RESUMEN
Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by progressive joint destruction associated with increased pro-inflammatory mediators. In inflammatory microenvironments, exogenous ATP (eATP) is hydrolyzed to adenosine, which exerts immunosuppressive effects, by the consecutive action of the ectonucleotidases CD39 and CD73. Mature B cells constitutively express both ectonucleotidases, converting these cells to potential suppressors. Here, we assessed CD39 and CD73 expression on B cells from treated or untreated patients with RA. Neither the frequency of CD73+CD39+ and CD73-CD39+ B cell subsets nor the levels of CD73 and CD39 expression on B cells from untreated or treated RA patients showed significant changes in comparison to healthy controls (HC). CpG+IL-2-stimulated B cells from HC or untreated RA patients increased their CD39 expression, and suppressed CD4+ and CD8+ T cell proliferation and intracellular TNF-production. A CD39 inhibitor significantly restored proliferation and TNF-producing capacity in CD4+ T cells, but not in CD8+ T cells, from HC and untreated RA patients, indicating that B cells from untreated RA patients conserved CD39-mediated regulatory function. Good responder patients to therapy (R-RA) exhibited an increased CD39 but not CD73 expression on B cells after treatment, while most of the non-responder (NR) patients showed a reduction in ectoenzyme expression. The positive changes of CD39 expression on B cells exhibited a negative correlation with disease activity and rheumatoid factor levels. Our results suggest modulating the ectoenzymes/ADO pathway as a potential therapy target for improving the course of RA.
Asunto(s)
Apirasa/metabolismo , Artritis Reumatoide/inmunología , Artritis Reumatoide/metabolismo , Linfocitos B/inmunología , Linfocitos B/metabolismo , Inmunomodulación , Adenosina/metabolismo , Apirasa/genética , Artritis Reumatoide/patología , Artritis Reumatoide/terapia , Linfocitos B Reguladores/inmunología , Linfocitos B Reguladores/metabolismo , Estudios de Casos y Controles , Citocinas/biosíntesis , Manejo de la Enfermedad , Susceptibilidad a Enfermedades , Expresión Génica , Humanos , Activación de Linfocitos , Recuento de Linfocitos , Linfocitos T/inmunología , Linfocitos T/metabolismo , Resultado del TratamientoRESUMEN
Solid tumors are infiltrated by immune cells where macrophages and senescent T cells are highly represented. Within the tumor microenvironment, a cross-talk between the infiltrating cells may occur conditioning the characteristic of the in situ immune response. Our previous work showed that tumors induce senescence of T cells, which are powerful suppressors of lympho-proliferation. In this study, we report that Tumor-Induced Senescent (TIS)-T cells may also modulate monocyte activation. To gain insight into this interaction, CD4+ or CD8+TIS-T or control-T cells were co-incubated with autologous monocytes under inflammatory conditions. After co-culture with CD4+ or CD8+TIS-T cells, CD14+ monocytes/macrophages (Mo/Ma) exhibit a higher expression of CD16+ cells and a reduced expression of CD206. These Mo/Ma produce nitric oxide and reactive oxygen species; however, TIS-T cells do not modify phagocyte capacity of Mo/Ma. TIS-T modulated-Mo/Ma show a higher production of pro-inflammatory cytokines (TNF, IL-1ß and IL-6) and angiogenic factors (MMP-9, VEGF-A and IL-8) and a lower IL-10 and IP-10 secretion than monocytes co-cultured with controls. The mediator(s) present in the supernatant of TIS-T cell/monocyte-macrophage co-cultures promote(s) tubulogenesis and tumor-cell survival. Monocyte-modulation induced by TIS-T cells requires cell-to-cell contact. Although CD4+ shows different behavior from CD8+TIS-T cells, blocking mAbs against T-cell immunoglobulin and mucin protein 3 and CD40 ligand reduce pro-inflammatory cytokines and angiogenic factors production, indicating that these molecules are involved in monocyte/macrophage modulation by TIS-T cells. Our results revealed a novel role for TIS-T cells in human monocyte/macrophage modulation, which may have deleterious consequences for tumor progression. This modulation should be considered to best tailor the immunotherapy against cancer.
