RESUMO
AI is rapidly becoming part of many aspects of daily life, with an impact that reaches all fields of research. We asked investigators to share their thoughts on how AI is changing immunology research, what is necessary to move forward, the potential and the pitfalls, and what will remain unchanged as the field journeys into a new era.
Assuntos
Alergia e Imunologia , Inteligência Artificial , Humanos , AnimaisRESUMO
CREATE (Consider, Read, Elucidate hypotheses, Analyze and interpret data, Think of the next Experiment) is a pedagogical approach for teaching and learning science through the rigorous analysis of primary scientific literature. This mini-review focuses on the tools, assignments, and in-class activities by which this strategy immerses students in the process of science and further challenges students to embody the intellectual activities of actual scientists. We highlight the innovative ways in which CREATE pedagogy encourages students to think deeply about science. Applying this strategy has been shown to promote student gains in cognitive and affective behaviors while also fostering the development of science process skills. Herein we also provide a case study of CREATE implementation, which provides a detailed perspective on the realities of teaching with this strategy. Finally, we offer insights gained through the study of this pedagogy at different types of institutions, courses and student populations to demonstrate how CREATE can be broadly applied in STEM education.
Assuntos
Biologia/educação , Criatividade , Estudantes , Ensino/educação , Pensamento , Universidades , Interpretação Estatística de Dados , Humanos , Pensamento/fisiologiaRESUMO
Nkx2.2 is a homeodomain-containing transcription factor essential for pancreatic islet cell specification. In this study we investigate the role of Nkx2.2 within the small intestine. We have determined that Nkx2.2 is expressed at the onset of intestinal epithelial cell differentiation in specific intestinal cell populations, including a subset of enteroendocrine cells. Similar to its role in the pancreatic islet, Nkx2.2 regulates cell fate choices within the intestinal enteroendocrine population; in the Nkx2.2 null mice, several hormone-producing enteroendocrine cell populations are absent or reduced and the ghrelin-producing cell population is upregulated. The remaining intestinal cell populations, including the paneth cells, goblet cells, and enterocytes appear to be unaffected by the loss of Nkx2.2. Furthermore, similar to the pancreatic islet, Nkx2.2 appears to function upstream of Pax6 in regulating intestinal cell fates; Pax6 mRNA and protein expression is decreased in the Nkx2.2 null mice. These studies identify a novel role for Nkx2.2 in intestinal endocrine cell development and reveal the regulatory similarities between cell type specification in the pancreatic islet and in the enteroendocrine population of the intestine.
Assuntos
Linhagem da Célula , Glândulas Endócrinas/citologia , Proteínas de Homeodomínio/fisiologia , Intestino Delgado/citologia , Intestino Delgado/fisiologia , Fatores de Transcrição/fisiologia , Animais , Diferenciação Celular , Glândulas Endócrinas/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Proteína Homeobox Nkx-2.2 , Camundongos , Proteínas de Peixe-ZebraRESUMO
INTRODUCTION: Clinical pathways (CPs) translate best available evidence to the local care context and intend to inform clinical decision-making, optimize care, and decrease variation. This article describes a CPs program improvement process at a free-standing academic children's hospital. Aims: (1) improve the pathway development process; (2) identify and address gaps; (3) strengthen measurement; (4) increase efficiency in cycle time to build a pathway; (5) increase multidisciplinary participation; (6) integrate into the electronic health record ; and (7) and increase pathway utilization. METHODS: We renovated the CP program using a structured, improvement process. A series of internal stakeholder and external colleague interviews informed the process. To improve the program, we developed and implemented different interventions. RESULTS: The streamlined process reduced the overall time for completion from a median of 15 to 5 months (measured from the date of first meeting with the clinical improvement team to approval), a 70% increase in efficiency. Between 1994 and 2015, the hospital had 33 CPs. There was a 78% increase in the total number of pathways after the renovation with 26 additional pathways. CONCLUSIONS: Renovation of the CP program led to early success through an improved development process, alleviation of programmatic gaps, inclusion of measures within each pathway, increased timely completion, multidisciplinary involvement, integration into the electronic health record, and improved utilization. Initial results are encouraging, and the lessons learned should be helpful to other programs. Further program development is ongoing, focusing on continued improvements in implementation and overall program measures.
