RESUMO
Cellular function in tissue is dependent on the local environment, requiring new methods for spatial mapping of biomolecules and cells in the tissue context1. The emergence of spatial transcriptomics has enabled genome-scale gene expression mapping2-5, but the ability to capture spatial epigenetic information of tissue at the cellular level and genome scale is lacking. Here we describe a method for spatially resolved chromatin accessibility profiling of tissue sections using next-generation sequencing (spatial-ATAC-seq) by combining in situ Tn5 transposition chemistry6 and microfluidic deterministic barcoding5. Profiling mouse embryos using spatial-ATAC-seq delineated tissue-region-specific epigenetic landscapes and identified gene regulators involved in the development of the central nervous system. Mapping the accessible genome in the mouse and human brain revealed the intricate arealization of brain regions. Applying spatial-ATAC-seq to tonsil tissue resolved the spatially distinct organization of immune cell types and states in lymphoid follicles and extrafollicular zones. This technology progresses spatial biology by enabling spatially resolved chromatin accessibility profiling to improve our understanding of cell identity, cell state and cell fate decision in relation to epigenetic underpinnings in development and disease.
Assuntos
Montagem e Desmontagem da Cromatina , Sequenciamento de Cromatina por Imunoprecipitação , Cromatina , Animais , Encéfalo/metabolismo , Diferenciação Celular , Linhagem da Célula , Cromatina/genética , Cromatina/metabolismo , Montagem e Desmontagem da Cromatina/genética , Sequenciamento de Cromatina por Imunoprecipitação/métodos , Epigenômica , Perfilação da Expressão Gênica , Genoma , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Camundongos , Tonsila Palatina/citologia , Tonsila Palatina/imunologiaRESUMO
Unlike other types of T helper (Th) responses, whether the development of Th2 cells requires instruction from particular subset of dendritic cells (DCs) remains unclear. By using an in vivo depletion approach, we have shown that DCs expressing CD301b were required for the generation of Th2 cells after subcutaneous immunization with ovalbumin (OVA) along with papain or alum. CD301b⺠DCs are distinct from epidermal or CD207⺠dermal DCs (DDCs) and were responsible for transporting antigen injected subcutaneously with Th2-type adjuvants. Transient depletion of CD301b⺠DCs resulted in less effective accumulation and decreased expression of CD69 by polyclonal CD4⺠T cells in the lymph node. Moreover, despite intact cell division and interferon-γ production, CD301b⺠DC depletion led to blunted interleukin-4 production by OVA-specific OT-II transgenic CD4⺠T cells and significantly impaired Th2 cell development upon infection with Nippostrongylus brasiliensis. These results reveal CD301b⺠DDCs as the key mediators of Th2 immunity.
Assuntos
Células Dendríticas/imunologia , Imunidade Celular , Lectinas Tipo C/imunologia , Pele/imunologia , Infecções por Strongylida/imunologia , Células Th2/imunologia , Alérgenos/imunologia , Compostos de Alúmen/administração & dosagem , Animais , Diferenciação Celular , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/parasitologia , Células Dendríticas/patologia , Regulação da Expressão Gênica , Interleucina-4/genética , Interleucina-4/imunologia , Lectinas Tipo C/genética , Camundongos , Camundongos Transgênicos , Nippostrongylus/imunologia , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Transdução de Sinais , Pele/efeitos dos fármacos , Pele/parasitologia , Pele/patologia , Infecções por Strongylida/parasitologia , Infecções por Strongylida/patologia , Células Th2/efeitos dos fármacos , Células Th2/parasitologia , Células Th2/patologiaRESUMO
In this study, we extended co-indexing of transcriptomes and epitopes (CITE) to the spatial dimension and demonstrated high-plex protein and whole transcriptome co-mapping. We profiled 189 proteins and whole transcriptome in multiple mouse tissue types with spatial CITE sequencing and then further applied the method to measure 273 proteins and transcriptome in human tissues, revealing spatially distinct germinal center reactions in tonsil and early immune activation in skin at the Coronavirus Disease 2019 mRNA vaccine injection site.
Assuntos
Análise de Célula Única , Transcriptoma , Animais , Camundongos , Humanos , Transcriptoma/genética , Epitopos , RNA Mensageiro , Perfilação da Expressão Gênica/métodosRESUMO
We present spatial-CITE-seq for high-plex protein and whole transcriptome co-mapping, which was firstly demonstrated for profiling 198 proteins and transcriptome in multiple mouse tissue types. It was then applied to human tissues to measure 283 proteins and transcriptome that revealed spatially distinct germinal center reaction in tonsil and early immune activation in skin at the COVID-19 mRNA vaccine injection site. Spatial-CITE-seq may find a range of applications in biomedical research.
