RESUMO
The objective of this study was to evaluate the frequency of CD4+ T cell subsets in peripheral blood mononuclear cells (PBMC), urine and renal tissue from patients with lupus nephritis (LN). PBMC and urinary cells were collected from 17 patients with active LN, 20 disease controls (DC) with primary glomerulonephritis and 10 healthy controls (HC) and were analysed by flow cytometry with markers for T helper type 1 (Th1), Th2, Th17 and regulatory T cells (Treg ) cells. T cell subsets were assessed by immunohistochemistry from LN biopsy specimens from 12 LN patients. T cell subtypes in PBMC were re-evaluated at 6 months of therapy. CD4+ T cells were decreased in PBMC in LN compared with DC and HC (P = 0·0001). No differences were observed in urinary CD4+ T cell subsets between LN and DC. The frequency of urinary Th17 cells was higher in patients with non-proliferative than in proliferative LN (P = 0·041). CD3+ and T-box 21 ( Tbet+) cells were found in glomeruli and interstitium of LN patients, while forkhead box protein 3 (FoxP3), retinoid-related orphan receptor gamma (ROR-γ) and GATA binding protein 3 (GATA-3) were present only in glomeruli. Th1 cells in PBMC were correlated negatively with urinary Th1 cells (Rho = -0·531; P = 0·028) and with Tbet in renal interstitium (Rho = -0·782; P = 0·004). At 6 months, LN patients showed an increase in Th17 cells in PBMC. In conclusion, the inverse association between Th1 cells from PBMC and urinary/renal tissue indicate a role for Th1 in LN pathophysiology. Urinary Th17 cells were associated with less severe LN, and Th17 increased in PBMC during therapy. Urinary CD4+ T cells were not different between LN and DC.
Assuntos
Nefrite Lúpica/imunologia , Nefrite Lúpica/metabolismo , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Adulto , Biomarcadores , Estudos de Casos e Controles , Feminino , Fator de Transcrição GATA3/metabolismo , Humanos , Imuno-Histoquímica , Imunofenotipagem , Nefrite Lúpica/patologia , Nefrite Lúpica/terapia , Contagem de Linfócitos , Masculino , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/patologia , Linfócitos T Auxiliares-Indutores/patologia , Linfócitos T Reguladores/patologia , Células Th1/imunologia , Células Th1/metabolismo , Células Th2/imunologia , Células Th2/metabolismoRESUMO
PURPOSE: To study the levels of pathogenic and non-pathogenic Th17 and Th22 cells in autoimmune thyroid disorders patients. Although Th17 cells seem to play an important role in the pathogenesis of thyroid autoimmune disorders, the specific subsets of these lymphocytes have not been analyzed in this condition. METHODS: We assessed the levels of Th17 (pathogenic and non-pathogenic) and Th22 cells in peripheral blood and thyroid glands of autoimmune thyroid disorders patients (n = 26, 16 with Graves' disease and 10 with Hashimoto's thyroiditis) and 15 healthy controls by multi-parametric flow cytometry and immunofluorescence microscopy. RESULTS: We found increased levels of pathogenic Th17 lymphocytes and Th22 cells in peripheral blood from autoimmune thyroid disorders patients. In addition, these cells were detected in thyroid glands from HT patients. Furthermore, we found significant correlations between the levels of these cells and disease activity, disease duration, and the presence of ophthalmopathy. CONCLUSIONS: The increased levels of pathogenic Th17 lymphocytes and Th22 cells in autoimmune thyroid disorders suggest their involvement in the pathogenesis of this condition.
