RESUMO
Desalination of seawater is becoming an important means to address the increasing scarcity of freshwater resources in the world. Seawater has been used as drinking water in the health, food, and medical fields and various beneficial effects have been suggested, although not confirmed. Given the presence of 63 minerals and trace elements in drinking desalinated seawater (63 DSW), we evaluated their effects on the behavior of tumorigenic and nontumorigenic cells through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and annexin-V-fluorescein isothiocyanate/propidium iodide staining. Our results showed that cell viability and proliferation in the presence of 63 DSW were significantly greater than in mineral water and in the presence of fetal bovine serum in a dose-dependent manner. Furthermore, 63 DSW showed no toxic effect on murine embryonic fibroblast (NIH-3T3) and murine melanoma (B16-F10) cells. In another assay, we also showed that pre-treatment of non-adherent THP-1 cells with 63 DSW reduces apoptosis incidence, suggesting a protective effect against cell death. We conclude that cell viability and proliferation were improved by the mineral components of 63 DSW and this effect can guide further studies on health effects associated with DSW consumption.
Assuntos
Água Potável/química , Minerais/farmacologia , Água do Mar/análise , Purificação da Água , Animais , Anexina A5/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Corantes/química , Fluoresceína-5-Isotiocianato/química , Camundongos , Células NIH 3T3 , Propídio/química , Água do Mar/química , Coloração e Rotulagem , Sais de Tetrazólio/química , Tiazóis/químicaRESUMO
The participation of B-1 cells in a murine model of spontaneous diabetes has been recently reported. Here, we describe the role of B-1 cells in streptozotocin (STZ) induced diabetes in mice. We demonstrated that XID (B-1 cell-deficient) mice are more susceptible to STZ treatment than WT mice, as evidenced by their higher blood glucose level in response to STZ. Unexpectedly, the XID mice that were i.p. transferred with purified B-1 cells, either before or after the STZ treatment, did not develop diabetes. These cell transfers provided long-lasting protection for the XID mice against STZ-induced diabetes, suggesting that B-1 cells play an important role in the experimental diabetes pathobiology. We also showed that B-1 cell culture supernatants were able to regulate the blood glucose level of the diabetic XID mice, and we identified insulin-producing cells when B-1 cells were differentiated in B-1 cell-derived phagocyte in vitro. These findings provide a novel role for B-1 cells in metabolic processes, presenting a new mechanism to explain the pathogenesis of diabetes and a possible therapeutical target.
Assuntos
Subpopulações de Linfócitos B/imunologia , Subpopulações de Linfócitos B/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/prevenção & controle , Insulina/biossíntese , Transferência Adotiva , Tirosina Quinase da Agamaglobulinemia , Animais , Subpopulações de Linfócitos B/efeitos dos fármacos , Glicemia/metabolismo , Diabetes Mellitus Experimental/etiologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Pâncreas/efeitos dos fármacos , Pâncreas/metabolismo , Pâncreas/patologia , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/imunologia , Estreptozocina/administração & dosagem , Estreptozocina/toxicidadeRESUMO
B-1 lymphocytes comprise a unique subset of B cells that differ phenotypically, ontogenetically and functionally from conventional B-2 cells. A frequent specificity of the antibody repertoire of peritoneal B-1 cells is phosphatidylcholine. Liposomes containing phosphatidylcholine have been studied as adjuvants and their interaction with dendritic cells and macrophages has been demonstrated. However, the role of B-1 cells in the adjuvanticity of liposomes composed of phosphatidylcholine has not been explored. In the present work, we studied the contribution of B-1 cells to the humoral response against ovalbumin (OVA) encapsulated into dipalmitoylphosphatidylcholine (DPPC) and cholesterol-containing liposomes. BALB/X-linked immunodeficient (xid) mice, which are deficient in B-1 cells, showed quantitative and qualitative differences in the anti-OVA antibody response compared with wild-type animals after immunization with these liposomes. The OVA-specific immune response was significantly increased in the BALB/xid mice when reconstituted with B-1 cells from naive BALB/c mice. Our results indicate the internalization of DPPC-containing liposomes by these cells and their migration from the peritoneal cavity to the spleen. Phosphatidylcholine significantly contributed to the immunogenicity of liposomes, as DPPC-containing liposomes more effectively stimulated the anti-OVA response compared with vesicles composed of dipalmitoylphosphatidylglycerol. In conclusion, we present evidence for a cognate interaction between B-1 cells and phosphatidylcholine liposomes, modulating the immune response to encapsulated antigens. This provides a novel targeting approach to assess the role of B-1 cells in humoral immunity.
