RESUMO
Successful sperm maturation and storage rely on a unique immunological balance that protects the male reproductive organs from invading pathogens and spermatozoa from a destructive autoimmune response. We previously characterized one subset of mononuclear phagocytes (MPs) in the murine epididymis, CX3CR1+ cells, emphasizing their different functional properties. This population partially overlaps with another subset of understudied heterogeneous MPs, the CD11c+ cells. In the present study, we analyzed the CD11c+ MPs for their immune phenotype, morphology, and antigen capturing and presenting abilities. Epididymides from CD11c-EYFP mice, which express enhanced yellow fluorescent protein (EYFP) in CD11c+ MPs, were divided into initial segment (IS), caput/corpus, and cauda regions. Flow cytometry analysis showed that CD11c+ MPs with a macrophage phenotype (CD64+ and F4/80+) were the most abundant in the IS, whereas those with a dendritic cell signature [CD64- major histocompatibility complex class II (MHCII)+] were more frequent in the cauda. Immunofluorescence revealed morphological and phenotypic differences between CD11c+ MPs in the regions examined. To assess the ability of CD11c+ cells to take up antigens, CD11c-EYFP mice were injected intravenously with ovalbumin. In the IS, MPs expressing macrophage markers were most active in taking up the antigens. A functional antigen-presenting coculture study was performed, whereby CD4+ T cells were activated after ovalbumin presentation by CD11c+ epididymal MPs. The results demonstrated that CD11c+ MPs in all regions were capable of capturing and presenting antigens. Together, this study defines a marked regional variation in epididymal antigen-presenting cells that could help us understand fertility and contraception but also has larger implications in inflammation and disease pathology.
Assuntos
Antígeno CD11c/metabolismo , Epididimo/metabolismo , Fagócitos/metabolismo , Animais , Biomarcadores/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Células Dendríticas/metabolismo , Fertilidade/fisiologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Transgênicos , Fenótipo , Espermatozoides/metabolismoRESUMO
Leptin, an adipose-secreted hormone, links metabolism and immunity. Our aim was to determine whether leptin affects the alloimmune response. We used an allogeneic skin transplant model as a means to analyze the allograft immune response in Lep(ob/ob) and wild-type mice. Leptin deficiency results in an increased frequency of Treg and Th2 cells and a prolonged graft survival. These effects of leptin deficiency indicate the importance of leptin and obesity in modulating the allograft immune responses. Our data suggest a possible explanation for the increased susceptibility of hyperleptinemic obese patients to acute and chronic graft rejection.
Assuntos
Sobrevivência de Enxerto/fisiologia , Leptina/fisiologia , Células Th2/imunologia , Animais , Citometria de Fluxo , Teste de Cultura Mista de Linfócitos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase , Transplante HomólogoRESUMO
In a recent workshop organized by the JDRF focused on the 'Identification and Utilization of Robust Biomarkers in Type1 Diabetes', leaders in the field of type 1 diabetes (T1D)/autoimmunity and assay technology came together from academia, government and industry to assess the current state of the field, evaluate available resources/technologies and identify gaps that need to be filled for moving the field of T1D research forward. The highlights of this workshop are discussed in this paper, as well as the proposal for a larger, planned consortium effort, incorporating a JDRF Biomarker Core, to foster collaboration and accelerate progress in this critically needed area of T1D research.
Assuntos
Autoimunidade/imunologia , Biomarcadores/análise , Diabetes Mellitus Tipo 1/imunologia , Humanos , Linfócitos T/imunologiaAssuntos
Autoantígenos , Diabetes Mellitus Tipo 1/diagnóstico , Lúpus Eritematoso Sistêmico/diagnóstico , Esclerose Múltipla/diagnóstico , Febre Reumática/diagnóstico , Autoantígenos/imunologia , Autoantígenos/isolamento & purificação , Linfócitos B/imunologia , Linfócitos B/patologia , Líquidos Corporais/imunologia , Técnicas de Laboratório Clínico/métodos , Diabetes Mellitus Tipo 1/imunologia , Humanos , Lúpus Eritematoso Sistêmico/imunologia , Esclerose Múltipla/imunologia , Febre Reumática/imunologia , Linfócitos T/imunologia , Linfócitos T/patologiaRESUMO
Dendritic cells (DCs) are professional antigen-presenting cells that control the generation of adaptive immunity. Consequently, DCs have a central role in the induction of protective immunity to pathogens and also in the pathogenic immune response responsible for the development and progression of autoimmune disorders. Thus the study of the molecular pathways that control DC development and function is likely to result in new strategies for the therapeutic manipulation of the immune response. In this review, we discuss the role and therapeutic value of DCs in autoimmune diseases, with a special focus on multiple sclerosis.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Autoimunidade , Sistema Nervoso Central/imunologia , Células Dendríticas/imunologia , Esclerose Múltipla/imunologia , Imunidade Adaptativa , Doenças Autoimunes/imunologia , Células Dendríticas/classificação , Humanos , Interleucina-27/imunologia , VacinaçãoRESUMO
