RESUMO
BACKGROUND: Induction of oral tolerance to haptens is an efficient way to prevent allergic contact dermatitis (ACD) in mice. Toll-like receptor (TLR)-mediated sensing of the microbiota contributes to gut homeostasis, yet whether it contributes to induction of oral tolerance has not been documented. OBJECTIVE: We examined whether oral tolerance to the contact sensitizer 2,4-dinitro-fluorobenzene (DNFB) depends on microbiota/TLRs and evaluated the role of TLR4 on the tolerogenic function of intestinal dendritic cells (DCs). METHODS: Oral tolerance was induced by DNFB gavage in germ-free and mice deficient in several TLRs. Tolerance was assessed by means of suppression of contact hypersensitivity and hapten-specific IFN-γ-producing effector T cells. The tolerogenic function of intestinal DCs was tested by adoptive transfer experiments, ex vivo hapten presentation, and forkhead box p3 regulatory T-cell conversion. RESULTS: Oral tolerance induced by DNFB gavage was impaired in germ-free mice and TLR4-deficient mice. Bone marrow chimeras revealed that TLR4 expression on hematopoietic cells was necessary for oral tolerance induction. TLR4 appeared to be essential for the ability of intestinal dendritic cells from DNFB-fed mice to inhibit ACD on adoptive transfer. Indeed, TLR4 conditioned the in vivo mobilization to mesenteric lymph nodes of intestinal migratory CD103+ DCs carrying oral DNFB, especially the CD103+CD11b+ DC subset expressing the vitamin A-converting enzyme retinaldehyde dehydrogenase and specialized in forkhead box p3-positive regulatory T-cell conversion. CONCLUSIONS: Our data demonstrate that TLR4 conditions induction of oral tolerance to DNFB through licensing tolerogenic gut DCs. Oral biotherapy with TLR4 ligands might be useful to potentiate oral tolerance to haptens and alleviate ACD in human subjects.
Assuntos
Células Dendríticas/imunologia , Dermatite Alérgica de Contato/imunologia , Microbioma Gastrointestinal/imunologia , Tolerância Imunológica , Intestinos/imunologia , Linfócitos T Reguladores/imunologia , Receptor 4 Toll-Like/imunologia , Animais , Células Dendríticas/patologia , Dermatite Alérgica de Contato/genética , Dermatite Alérgica de Contato/patologia , Dinitrofluorbenzeno/toxicidade , Interferon gama/genética , Interferon gama/imunologia , Intestinos/patologia , Camundongos , Camundongos Knockout , Linfócitos T Reguladores/patologia , Receptor 4 Toll-Like/genéticaRESUMO
BACKGROUND & AIMS: The liver receives blood from the gastrointestinal tract through the portal vein, and thereby is exposed continuously to dietary antigens and commensal bacteria. Alcoholic liver disease (ALD) is associated with intestinal dysbiosis, increased intestinal permeability, release of microbes into the portal circulation, and increased serum levels and liver deposits of IgA. We characterized B-cell production of IgA in livers of mice at homeostasis, after oral immunization, in a mouse model of ALD and in human liver samples. METHODS: We performed studies with Balb/c and C57BL/6-Ly5.1 mice, as well as transgenic mice (quasimonoclonal, activation-induced [cytidine] deaminase-Cre-tamoxifen-dependent estrogen receptor 2 [ERT2], Blimp-1-green fluorescent protein [GFP]). C57BL/6-Ly5.1 mice were fed chronic plus binge ethanol to create a model of ALD. Some mice also were given repeated injections of FTY720, which prevents egress of IgA-secreting cells from Peyer's patches. We obtained nontumor liver tissues from patients with colorectal carcinoma undergoing surgery for liver metastases or hepatocellular carcinoma. B cells were isolated from mouse and human liver tissues and analyzed by flow cytometry and enzyme-linked ImmunoSpot (ELISpot). In wild-type and transgenic mice, we traced newly generated IgA-secreting cells at steady state and after oral immunization with 4-hydroxy-3-nitrophenylacetyl (NP)-Ficoll or cholera toxin. IgA responses were also evaluated in our model of ALD. RESULTS: Livers of control mice contained proliferative plasmablasts that originated from Peyer's patches and produced IgAs reactive to commensal bacteria. After oral immunization with cholera toxin or a thymus-independent antigen, a substantial number of antigen-specific IgA-secreting cells was found in the liver. Mice fed ethanol had features of hepatitis and increased numbers of IgA-secreting cells in liver, compared with mice given control diets, as well as higher levels of serum IgA and IgA deposits in liver sinusoids. Injection of FTY720 during ethanol feeding reduced liver and serum levels of IgA and IgA deposits in liver and prevented liver injury. Human liver tissues contained a significant proportion of IgA-producing plasma cells that shared phenotypic and functional attributes with those from mouse liver, including reactivity to commensal bacteria. CONCLUSIONS: Based on studies of mice and human liver tissues, we found the liver to be a site of IgA production by B cells, derived from gut-associated lymphoid tissues. These IgAs react with commensal bacteria and oral antigens. Livers from mice with ethanol-induced injury contain increased numbers of IgA-secreting cells and have IgA deposits in sinusoids. IgAs in the liver could mediate clearance of gut-derived antigens that arrive through portal circulation at homeostasis and protect these organs from pathogens.