Assuntos
Antígenos/imunologia , Subpopulações de Linfócitos B/imunologia , Adjuvantes Imunológicos , Animais , Anticorpos/imunologia , Formação de Anticorpos/imunologia , Especificidade de Anticorpos , Antígenos/química , Subpopulações de Linfócitos B/metabolismo , Movimento Celular , Feminino , Imunização , Lipossomos , Camundongos , Ovalbumina/imunologia , Fosfatidilcolinas/química , Fosfatidilcolinas/imunologia , Baço/imunologiaRESUMO
Experimental autoimmune encephalomyelitis (EAE) has been widely employed as a model to study multiple sclerosis (MS) and indeed has allowed some important advances in our comprehension of MS pathogenesis. Several pieces of evidence suggest that infiltrating Th1 and Th17 lymphocytes are important players leading to CNS demyelination and lesion during the peak of murine EAE. Subsequently, effector T cell responses rapidly decline and the recovery phase of the disease strongly correlates with the expression of anti-inflammatory cytokines and the enrichment of Foxp3+ regulatory T (Treg) cells within the target organ. However, the mechanisms leading to the increased presence of Treg cells and to the remission phase of the disease are still poorly understood. Recent researches demonstrated that chemically induced amino-acid starvation response might suppress CNS immune activity. Here we verified an important participation of the general control nonrepressible 2 (GCN2), a key regulator kinase of the amino-acid starvation response, in the development of the remission phase of EAE in C57BL/6 mice. By immunizing wild type C57BL/6 (WT) and GCN2 knock-out mice (GCN2 KO) with myelin oligodendrocyte glycoprotein peptide (MOG35-55), it was noticed that GCN2 KO mice did not develop the remission phase of the disease and this was associated with higher levels of CNS inflammation and increased presence of effector T cells (Th1/Th17). These animals also showed lower frequency of Treg cells within the CNS as compared to WT animals. Higher expression of indoleamine 2,3-dioxygenase (IDO) and higher frequency of plasmacytoid dendritic cells (pDCs) were found at the peak of the disease in the CNS of WT animals. Our results suggest that the GCN2 kinase-dependent sensing of IDO activity represents an important trigger to the EAE remission phase. The IDO-mediated immunoregulatory events may include the arresting of effector T cell responses and the differentiation/expansion of Treg cells within the target organ.
Assuntos
Encefalomielite Autoimune Experimental/enzimologia , Proteínas Serina-Treonina Quinases/fisiologia , Animais , Citocinas/metabolismo , Encefalomielite Autoimune Experimental/imunologia , Fatores de Transcrição Forkhead/metabolismo , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas Serina-Treonina Quinases/genética , Remissão Espontânea , Medula Espinal/patologia , Células Th1/metabolismo , Células Th17/metabolismoRESUMO
New Zealand Black X New Zealand White F1 [(NZB/NZW)F1] mice develop an autoimmune condition with similarities to human systemic lupus erythematosus (SLE). In this study, we demonstrate that B-1 cells, which have previously been reported to be involved in several autoimmune diseases, have altered gene expression in these mice. RNA was extracted from purified B-1 cells of disease-free C57BL/6 mice and lupus-prone (NZB/NZW)F1 mice. Gene expression was analysed using DNA microarray techniques and validated by real time reverse transcriptase polymerase chain reaction (RT-PCR). In (NZB/NZW)F1 mice, some genes had altered expression patterns compared to disease-free controls. Specifically, the upregulation of Ifitm1, Pvrl2 and Ifi202b and downregulation of Trp53bp1 mRNA were observed in (NZB/NZW)F1 mice. These genes are known to be associated with autoimmune diseases. This pattern of gene expression in B-1 cells could understanding of the pathogenesis of SLE. Thus, it is reasonable to hypothesise that the altered gene expression observed in B-1 cells in our experimental model is important for SLE prognosis and therapy, and these implications are discussed herein.