Assuntos
Autoimunidade , Doença de Graves/imunologia , Doença de Hashimoto/imunologia , Linfocitose/etiologia , Linfócitos T Auxiliares-Indutores/imunologia , Células Th17/imunologia , Glândula Tireoide/imunologia , Adulto , Autoanticorpos/análise , Células Cultivadas , Feminino , Citometria de Fluxo , Doença de Graves/metabolismo , Doença de Graves/patologia , Doença de Graves/fisiopatologia , Oftalmopatia de Graves/etiologia , Doença de Hashimoto/metabolismo , Doença de Hashimoto/patologia , Doença de Hashimoto/fisiopatologia , Humanos , Iodo/urina , Ativação Linfocitária , Contagem de Linfócitos , Masculino , Microscopia de Fluorescência , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Linfócitos T Auxiliares-Indutores/metabolismo , Linfócitos T Auxiliares-Indutores/patologia , Células Th17/metabolismo , Células Th17/patologia , Glândula Tireoide/metabolismo , Glândula Tireoide/patologia , Glândula Tireoide/fisiopatologia , Hormônios Tireóideos/sangueRESUMO
CD4+ T follicular helper cells (TFH) were assessed in adult patients with common variable immune deficiency (CVID) classified according to the presence of granulomatous disease (GD), autoimmunity (AI), or both GD and AI (Group I) or the absence of AI and GD (Group II). TFH lymphocytes were characterized by expression of CXCR5 and PD-1. TFH were higher (in both absolute number and percentage) in Group I than in Group II CVID patients and normal controls (N). Within CXCR5+CD4+ T cells, the percentage of PD-1 (+) was higher and that of CCR7 (+) was lower in Group I than in Group II and N. The percentages of Treg and TFH reg were similar in both CVID groups and in N. TFH responded to stimulation increasing the expression of the costimulatory molecules CD40L and ICOS as did N. After submitogenic PHA+IL-2 stimulation, intracellular expression of TFH cytokines (IL-10, IL-21) was higher than N in Group I, and IL-4 was higher than N in Group II. These results suggest that TFH are functional in CVID and highlight the association of increased circulating TFH with AI and GD manifestations.
Assuntos
Imunodeficiência de Variável Comum/imunologia , Regulação da Expressão Gênica/imunologia , Doença Granulomatosa Crônica/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Reguladores/imunologia , Adulto , Idoso , Autoimunidade , Ligante de CD40/genética , Ligante de CD40/imunologia , Estudos de Casos e Controles , Imunodeficiência de Variável Comum/complicações , Imunodeficiência de Variável Comum/genética , Imunodeficiência de Variável Comum/patologia , Feminino , Doença Granulomatosa Crônica/complicações , Doença Granulomatosa Crônica/genética , Doença Granulomatosa Crônica/patologia , Humanos , Proteína Coestimuladora de Linfócitos T Induzíveis/genética , Proteína Coestimuladora de Linfócitos T Induzíveis/imunologia , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-2/farmacologia , Interleucina-4/genética , Interleucina-4/imunologia , Interleucinas/genética , Interleucinas/imunologia , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Fito-Hemaglutininas/farmacologia , Cultura Primária de Células , Receptor de Morte Celular Programada 1/genética , Receptor de Morte Celular Programada 1/imunologia , Receptores CCR7/genética , Receptores CCR7/imunologia , Receptores CXCR5/genética , Receptores CXCR5/imunologia , Transdução de Sinais , Linfócitos T Auxiliares-Indutores/efeitos dos fármacos , Linfócitos T Auxiliares-Indutores/patologia , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/patologiaRESUMO
We conducted a genome-wide scan for visceral leishmaniasis in mixed-breed dogs from a highly endemic area in Brazil using 149,648 single nucleotide polymorphism (SNP) markers genotyped in 20 cases and 28 controls. Using a mixed model approach, we found two candidate loci on canine autosomes 1 and 2. The positional association on chromosome 2 mapped to a predicted DNAse sensitive site in CD14+ monocytes that serve as a cis-regulatory element for the expression of interleukin alpha receptors 2 (IL2RA) and 15 (IL15RA). Both interleukins were previously found to lead to protective T helper 1 cell (Th1) response against Leishmania spp. in humans and mice. The associated marker on chromosome 1 was located between two predicted transcription factor binding sites regulating the expression of the transducin-like enhancer of split 1 gene (TLE1), an important player in Notch signaling. This pathway is critical for macrophage activity and CD4+ T cell differentiation into Th1 and T helper 2. Together, these findings suggest that the human and mouse model for protective response against Leishmania spp., which involves Th1 and macrophage modulation by interleukins 2, 15, gamma interferon and Notch signaling, may also hold for the canine model.