Asunto(s)
Linfocitos T CD4-Positivos/metabolismo , Ligando de CD40/metabolismo , Linfocitos T CD8-positivos/metabolismo , Citocinas/biosíntesis , Proteínas de la Membrana/metabolismo , Monocitos/metabolismo , Linfocitos T CD4-Positivos/citología , Ligando de CD40/genética , Linfocitos T CD8-positivos/citología , Comunicación Celular , Supervivencia Celular , Senescencia Celular , Técnicas de Cocultivo , Citocinas/metabolismo , Proteínas Ligadas a GPI/genética , Proteínas Ligadas a GPI/metabolismo , Regulación de la Expresión Génica , Células HeLa , Receptor 2 Celular del Virus de la Hepatitis A , Humanos , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Receptor de Manosa , Lectinas de Unión a Manosa/genética , Lectinas de Unión a Manosa/metabolismo , Metaloproteinasa 9 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/metabolismo , Proteínas de la Membrana/genética , Monocitos/citología , Óxido Nítrico/metabolismo , Cultivo Primario de Células , Especies Reactivas de Oxígeno/metabolismo , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Receptores de IgG/genética , Receptores de IgG/metabolismo , Transducción de Señal , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Helminth parasite infections are associated with predominant Th2-type cytokine responses, and parasite glycoconjugates have been recognized as partially responsible for such immune bias. It has been proved that Echinococcus granulosus evokes a Th2-type cytokine pattern characterized by a high production of IL-4, IL-5, IL-6 and IL-10, and no or mild IFN-γ levels in animal models and in patients with cystic echinococcosis, respectively. Here, we show that E4(+) (a glycoconjugate-enriched fraction from E. granulosus protoscolex) stimulated the secretion of a high concentration of IL-6, followed by IL-10 and TNF-α by normal peritoneal B cells. We determined that E4(+) bound to the surface of peritoneal B cells and induced their activation and, also, triggered the differentiation of peritoneal B cells into IgM-, IgG2b- and IgG3-secreting cells in a T-independent way. Interestingly, the IgM released by E4(+) -stimulated peritoneal B cells from normal mice recognized protoscolex antigens. Results showed that, after the encounter with antigens from E. granulosus protoscolex, peritoneal B cells are a source of Th2-type cytokines and polyclonal antibodies, some of which recognize parasite antigens, suggesting that peritoneal B cells can condition the outcome of the infection.
Asunto(s)
Anticuerpos Antihelmínticos/biosíntesis , Linfocitos B/inmunología , Equinococosis/inmunología , Echinococcus granulosus/inmunología , Glicoconjugados/inmunología , Animales , Anticuerpos Antihelmínticos/inmunología , Antígenos Helmínticos/inmunología , Linfocitos B/metabolismo , Diferenciación Celular , Células Cultivadas , Equinococosis/parasitología , Echinococcus granulosus/metabolismo , Femenino , Interacciones Huésped-Parásitos , Interleucina-10/biosíntesis , Interleucina-10/inmunología , Interleucina-6/biosíntesis , Interleucina-6/inmunología , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Peritoneo/inmunología , Factor de Necrosis Tumoral alfa/biosíntesis , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Trypanosoma cruzi, the causative agent of Chagas' disease, may sabotage humoral response by affecting B cells at the different stages of its development. The present review highlights the contributions of our laboratory in understanding how T. cruzi hinders B-cell generation and B-cell expansion limiting host defence and favouring its chronic establishment. We discuss how homoeostatic mechanisms can be triggered to control exacerbated B-cell proliferation that favour T. cruzi infection by eliminating parasite-specific B cells. Specific targeting of evasion mechanisms displayed in T. cruzi infection, as in vivo Fas/FasL blockade or Gal-3 expression inhibition, allowed us to modulate B-cell responses enhancing the anti-parasite humoral immune response. A comprehensive understanding of the biology of the B cell in health and disease is strictly required to devise immunointervention strategies aimed at enhancing protective immune responses during infections.