Assuntos
Linfócitos B/imunologia , Lúpus Eritematoso Sistêmico/genética , Animais , Linfócitos T CD4-Positivos/imunologia , Modelos Animais de Doenças , Feminino , Perfilação da Expressão Gênica , Lúpus Eritematoso Sistêmico/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Análise de Sequência com Séries de OligonucleotídeosRESUMO
The protein S100A9 plays a key role in the control of inflammatory response. The C-terminus of the murine S100A9 protein (mS100A9p) downregulates the spreading and phagocytic activity of adherent peritoneal cells. Murine peritoneal cells are constituted by macrophages and B-1 cells, and the latter exert an inhibitory effect on macrophage functions by secreting interleukin- (IL-) 10. Here, we investigated the influence of B-1 cells on the inhibitory effect evoked by mS100A9p on macrophages. mS100A9p did not alter spreading and phagocytosis either by peritoneal macrophages obtained from mice deprived of B-1 cells or by bone marrow-derived macrophages (BMDMÏ). Nevertheless, when BMDMÏ were cocultivated by direct or indirect contact with B-1 cells treated with mS100A9p, the phagocytosis by BMDMÏ was decreased, showing that the effect of mS100A9p on macrophages was modulated by B-1 cells and/or their secretory compounds. Furthermore, the inhibitory action of mS100A9p on phagocytosis by adherent peritoneal cells was abolished in cells obtained from IL-10 knockout mice. Taken together, the results show that mS100A9p has no direct inhibitory effect on macrophages; however, mS100A9p modulates B-1 cells, which in turn downregulates macrophages, at least in part, via IL-10. These data contribute to the characterization of S100A9 functions involving B-1 cells in the regulation of the inflammatory process.
Assuntos
Calgranulina B/química , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Peptídeos/farmacologia , Animais , Subpopulações de Linfócitos B/efeitos dos fármacos , Subpopulações de Linfócitos B/metabolismo , Linhagem Celular , Células Cultivadas , Interleucina-10/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Peptídeos/química , Fagocitose/efeitos dos fármacosRESUMO
Paracoccidioidomycosis (PCM) is a systemic granulomatous disease, endemic in Latin America, caused by the thermal dimorphic fungus Paracoccidioides brasiliensis. Although some fungal antigens have already been characterized and used for serological diagnosis, cross-reactions have been frequently observed. Thus, the examination of fungal forms in clinical specimens or isolation of P. brasiliensis by culture is still the most frequent method for the diagnosis of this mycosis. In this study, a random peptide phage display library was used to select mimotopes of P. brasiliensis, which were employed as antigens in an indirect enzyme-linked immunosorbent assay. The protective monoclonal antibody against experimental PCM (anti-gp75) was used as molecular target to screen a phage display library. That approach led to a synthetic peptide named P2, which was synthesized and tested against PCM patients' sera to check whether it was recognized. There was significant recognition of P2 by sera of untreated PCM patients when compared with normal human sera. Sera from treated PCM group, patients with other mycosis or co-infected with HIV had much lower recognition of P2 than untreated patient group. The test showed a sensitivity of 100 and 94.59% of specificity in relation to human sera control. These data indicate a potential use of P2 as diagnostic tool in PCM. Its application for serological diagnosis of PCM may contribute to the development and standardization of simpler, faster and highly reproducible immunodiagnostic tests at low cost.
Assuntos
Anticorpos Antifúngicos/sangue , Micologia/métodos , Paracoccidioidomicose/diagnóstico , Peptídeos , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Biblioteca de Peptídeos , Peptídeos/isolamento & purificação , Sensibilidade e Especificidade , Testes Sorológicos/métodosRESUMO
Glycoprotein gp70 is an important intracellular antigen from Paracoccidioides brasiliensis that elicits both humoral and cellular immune responses. Herein, the PbGP70 gene cloning from isolate Pb18 using internal peptide sequence information is reported. The deduced protein sequence bears two N-glycosylation sites, antigenic sites and two mouse T-cell epitopes. Anti-recombinant gp70 (rPbgp70) polyclonal antibodies reacted with a 70-kDa component in total cell extract of P. brasiliensis, while MAbC5F11 and paracoccidioidomycosis patients' sera recognized rPbgp70. Confocal microscopy with anti-rPbgp70 and MAbC5F11 showed intense staining and cytoplasmatic co-localization. The protein sequence belongs to the flavoprotein monooxygenase family which groups important anti-oxidative bioactive compounds. We found increased PbGP70 transcript accumulation under oxidative stress induced by H(2)O(2), during fungal growth and in macrophage phagocyted/bound yeasts. Therefore, gp70 might play a dual role in P. brasiliensis by both eliciting immune cellular and humoral responses in the host and protecting the fungus from oxidative stress generated by phagocytic cells.