RESUMO
Breach of B cell tolerance is central to the pathogenesis of systemic lupus erythematosus (SLE). However, how B cell tolerance is subverted in human SLE is poorly understood due to difficulties in identifying relevant autoreactive B cells and in obtaining lymphoid tissue. We have circumvented these limitations by using tonsil biopsies to study autoreactive B cells (9G4 B cells), whose regulation is abnormal in SLE. Here we show that 9G4 B cells are physiologically excluded during the early stages of the GC reaction before acquiring a centroblast phenotype. Furthermore, we provide evidence to indicate that an anergic response to B cell receptor stimulation may be responsible for such behavior. In contrast, in SLE, 9G4 B cells progressed unimpeded through this checkpoint, successfully participated in GC reactions, and expanded within the post-GC IgG memory and plasma cell compartments. The faulty regulation of 9G4 B cells was not shared by RA patients. To our knowledge, this work represents the first comparative analysis of the fate of a specific autoreactive human B cell population. The results identify a defective tolerance checkpoint that appears to be specific for human SLE.
Assuntos
Subpopulações de Linfócitos B/imunologia , Deleção Clonal/imunologia , Centro Germinativo/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Artrite Reumatoide/imunologia , Autoanticorpos/biossíntese , Autoanticorpos/fisiologia , Subpopulações de Linfócitos B/citologia , Subpopulações de Linfócitos B/patologia , Linhagem Celular , Proliferação de Células , Pré-Escolar , Anergia Clonal/imunologia , Feminino , Centro Germinativo/citologia , Centro Germinativo/patologia , Humanos , Imunoglobulina G/biossíntese , Imunoglobulina G/fisiologia , Imunoglobulina M/biossíntese , Memória Imunológica/fisiologia , Imunofenotipagem , Lúpus Eritematoso Sistêmico/patologia , Baço/citologia , Baço/imunologiaRESUMO
Emerging evidence indicates that in addition to their well-characterized role in antigen presentation, MHC II molecules transmit signals that induce death of APCs. Appropriately timed APC death is important for prevention of autoimmunity. Though the exact mechanism of MHC II-mediated cell death signaling is unknown, the response appears independent of caspase activation and does not involve Fas-FasL interaction. Here we investigated MHC II structural requirements for mediation of cell death signaling in a murine B cell lymphoma. We found that neither the transmembrane spanning regions nor the cytoplasmic tails of MHC II, which are required for MHC II-mediated cAMP production and PKC activation, are required for the death response. However, mutations in the connecting peptide region of MHC II alpha chain (alphaCP), but not the beta chain (betaCP), resulted in significant impairment of the death response. The alphaCP mutant was also unable to mediate calcium mobilization responses, and did not associate with Igalpha/beta. Knock-down of Igbeta by shRNA eliminated the MHC II-mediated calcium response but not cell death. We propose that MHC II mediates cell death signaling via association with an undefined cell surface protein(s), whose interaction is partially dependent on alphaCP region.
Assuntos
Antígenos CD79/imunologia , Sinalização do Cálcio , Cálcio/metabolismo , Antígenos de Histocompatibilidade Classe II/química , Antígenos de Histocompatibilidade Classe II/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Antígenos CD79/química , Antígenos CD79/genética , Morte Celular , Linhagem Celular Tumoral , Camundongos , Dados de Sequência MolecularRESUMO
All cells continually survey their environment and make decisions based on cues encountered. This requires specific receptors that detect such cues, then transduce signals that initiate the appropriate responses. B lymphocytes provide an archetypal model for such 'adaptive' cellular responses, where signals transmitted by the B cell Ag-receptor (BCR) influence not only cellular selection, maturation, and survival, but are imperative in generating the ultimate effector function of B cells, i.e. antibody production. While other extracellular stimuli and their cognate receptor signals can also influence B cell development, BCR-mediated signals and the way in which they are integrated and regulated are paramount in defining the cell's physiological fate.
Assuntos
Linfócitos B/fisiologia , Receptores de Antígenos de Linfócitos B/fisiologia , Transdução de Sinais/fisiologia , Animais , Linfócitos B/imunologia , Humanos , Receptores de Antígenos de Linfócitos B/imunologia , Transdução de Sinais/imunologiaRESUMO
Human memory B cells comprise isotype-switched and nonswitched cells with both subsets displaying somatic hypermutation. In addition to somatic hypermutation, CD27 expression has also been considered a universal memory B cell marker. We describe a new population of memory B cells containing isotype-switched (IgG and IgA) and IgM-only cells and lacking expression of CD27 and IgD. These cells are present in peripheral blood and tonsils of healthy subjects and display a degree of hypermutation comparable to CD27+ nonswitched memory cells. As conventional memory cells, they proliferate in response to CpG DNA and fail to extrude rhodamine. In contrast to other recently described CD27-negative (CD27neg) memory B cells, they lack expression of FcRH4 and recirculate in the peripheral blood. Although CD27neg memory cells are relatively scarce in healthy subjects, they are substantially increased in systemic lupus erythematosus (SLE) patients in whom they frequently represent a large fraction of all memory B cells. Yet, their frequency is normal in patients with rheumatoid arthritis or chronic hepatitis C. In SLE, an increased frequency of CD27neg memory cells is significantly associated with higher disease activity index, a history of nephritis, and disease-specific autoantibodies (anti-dsDNA, anti-Smith (Sm), anti-ribonucleoprotein (RNP), and 9G4). These findings enhance our understanding of the B cell diversification pathways and provide mechanistic insight into the immunopathogenesis of SLE.