Assuntos
Antígenos/imunologia , Hepatócitos/metabolismo , Imunoglobulina A Secretora/biossíntese , Intestinos/imunologia , Fígado/citologia , Nódulos Linfáticos Agregados/imunologia , Animais , Linfócitos B/imunologia , Hepatócitos/imunologia , Humanos , Fígado/imunologia , Fígado/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BLRESUMO
Intestinal DCs orchestrate gut immune homeostasis by dampening proinflammatory T-cell responses and inducing anti-inflammatory IgA responses. Although no specific DC subset has been strictly assigned so far to govern IgA response, some candidate subsets emerge. In particular, plasmacytoid DCs (pDCs), which notoriously promote anti-viral immunity and T-cell tolerance to innocuous antigens (Ags), contribute to IgA induction in response to intestinal viral infection and promote T-cell-independent IgA responses in vitro. Here, using two transgenic mouse models, we show that neither short-term nor long-term pDC depletion alters IgA class switch recombination in Peyer's patches and frequency of IgA plasma cells in intestinal mucosa at steady state, even in the absence of T-cell help. In addition, pDCs are dispensable for induction of intestinal IgA plasma cells in response to oral immunization with T-cell-dependent or T-cell-independent Ags, and are not required for proliferation and IgA switch of Ag-specific B cells in GALT. These results show that pDCs are dispensable for noninfectious IgA responses, and suggest that various DC subsets may play redundant roles in the control of intestinal IgA responses.
Assuntos
Linfócitos B/imunologia , Células Dendríticas/imunologia , Imunoglobulina A/metabolismo , Mucosa Intestinal/imunologia , Plasmócitos/imunologia , Animais , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Homeostase , Humanos , Tolerância Imunológica , Imunização , Switching de Imunoglobulina , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Linfócitos T/imunologia , Fator de Transcrição 4RESUMO
Oral tolerance prevents oral sensitization to dietary antigens (Ags), including proteins and haptens, and development of delayed-type hypersensitivity (DTH) responses. We showed here that plasmacytoid dendritic cells (pDCs) prevented oral T cell priming and were responsible for systemic tolerance to CD4(+) and CD8(+) T cell-mediated DTH responses induced by Ag feeding. Systemic depletion of pDCs prevented induction of tolerance by antigen feeding. Transfer of oral Ag-loaded liver pDCs to naive recipient mice induced Ag-specific suppression of CD4(+) and CD8(+) T cell responses to protein and hapten, respectively. Liver is a site of oral Ag presentation, and pDCs appeared to induce anergy or deletion of Ag-specific T cells in the liver relatively rapidly via a CD4(+) T cell-independent mechanism. These data demonstrate that oral tolerance relies on Ag presentation by pDC to T cells and suggest that pDC could represent a key therapeutic target for intestinal and systemic inflammatory diseases.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Células Dendríticas/imunologia , Hipersensibilidade Tardia/imunologia , Tolerância Imunológica , Mucosa Bucal/imunologia , Animais , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Células Dendríticas/metabolismo , Dinitrofluorbenzeno/imunologia , Feminino , Hipersensibilidade Tardia/metabolismo , Fígado/imunologia , Linfonodos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Baço/imunologiaRESUMO
Patrolling Ly6C(-) monocytes are blood-circulating cells that play a role in inflammation and in the defense against pathogens. Here, we show that similar to natural killer (NK) cells, patrolling monocytes express high levels of S1PR5, a G-coupled receptor for sphingosine-1 phosphate. We found that S1pr5(-/-) mice lack peripheral Ly6C(-) monocytes but have a normal number of these cells in the bone marrow (BM). Various lines of evidence exclude a direct contribution of S1PR5 in the survival of Ly6C(-) monocytes at the periphery. Rather, our data support a role for S1PR5 in the egress of Ly6C(-) monocytes from the BM. In particular, we observed a reduced frequency of patrolling monocytes in BM sinusoids of S1PR5 KO mice. Unexpectedly, S1P was not a chemoattractant for patrolling monocytes and had no significant effect on their viability in vitro. Moreover, the disruption of S1P gradients in vivo did not alter Ly6C(-) monocyte trafficking and viability. These data suggest that S1PR5 regulates the trafficking of monocytes via a mechanism independent of S1P gradients.