Assuntos
Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Paracoccidioides/enzimologia , Paracoccidioides/genética , Sequência de Aminoácidos , Animais , Southern Blotting , Células Cultivadas , Flavoproteínas/metabolismo , Proteínas Fúngicas/química , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Regulação Fúngica da Expressão Gênica/genética , Glicoproteínas/química , Glicoproteínas/genética , Glicoproteínas/metabolismo , Peróxido de Hidrogênio/farmacologia , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Paracoccidioides/efeitos dos fármacos , Paracoccidioides/crescimento & desenvolvimento , Homologia de Sequência de AminoácidosRESUMO
We previously demonstrated that B-1b cells can undergo differentiation to acquire a mononuclear phagocyte phenotype upon attachment to substrate in vitro. Here we followed the expression of surface markers and transcription factors during this differentiation. B-1b cells spontaneously express both myeloid and lymphoid restricted transcription factors. When induced to differentiate into a phagocyte, the lymphoid genes E box protein (E2A), early B-cell factor (EBF), paired box 5 (Pax5) are down-modulated, while expression of genes related to myeloid commitment is sustained. Furthermore, B-1b cell-derived phagocytes (B-1CDPs) decrease immunoglobulin M (IgM) expression but retain the expression of the heavy chain variable gene VH11 or VH12, an immunoglobulin gene rearrangement predominantly expressed by B-1 cells. The maintenance of lymphoid characteristics in B-1CDPs characterizes a unique type of phagocyte, not related to monocyte-derived macrophages.
Assuntos
Subpopulações de Linfócitos B/imunologia , Linfócitos B/imunologia , Fagócitos/imunologia , Fatores de Transcrição/metabolismo , Animais , Subpopulações de Linfócitos B/metabolismo , Linfócitos B/metabolismo , Diferenciação Celular/imunologia , Células Cultivadas , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica , Imunoglobulina M/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células Mieloides/imunologia , Fagocitose/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Baço/imunologiaRESUMO
Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by Paracoccidioides brasiliensis (Pb), a thermal dimorphic fungus. Its major antigen is a 43-kDa glycoprotein. Gp43 embodies different functions: it participates in evasion mechanisms during the installation of primary infection, stimulates granuloma-like formation in vitro and presents T-cell epitopes that induce protective response against the fungus. Here, we investigated epitopes from gp43 inhibitory of both, macrophage functions and inflammatory reaction. Different gp43 peptides, spanning the entire sequence of the molecule, were added to cultures of bone marrow-derived macrophages. After challenge with zymosan or Pb cells, phagocytic indexes were measured. Peptides expressed on the molecule surface were determined by graphic analysis using the Protean module; DNAstar Inc. Two peptides which decreased phagocytic index and were expressed at the surface of the molecule, P4 and P23, were selected for further studies. It was shown that both inhibited the release of NO by zymosan stimulated macrophages while enhanced release of H(2)O(2). The release of TNF-alpha in culture supernatants from in vitro phagocytic tests showed different response depending of P4 concentration (data not shown). In vivo assays with Mycobacterium bovis - bacillus Calmette-Guérin (BCG) or Pb cells demonstrated that these peptides presented non-specific and specific anti-inflammatory properties.
Assuntos
Antígenos de Fungos/química , Proteínas Fúngicas/química , Glicoproteínas/química , Macrófagos/efeitos dos fármacos , Paracoccidioides/imunologia , Peptídeos/farmacologia , Animais , Antígenos de Fungos/farmacologia , Células Cultivadas , Feminino , Proteínas Fúngicas/farmacologia , Glicoproteínas/farmacologia , Inflamação/tratamento farmacológico , Macrófagos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Paracoccidioides/química , Paracoccidioidomicose/microbiologia , Peptídeos/síntese química , Peptídeos/química , Fagocitose/efeitos dos fármacosRESUMO
Characterization of the origin, properties, functions and fate of cells is a fundamental task for the understanding of physiological and pathological phenomena. Despite the bulk of knowledge concerning the diverse characteristics of mammalian cells, some of them, such as B-1 cells, are still poorly understood. Here we report the results obtained in our laboratory on these cells in the last 10 years. After showing that B-1 cells could be cultured and amplified in vitro, a series of experiments were performed with these cells. They showed that B1 cells reside mostly in the peritoneal and pleural cavities, migrate to distant inflammatory foci, coalesce to form giant cells and participate in granuloma formation, both in vitro and in vivo. They are also able to present antigens to immunologically responsive cells and are endowed with regulatory properties. Further, we have also shown that these cells facilitate different types of infection as well as tumor growth and spreading. These data are presently reviewed pointing to a pivotal role that these cells may play in innate and acquired immunity.