RESUMO
OBJECTIVE: Epstein-Barr virus (EBV) infection has been linked to systemic lupus erythematosus (SLE), as demonstrated by the presence of increased seroprevalence and elevated viral loads, but the mechanism of this linkage has not been elucidated. Increased interferon-alpha (IFNalpha) levels and signatures, which are associated with innate immune responses, have been found in patients with SLE. Plasmacytoid dendritic cells (PDCs) are innate immune cells that mediate viral immunity by producing large quantities of IFNalpha, but the role they play during infection with EBV remains unclear. To address this issue, we investigated the ability of EBV to promote IFNalpha production by PDCs in healthy subjects. METHODS: Human PDCs were sorted and cultured in the presence of EBV, EBV-encoded RNA, and EBV double-stranded DNA. IFNalpha production by PDCs was measured by enzyme-linked immunosorbent assay, with the activation of these cells measured by flow cytometry. RESULTS: We found that EBV DNA and RNA promoted IFNalpha production by human PDCs through engagement of Toll-like receptor 9 (TLR-9) and TLR-7, respectively, with the initial viral recognition by PDCs mediated by binding to class II major histocompatibility complex (MHC) molecules. CONCLUSION: These data demonstrate that class II MHC-specific engagement by virus, with subsequent viral nucleic acid recognition, mediates IFNalpha production by PDCs. Our results suggest that elevated levels of IFNalpha found in SLE patients may be a result of aberrantly controlled chronic viral infection.
Assuntos
Células Dendríticas/imunologia , Herpesvirus Humano 4/imunologia , Interferon-alfa/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Antígenos Nucleares/imunologia , Células Dendríticas/citologia , Células Dendríticas/virologia , Relação Dose-Resposta Imunológica , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Imunidade Humoral/imunologia , Lúpus Eritematoso Sistêmico/virologia , Complexo Principal de Histocompatibilidade/imunologia , Estatísticas não Paramétricas , Receptor 7 Toll-Like/imunologia , Receptor Toll-Like 9/imunologiaRESUMO
Cross-linking of high-affinity immunoglobulin E (IgE) results in the life-threatening allergic reaction anaphylaxis. Yet the cellular mechanisms that induce B cells to produce IgE in response to allergens remain poorly understood. T follicular helper (TFH) cells direct the affinity and isotype of antibodies produced by B cells. Although TFH cell-derived interleukin-4 (IL-4) is necessary for IgE production, it is not sufficient. We report a rare population of IL-13-producing TFH cells present in mice and humans with IgE to allergens, but not when allergen-specific IgE was absent or only low-affinity. These "TFH13" cells have an unusual cytokine profile (IL-13hiIL-4hiIL-5hiIL-21lo) and coexpress the transcription factors BCL6 and GATA3. TFH13 cells are required for production of high- but not low-affinity IgE and subsequent allergen-induced anaphylaxis. Blocking TFH13 cells may represent an alternative therapeutic target to ameliorate anaphylaxis.
Assuntos
Anafilaxia/imunologia , Imunoglobulina E/imunologia , Interleucina-13/metabolismo , Linfócitos T Auxiliares-Indutores/imunologia , Adolescente , Animais , Criança , Fator de Transcrição GATA3/metabolismo , Fatores de Troca do Nucleotídeo Guanina/genética , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Humanos , Interleucina-13/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Proteínas Proto-Oncogênicas c-bcl-6/metabolismoRESUMO
Tissue repair is a subset of a broad repertoire of interleukin-4 (IL-4)- and IL-13-dependent host responses during helminth infection. Here we show that IL-4 or IL-13 alone was not sufficient, but IL-4 or IL-13 together with apoptotic cells induced the tissue repair program in macrophages. Genetic ablation of sensors of apoptotic cells impaired the proliferation of tissue-resident macrophages and the induction of anti-inflammatory and tissue repair genes in the lungs after helminth infection or in the gut after induction of colitis. By contrast, the recognition of apoptotic cells was dispensable for cytokine-dependent induction of pattern recognition receptor, cell adhesion, or chemotaxis genes in macrophages. Detection of apoptotic cells can therefore spatially compartmentalize or prevent premature or ectopic activity of pleiotropic, soluble cytokines such as IL-4 or IL-13.