Assuntos
Antígenos Ly/metabolismo , Medula Óssea/imunologia , Monócitos/imunologia , Receptores de Lisoesfingolipídeo/metabolismo , Animais , Circulação Sanguínea , Movimento Celular/imunologia , Sobrevivência Celular , Células Cultivadas , Feminino , Homeostase , Vigilância Imunológica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores de Lisoesfingolipídeo/genéticaRESUMO
Absent in peripheral tissues during homeostasis, human plasmacytoid dendritic cells (pDCs) are described in inflamed skin or mucosa. Here, we report that, unlike blood pDCs, a subset of tonsil pDCs express functional CCR6 and CCR10, and their respective ligands CCL20 and CCL27are detected in inflamed epithelia contacting blood dendritic cell antigen 2(+) pDCs. Moreover, pDCs are recruited to imiquimod-treated skin tumors in WT but not CCR6-deficient mice, and competitive adoptive transfers reveal that CCR6-deficient pDCs are impaired in homing to inflamed skin tumors after intravenous transfer. On IL-3 culture, CCR6 and CCR10 expression is induced on human blood pDCs that become responsive to CCL20 and CCL27/CCL28, respectively. Interestingly, unlike myeloid DC, blood pDCs initially up-regulate CCR7 expression and CCL19 responsiveness on IL-3 ± CpG-B and then acquire functional CCR6 and CCR10. Finally, IL-3-differentiated CCR6(+) CCR10(+) pDCs secrete high levels of IFN-α in response to virus. Overall, we propose an unexpected pDCs migratory model that may best apply for mucosal-associated lymphoid tissues. After CCR7-mediated extravasation into lymphoid tissues draining inflamed epithelia, blood pDCs may be instructed to up-regulate CCR6 and/or CCR10 allowing their homing into inflamed epithelia (in mucosae or skin). At this site, pDCs can then produce IFN-α contributing to pathogen clearance and/or local inflammation.
Assuntos
Células Dendríticas/imunologia , Inflamação/imunologia , Receptores CCR10/metabolismo , Receptores CCR6/metabolismo , Transferência Adotiva , Animais , Diferenciação Celular/imunologia , Movimento Celular/imunologia , Quimiocina CCL19/farmacologia , Quimiocina CCL20/farmacologia , Células Dendríticas/patologia , Epitélio/imunologia , Epitélio/patologia , Feminino , Humanos , Inflamação/patologia , Interferon-alfa/biossíntese , Interleucina-3/farmacologia , Ligantes , Tecido Linfoide/imunologia , Tecido Linfoide/patologia , Melanoma Experimental/imunologia , Melanoma Experimental/patologia , Glicoproteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Modelos Imunológicos , Tonsila Palatina/citologia , Tonsila Palatina/imunologia , Receptores CCR6/deficiência , Receptores CCR6/genética , Receptor 7 Toll-Like/metabolismoRESUMO
The vast spectrum of clinical features of COVID-19 keeps challenging scientists and clinicians. Low resistance to infection might result in long-term viral persistence, but the underlying mechanisms remain unclear. Here, we studied the immune response of immunocompetent COVID-19 patients with prolonged SARS-CoV-2 infection by immunophenotyping, cytokine and serological analysis. Despite viral loads and symptoms comparable to regular mildly symptomatic patients, long-term carriers displayed weaker systemic IFN-I responses and fewer circulating pDCs and NK cells at disease onset. Type 1 cytokines remained low, while type-3 cytokines were in turn enhanced. Of interest, we observed no defects in antigen-specific cytotoxic T cell responses, and circulating antibodies displayed higher affinity against different variants of SARS-CoV-2 Spike protein in these patients. The identification of distinct immune responses in long-term carriers adds up to our understanding of essential host protective mechanisms to ensure tissue damage control despite prolonged viral infection.
RESUMO
Background and aims: We aimed to analyze circulating CD4+ T cell subsets and cytokines during the course of Crohn's disease (CD). Methods and results: CD4+ T cell subsets, ultrasensitive C-reactive protein (usCRP), and various serum cytokines (IL-6, IL-8, IL-10, IL-13, IL-17A, IL-23, TNFα, IFNγ, and TGFß) were prospectively monitored every 3 months for 1 year, using multicolor flow cytometry and an ultrasensitive Erenna method in CD patients in remission at inclusion. Relapse occurred in 35 out of the 113 consecutive patients (31%). For patients in remission within 4 months prior to relapse and at the time of relapse, there was no significant difference in Th1, Th17, Treg, and double-positive CD4+ T cell subsets co-expressing either IFNγ and FOXP3, IL-17A and FOXP3, or IFNγ and IL-17A. On the contrary, in patients who remained in remission, the mean frequency and number of double-positive IL-17A+FOXP3+ CD4+ T cells and the level of usCRP were significantly higher (p ≤ 0.01) 1 to 4 months prior to relapse. At the time of relapse, only the IL-6 and usCRP levels were significantly higher (p ≤ 0.001) compared with those patients in remission. On multivariate analysis, a high number of double-positive IL-17A+FOXP3+ CD4+ T cells (≥1.4 cells/mm3) and elevated serum usCRP (≥3.44 mg/L) were two independent factors associated with risk of relapse. Conclusions: Detection of circulating double-positive FOXP3+IL-17A+ CD4+ T cell subsets supports that T cell plasticity may reflect the inflammatory context of Crohn's disease. Whether this subset contributes to the pathogenesis of CD relapse needs further studies.