Assuntos
Linfócitos B/imunologia , Granuloma/imunologia , Inflamação/imunologia , Neoplasias/imunologia , Fagócitos/imunologia , Animais , Células Apresentadoras de Antígenos/imunologia , Linfócitos B/citologia , Imunidade Celular , CamundongosRESUMO
BACKGROUND: Diseases caused by melanized fungi include mycetoma, chromoblastomycosis and phaeohyphomycosis. This broad clinical spectrum depends on the dynamic interactions between etiologic agent and host. The immune status of the host influences on the development of the disease, as, an exemple. phaeohyphomicosis is more frequently observed in immunocompromised patients. OBJECTIVES: Examine the histological inflammatory response induced by Fonsecaea pedrosoi in several different strains of mice (BALB/c, C57BL/6, Nude and SCID, and reconstituted Nude). METHODS: Fonsecaea pedrosoi was cultivated on agar gel and a fragment of this gel was implanted subcutaneously in the abdominal region of female adult mice. After infection has been obtained, tissue fragment was studied histopathologically. RESULTS: There were significant changes across the strains, with the nodular lesion more persistent in Nude and SCID mice, whereas in immunocompetent mice the lesion progressed to ulceration and healing. The histopathological analysis showed a significant acute inflammatory reaction which consisted mainly of neutrophils in the initial phase that was subsequently followed by a tuberculoid type granuloma in immunocompetent mice. STUDY LIMITATIONS: There is no a suitable animal model for chromoblastomycosis. CONCLUSIONS: The neutrophilic infiltration had an important role in the containment of infection to prevent fungal spreading, including in immunodeficient mice. The fungal elimination was dependent on T lymphocytes. The re-exposure of C57BL/6 mice to Fonsecaea pedrosoi caused a delay in resolving the infection, and appearance of muriform cells, which may indicate that re-exposure to fungi, might lead to chronicity of infection.
Assuntos
Ascomicetos , Dermatomicoses/imunologia , Imunocompetência , Inflamação/imunologia , Inflamação/microbiologia , Animais , Contagem de Células Sanguíneas , Cromoblastomicose/imunologia , Cromoblastomicose/patologia , Doença Crônica , Dermatomicoses/patologia , Modelos Animais de Doenças , Feminino , Inflamação/patologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Nus , Camundongos SCID , Neutrófilos , Especificidade da Espécie , Fatores de TempoRESUMO
Here, we have investigated the possible effect of B-1 cells on the activity of peritoneal macrophages in E. cuniculi infection. In the presence of B-1 cells, peritoneal macrophages had an M1 profile with showed increased phagocytic capacity and index, associated with the intense microbicidal activity and a higher percentage of apoptotic death. The absence of B-1 cells was associated with a predominance of the M2 macrophages, reduced phagocytic capacity and index and microbicidal activity, increased pro-inflammatory and anti-inflammatory cytokines production, and higher percentual of necrosis death. In addition, in the M2 macrophages, spore of phagocytic E. cuniculi with polar tubular extrusion was observed, which is an important mechanism of evasion of the immune response. The results showed the importance of B-1 cells in the modulation of macrophage function against E. cuniculi infection, increasing microbicidal activity, and reducing the fungal mechanisms involved in the evasion of the immune response.