Assuntos
Interleucina-13/imunologia , Interleucina-4/imunologia , Macrófagos/imunologia , Nippostrongylus/fisiologia , Regeneração , Animais , Apoptose , Inflamação/induzido quimicamente , Inflamação/patologia , Camundongos , Infecções por Strongylida/imunologia , TioglicolatosRESUMO
Following infection, T cells differentiate into a heterogeneous population of effector T cells that can mediate pathogen clearance. A subset of these effector T cells possesses the ability to survive long term and mature into memory T cells that can provide long-term immunity. Understanding the signals that regulate the development of memory T cells is crucial to efforts to design vaccines capable of eliciting T cell-based immunity. CD4(+) T cells are essential in the formation of protective memory CD8(+) T cells following infection or immunization. However, until recently, the mechanisms by which CD4(+) T cells act to support memory CD8(+) T cell development following infection were unclear. Here, we discuss recent studies that provide insight into the multifaceted role of CD4(+) T cells in the regulation of memory CD8(+) T cell differentiation.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Memória Imunológica/imunologia , Infecções/imunologia , Vacinação , Humanos , ImunizaçãoRESUMO
Host responses against metazoan parasites or an array of environmental substances elicit type 2 immunity. Despite its protective function, type 2 immunity also drives allergic diseases. The mechanisms that regulate the magnitude of the type 2 response remain largely unknown. Here, we show that genetic ablation of a receptor tyrosine kinase encoded byTyro3in mice or the functional neutralization of its ortholog in human dendritic cells resulted in enhanced type 2 immunity. Furthermore, the TYRO3 agonist PROS1 was induced in T cells by the quintessential type 2 cytokine, interleukin-4. T cell-specificPros1knockouts phenocopied the loss ofTyro3 Thus, a PROS1-mediated feedback from adaptive immunity engages a rheostat, TYRO3, on innate immune cells to limit the intensity of type 2 responses.
Assuntos
Imunidade Adaptativa/genética , Asma/imunologia , Interações Hospedeiro-Parasita/imunologia , Imunidade Inata/genética , Receptores Proteína Tirosina Quinases/fisiologia , Animais , Asma/genética , Proteínas Sanguíneas/antagonistas & inibidores , Proteínas Sanguíneas/genética , Proteínas Sanguíneas/metabolismo , Células Dendríticas/imunologia , Modelos Animais de Doenças , Técnicas de Inativação de Genes , Interações Hospedeiro-Parasita/genética , Humanos , Interleucina-4/imunologia , Interleucina-4/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Nippostrongylus/imunologia , Proteína S , Pyroglyphidae/imunologia , Receptores Proteína Tirosina Quinases/genética , Infecções por Strongylida/imunologia , Linfócitos T/imunologiaRESUMO
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by loss of T cell tolerance to nuclear antigens. Studies in mice and humans have demonstrated that T cells from individuals with lupus are abnormal. Here, we review the known T cell defects in lupus and their possible biochemical nature, genetic causes, and significance for lupus pathogenesis.
Assuntos
Lúpus Eritematoso Sistêmico/imunologia , Linfócitos T/imunologia , Animais , Humanos , Lúpus Eritematoso Sistêmico/genética , CamundongosRESUMO
T follicular helper (T(FH)) cells select high-affinity, antibody-producing B cells for clonal expansion in germinal centers (GCs), but the nature of their interaction is not well defined. Using intravital imaging, we found that selection is mediated by large but transient contacts between T(FH) and GC B cells presenting the highest levels of cognate peptide bound to major histocompatibility complex II. These interactions elicited transient and sustained increases in T(FH) intracellular free calcium (Ca(2+)) that were associated with T(FH) cell coexpression of the cytokines interleukin-4 and -21. However, increased intracellular Ca(2+) did not arrest TFH cell migration. Instead, T(FH) cells remained motile and continually scanned the surface of many GC B cells, forming short-lived contacts that induced selection through further repeated transient elevations in intracellular Ca(2+).
Assuntos
Linfócitos B/imunologia , Sinalização do Cálcio/imunologia , Centro Germinativo/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Animais , Proteínas de Fluorescência Verde/metabolismo , Interleucina-4/imunologia , Interleucinas/imunologia , Camundongos , Camundongos Knockout , Imagem MolecularRESUMO
Follicular helper T (T(FH)) cells are essential for B-cell maturation and immunoglobulin production after immunization with thymus-dependent antigens. Nevertheless, the development and function of T(FH) cells have been less clearly defined than classic CD4(+) effector T-cell subsets, including T-helper-1 (T(H)1), T(H)2 and T(H)17 cells. As such, our understanding of the genesis of T(FH) cells in humans and their role in the development of autoimmunity remains incomplete. However, evidence from animal models of systemic lupus erythematosus (SLE) and patients with systemic autoimmune diseases suggests that these cells are necessary for pathogenic autoantibody production, in a manner analogous to their role in promotion of B-cell maturation during normal immune responses. In this Review, I discuss the findings that have increased our knowledge of T(FH)-cell development and function in normal and aberrant immune responses. Such information might improve our understanding of autoimmune diseases, such as SLE, and highlights the potential of T(FH) cells as therapeutic targets in these diseases.