OBJECTIVE: Multiple sclerosis (MS) is characterized by the local production of antibodies in the CNS and the presence of oligoclonal bands in the CSF. Antigen arrays allow the study of antibody reactivity against a large number of antigens using small volumes of fluid with greater sensitivity than ELISA. We investigated whether there were unique autoantibodies in the CSF of patients with MS as measured by antigen arrays and whether these antibodies differed from those in serum. METHODS: We used antigen arrays to analyze the reactivity of antibodies in matched serum and CSF samples of 20 patients with untreated relapsing-remitting MS (RRMS), 26 methylprednisolone-treated patients with RRMS, and 20 control patients with other noninflammatory neurologic conditions (ONDs) against 334 different antigens including heat shock proteins, lipids, and myelin antigens. RESULTS: We found different antibody signatures in matched CSF and serum samples The targets of these antibodies included epitopes of the myelin antigens CNP, MBP, MOBP, MOG, and PLP (59%), HSP60 and HSP70 (38%), and the 68-kD neurofilament (3%). The antibody response in patients with MS was heterogeneous; CSF antibodies in individual patients reacted with different autoantigens. These autoantibodies were locally synthesized in the CNS and were of the immunoglobulin G class. Finally, we found that treatment with steroids decreased autoantibody reactivity, epitope spreading, and intrathecal autoantibody synthesis. CONCLUSIONS: These studies provide a new avenue to investigate the local antibody response in the CNS, which may serve as a biomarker to monitor both disease progression and response to therapy in MS.
Assuntos
Autoanticorpos/imunologia , Esclerose Múltipla Recidivante-Remitente/imunologia , Análise Serial de Proteínas , Adulto , Autoanticorpos/sangue , Autoanticorpos/líquido cefalorraquidiano , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla Recidivante-Remitente/sangue , Esclerose Múltipla Recidivante-Remitente/líquido cefalorraquidiano , Bainha de Mielina/imunologia , Bandas Oligoclonais/sangue , Bandas Oligoclonais/líquido cefalorraquidiano , Bandas Oligoclonais/imunologiaRESUMO
BACKGROUND: Natalizumab is an antibody directed against integrin alpha4 that reduces disease activity in patients with multiple sclerosis (MS) by blocking migration of T and B cells into the CNS. The goal of this study was to characterize the effects of natalizumab treatment on cytokine production and expression of activation markers, costimulatory molecules, and trafficking determinants on CD4+ and CD8+ T cells. METHODS: In a longitudinal study, we investigated the expression of surface makers and cytokine expression on peripheral blood lymphocytes from 28 patients with MS who started natalizumab treatment and were followed for 1 year. A mixed effects model was used to compare pretreatment to on-treatment measurements. RESULTS: The frequency of CD4+ T cells producing interferon-gamma, tumor necrosis factor, and interleukin (IL)-17 upon anti-CD3 stimulation increased 6 months after initiation of natalizumab treatment and remained elevated throughout the follow-up. The frequency of CD4+ T cells expressing CD25, HLA-DR, and CCR6 ex vivo was increased at one or more time points during treatment. Among CD8+ T cells, the frequency of cells producing IL-2 and IL-17 after stimulation was increased during natalizumab treatment, as was the frequency of CD8+ T cells expressing CD58 and CCR5 ex vivo. The increase in the frequency of activated cells could not be replicated by in vitro exposure to natalizumab. CONCLUSION: Natalizumab treatment increases the percentage of activated leukocytes producing proinflammatory cytokines in blood, presumably due to sequestration of activated cells in the peripheral circulation.
Assuntos
Anticorpos Monoclonais/farmacologia , Inibição de Migração Celular/efeitos dos fármacos , Inflamação/tratamento farmacológico , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Linfócitos T/efeitos dos fármacos , Adulto , Anticorpos Monoclonais Humanizados , Antígenos de Superfície/efeitos dos fármacos , Antígenos de Superfície/metabolismo , Biomarcadores/análise , Biomarcadores/sangue , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Inibição de Migração Celular/imunologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Quimiotaxia de Leucócito/imunologia , Citocinas/sangue , Citocinas/efeitos dos fármacos , Citocinas/metabolismo , Feminino , Humanos , Imunossupressores/farmacologia , Inflamação/imunologia , Inflamação/fisiopatologia , Estudos Longitudinais , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla Recidivante-Remitente/sangue , Esclerose Múltipla Recidivante-Remitente/imunologia , Natalizumab , Linfócitos T/imunologia , Linfócitos T/metabolismo , Adulto JovemRESUMO
T-cell vaccination (TCV) controls pathogenic autoimmune T-cell responses via two different regulatory cell populations: anti-idiotypic and anti-ergotypic T cells. Anti-idiotypic T cells recognize clone-specific determinants, like the CDR3 region of the T-cell receptor. Anti-ergotypic T cells recognize antigenic determinants derived from activation markers, which are upregulated by activated T cells, like CD25. In this review, we analyse the different components of the anti-ergotypic response: (1) the target T cells, which can be CD8+ or CD4+ T cells that express TCRalphabeta or TCRgammadelta; (2) the ergotope, which can be a T cell-restricted ergotope not expressed by other cell types or a widely expressed, shared ergotope and (3) the anti-ergotypic T cells, which are detectable in the naive immune system, but whose numbers can be expanded during the induction of an immune response against, or as a result of TCV or specific, anti-ergotypic vaccination. Finally, we discuss possible interactions between anti-ergotypic regulators and other regulatory T cells. We propose that the expression of major histocompatibility complex class II molecules by regulatory CD4+CD25+ T cells may make possible the cross-regulation of anti-ergotypic and CD4+CD25+ regulatory T cells, fine-tuning immunoregulation in the mature immune system.