Assuntos
Subpopulações de Linfócitos B , Encephalitozoon cuniculi/imunologia , Encefalitozoonose/imunologia , Encefalitozoonose/patologia , Macrófagos Peritoneais/imunologia , Animais , Apoptose , Células Cultivadas , Feminino , Macrófagos Peritoneais/microbiologia , Camundongos Endogâmicos BALB C , Fagocitose/imunologia , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/imunologia , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/genéticaRESUMO
The role of natural killer (NK) T cells in the development of lupus-like disease in mice is still controversial. We treated NZB/W mice with anti-NK1.1 monoclonal antibodies (mAbs) and our results revealed that administration of either an irrelevant immunoglobulin G2a (IgG2a) mAb or an IgG2a anti-NK1.1 mAb increased the production of anti-dsDNA antibodies in young NZB/W mice. However, the continuous administration of an anti-NK1.1 mAb protected aged NZB/W mice from glomerular injury, leading to prolonged survival and stabilization of the proteinuria. Conversely, the administration of the control IgG2a mAb led to an aggravation of the lupus-like disease. Augmented titres of anti-dsDNA in NZB/W mice, upon IgG2a administration, correlated with the production of BAFF/BLyS by dendritic, B and T cells. Treatment with an anti-NK1.1 mAb reduced the levels of interleukin-16, produced by T cells, in spleen cell culture supernatants from aged NZB/W. Adoptive transfer of NK T cells from aged to young NZB/W accelerated the production of anti-dsDNA in recipient NZB/W mice, suggesting that NK T cells from aged NZB/W are endowed with a B-cell helper activity. In vitro studies, using purified NK T cells from aged NZB/W, showed that these cells provided helper B-cell activity for the production of anti-dsDNA. We concluded that NK T cells are involved in the progression of lupus-like disease in mature NZB/W mice and that immunoglobulin of the IgG2a isotype has an enhancing effect on antibody synthesis due to the induction of BAFF/BLyS, and therefore have a deleterious effect in the NZB/W mouse physiology.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Fator Ativador de Células B/biossíntese , Imunoglobulina G/uso terapêutico , Nefrite Lúpica/prevenção & controle , Envelhecimento/imunologia , Animais , Anticorpos Antinucleares/biossíntese , Anticorpos Monoclonais/imunologia , Antígenos Ly/imunologia , Células Cultivadas , Progressão da Doença , Feminino , Imunoglobulina G/imunologia , Interleucina-16/biossíntese , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/transplante , Lipopolissacarídeos/imunologia , Fígado/imunologia , Nefrite Lúpica/imunologia , Camundongos , Camundongos Endogâmicos , Subfamília B de Receptores Semelhantes a Lectina de Células NK/imunologia , Índice de Gravidade de Doença , Baço/imunologia , Linfócitos T Auxiliares-Indutores/imunologiaRESUMO
Increasing evidence indicates that tumors require a constant influx of myelomonocytic cells to support their malignant behavior. This is caused by tumor-derived factors, which recruit and induce functional differentiation of myelomonocytic cells, most of which are macrophages. Although myeloid lineages are the classical precursors of macrophages, B-lymphoid lineages such as B-1 cells, a subset of B-lymphocytes found predominantly in pleural and peritoneal cavities, are also able to migrate to inflammatory sites and differentiate into mononuclear phagocytes exhibiting macrophage-like phenotypes. Here we examined the interplay of B-1 cells and tumor cells, and checked whether this interaction provides signals to influence melanoma cells metastases. Using in vitro coculture experiments we showed that B16, a murine melanoma cell line, and B-1 cells physically interact. Moreover, interaction of B16 with B-1 cells leads to up-regulation of metastasis-related gene expression (MMP-9 and CXCR-4), increasing its metastatic potential, as revealed by experimental metastases assays in vivo. We also provide evidence that B16 cells exhibit markedly up-regulated phosphorylation of the extracellular signal-regulated kinase (ERK) when cocultured with B-1 cells. Inhibition of ERK phosphorylation induced by B-1 cells with inhibitors of MEK1/2 strongly suppressed the induction of MMP-9 and CXCR-4 mRNA expression and impaired the increased metastatic behavior of B16. In addition, constitutive levels of ERK1/2 phosphorylation in B-1 cells are necessary for their commitment to affect the metastatic potential of B16 cells. Our findings show for the first time that B-1 lymphocytes can contribute to tumor cell properties required for invasiveness during metastatic spread.