Assuntos
Doença de Crohn , Interleucina-17 , Humanos , Interleucina-17/metabolismo , Doença de Crohn/patologia , Citocinas/metabolismo , Interleucina-6/metabolismo , Subpopulações de Linfócitos T/metabolismo , Células Th17/metabolismo , Fatores de Transcrição Forkhead/metabolismo , RecidivaRESUMO
BACKGROUND: Regulatory T cells contribute to peripheral immune tolerance, yet their ability to control immediate-type hypersensitivity (ITH) reactions involved in IgE-mediated food allergy is still poorly documented. OBJECTIVES: We investigated in mice whether CD4+CD25+ regulatory T cells could control ITH to ß-lactoglobulin (BLG), a major allergen in cow's milk. METHODS: C3H/HeOuJ mice were sensitized by repeated oral gavage with BLG plus cholera toxin as adjuvant and orally challenged with BLG alone to elicit allergic symptoms. Mice were treated with the anti-CD25 mAb (PC61) before sensitization. Oral sensitization (afferent phase of ITH) was assessed by production of BLG-specific serum antibodies and Th1/Th2-type cytokines by specific CD4+ T cells in mesenteric lymph nodes. ITH was elicited by oral BLG challenge (efferent phase of ITH) and we monitored symptom scores, numbers and function of intestinal mast cells and serum level of the mucosal mast cell protease mMCP-1. RESULTS: Upon oral BLG challenge, orally sensitized mice developed only mild clinical signs. Anti-CD25 mAb-treated mice exhibited enhancement of both BLG-specific CD4+ T cell priming with IL-4, IL-5, IL-13 and IFN-γ production and total IgE, and BLG-specific IgE, IgG1 and IgG2a in serum. Anti-CD25 mAb treatment caused more severe symptoms upon BLG challenge, which correlated with enhanced serum levels of the mucosal mast cell protease mMCP-1. CONCLUSIONS: These data document that constitutive CD4+CD25+ regulatory T cells alleviate clinical signs of ITH to dietary BLG by modulating the priming of BLG-specific T and B cell responses during oral sensitization and enhancing mast cell degranulation.
Assuntos
Hipersensibilidade Alimentar/imunologia , Lactoglobulinas/imunologia , Linfócitos T Reguladores/imunologia , Administração Oral , Animais , Anticorpos/imunologia , Anticorpos/farmacologia , Citocinas/imunologia , Feminino , Imunização , Subunidade alfa de Receptor de Interleucina-2/antagonistas & inibidores , Intestinos/imunologia , Lactoglobulinas/administração & dosagem , Mastócitos/efeitos dos fármacos , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos C3H , Linfócitos T Reguladores/efeitos dos fármacos , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th2/efeitos dos fármacos , Células Th2/imunologiaRESUMO
Evidence that CD4(+) regulatory T cells can control Ag-specific CD8(+) T cell-mediated colitis in immunocompetent mice is poorly documented. To examine the potential of CD4(+) T cells to control colitis, we used our model of CD8(+) T cell-mediated colitis induced by intracolonic sensitization followed by challenge with the hapten 2,4-dinitrobenzene sulfonic acid. The defect of CD4(+) T cells in MHC class II-deficient (Abeta(degrees/degrees)) mice allowed priming of 2,4-dinitrobenzene sulfonic acid-specific IFN-gamma-producing CD8 colitogenic effectors and development of colitis in the otherwise resistant C57BL/6 strain. Cotransfer experiments in RAG2(degrees/degrees) mice and ex vivo studies showed that CD4(+)CD25(+) T cells completely prevented CD8(+) T cell-mediated colitis and controlled CD8(+) T cell activation, respectively. In the susceptible BALB/c strain, Ab depletion revealed that lack of CD4(+) regulatory T cells resulted in 1) acute colitis elicited by a suboptimal dose of hapten challenge and 2) more severe relapsing episodes of colitis induced by effector/memory CD8(+) T cell-mediated colitis at an optimal dose of hapten challenge, even when CD4 depletion was performed just before the second challenge. Oral administration of the probiotic strain Lactobacillus casei DN-114 001 alleviated colitis and increased the suppressive function of Foxp3(+)CD4(+) regulatory T cells of colon lamina propria. These data demonstrate that CD4(+) regulatory T cells exert a protective effect on colitis by controlling colitogenic effector/memory CD8(+) T cells during the effector (symptomatic) phase of acute and relapsing colitis, respectively. Probiotics with natural adjuvant effects on mucosal regulatory T cells may represent a valuable approach to alleviate the colitogenic effect of Tc1-type CD8(+) effectors.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Colite/imunologia , Colite/microbiologia , Lacticaseibacillus casei/imunologia , Animais , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD8-Positivos/patologia , Células Cultivadas , Técnicas de Cocultura , Colite/prevenção & controle , Predisposição Genética para Doença , Imunidade Inata , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , RecidivaRESUMO