Asunto(s)
Subgrupos de Linfocitos B/inmunología , Subgrupos de Linfocitos B/patología , Enfermedad de Chagas/inmunología , Enfermedad de Chagas/parasitología , Trypanosoma cruzi/crecimiento & desarrollo , Trypanosoma cruzi/inmunología , Animales , Subgrupos de Linfocitos B/parasitología , Diferenciación Celular/inmunología , Enfermedad de Chagas/patología , HumanosRESUMEN
During ageing, autoimmune disorders and the higher susceptibility to infectious have been associated with alterations in the humoral immune response. We report that splenic B lymphocytes from aged mice exhibit lower level of apoptosis induced by B-cell antigen receptor (BCR) ligation in vitro. Respect to B cells from young mice the anti-mu stimulated aged B cells show similar Bcl-2 and Bcl-xL expression but differential kinetic of A1 degradation and a higher level of cFLIP and FAIM. Even though B cells from aged mice show minor Fas expression they exhibit the same susceptibility to anti-Fas induced apoptosis. Aged B cells also present upon BCR stimulation, a higher proliferative response and similar level of activation markers expression than B cells from young mice. These data agree with the observation that aged mice exhibit an increment of T2 and mature B cell subset which rapidly enters cell cycle upon BCR engagement. The diminished apoptosis after activation in aged mice could compromise homeostatic mechanism allowing the persistence of self and non-self antigen specific B cells.
Asunto(s)
Envejecimiento/inmunología , Antígenos/inmunología , Subgrupos de Linfocitos B/patología , Receptores de Antígenos de Linfocitos B/inmunología , Transducción de Señal , Regulación hacia Arriba , Animales , Apoptosis , Proteínas Reguladoras de la Apoptosis/análisis , Biomarcadores/análisis , Western Blotting/métodos , Proteína Reguladora de Apoptosis Similar a CASP8 y FADD , Supervivencia Celular , Células Cultivadas , Electroforesis en Gel de Poliacrilamida , Femenino , Citometría de Flujo , Proteínas Inhibidoras de la Apoptosis/análisis , Péptidos y Proteínas de Señalización Intracelular/análisis , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Receptor fas/inmunologíaRESUMEN
Recent evidence has implicated galectins, a family of evolutionarily conserved carbohydrate-binding proteins, as regulators of immune cell homeostasis and host-pathogen interactions. Galectins operate at different levels of innate and adaptive immune responses, by modulating cell survival and cell activation or by influencing the Th1/Th2 cytokine balance. Furthermore, galectins may contribute to host-pathogen recognition and may serve as receptors for specific interactions of pathogens with their insect vectors. Here we will explore the influence of galectins in immunological processes relevant to microbial infection and will summarize exciting recent work related to the specific interactions between galectins and their glycoconjugate ligands as critical determinants of pathogen recognition. Understanding the role of galectin-sugar interactions during the course of microbial infections might contribute to defining novel targets for disease prevention and immune intervention.
Asunto(s)
Galectinas/inmunología , Infecciones/inmunología , Animales , Apoptosis , Supervivencia Celular , Citocinas/inmunología , Citocinas/metabolismo , Galectinas/química , Galectinas/metabolismo , Glicoconjugados/metabolismo , Homeostasis , Humanos , Inmunidad Innata , Insectos Vectores/inmunología , Activación de Linfocitos , Linfocitos/inmunologíaRESUMEN
In the present work, we demonstrate that interleukin (IL)-4 is able to rescue B cells from Trypanosoma cruzi-infected mice, counteracting the strong apoptotic signals that these cells received in vivo. We have observed that IL-4 restrains the apoptosis of immunoglobulin (Ig)M(+) and IgG(+) B cells from infected and normal mice without inducing them to proliferate. In addition, IL-4 does not modify the quantity or quality of the antibodies secreted by B cells from infected mice, as it blocks their terminal differentiation to plasma cells and favors memory pathway. It is interesting that the protective effect of IL-4 over B cells from infected mice is mediated, at least partly, by the down-regulation of Fas ligand (FasL) expression, which leads to interference in the apoptosis executed by these B cells through the Fas/FasL death pathway. Accordingly, a marked up-regulation of the "FasL gene repressor" class II transactivator was observed, suggesting that this would be one mechanism underlying the IL-4-mediated FasL down-regulation.