Assuntos
Linfócitos B/fisiologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Sistema de Sinalização das MAP Quinases , Melanoma/enzimologia , Animais , Feminino , Melanoma/patologia , Camundongos , Camundongos Endogâmicos BALB C , Metástase Neoplásica , Fosforilação , Células Tumorais Cultivadas , Regulação para CimaRESUMO
Protective immunity in paracoccidioidomycosis is mainly mediated by cellular immunity. The role of B cells in this disease, in particular B-1 cells, is poorly understood. The aim of this study was to characterize the participation of B-1 cells in resistance or susceptibility of BALB/c and BALB/Xid mice to P. brasiliensis (Pb) pulmonary infection. BALB/Xid, which lacks B-1 cells, exhibited higher resistance to infection when compared with BALB/c mice. However, adoptive transfer of B-1 cells to BALB/Xid mice drastically increased the susceptibility of these animals to Pb infection. The fungal burden in BALB/c and B-1-reconstituted BALB/Xid was significantly higher as compared to BALB/Xid strain. Compact, well-organized granulomas were observed in the lungs of BALB/Xid mice, whereas large lesions with necrotic center with a plethora of fungi developed in BALB/c mice. It was also shown that B-1 cells impair phagocytosis of Pb by macrophages in vitro via secretion of IL-10, which was increased upon stimulation with a purified Pb antigen, gp43. Finally, in vivo blockade of IL-10 led to a better control of infection by the highly susceptible B10.A mouse. These findings suggest that B-1 cells play a major role in resistance/susceptibility to Pb infection in murine models, most likely via production of IL-10.
Assuntos
Linfócitos B/imunologia , Interleucina-10/imunologia , Paracoccidioides/imunologia , Paracoccidioidomicose/imunologia , Transferência Adotiva , Animais , Contagem de Colônia Microbiana , Suscetibilidade a Doenças , Granuloma/microbiologia , Granuloma/patologia , Interleucina-10/antagonistas & inibidores , Interleucina-10/metabolismo , Pulmão/microbiologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fagocitose/imunologia , Pneumonia/imunologia , Pneumonia/microbiologiaRESUMO
Paracoccidioides brasiliensis is a thermo-dimorphic fungus responsible for paracoccidioidomycosis (PCM), a systemic granulomatous mycosis prevalent in Latin America. The fungus releases many antigens which may be transiently bound to its cell surface. Some of them may show enzymatic functions essential for maintaining many cell processes and survival of the microorganism at different conditions. In this study, we report the characterization of a secreted 75kDa protein from P. brasiliensis with phosphatase activity. Biologic function of the molecule was demonstrated using two specific mAbs produced and characterized as IgM and IgG isotypes. Confocal microscopy and flow cytometry analysis demonstrated that both mAbs recognized the protein on the fungus surface, mainly in its budding sites. In vitro experiments showed that fungal growth was inhibited by blocking the protein with mAbs. In addition, opsonized yeast cells with both mAbs facilitated phagocytosis by murine peritoneal macrophages. Passive immunization using mAbs before P. brasiliensis mice infection reduced colony-forming units (CFU) in the lungs as compared with controls. Histopathology showed smaller inflammation, absence of yeast cells and no granuloma formation.
Assuntos
Fosfatase Ácida/imunologia , Anticorpos Monoclonais/imunologia , Proteínas Fúngicas/imunologia , Imunização Passiva , Proteínas de Membrana/imunologia , Paracoccidioides/crescimento & desenvolvimento , Paracoccidioidomicose/imunologia , Fosfatase Ácida/metabolismo , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/biossíntese , Antígenos de Fungos/imunologia , Antígenos de Fungos/metabolismo , Proteínas Fúngicas/metabolismo , Macrófagos Peritoneais/microbiologia , Masculino , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Paracoccidioides/imunologia , Paracoccidioidomicose/microbiologia , Paracoccidioidomicose/prevenção & controle , FagocitoseRESUMO
Since B-1 cells were first described, their origin and function remain controversial. Given the ability to produce natural antibodies and large amounts of IL-10, there is a consensus about their role in innate immunity. More recently, however, B-1 cells have been associated to adaptive immunity as well, due to the demonstration of immunological memory and antigen presentation capability. Here we demonstrate that adoptive transfer of pre-sensitized B-1b cells (obtained from OVA-sensitized mice) to naïve B-1 deficient animals, drastically affects the ability of transplanted animals to mount an adaptive response upon immunization with OVA. In contrast to naïve B-1 populated mice, mice transplanted with sensitized B-1 exhibit lower anti-OVA antibody levels, milder footpad swelling in response to OVA subcutaneous injection and reduced granulomatous reaction to OVA-coated beads. Moreover, we show that these pre-sensitized B-1 cells, when acting as APCs, induce poor T cell proliferation in vitro when compared with macrophages or B-1 cells obtained from naïve mice. This property may be due in part to insufficient expression of the co-stimulatory molecule CD86, necessary for optimal antigen presentation. In conclusion, our data suggest a novel role for B-1 cells as part of suppressor mechanisms in the immune system.