BACKGROUND: CD4(+)CD25(+) regulatory T (Treg) cells are involved in the downmodulation of numerous immune responses to pathogens, tumors, or allergens. OBJECTIVE: In this study, we further characterized the nature of Treg cells that control skin inflammatory reactions to haptens. METHODS: In a model of contact hypersensitivity to 2,4-dinitro-fluorobenzene, we have investigated the phenotype, the specificity, and the origin of Treg cells that modulate the priming of effector CD8(+) T cells responsible for the development of the pathology. RESULTS: 2,4-Dinitrofluorobenzene immunization induced a population of CD4(+)CD25(+) Treg cells that controlled CD8(+) T-cell effector responses in a hapten-specific manner in vivo. High levels of inducible costimulator (ICOS) expression defined a population of CD4(+)CD25(+)FoxP3(+) (forkhead box protein 3) Treg cells that presented superior suppressive activity. Importantly, ICOS(+) Treg cells were distinguishable from all other FoxP3(+) Treg cells by the expression of IL-10, IL-17, and IFN-gamma. Hapten-specific Treg cells proliferating in response to their cognate antigen in vivo predominantly displayed a CD25(+)FoxP3(+)ICOS(+) phenotype. By using reporter mice, we showed that ICOS(+) Treg cells derived from the expansion of natural CD4(+)FoxP3(+) Treg cells rather than generation of adaptive Treg cells. Furthermore, the generation of ICOS(+) Treg cells depended on innate cells rather than the effector CD8(+) T-cell population. CONCLUSION: Taken together, our data show that a population of CD4(+)CD25(+)FoxP3(+) T cells upregulates ICOS on in vivo sensitization and specifically suppresses hapten-reactive CD8(+) T cells both in vivo and in vitro.
Assuntos
Antígenos de Diferenciação de Linfócitos T/imunologia , Dermatite Alérgica de Contato/imunologia , Linfócitos T Reguladores/imunologia , Células Th1/imunologia , Regulação para Cima/imunologia , Animais , Antígenos de Diferenciação de Linfócitos T/biossíntese , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Proliferação de Células/efeitos dos fármacos , Citocinas/biossíntese , Citocinas/imunologia , Dermatite Alérgica de Contato/metabolismo , Dinitrofluorbenzeno/efeitos adversos , Dinitrofluorbenzeno/farmacologia , Modelos Animais de Doenças , Fatores de Transcrição Forkhead/biossíntese , Fatores de Transcrição Forkhead/imunologia , Haptenos/efeitos adversos , Haptenos/farmacologia , Imunidade Inata/efeitos dos fármacos , Imunidade Inata/imunologia , Proteína Coestimuladora de Linfócitos T Induzíveis , Camundongos , Linfócitos T Reguladores/metabolismo , Células Th1/metabolismo , Regulação para Cima/efeitos dos fármacosRESUMO
BACKGROUND & AIMS: Orally induced tolerance to environmental allergens prevents deleterious, systemic, delayed-type hypersensitivity responses via immune suppression mechanisms believed to include either anergy/deletion of specific effector T cells or active suppression by CD4(+)CD25(+) regulatory T cells (Tregs). The aim of this study was to investigate whether and how antigen (Ag) penetration through the gut orchestrates these 2 distinct mechanisms. METHODS: Using a model of allergic contact dermatitis (ACD) mediated by hapten-specific cytolytic CD8(+) T cells and a T-cell transfer model of contact hypersensitivity in CD3epsilon-deficient mice, we studied the outcome of Ag gavage on CD8(+) effectors and Tregs. RESULTS: Full protection from ACD by gavage with the relevant allergen required 2 coordinated events taking place first in gut-associated lymphoid tissues and then systemically. Allergen gavage induced deletion by plasmacytoid dendritic cells of a large fraction of Ag-specific CD8(+) T cells in liver and mesenteric lymph nodes and also triggered the suppressive function of Treg of secondary lymphoid organs. Residual Ag-specific CD8(+) T cells conditioned during this mucosal step are fully susceptible to suppression by activated Treg, which completely prevented their differentiation into ACD effectors, upon re-exposure to the allergen via the skin. CONCLUSIONS: Thus, oral tolerance initiated in gut-associated lymphoid tissues by the plasmacytoid dendritic cells-mediated deletion of Ag-specific T cells is completed systemically by CD4(+)CD25(+) T cells. Biotherapies able to increase the susceptibility of effector T cells to the suppressive function of Treg may be valuable for the treatment of autoimmune and inflammatory diseases.