Asunto(s)
Apoptosis/efectos de los fármacos , Linfocitos B/parasitología , Interleucina-4/farmacología , Proteínas Nucleares , Trypanosoma cruzi , Animales , Linfocitos B/citología , Linfocitos B/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Enfermedad de Chagas , Proteína Ligando Fas , Regulación de la Expresión Génica , Genes MHC Clase II , Glicoproteínas de Membrana/efectos de los fármacos , Glicoproteínas de Membrana/fisiología , Ratones , Transactivadores/biosíntesis , Transactivadores/efectos de los fármacosRESUMEN
It has been proposed that Trypanosoma cruzi, the aetiologic agent of Chagas' disease, produces mitogenic substances responsible for the polyclonal B-cell activation observed during the acute phase of the infection. Isolation and characterization of the molecules involved in the induction of polyclonal activation observed during infectious diseases have posed a great challenge for the immunologist over the last decade. In this work we report that a 33 kD protein obtained from an alkaline fraction of T. cruzi epimastigotes (FI) stimulates proliferation and promotes differentiation into antibody-secreting cells of normal murine B cells in a T-cell independent manner. By flow cytometry we also found that the 33 kDa protein induces an increase in the expression of MHC class II and B7.2 but not B7.1 molecules on the B-cell surface. Sequencing by mass spectrometry identified the T. cruzi 33 kD protein as hypothetical oxidoreductase, a member of the aldo/ketoreductase family. In this report we demonstrate that this protein is also present in the infective bloodstream trypomastigote form of the parasite and was identified as T. cruzi mitochondrial malate dehydrogenase (mMDH) by enzyme activity and by Western blotting using a specific mMDH polyclonal antiserum. The biologic relevance of mMDH-induced polyclonal activation concerning T. cruzi infection is discussed.
Asunto(s)
Antígenos de Protozoos/inmunología , Linfocitos B/inmunología , Enfermedad de Chagas/inmunología , Activación de Linfocitos/inmunología , Malato Deshidrogenasa/inmunología , Trypanosoma cruzi/inmunología , Animales , Formación de Anticuerpos , Linfocitos B/parasitología , Ratones , Ratones Endogámicos BALB C , Mitocondrias/enzimología , Mitocondrias/inmunología , Trypanosoma cruzi/enzimología , Trypanosoma cruzi/ultraestructuraRESUMEN
Galectin-1 is a beta-galactoside-binding protein with potent anti-inflammatory and immunoregulatory effects. However, its expression and function have not been assessed in the context of an infectious disease. The present study documents, for the first time, the regulated expression of galectin-1 in the context of an infectious process and its influence in the modulation of macrophage microbicidal activity and survival. A biphasic modulation in parasite replication and cell viability was observed when macrophages isolated from Trypanosoma cruzi-infected mice were exposed to increasing concentrations of galectin-1. While low concentrations of this protein increased parasite replication and did not affect macrophage survival, higher inflammatory doses of galectin-1 were able to commit cells to apoptosis and inhibited parasite replication. Furthermore, galectin-1 at its lowest concentration was able to down-regulate critical mediators for parasite killing, such as interleukin 12 (IL-12) and nitric oxide, while it did not affect IL-10 secretion. Finally, endogenous galectin-1 was found to be up-regulated and secreted by the J774 macrophage cell line cultured in the presence of trypomastigotes. This result was extended in vivo by Western blot analysis, flow cytometry, and reverse transcription-PCR using macrophages isolated from T. cruzi-infected mice. This study documents the first association between galectin-1's immunoregulatory properties and its role in infection and provides new clues to the understanding of the mechanisms implicated in host-parasite interactions during Chagas' disease and other parasite infections.
Asunto(s)
Adyuvantes Inmunológicos , Hemaglutininas/genética , Macrófagos/inmunología , Trypanosoma cruzi/fisiología , Regulación hacia Arriba , Animales , Apoptosis/inmunología , Relación Dosis-Respuesta a Droga , Galectina 1 , Hemaglutininas/farmacología , Interleucina-10/biosíntesis , Interleucina-12/biosíntesis , Macrófagos/efectos de los fármacos , Macrófagos/parasitología , Ratones , Óxido Nítrico/biosíntesis , Conejos , Trypanosoma cruzi/crecimiento & desarrolloRESUMEN
Galectin-1 (GAL-1), a highly conserved beta-galactoside-binding protein, has shown immunomodulatory properties. In this study, we investigated the regulation of GAL-1 expression within the B-cell compartment using Trypanosoma cruzi infection as a natural model of in vivo B-cell activation. GAL-1 was found to be expressed on activated B cells from T. cruzi-infected mice, mainly localized at the cytosolic compartment. Expression of this protein was found to be modulated according to the activation state of the cells, revealing a significant increase in stimulated B cells that received signals via cross-linking of the B-cell receptor and CD40. It was found that GAL-1 was secreted by B cells to the extracellular milieu upon activation. Finally, purified GAL-1 produced by activated B cells induced apoptosis of T cells but not B cells and also influenced interferon-gamma cytokine production. Hence, the present study describes a potential mechanism by which B cells can regulate T-cell function and survival.