Assuntos
Subpopulações de Linfócitos B/imunologia , Hipersensibilidade/imunologia , Tolerância Imunológica , Linfócitos T/imunologia , Transferência Adotiva , Animais , Subpopulações de Linfócitos B/citologia , Subpopulações de Linfócitos B/transplante , Proliferação de Células , Imunidade Ativa , Imunidade Inata , Imunização , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Mutantes , Ovalbumina/imunologiaRESUMO
Encephalitozoon cuniculi is an opportunist intracellular pathogen of mammals. The adaptive immune response is essential to eliminate E. cuniculi, but evidence is mounting that the response initiated by the innate immune response may ultimately define whether or not the parasite can survive. B-1 cells may act as antigen-presenting cells or differentiate into phagocytes, playing different roles in many infection models. However, the role of these cells in the dynamics of Encephalitozoon sp. infections is still unknown. To investigate the role of B-1 cells in E. cuniculi infection, BALB/c and BALB/c XID (B-1 cells deficient) mice were infected with E. cuniculi spores. Cytometric analyses of peritoneal cells showed that B-1 cells and macrophages increased significantly in infected BALB/c mice compared to uninfected controls. Despite the increase in the number of CD4+ and CD8+ lymphocytes in XID mice, these animals were more susceptible to infection as evidenced histologically with more prominent inflammatory lesions and parasite burden. Pro-inflammatory cytokines increased in both infected BALB/c and BALB/c XID mice. To confirm B-1 cells role in encephalitozoonosis, we adoptively transferred B-1 cells to BALB/c XID mice and this group showed few symptoms and microscopic lesions, associated with an increased in cytokines. Together, these results suggest that B-1 cells may increase the resistance of BALB/c mice to encephalitozoonosis, evidencing for the first time the important role of B-1 lymphocytes in the control of microsporidia infection.
Assuntos
Subpopulações de Linfócitos B/imunologia , Subpopulações de Linfócitos B/metabolismo , Suscetibilidade a Doenças , Encefalitozoonose/imunologia , Encefalitozoonose/metabolismo , Interações Hospedeiro-Patógeno/imunologia , Animais , Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Encephalitozoon cuniculi/imunologia , Encefalitozoonose/microbiologia , Encefalitozoonose/patologia , Feminino , Contagem de Linfócitos , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismoRESUMO
B-1 cells are a subtype of B cells with peculiar characteristics. These cells are distinct from B-2 lymphocytes in their morphology, ontogeny, tissue distribution, and phenotypic functional features. B-1 cells can participate in the immune response in several ways, for example, by being recruited to inflammatory foci, producing large amounts of IL-10 cytokine, and differentiating into IgM-secreting cells or phagocytes. Nevertheless, the role of B-1 cells in the pathogenesis of experimental leishmaniasis has not been fully elucidated. Here we evaluated the role of B-1 cells in Leishmania ( L.) amazonensis infection using X-linked immunodeficient (XID) mice that possess a mutation in Bruton's tyrosine kinase (Btk) that leads to a reduced number of B-1 cells. The course of infection and the corresponding immune response were analyzed in infected mice. XID mice showed an increase in parasite number in paws, lymph nodes, and spleen compared to BALB/c infected controls. Infected XID mice had higher IL-10 levels and lower anti- Leishmania IgM. The adoptive transfer of peritoneal B-1 cells into XID mice restored peritoneal B-1 cells and parasite burden in the footpad in a pattern similar to that observed in the BALB/c controls at 10 wk. Our results demonstrate the higher susceptibility of XID mice to infection with L. ( L.) amazonensis compared to controls. In addition, we show that the presence of B-1 cells contributes to improved animal resistance to parasites, suggesting that these cells are involved in the control of cutaneous infection caused by L. ( L.) amazonensis.