Assuntos
Estudo de Associação Genômica Ampla , Subunidade alfa de Receptor de Interleucina-2/genética , Leishmaniose Visceral/genética , Receptores de Interleucina-15/genética , Animais , Brasil , Linfócitos T CD4-Positivos/metabolismo , Diferenciação Celular/genética , Proteínas Correpressoras , Cães , Genótipo , Humanos , Leishmania infantum/patogenicidade , Leishmaniose Visceral/patologia , Leishmaniose Visceral/veterinária , Polimorfismo de Nucleotídeo Único , Receptores Notch/genética , Proteínas Repressoras/genética , Transdução de Sinais , Linfócitos T Auxiliares-Indutores/metabolismo , Linfócitos T Auxiliares-Indutores/patologiaRESUMO
BACKGROUND: Allergic asthma is a chronic pulmonary disease characterized by a Th2 inflammatory response. The modulation of a Th2 immune response based on immune deviation to a Th1 pattern or induction and migration of regulatory T cells to the lungs constitutes one of the major therapeutic approaches that is being investigated for the treatment of allergic asthma. The potentials of Mycobacterium leprae 65-kD heat-shock protein or Toll-like receptor 9 ligand (CpG oligodeoxynucleotides) as immune modulators for the treatment of airway allergic disease have been studied individually. OBJECTIVE: Mycobacterial protein combined with CpG was used as immunotherapy for airway allergy. METHODS: Using an ovalbumin-induced asthma model, mice were sensitized and challenged, and then treated with mycobacterial heat-shock protein (Hsp65) combined with CpG. RESULTS: The treatment of mice with established allergy led to the attenuation of eosinophilia, Th2 cytokines and airway hyperresponsiveness. Hsp65 plus CpG treatment also induced an increase in OVA-specific IFN-γ levels and in the frequency of lung inflammatory monocytes. Moreover, we show that the reduction of eosinophilia and the recruitment of inflammatory monocytes to the lungs required early triggering of TLR9, IFN-γ and CCR2 by immunotherapy components. CONCLUSION: In addition to immune deviation to a Th1 response in the modulation of Th2 allergic inflammation, our findings also attribute an important role to the innate response mediated by TLR9, associated with the recruitment of CCR2-dependent monocytes. CLINICAL RELEVANCE: Our findings show that the Hsp65/CpG treatment is a promising strategy for consideration in translational studies.
Assuntos
Asma/tratamento farmacológico , Proteínas de Bactérias/farmacologia , Chaperonina 60/farmacologia , Interferon gama/imunologia , Mycobacterium leprae , Oligodesoxirribonucleotídeos/farmacologia , Receptores CCR2/imunologia , Transdução de Sinais/efeitos dos fármacos , Receptor Toll-Like 9/imunologia , Animais , Asma/genética , Asma/imunologia , Imunoterapia , Interferon gama/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Receptores CCR2/genética , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/patologia , Receptor Toll-Like 9/genéticaRESUMO
Angioimmunoblastic T-cell lymphoma (AITL) is frequently associated with skin lesions, but epidermotropic cutaneous involvement has never been described. A 37-year-old man presented with erythematous and pruriginous plaques, clinically suggestive of mycosis fungoides, distributed all over the body, 3 weeks after the last line of a polychemotherapy, given for an AITL diagnosed 1 year earlier on a lymph node biopsy. Skin biopsy showed an epidermotropic CD4(+) T-cell lymphoma, so that a diagnosis of mycosis fungoides was first proposed. Further investigations showed that atypical lymphocytes strongly expressed CD10 and markers of follicular helper T cells (T(FH) ) including PD1, BCL-6 and CXCL13. The diagnosis of an unusual epidermotropic cutaneous localization of the AITL was finally made, supported by the presence of the same T-cell clone in the initial lymph node biopsy and the skin. We therefore recommend performing markers of T(FH) cells in patients with unusual epidermotropic cutaneous T-cell lymphomas, particularly if they have any clinical features suggestive of AITL.