Asunto(s)
Apoptosis/inmunología , Linfocitos B/inmunología , Hemaglutininas/biosíntesis , Activación de Linfocitos/fisiología , Linfocitos T Reguladores/inmunología , Animales , Anticuerpos/inmunología , Anticuerpos/farmacología , Linfocitos B/efectos de los fármacos , Linfocitos B/metabolismo , Antígenos CD40/inmunología , Enfermedad de Chagas/inmunología , Enfermedad de Chagas/metabolismo , Citocinas/biosíntesis , Galectina 1 , Hemaglutininas/inmunología , Lipopolisacáridos/farmacología , Ratones , Linfocitos T Reguladores/citología , Linfocitos T Reguladores/metabolismo , Trypanosoma cruziRESUMEN
Human very-low-density lipoprotein (VLDL) inhibits DNA synthesis in lymphocytes activated by the nonspecific mitogen concanavalin A (Con A). We studied the effects of VLDL on lymphocyte activation (IL-2 receptor expression), cell cycle progression, and production of IL-2 and of IL-4 (a proinflammatory and an anti-inflammatory interleukin, respectively) to understand why an atherogenic lipoprotein inhibits cell proliferation. After 48 h of stimulation with the mitogen, VLDL decreased the population of cells bearing IL-2 receptor and the population of T-cells that progress through the cell cycle, increasing the population of T-cells in G(0)/G(1). Cells cultured in the presence of Con A and VLDL produced higher levels of IL-2 and lower levels of IL-4 than cells cultured without VLDL. These results suggest that VLDL inhibits lymphocyte proliferation by reducing IL-2 receptor and enhancing the levels of IL-2. Probably, one atherogenic effect of VLDL is to modulate the cytokine secretion profile of lymphocytes to a predominantly proinflammatory response.
Asunto(s)
Citocinas/biosíntesis , ADN/biosíntesis , Lipoproteínas VLDL/farmacología , Activación de Linfocitos/fisiología , Linfocitos T/inmunología , Animales , Células Cultivadas , Concanavalina A/farmacología , Citocinas/metabolismo , Citometría de Flujo , Humanos , Inmunofenotipificación , Inflamación , Interleucina-2/biosíntesis , Interleucina-4/biosíntesis , Lipoproteínas VLDL/sangre , Lipoproteínas VLDL/aislamiento & purificación , Activación de Linfocitos/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Albúmina Sérica/farmacología , Bazo/inmunología , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Timidina/metabolismoRESUMEN
Acute infection with Trypanosoma cruzi is characterized by multiple manifestations of immunosuppression of both cellular and humoral responses. B cells isolated at the acute stage of infection have shown marked impairment in their response to polyclonal activators in vitro. The present work aims at studying the B cell compartment in the context of acute T. cruzi infection to provide evidence for B cell activation, spontaneous apoptosis and arrest of the cell cycle upon mitogenic stimulation as a mechanism underlying B cell hyporesponse. We found that B cells from acutely infected mice, which fail to respond to the mitogen LPS, showed spontaneous proliferation and production of IgM, indicating a high level of B cell activation. Furthermore, these activated B cells also exhibited an increase in Fas expression and apoptosis in cultures without an exogenous stimulus. On the other hand, B cells from early acute and chronic infected mice did not present activation or apoptosis, and were able to respond properly to the mitogen. Upon in vitro stimulation with LPS, B cells from hyporesponder mice failed to progress through the cell cycle (G0/G1 arrest), nor did they increase the levels of apoptosis. These results indicate that B cell apoptosis and cell cycle arrest could be the mechanisms that control intense B cell expansion, but at the same time could be delaying the emergence of a specific immune response against the parasite.