Assuntos
Células Dendríticas/imunologia , Tolerância Imunológica , Linfócitos T Reguladores/imunologia , Administração Oral , Transferência Adotiva , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Dermatite de Contato/imunologia , Dinitrofluorbenzeno/administração & dosagem , Dinitrofluorbenzeno/imunologia , Feminino , Haptenos/administração & dosagem , Haptenos/imunologia , Imunização , Interleucina-10/imunologia , Subunidade alfa de Receptor de Interleucina-2/análise , Fígado/imunologia , Linfonodos/imunologia , Subpopulações de Linfócitos , Mesentério , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Mucosae and skin are exposed to environmental antigens and are natural entry routes for most infectious agents. To maintain immunological tolerance and ensure protective immunity against pathogens, epithelial surfaces are surveyed permanently by antigen-presenting dendritic cells (DCs). Many DC subsets have been described in epithelial tissues, depending on the inflammatory state and the type of epithelium. Identification of the DC subset able to induce cytotoxic CD8+ T cells against antigens delivered via mucosae or skin, is a major issue for the development of efficient anti-infectious and anti-tumoral vaccines. Until recently, it was commonly accepted that Langerhans cells (LC), the prototype of immature DCs residing in skin and certain mucosae, can capture and process antigens and, in response to danger signals, undergo a maturation program allowing their migration to the draining lymph nodes for priming of naïve T cells. This concept likely needs to be revisited. Recent evidence from animal models revealed that resident epithelial tissue DCs, including LCs, do not play a direct role in T cell priming, but may contribute to maintenance of peripheral tolerance. Alternatively, DCs newly recruited into muco-cutaneous tissues exposed to pro-inflammatory stimuli are responsible for efficient priming and differentiation of CD8+ T cells into cytolytic effectors. These DC originate from blood monocytes and can cross-present protein antigens to CD8+ T cells, which subsequently give rise to specific CTL effectors. Remarkably, components derived from bacteria, virus and chemicals capable to enhance CCL20 production in epithelia, promote CCR6-dependent DC recruitment and behave as adjuvants allowing for cross-primed CD8+ CTL. These advances in the dynamic and function of epithelial tissue DC provide a rationale for the screening of novel CD8+ T cell adjuvants and the design of novel mucosal and skin vaccines.
Assuntos
Células Dendríticas/imunologia , Células de Langerhans/imunologia , Mucosa/imunologia , Pele/imunologia , Linfócitos T Citotóxicos/imunologia , Vacinação , Vacinas/imunologia , Animais , Apresentação de Antígeno/imunologia , Antígenos/administração & dosagem , Antígenos/imunologia , Diferenciação Celular , Movimento Celular , Citotoxicidade Imunológica/imunologia , Células-Tronco Hematopoéticas/citologia , Humanos , Tolerância Imunológica/imunologia , Inflamação/imunologia , Camundongos , Modelos Imunológicos , Vacinas/administração & dosagemRESUMO
BACKGROUND AND AIMS: Regulatory Foxp3+CD4+ T cells [Tregs] have been implicated in the control of colitis in T-cell transfer models, yet their ability to regulate colitis induced by innate immunity and the impact of gut inflammation on their fate and function have been poorly documented. METHODS: Colitis was induced by dextran sodium sulphate in DEREG transgenic mice. Tregs ablation and transfer experiments showd that Tregs could limit the severity of colitis in B6 mice. RESULTS: Gut inflammation resulted in increased number of Tregs in mesenteric lymph nodes [MLN] and colon lamina propria [LP], although their frequency decreased due to massive concomitant leukocyte infiltration. This coincided at both sites with a dramatic increase in Ki67+ Tregs which retained proliferative capacity. Gut inflammation resulted in enhanced suppressive function of Tregs in colon lamina propria and neuropillin-1- [NRP1-] Treg in MLN. Real-time polymerase chain reaction analysis and flow cytometry [using IL10-egfp-reporter mice] showed that compared with NRP1+ Treg, NRP1- Treg express higher levels of IL-10 transcripts and were enriched in IL10-expressing cells both in the steady state and during colitis. Moreover, Treg conversion in vivo from from naïve CD4+ T cells or Treg precursors was impaired in colitic mice. Finally, gut inflammation caused a decrease in intestinal dendritic cells, affecting both CD103+CD11b+ and CD103+CD11b- subsets and affected their Treg conversion capacity. CONCLUSIONS: Together, our data indicate that non-specific colon inflammation triggers proliferation and suppressive function of Tregs in the lamina propria and MLN, but impairs their de novo conversion from CD4+ T cells by intestinal dendritic cells.