Assuntos
Subpopulações de Linfócitos B/imunologia , Subpopulações de Linfócitos B/metabolismo , Memória Imunológica , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/patologia , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/biossíntese , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/deficiência , Autoanticorpos/biossíntese , Autoanticorpos/sangue , Subpopulações de Linfócitos B/patologia , Biomarcadores/sangue , Proliferação de Células , Feminino , Humanos , Imunoglobulina D/biossíntese , Imunoglobulina D/deficiência , Imunoglobulina D/genética , Memória Imunológica/genética , Imunofenotipagem , Lúpus Eritematoso Sistêmico/genética , Masculino , Proteínas de Membrana/biossíntese , Proteínas de Membrana/deficiência , Proteínas de Membrana/genética , Tonsila Palatina/imunologia , Tonsila Palatina/metabolismo , Índice de Gravidade de Doença , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/genéticaRESUMO
PURPOSE OF REVIEW: The purpose of this review is to inform the scientific community of the most recent findings surrounding B cell receptor signaling function in human systemic lupus erythematosus and how altered B cell signaling may explain the characteristic hyperactivity of B cells in active disease and contribute to its pathogenesis. RECENT FINDINGS: B cell receptor signaling is abnormal in patients with active systemic lupus erythematosus as demonstrated by increased calcium flux and global B cell hyperactivity. Altered signaling has been explained by a variety of factors such as defective FcgammaRIIB signaling, decreased expression of the protein tyrosine kinase Lyn, and increased serum levels of B lymphocyte stimulator. SUMMARY: The studies reviewed suggest that B cells from systemic lupus erythematosus patients display molecular signaling defects that most likely contribute to pathogenesis of the disease and explain the characteristic hyperactivity of B cells in active disease.
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Linfócitos B/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Ativação Linfocitária/imunologia , Receptores de Antígenos de Linfócitos B/imunologia , Transdução de Sinais/imunologia , Antígenos CD/genética , Antígenos CD/imunologia , Linfócitos B/patologia , Humanos , Interleucina-1/imunologia , Lúpus Eritematoso Sistêmico/genética , Lúpus Eritematoso Sistêmico/patologia , Ativação Linfocitária/genética , Receptores de Antígenos de Linfócitos B/genética , Receptores de IgG/genética , Receptores de IgG/imunologia , Transdução de Sinais/genética , Quinases da Família src/genética , Quinases da Família src/imunologiaRESUMO
The pancreatic islet is necessary for maintaining glucose homeostasis. Within the pancreatic islet, the homeodomain protein Nkx2.2 is essential for the differentiation of all insulin-producing beta cells and a subset of glucagon-producing alpha cells (1). Mice lacking Nkx2.2 have relatively normal sized islets, but a large number of cells within the mutant islet fail to produce any of the four major islet hormones. In this study we demonstrate that Nkx2.2 mutant endocrine cells have been replaced by cells that produce ghrelin, an appetite-promoting peptide predominantly found in the stomach. Intriguingly, normal mouse pancreas also contains a small population of ghrelin-producing cells, defining a new islet "epsilon" cell population. The expansion of ghrelin-producing cells at the expense of beta cells may be a general phenomenon, because we demonstrate that Pax4 mutant mice display a similar phenotype. We propose that insulin and ghrelin cells share a common progenitor and that Nkx2.2 and Pax4 are required to specify or maintain differentiation of the beta cell fate. This finding also suggests that there is a genetic component underlying the balance between insulin and ghrelin in regulating glucose metabolism.