Asunto(s)
Apoptosis/inmunología , Linfocitos B/inmunología , Enfermedad de Chagas/inmunología , Terapia de Inmunosupresión , Lipopolisacáridos/inmunología , Trypanosoma cruzi , Animales , Linfocitos B/patología , División Celular/inmunología , Enfermedad de Chagas/patología , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB CRESUMEN
The aim of this work was to study the antibody response of acute and chronic chagasic patients against a Trypanosoma cruzi alkaline fraction (FI) in comparison with the reactivity against a T. cruzi acidic antigen, the main cystein proteinase of the parasite named cruzipain, and "natural" antigens. FI-specific antibodies were detected only during the acute phase of the infection and IgM was the main isotype produced, whereas cruzipain-specific antibodies were detected during all phases of the infection. By means of immunoblot and sequencing analysis we identified a 47-kDa FI proteic band recognized by IgM from acute chagasic patients as the T. cruzi glutamate dehydrogenase (GluDH). Furthermore, the antibody response against isolated GluDH showed similar characteristics as the one against FI. We also observed a strict association between the reactivity of IgM against FI and GluDH and IgM natural antibodies. However, reactivity against these alkaline antigens was not modified after absorption of natural antibodies in sera from acute chagasic patients, indicating that these parasite antigens are not recognized by the polyspecific natural antibodies. The most important goal of this report is that for the first time the T. cruzi antigen isoelectric point has been associated with its ability to trigger immunological memory, raising a novel antigen property that should be considered in the selection of antigens used in Chagas' disease diagnostic test and in the design of a vaccine against T. cruzi infection.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Enfermedad de Chagas/sangre , Trypanosoma cruzi/inmunología , Enfermedad Aguda , Secuencia de Aminoácidos , Animales , Antígenos de Protozoos/sangre , Autoanticuerpos/sangre , Niño , Preescolar , Enfermedad Crónica , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoglobulina M/inmunología , LactanteRESUMEN
We have previously reported that a cytosolic alkaline fraction (FI) obtained from epimastigotes of Trypanosoma cruzi promotes the activation, proliferation and differentiation of normal murine B cells into antibody-secreting plasmocytes. Neither the mechanism nor the cells involved in the FI-induced polyclonal B-cell activation were established. In this work we report that accessory cells are required for FI-induced polyclonal B-cell activation as no proliferative responses were obtained following treatment of normal spleen mononuclear cells (NSMC) with L-leucine methyl ester. Furthermore, FI did not induce the expression of CD25 on T cells and it promoted the proliferation of a T-cell-depleted population, indicating that it acts in a T-independent manner. We observed that NSMC were stimulated in vitro by FI-released cytokines, such as interleukin (IL)-4, IL-6 and IL-10, which are involved in B-cell proliferation and differentiation. Interestingly, while significant amounts of interferon-gamma (IFN-gamma) were found in culture supernatants we did not observe detectable levels of IL-2. Additionally, we found that B-cell receptor (BCR) and major histocompatibility complex (MHC) class II antigens were involved in the proliferative response induced by FI because antibodies directed against cell-surface immunoglobulin M (IgM), CD45 and MHC class II molecules inhibited the FI-induced B-cell proliferation. CD40 ligand (CD40L) did not participate in such a phenomenon.
Asunto(s)
Antígenos de Protozoos/inmunología , Linfocitos B/inmunología , Receptores de Antígenos de Linfocitos B/inmunología , Linfocitos T/inmunología , Trypanosoma cruzi/inmunología , Animales , Ligando de CD40 , División Celular , Antígenos de Histocompatibilidad Clase II/inmunología , Cadenas mu de Inmunoglobulina/inmunología , Antígenos Comunes de Leucocito/inmunología , Activación de Linfocitos/inmunología , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos BALB C , Receptores de Interleucina-2/biosíntesis , Bazo/citología , Bazo/inmunologíaRESUMEN
This paper shows that human antibodies specific for exoantigens of pI 4.5 (Eas 4.5), released by the blood forms of the parasite, obtained from chagasic patients sera by immunoabsorption react with cruzipain, the major cysteinyl proteinase of Trypanosoma cruzi. Sera from mice immunized with Eas 4.5 also recognize cruzipain. In addition, mouse antisera to cruzipain were reactive with Eas 4.5 as well as with total antigens excreted by culture-trypomastigotes. This reactivity was inhibited by cruzipain as revealed by enzyme-linked immunosorbent assay (ELISA). Furthermore, it was observed by immunoblot that the exoantigens recognized by mouse antisera to cruzipain have molecular weights between 50 and 60 kDa and human antibodies specific for Eas 4.5 recognize cruzipain with apparent molecular weight of 50 kDa. These findings suggest the presence of cruzipain in Eas and the subsequent release of this enzyme by the parasite.