Assuntos
Linfócitos T CD4-Positivos/fisiologia , Colite/fisiopatologia , Fatores de Transcrição Forkhead/fisiologia , Mucosa Intestinal/citologia , Linfócitos T Reguladores/fisiologia , Animais , Proliferação de Células/fisiologia , Colite/imunologia , Feminino , Mucosa Intestinal/imunologia , Mucosa Intestinal/fisiopatologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Mucosa/citologia , Mucosa/imunologia , Mucosa/fisiopatologiaRESUMO
Allergic contact dermatitis (ACD) to strong experimental haptens is mediated by specific CD8+ T cells. Here, we show that similar mechanisms occur for weak haptens, which comprise the vast majority of chemicals responsible for human ACD. We used a model of ACD, that is, the contact hypersensitivity reaction, to test for the allergenicity of three weak haptens involved in fragrance allergy. ACD to weak haptens could not be induced in normal mice. In contrast, mice acutely depleted in CD4+ T cells developed a typical ACD reaction to the three weak fragrance allergens that peaked 24 hours after challenge. Priming of CD8+ T cells was observed in draining lymph nodes 5 days after sensitization and development of ACD was associated with the infiltration of activated CD8+ T cells in the challenged skin. CD8+ T cells were effectors of the ACD reaction as in vivo treatment with depleting anti-CD8 mAbs abrogated the ACD responses and as purified CD8+ T cells could adoptively transfer ACD to naive recipients. In conclusion, our data demonstrate a dominant role of CD8+ T cells as initiators of ACD to weak haptens, and suggest that CD8+ T cells may represent potential targets for preventing or treating ACD.
Assuntos
Alérgenos/toxicidade , Linfócitos T CD8-Positivos/imunologia , Dermatite Alérgica de Contato/imunologia , Haptenos/toxicidade , Animais , Anticorpos Monoclonais/farmacologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Dermatite Alérgica de Contato/patologia , Linfonodos/citologia , Linfonodos/imunologia , Depleção Linfocítica , Camundongos , Camundongos Endogâmicos C57BL , Pele/efeitos dos fármacos , Pele/imunologia , Pele/patologiaRESUMO
The IV meeting of the European Mucosal Immunology Group, held October 8-10, 2004, in Lyon, gathered fundamental and clinical research scientists to discuss the most recent updates on basic and clinical aspects of mucosal immunology. The meeting was focused on innate and acquired immune mechanisms underlying handling and immune recognition of commensals, allergens, and pathogens by the mucosal immune system and its outcome in health and disease as well as for vaccine development. The scientific program featured five topics of growing interest for fundamental research scientists and clinicians, including the role of commensal bacteria in mucosal immunity; function of dendritic cells in infection, inflammation, and tolerance; control of mucosal inflammation by regulatory T cells; novel routes and adjuvants for mucosal vaccines; and mucosal immunity against HIV infection and vaccination strategies.
Assuntos
Infecções por HIV/imunologia , HIV/imunologia , Imunidade nas Mucosas/imunologia , Animais , Controle de Doenças Transmissíveis , Doenças Transmissíveis/imunologia , Europa (Continente) , Infecções por HIV/prevenção & controle , Humanos , Inflamação/imunologia , Sociedades Científicas , Vacinas Virais/imunologiaRESUMO
Allergic contact dermatitis (ACD) is a skin inflammatory disease mediated by activation of CD8+ cytotoxic T cells specific for haptens in contact with the skin. CD4+ T cells behave as both regulatory and tolerogenic cells since they down-regulate the skin inflammation in patients with ACD (regulation) and prevent the development of eczema (tolerance) in normal individuals. Thus, ACD corresponds to a breakdown of immune tolerance to haptens in contact with the skin. Several regulatory CD4+ T cell subsets (Treg), especially CD4+CD25+ natural Treg cells, are involved in immunological tolerance and regulation to haptens through the production of the immunosuppressive cytokines IL-10 and TGF-beta. Ongoing strategies to re-induce immune tolerance to haptens in patients with eczema include improvement of existing methods of tolerance induction (oral tolerance, low dose tolerance, allergen-specific immunotherapy, UV-induced tolerance) as well as development of new drugs able to activate IL-10 producing Treg cells in vivo. Ongoing and future progress in this area will open up new avenues for treatment of eczema and more generally autoimmune and allergic diseases resulting from a breakdown of tolerance to autoantigens and allergens, respectively.
