a vast array of M. tuberculosis epitopes are compartmentalized at the site of disease in anergic patients but appear in peripheral blood after chemotherapy.
Asunto(s)
Líquidos Corporales/inmunología , Linfocitos T CD4-Positivos/inmunología , Anergia Clonal , Tuberculosis Pulmonar/inmunología , Antígenos Bacterianos/inmunología , Humanos , Lipoproteínas/inmunología , Tuberculina/inmunología , Tuberculosis/tratamiento farmacológico , Tuberculosis/inmunología , Tuberculosis Pulmonar/tratamiento farmacológicoRESUMEN
A study aiming to assess the clinical tolerance of a special follow-up soybean milk formula (Neo-Soyal 2) in children with cow's milk protein intolerance or allergy (CMPI/CMPA) has been conducted in 17 infants and children (9 girls and 8 boys) aged from 6 months to 3 years. All the patients had a past history suggestive of CMPI/CMPA and were fed a restricted hypoallergenic diet with a semi-elemental formula based on protein hydrolysates. At the time of initiation of the study, all the patients were asymptomatic with normal growth while on a semi-elemental diet. The formula with protein hydrolysates was substituted by the test milk (Neo-Soyal 2) and blood samples were taken before and after 2 months. The clinical tolerance to the follow-up soybean formula was considered excellent in 16 children who remained asymptomatic with normal growth velocity. Only one patient developed a severe diarrhoea within 72 hours after introduction of the soybean formula which was stopped. However, after one month he was able to tolerate a certain amount of the soybean milk (240 ml) without symptoms. For all the patients studied, there was no change in the nutritional parameters measured in the blood before and after 2 months of the trial. Likewise, the plasma amino acid levels determined in 2 patients remained unchanged before and after the experimental period. These data indicate that the soybean milk formula used here (Neo-Soyal 2) can be safely given as a substitute of semi-elemental diets during the follow-up of children with CMPI/CMPA after the age of 6 months.