Asunto(s)
Anticuerpos Antiprotozoarios/análisis , Enfermedad de Chagas/inmunología , Cisteína Endopeptidasas/inmunología , Trypanosoma cruzi/inmunología , Animales , Western Blotting , Enfermedad de Chagas/sangre , Ensayo de Inmunoadsorción Enzimática , Humanos , Técnicas de Inmunoadsorción , Ratones , Proteínas ProtozoariasRESUMEN
Several reports have described polyclonal activation in mice acutely infected with Trypanosoma cruzi. The aim of this work was to analyse the participation of one T. cruzi antigenic fraction in this immunological event. The antigen selected was FI, an antigenic fraction of pI 7-9 obtained from T. cruzi cytosol separated by isoelectricfocusing. FI is constituted by molecules with molecular weights of around 60 and 20 KDa. The authors assayed the ability of this antigenic fraction to induce polyclonal activation of spleen mononuclear cells from normal (NSMC) BALB/c mice. NSMC showed a marked lymphoproliferative response measured by 3H-thymidine incorporation after 3 days of culture in presence of FI. The values reached by FI-stimulated cells were 10 times higher than the controls (non-stimulated cells). This effect was dose-dependent. Furthermore, the authors observed that a purified T-cell population in the presence of adherent cells was unaffected by FI. Additionally, in a culture of NSMC, FI stimulated the proliferation of B cells as observed by the increase of the percentage of B220+ cells determined by FACS using FITC-conjugated anti-mouse B220. The authors noticed that the percentage of B220+Ly1+(CD5) populations in the presence of FI did not change with respect to the control (non-stimulated cells), indicating that FI expanded both conventional and CD5+ B cells. The isotypic pattern of the antibodies produced after 6 days of culture of NSMC in the presence of FI was predominantly IgM, which reacted with highly conserved antigens such as actin, myosin, myoglobin, thyroglobulin and carbonic anhydrase, but did not react with FI. A slight increase of IgG1 and IgG3 with respect to the control was observed but no changes on the levels of IgG2 was noticed. These results indicate that FI promotes activation, proliferation and differentiation in antibody-secreting cells of normal murine B lymphocytes.
Asunto(s)
Antígenos de Protozoos/inmunología , Linfocitos B/inmunología , Activación de Linfocitos , Trypanosoma cruzi/inmunología , Animales , Formación de Anticuerpos , Ratones , Ratones Endogámicos BALB C , Linfocitos T/inmunologíaRESUMEN
The purpose of these studies was to analyze the role of different immune cell populations in the immune response against Trypanosoma cruzi antigens in aged mice. Mice of different ages (3 and 12 months old) were immunized i.d. with S-105 plus Bordetella pertussis as adjuvant and we compared the activities of the lymph node cells taken from 3- and 12-month-old donor animals to transfer DTH or antibody production to 3-month-old recipients. This study revealed that adherent and non-adherent immune lymph node cells of aged donor animals did not transfer response against the foreign antigen (S-105) whereas 3-month-old non-adherent lymph node cells transferred a DTH response as well as helped the specific antibody production. When total lymph node cells from 3- and 12-month-old mice were mixed, we observed an inhibition of S-105 transferred response indicating a suppressive effect of aged cells on the 3-month-old mice cells. Furthermore, we analyzed the participation of antigen-presenting cells (APC) in the immune response changes related to the previously described aged mice. Peritoneal cavities cells (PC), pulsed in vivo with S-105, obtained from 3- and 12-month-old mice were transferred to normal recipients and a DTH response to S-105 was studied. We observed that the DTH response was lower in the recipients of aged PC with respect to recipients of young PC. The results suggest that APC from aged mice are involved in controlling the cellular immune response to S-105. Age-related changes in immune T cell and APC are discussed in the context of these observations.