Assuntos
Autoantígenos/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T/imunologia , Animais , Epitopos de Linfócito T/fisiologia , Humanos , Ativação Linfocitária , Camundongos , Ratos , Receptores de Antígenos de Linfócitos T alfa-beta/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Subpopulações de Linfócitos T/metabolismo , Linfócitos T/metabolismo , VacinaçãoRESUMO
Traditionally, immunologic diagnosis has been based on an attempt to correlate each disease with a specific immune reactivity, such as an antibody or a T-cell response to a single antigen specific for the disease entity. The state of the body, however, appears to be encoded by the immune system in collectives of reactivities and not by single reactivities. Here we describe our use of microarray technology and informatics to develop an antigen chip capable of detecting global patterns of antibodies binding to hundreds of antigens simultaneously. The patterns fashion diagnostic signatures.
Assuntos
Antígenos/imunologia , Análise Serial de Proteínas/métodos , Animais , Antígenos/metabolismo , Autoanticorpos/imunologia , Autoanticorpos/metabolismo , Biologia Computacional/métodos , Humanos , Linfócitos T/imunologiaRESUMO
Autoantibodies are commonly found in healthy individuals and strains of mice that are not prone to autoimmunity. The present study was undertaken to identify self antigens recognized by serum autoantibodies from unimmunized mice of two strains: NOD mice prone to spontaneously develop autoimmune diabetes and C57BL/6 mice known to be relatively resistant to autoimmune disease. IgM and IgG autoantibodies detected in the sera of NOD and C57BL/6 mice manifested different patterns of reactivity. The IgM autoantibodies from C57BL/6 serum reacted with more self antigens and showed higher OD values than the IgM autoantibodies from NOD mice. In contrast, the IgG autoantibodies from NOD serum reacted with more antigens and displayed higher OD readings than did IgG autoantibodies from C57BL/6 mice. Among the antigens recognized by the autoantibodies, particularly of the IgG class, were self antigens known to induce experimental autoimmune diseases in NOD and C57BL/6 mice. In addition, IgG autoantibodies from NOD mice reacted with self antigens reported to mark the spontaneous autoimmune diabetes that characterizes this strain of mice. These results suggest that naturally occurring IgG autoantibodies reflect susceptibility to induction of specific autoimmune diseases. In addition, the results suggest that IgM autoantibodies may by associated with mechanisms that might prevent autoimmune disease.
Assuntos
Autoanticorpos/sangue , Diabetes Mellitus Tipo 1/imunologia , Camundongos Endogâmicos C57BL/imunologia , Camundongos Endogâmicos NOD/imunologia , Animais , Autoanticorpos/biossíntese , Diabetes Mellitus Tipo 1/genética , Feminino , Predisposição Genética para Doença , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Camundongos , Camundongos Endogâmicos NOD/genéticaRESUMO
Nonobese diabetic (NOD) mice develop insulitis and diabetes through a process involving autoimmunity to the 60-kDa heat shock protein (HSP60). Treatment of NOD mice with HSP60 or with peptides derived from HSP60 inhibits this diabetogenic process. We now report that NOD diabetes can be inhibited by vaccination with a DNA construct encoding human HSP60, with the pcDNA3 empty vector, or with an oligonucleotide containing the CpG motif. Prevention of diabetes was associated with a decrease in the degree of insulitis and with down-regulation of spontaneous proliferative T cell responses to HSP60 and its peptide p277. Moreover, both the pcDNA3 vector and the CpG oligonucleotide induced specific Abs, primarily of the IgG2b isotype, to HSP60 and p277, and not to other islet Ags (glutamic acid decarboxylase or insulin) or to an unrelated recombinant Ag expressed in bacteria (GST). The IgG2b isotype of the specific Abs together with the decrease in T cell proliferative responses indicate a shift of the autoimmune process to a Th2 type in treated mice. These results suggest that immunostimulation by bacterial DNA motifs can modulate spontaneous HSP60 autoimmunity and inhibit NOD diabetes.