Assuntos
Dermatite Alérgica de Contato/terapia , Tolerância Imunológica , Terapia de Imunossupressão/métodos , Alérgenos/efeitos adversos , Alérgenos/uso terapêutico , Animais , Citocinas/fisiologia , Dermatite Alérgica de Contato/imunologia , Dermatite Alérgica de Contato/fisiopatologia , Dessensibilização Imunológica , Haptenos/imunologia , Haptenos/uso terapêutico , Humanos , Interleucina-10/fisiologia , Interleucina-2/uso terapêutico , Modelos Animais , Modelos Biológicos , Terapia PUVA , Subpopulações de Linfócitos T/imunologia , Fator de Crescimento Transformador beta/fisiologiaRESUMO
Pathophysiological characteristics of these two types of eczema, both involve antigen specific effector T lymphocytes and down-regulatory CD4+ T cells (Treg). These cells are responsible for the cutaneous inflammatory response through the production of inflammatory cytokines and the destruction of keratinocytes by apoptosis. The main difference between these two types of disease is the nature of the environmental antigen. Indeed, protein allergens are involved in cases of atopic dermatitis whereas non protein chemicals (calles haptens) are responsible for allergic contact dermatitis. The skin lesions of eczema follow the activation of antigen specific T lymphocytes within the skin, leading to the production of inflammatory cytokines and to the destruction of keratinocytes by apoptosis. CD4+ Treg cells are endowed with down-regulatory and tolerogenic functions since they limit the skin inflammation in patients and prevent sensitization and induction of eczema in normal individuals. Eczema should therefore be considered as inflammatory dermatoses caused by the loss of immune tolerance to environmental antigens. Different strategies capable of restoring immune tolerance could avoid eczema outbursts and/or induce prolonged remissions of the disease.
Assuntos
Eczema/epidemiologia , Eczema/fisiopatologia , Eczema/imunologia , Humanos , Linfócitos T/imunologiaRESUMO
BACKGROUND AND AIMS: Evidence from mouse colitis models indicates that cytotoxic CD8+ T cells [CTL] play a key role in the initiation of gut lesions. We investigated whether changes in CD8+ CTL in blood or lamina propria [LP] of the neoterminal ileum were associated with postoperative endoscopic recurrence of Crohn's disease [CD]. METHODS: A total of 37 CD patients with ileocolonic resection were endoscopically followed up at 6 and 12 months post-surgery. CD8+ T cells were analysed by flow cytometry in blood and ileal LP. RESULTS: Granzyme B- and perforin-producing CD8+ T cells were significantly increased at 6 months in blood and in ileum LP in patients with endoscopic recurrence, as compared with those in remission. At a cutoff point of 45% of CD8+ CTL, the overall accuracies of the frequency of blood granzyme B+ or perforin+ CD8+ T cells to identify patients with postoperative endoscopic recurrence were 77% and 83%, respectively. Interestingly, patients with endoscopic recurrence at 12 months were those showing the highest mucosal CD8+ CTL frequency at 6 months, while still in remission. CONCLUSIONS: Enrichment of cytotoxic CD8+ T cells in blood and ileal mucosa coincides with CD postoperative endoscopic recurrence. This underscores that CD8 CTL may play a pathophysiological role in the initiation of gut lesions during CD.
Assuntos
Colectomia , Doença de Crohn/imunologia , Doença de Crohn/cirurgia , Íleo/imunologia , Mucosa Intestinal/imunologia , Linfócitos T Citotóxicos/metabolismo , Adulto , Idoso , Estudos de Casos e Controles , Colonoscopia , Doença de Crohn/diagnóstico por imagem , Feminino , Seguimentos , Humanos , Íleo/cirurgia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Recidiva , Resultado do TratamentoRESUMO
Development of vaccines able to induce mucosal immunity in the genital and gastrointestinal tracts is a major challenge to counter sexually transmitted pathogens such as HIV-1 and HSV-2. Herein, we showed that intradermal (ID) immunisation with sub-unit vaccine antigens (i.e., HIV-1 gp140 and HSV-2 gD) delivered with Poly(I:C) or CpG1668 as adjuvant induces long-lasting virus-specific immunoglobulin (Ig)-G and IgA antibodies in the vagina and feces. Poly(I:C)-supplemented sub-unit viral vaccines caused minimal skin reactogenicity at variance to those containing CpG1668, promoted a delayed-type hypersensitivity (DTH) to the vaccine and protected mice from genital and neurological symptoms after a lethal vaginal HSV-2 challenge. Interestingly, Poly(I:C12U) (Ampligen), a Poly(I:C) structural analogue that binds to TLR3 but not MDA-5, promoted robust mucosal and systemic IgG antibodies, a weak skin DTH to the vaccine but not IgA responses and failed to confer protection against HSV-2 infection. Moreover, Poly(I:C) was far superior to Poly(I:C12U) at inducing prompt and robust upregulation of IFNß transcripts in lymph nodes draining the injection site. These data illustrate that ID vaccination with glycoproteins and Poly(I:C) as adjuvant promotes long-lasting mucosal immunity and protection from genital HSV-2 infection, with an acceptable skin reactogenicity profile. The ID route thus appears to be an unexpected inductive site for mucosal immunity and anti-viral protection suitable for sub-unit vaccines. This works further highlights that TLR3/MDA5 agonists such as Poly(I:C) may be valuable adjuvants for ID vaccination against sexually transmitted diseases.