Assuntos
Autoimunidade/imunologia , Imunidade/imunologia , Linfonodos/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Animais , Autoanticorpos/biossíntese , Autoanticorpos/imunologia , Linfócitos B/imunologia , Comunicação Celular/imunologia , Modelos Animais de Doenças , Humanos , Lúpus Eritematoso Sistêmico/imunologia , CamundongosRESUMO
The autoimmune disease systemic lupus erythematosus (SLE) results from an inability of the immune system to discriminate between certain self-antigens and foreign ones. The most common treatment of SLE involves the use of immunosuppressive drugs to reduce inflammation, but these therapies have serious side effects. Three recent papers in Science Translational Medicine redirect focus on neutrophils, platelets, and interferon-α in the pathogenesis of SLE and reinforce the notion that researchers should seek to discover and devise combination therapies that target these processes.
Assuntos
Lúpus Eritematoso Sistêmico/etiologia , Lúpus Eritematoso Sistêmico/imunologia , Autoanticorpos/biossíntese , Plaquetas/imunologia , Humanos , Interferon-alfa/biossíntese , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/terapia , Modelos Imunológicos , Neutrófilos/imunologia , Ativação PlaquetáriaRESUMO
To address the complex chronic effector properties of interleukin (IL)-10, we generated transgenic mice in which IL-10 was overexpressed in the lung. In these mice, IL-10 inhibited endotoxin-induced tumor necrosis factor production and neutrophil accumulation. IL-10 also caused mucus metaplasia, B and T cell-rich inflammation, and subepithelial fibrosis and augmented the levels of mRNA encoding Gob-5, mucins, and IL-13. In mice bred to have null mutations of IL-13, IL-4R(alpha), or STAT-6, transgenic IL-10 did not induce mucus metaplasia but did induce inflammation and fibrosis. IL-10 was also a critical mucin regulator of virus-induced mucus metaplasia. Thus, IL-10, although inhibiting lipopolysaccharide-induced inflammation, also causes mucus metaplasia, tissue inflammation, and airway fibrosis. These responses are mediated by multiple mechanisms with mucus metaplasia being dependent on and the inflammation and fibrosis being independent of an IL-13/IL-4R(alpha)/STAT-6 activation pathway.
Assuntos
Inflamação/genética , Interleucina-10/genética , Interleucina-13/metabolismo , Pulmão/metabolismo , Animais , Sequência de Bases , Canais de Cloreto/genética , Clonagem Molecular , Primers do DNA , Imunofluorescência , Regulação da Expressão Gênica , Hibridização In Situ , Interleucina-10/metabolismo , Pulmão/patologia , Camundongos , Camundongos Transgênicos , Dados de Sequência Molecular , Mucoproteínas/genética , Mucosa/patologia , Fenótipo , Reação em Cadeia da Polimerase , Receptores de Interleucina-4/fisiologia , Fator de Transcrição STAT6 , Transativadores/fisiologiaRESUMO
Signal transducer and activator of transcription 3 (STAT3) is a key transcriptional mediator for many cytokines and is essential for normal embryonic development. We have generated a unique strain of mice with tissue-specific disruption of STAT3 in bone marrow cells during hematopoiesis. This specific STAT3 deletion causes death of these mice within 4-6 weeks after birth with Crohn's disease-like pathogenesis in both the small and large intestine, including segmental inflammatory cell infiltration, ulceration, bowel wall thickening, and granuloma formation. Deletion of STAT3 causes significantly increased cell autonomous proliferation of cells of the myeloid lineage, both in vivo and in vitro. Most importantly, Stat3 deletion during hematopoiesis causes overly pseudoactivated innate immune responses. Although inflammatory cytokines, including tumor necrosis factor alpha and IFN-gamma, are overly produced in these mice, the NAPDH oxidase activity, which is involved in antimicrobial and innate immune responses, is inhibited. The signaling responses to lipopolysaccharide are changed in the absence of STAT3, leading to enhanced NF-kappa B activation. Our results suggest a model in which STAT3 has critical roles in the development and regulation of innate immunity, and deletion of STAT3 during hematopoiesis results in abnormalities in myeloid cells and causes Crohn's disease-like pathogenesis.