Asunto(s)
Envejecimiento/fisiología , Células Presentadoras de Antígenos/fisiología , Antígenos de Protozoos/inmunología , Inmunización , Ganglios Linfáticos/fisiología , Trypanosoma cruzi/inmunología , Traslado Adoptivo , Animales , Formación de Anticuerpos , Femenino , Ganglios Linfáticos/citología , Masculino , Ratones , Ratones Endogámicos BALB CRESUMEN
Infection with Trypanosoma cruzi is characterized by hyporesponsiveness of the immune system during the acute phase of infection. To better understand the immunological mechanisms affected by T. cruzi, we studied if a reduced T cell proliferative response could originate from an inability of T cells to proliferate or a functional deficiency at the level of accessory cells (AC). The inhibitory effect exerted by T. cruzi was during the induction phase of the lymphoproliferative response, suggesting the participation of AC in the hyporesponse. Then we further investigated the potential of the parasite to interfere with accessory cell-dependent and -independent pathways of human T cell proliferation. Peripheral blood mononuclear cells and peripheral blood lymphocytes from healthy individuals, enriched for T cells, were analysed with regard to their proliferative capacity using: phytohaemagglutinin, immobilized anti-CD3 monoclonal antibody (MoAb) and MoAb to the CD28 antigen, anti-CD3 MoAb and recombinant IL-2 and anti-CD3 MoAb plus phorbol myristate acetate in the presence of parasites. Significant suppression of the proliferative response was caused by the parasite only when AC were present. The parasite markedly reduced the surface expression of HLA-DR and CD11b antigens, key molecules in PHA-induced proliferation. Addition of indomethacin to the culture failed to reverse the inhibitory effect of the parasites, suggesting that prostaglandin E2 was not involved. These data suggest that AC in contact with T. cruzi become incompetent as antigen presenting cell because they are unable to induce a normal proliferative response in T lymphocytes.
Asunto(s)
Células Presentadoras de Antígenos/inmunología , Activación de Linfocitos , Linfocitos T/inmunología , Trypanosoma cruzi/inmunología , Animales , Comunicación Celular/inmunología , Enfermedad de Chagas/inmunología , Inhibidores de la Ciclooxigenasa/farmacología , Antígenos HLA-DR/metabolismo , Interacciones Huésped-Parásitos/inmunología , Humanos , Tolerancia Inmunológica/efectos de los fármacos , Técnicas In Vitro , Indometacina/farmacología , Activación de Linfocitos/efectos de los fármacos , Antígeno de Macrófago-1/metabolismo , Ratones , Ratones Endogámicos BALB CRESUMEN
Exoantigens of pI 4.5 (Ea 4.5) of T. cruzi released to the circulation of infected mice are able to induce partial protective immune response in mice (F. Cerbán et al. 1991, International Archives of Allergy and Applied Immunology 96, 35-40). In order to analyze the participation of cellular immunity in the parasitemia control, we i.d. immunized mice with Ea 4.5 plus Bordetella pertussis as adjuvant. The role of immune cells in protective immunity was examined by adoptive transfer experiments. The immune lymph node cells (LNC) transferred the capacity to control the parasitemia, since it was observed that the normal recipients of immune LNC, which were afterward infected, presented a significant decrease in parasite levels with respect to the animals receiving LNC from control mice. This capacity was absent in the spleen cells. In addition, polystyrene nonadherent cells from immune LNC transferred the capacity to control T. cruzi infection. It was observed that Ig+ cells and enriched T cells from immunized mice are able to control the parasitemia. To define epitopes of Ea 4.5 able to stimulate protective immunity, the levels of parasitemia were examined in mice immunized with Ea 4.5 untreated or treated with sodium metaperiodate. These animals presented similar levels of parasitemia and in both cases they are significantly lower than the parasitemias of the control animals, suggesting that the most relevant epitopes for the protective immune response that control the beginning of the infection are not carbohydrates. Later, on Day 30 postinfection only the animals immunized with untreated Ea 4.5 maintained a significant decrease in parasite levels.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Antígenos de Protozoos/inmunología , Enfermedad de Chagas/prevención & control , Inmunoterapia Adoptiva , Ganglios Linfáticos/inmunología , Trypanosoma cruzi/inmunología , Animales , Linfocitos B/inmunología , Enfermedad de Chagas/inmunología , Epítopos/inmunología , Inmunidad Celular , Inmunización , Ganglios Linfáticos/citología , Ratones , Ratones Endogámicos BALB C , Linfocitos T/inmunologíaRESUMEN
This paper shows that a polyclonal monospecific rabbit antiserum to cruzipain, the major T. cruzi cystein proteinase, cross-reacts with a cytosol acidic antigen (F IV) isolated from the epimastigote stage of the same parasite. In addition, antibodies specific for F IV purified from chagasic patient sera or murine anti F IV sera, also react with cruzipain. This was demonstrated by ELISA, DOT-ELISA, native and electrophoretic Immunoblot. These findings suggest that F IV contains an antigen immunologically cross-reactive with cruzipain.