Assuntos
Proteínas de Homeodomínio/genética , Insulina/metabolismo , Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/fisiologia , Pâncreas/embriologia , Hormônios Peptídicos/metabolismo , Fatores de Transcrição/deficiência , Fatores de Transcrição/genética , Amilases/genética , Animais , DNA Complementar/genética , Regulação da Expressão Gênica no Desenvolvimento , Grelina , Glucagon/genética , Hormônio do Crescimento/genética , Hormônio do Crescimento/metabolismo , Heterozigoto , Proteína Homeobox Nkx-2.2 , Proteínas de Homeodomínio/fisiologia , Insulina/genética , Secreção de Insulina , Camundongos , Camundongos Knockout , Análise de Sequência com Séries de Oligonucleotídeos , Fatores de Transcrição Box Pareados , Hormônios Peptídicos/genética , Recombinação Genética , Fatores de Transcrição/fisiologia , Proteínas de Peixe-ZebraRESUMO
Anti-lymphocyte autoantibodies are a well-recognized component of the autoimmune repertoire in human systemic lupus erythematosus (SLE) and have been postulated to have pathogenic consequences. Early studies indicated that IgM anti-lymphocyte autoantibodies mainly recognized T cells and identified CD45, a protein tyrosine phosphatase of central significance in the modulation of lymphocyte function, as the main antigenic target on T cells. However, more recent work indicates that lupus autoantibodies can also recognize B cells and that CD45 may also represent their antigenic target. In particular, IgM Abs encoded by V(H)4.34 appear to have special tropism for B cells, and strong, but indirect evidence suggests that they may recognize a B cell-specific CD45 isoform. Because V(H)4.34 Abs are greatly expanded in SLE, in the present study we investigated the antigenic reactivity of lupus sera V(H)4.34 IgG Abs and addressed their contribution to the anti-lymphocyte autoantibody repertoire in this disease. Our biochemical studies conclusively demonstrate that lupus IgG V(H)4.34 Abs target a developmentally regulated B220-specific glycoform of CD45, and more specifically, an N-linked N-acetyllactosamine determinant preferentially expressed on naive B cells that is sterically masked by sialic acid on B220-positive memory B cells. Strikingly, our data also indicate that this reactivity in SLE sera is restricted to V(H)4.34 Abs and can be eliminated by depleting these Abs. Overall, our data indicate that V(H)4.34 Abs represent a major component of the lupus IgG autoantibody repertoire and suggest that the carbohydrate moiety they recognize may act as a selecting Ag in SLE.
Assuntos
Autoanticorpos/metabolismo , Linfócitos B/imunologia , Sítios de Ligação de Anticorpos , Imunoglobulina G/metabolismo , Cadeias Pesadas de Imunoglobulinas/metabolismo , Região Variável de Imunoglobulina/metabolismo , Antígenos Comuns de Leucócito/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Autoanticorpos/sangue , Linfócitos B/metabolismo , Configuração de Carboidratos , Membrana Celular/imunologia , Membrana Celular/metabolismo , Criança , Pré-Escolar , Humanos , Soros Imunes/metabolismo , Imunoglobulina G/sangue , Cadeias Pesadas de Imunoglobulinas/sangue , Região Variável de Imunoglobulina/sangue , Interfase/imunologia , Antígenos Comuns de Leucócito/metabolismo , Lúpus Eritematoso Sistêmico/sangue , Peso Molecular , Tonsila Palatina/citologia , Isoformas de Proteínas/imunologia , Isoformas de Proteínas/metabolismoRESUMO
OBJECTIVE: B lymphocyte depletion has recently emerged as a promising approach to the treatment of systemic lupus erythematosus (SLE). As part of a phase I/II dose-ranging trial of rituximab in the treatment of SLE, we evaluated the fate of discrete B cell subsets in the setting of selective depletion by anti-CD20 monoclonal antibody and during the B cell recovery phase. METHODS: B cell depletion and phenotype were examined by flow cytometry of peripheral blood mononuclear cells for CD19, CD20, CD27, IgD, and CD38 expression. Changes in autoreactive B lymphocytes and plasma cells were assessed by determination of serum autoantibody levels (anti-double-stranded DNA and VH4.34) and by direct monitoring of a unique autoreactive B cell population bearing surface antibodies whose heavy chain is encoded by the VH4.34 gene segment. RESULTS: Compared with normal controls, SLE patients displayed several abnormalities in peripheral B cell homeostasis at baseline, including naive lymphopenia, expansion of a CD27-,IgD- (double negative) population, and expansion of circulating plasmablasts. Remarkably, these abnormalities resolved after effective B cell depletion with rituximab and immune reconstitution. The frequency of autoreactive VH4.34 memory B cells also decreased 1 year posttreatment, despite the presence of low levels of residual memory B cells at the point of maximal B cell depletion and persistently elevated serum autoantibody titers in most patients. CONCLUSION: This study is the first to show evidence that in SLE, specific B cell depletion therapy with rituximab dramatically improves abnormalities in B cell homeostasis and tolerance that are characteristic of this disease. The persistence of elevated autoantibody titers may reflect the presence of low levels of residual autoreactive memory B cells and/or long-lived autoreactive plasma cells.