Assuntos
Linfadenopatia Imunoblástica/diagnóstico , Linfoma de Células T Periférico/diagnóstico , Micose Fungoide/diagnóstico , Neoplasias Cutâneas/diagnóstico , Pele/patologia , Adulto , Biomarcadores Tumorais/metabolismo , Células Clonais , Diagnóstico Diferencial , Humanos , Linfadenopatia Imunoblástica/metabolismo , Linfonodos/patologia , Linfoma de Células T Periférico/metabolismo , Masculino , Neprilisina/metabolismo , Recidiva , Pele/metabolismo , Neoplasias Cutâneas/metabolismo , Linfócitos T Auxiliares-Indutores/metabolismo , Linfócitos T Auxiliares-Indutores/patologiaRESUMO
OBJECTIVES: To assess immunological variables, T-cell apoptosis and oxidative stress markers in the peripheral blood and peritoneal fluid of women with (WEN) and without (WWE) endometriosis. DESIGN: Observational and transverse case-control study. SETTING: National Institute of Perinatology, Mexico City, Mexico. POPULATION AND SAMPLE: Peripheral blood and peritoneal fluid obtained from 30 WWE and 32 WEN. METHODS: Blood was drawn before surgery and peritoneal fluid was collected during surgery but before any surgical procedure had been carried out. Flow cytometry, spectrophotometry, high-performance liquid chromatography and multiplex immunoassay analyses were performed. MAIN OUTCOME MEASURES Peripheral and peritoneal lymphocyte subpopulations (CD3(+), CD4(+) CD3(+), CD8(+) CD3(+), CD16(+) CD56(+), human leucocyte antigen-DR(+) CD3(+) and CD19(+)), intracellular CD4(+) CD3(+) and CD8(+) CD3(+) cytokine synthesis (interleukin-2 [IL-2] and interferon-γ [IFN-γ]), CD3(+) apoptosis, malondialdehyde and ascorbate concentrations and peritoneal cytokine concentrations. RESULTS: No differences were found in peripheral and peritoneal lymphocyte subsets between the groups. Peritoneal T lymphocytes from WEN produced less IL-2 and IFN-γ than those from WWE. Peritoneal malondialdehyde concentrations were higher and ascorbate concentrations were lower in WEN than in WWE. Higher peritoneal concentrations of pro-inflammatory cytokines (IL-1ß, tumour necrosis factor-α and IL-6) and chemokines (IL-10, IL-8, eotaxin, vascular endothelial growth factor, monocyte chemotactic protein-1 and regulated upon activation, normal T-cell expressed, and secreted) and lower concentrations of IFN-γ, IL-1 receptor antagonist and IL-15 were found in WEN. No statistical differences were found in IL-2, IL-4, IL-12 and IL-13 concentrations. CONCLUSION: The alterations observed in WEN were associated with a diminished peritoneal T helper type 1 immune response. Pro-inflammatory, chemotactic, angiogenic and oxidative stress markers were altered in the peritoneal milieu of WEN. These changes appeared to contribute to the peritoneal immune alterations found.
Assuntos
Apoptose/imunologia , Endometriose/imunologia , Estresse Oxidativo/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Adulto , Ácido Ascórbico/metabolismo , Estudos de Casos e Controles , Citocinas/metabolismo , Feminino , Citometria de Fluxo , Humanos , Imunidade Celular , Imunoensaio , Malondialdeído/metabolismo , Subpopulações de Linfócitos T , Linfócitos T Auxiliares-Indutores/patologiaRESUMO
INTRODUCTION: To evaluate the microscopic characteristics and densities (per mm(2) ) of tryptase(+) mast cells, CD4(+) T helper lymphocytes, CD45RO(+) memory T lymphocytes, foxp3(+) T regulatory lymphocytes, CD20(+) B lymphocytes, CD68(+) macrophages, and CD31(+) blood vessels in human dental pulpitis (n=38) and healthy pulpal tissue (n=6). METHODOLOGY: The pulps of 38 human teeth with a clinical diagnosis of irreversible pulpitis were removed by pulpectomy. The pulp tissue was immersed in 10% buffered formalin for evaluation using light microscopy. Tryptase, CD4, CD45RO, foxp3, CD20, CD68, and CD31 expressions were analysed using immunohistochemistry; other microscopic features, such as intensity of inflammatory infiltrate and collagen deposition, were evaluated using haematoxylin and eosin stain. Wilcoxon and Mann-Whitney tests were used for statistical analysis. The significance level was set at α=5%. RESULTS: Two microscopic patterns of pulpitis were found: group 1 (G1) (n=15) had an intense inflammatory infiltrate and mild collagen deposition; conversely, group 2 (G2) (n=23) had a scarce inflammatory infiltrate and intense collagen deposition. The numbers of CD68(+) macrophages (P=0.004) and CD20(+) B (P=0.068) lymphocytes and the density of blood vessels (P=0.002) were higher in G1 than in G2. However, a similar number of CD4(+) and CD45RO(+) T lymphocytes was found in both groups (P>0.05). When present, tryptase(+) mast cells were equally distributed in G1 and G2, whereas foxp3(+) T regulatory lymphocytes were detected in 59% and 14% of the samples of G1 and G2. Controls exhibited lower numbers of foxp3, tryptase, CD4, CD45RO, CD68 and CD20 positive cells than G1 and G2. CONCLUSIONS: Irreversible pulpitis had distinct microscopic features with important quantitative and qualitative differences in inflammatory cell infiltration.
Assuntos
Linfócitos/classificação , Macrófagos/patologia , Mastócitos/patologia , Pulpite/patologia , Antígenos CD/análise , Antígenos CD20/análise , Antígenos de Diferenciação Mielomonocítica/análise , Linfócitos B/patologia , Antígenos CD4/análise , Calcinose/imunologia , Calcinose/patologia , Contagem de Células , Colágeno/análise , Polpa Dentária/irrigação sanguínea , Fatores de Transcrição Forkhead/análise , Humanos , Imuno-Histoquímica , Memória Imunológica/imunologia , Antígenos Comuns de Leucócito/análise , Linfócitos/imunologia , Macrófagos/imunologia , Mastócitos/imunologia , Microvasos/imunologia , Microvasos/patologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , Pulpite/imunologia , Subpopulações de Linfócitos T/classificação , Linfócitos T Auxiliares-Indutores/patologia , Linfócitos T Reguladores/patologia , Triptases/análiseRESUMO
Multiple sclerosis (MS) is an autoimmune disease characterized by inflammatory immune response directed against myelin antigens of the central nervous system. In its murine model, EAE, Th17 cells play an important role in disease pathogenesis. These cells can induce blood-brain barrier disruption and CNS immune cells activation, due to the capacity to secrete high levels of IL-17 and IL-22 in an IL-6+TGF-ß dependent manner. Thus, using the oral tolerance model, by which 200 µg of MOG 35-55 is given orally to C57BL/6 mice prior to immunization, we showed that the percentage of Th17 cells as well as IL-17 secretion is reduced both in the periphery and also in the CNS of orally tolerated animals. Altogether, our data corroborates with the pathogenic role of IL-17 and IFN-γ in EAE, as its reduction after oral tolerance, leads to an overall reduction of pro-inflammatory cytokines, such as IL-1α, IL-6, IL-9, IL-12p70 and the chemokines MIP-1ß, RANTES, Eotaxin and KC in the CNS. It is noteworthy that this was associated to an increase in IL-10 levels. Thus, our data clearly show that disease suppression after oral tolerance induction, correlates with reduction in target organ inflammation, that may be caused by a reduced Th1/Th17 response.
Assuntos
Alérgenos/administração & dosagem , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/patologia , Tolerância Imunológica , Interleucina-17/antagonistas & inibidores , Depleção Linfocítica , Proteínas do Tecido Nervoso/administração & dosagem , Linfócitos T Auxiliares-Indutores/imunologia , Administração Oral , Alérgenos/imunologia , Sequência de Aminoácidos , Animais , Encefalomielite Autoimune Experimental/terapia , Glicoproteínas/administração & dosagem , Glicoproteínas/imunologia , Glicoproteínas/uso terapêutico , Imunossupressores/administração & dosagem , Imunossupressores/imunologia , Imunossupressores/uso terapêutico , Inflamação/imunologia , Inflamação/patologia , Inflamação/terapia , Interleucina-17/metabolismo , Depleção Linfocítica/métodos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Glicoproteína Mielina-Oligodendrócito , Proteínas do Tecido Nervoso/imunologia , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/imunologia , Fragmentos de Peptídeos/uso terapêutico , Linfócitos T Auxiliares-Indutores/patologiaRESUMO
Regulated glycosylation controls T cell processes, including activation, differentiation and homing by creating or masking ligands for endogenous lectins. Here we show that stimuli promoting T helper type 1 (TH1), TH2 or interleukin 17-producing T helper (TH-17) differentiation can differentially regulate the glycosylation pattern of T helper cells and modulate their susceptibility to galectin-1, a glycan-binding protein with anti-inflammatory activity. Although TH1- and TH-17-differentiated cells expressed the repertoire of cell surface glycans critical for galectin-1-induced cell death, TH2 cells were protected from galectin-1 through differential sialylation of cell surface glycoproteins. Consistent with those findings, galectin-1-deficient mice developed greater TH1 and TH-17 responses and enhanced susceptibility to autoimmune neuroinflammation. Our findings identify a molecular link among differential glycosylation of T helper cells, susceptibility to cell death and termination of the inflammatory response.