Intestinal damage is required for the pro-inflammatory differentiation of commensal CBir1-specific T cells.

Commensal-specific clusters of differentiation (CD)4+ T cells are expanded in patients with inflammatory bowel disease (IBD) compared to healthy individuals. How and where commensal-specific CD4+ T cells get activated is yet to be fully understood. We used CBir1 TCR-transgenic CD4+ T cells, specific to a commensal bacterial antigen, and different mouse models of IBD to characterize the dynamics of commensal-specific CD4+ T-cells activation. We found that CBir1 T cells proliferate following intestinal damage and cognate antigen presentation is mediated by CD11c+ cells in the colon-draining mesenteric lymph nodes. Using assay for transposase-accessible chromatin sequencing and flow cytometry, we showed that activated CBir1 T cells preferentially acquire an effector rather than regulatory phenotype, which is plastic over time. Moreover, CBir1 T cells, while insufficient to initiate intestinal inflammation, contributed to worse disease outcomes in the presence of other CD4+ T cells. Our results suggest that the commensal-specific T-cell responses observed in IBD exacerbate rather than initiate disease.

Intestinal helminth infection transforms the CD4+ T cell composition of the skin.

Intestinal helminth parasites can alter immune responses to vaccines, other infections, allergens and autoantigens, implying effects on host immune responses in distal barrier tissues. We herein show that the skin of C57BL/6 mice infected with the strictly intestinal nematode Heligmosomoides polygyrus contain higher numbers of CD4+ T cells compared to the skin of uninfected controls. Accumulated CD4+ T cells were H. polygyrus-specific TH2 cells that skewed the skin CD4+ T cell composition towards a higher TH2/TH1 ratio which persisted after worm expulsion. Accumulation of TH2 cells in the skin was associated with increased expression of the skin-homing chemokine receptors CCR4 and CCR10 on CD4+ T cells in the blood and mesenteric lymph nodes draining the infected intestine and was abolished by FTY720 treatment during infection, indicating gut-to-skin trafficking of cells. Remarkably, skin TH2 accumulation was associated with impaired capacity to initiate IFN-γ recall responses and develop skin-resident memory cells to mycobacterial antigens, both during infection and months after deworming therapy. In conclusion, we show that infection by a strictly intestinal helminth has long-term effects on immune cell composition and local immune responses to unrelated antigens in the skin, revealing a novel process for T cell colonisation and worm-mediated immunosuppression in this organ.

Liver X receptor regulates Th17 and RORγt+ Treg cells by distinct mechanisms.

The gastrointestinal microenvironment, dominated by dietary compounds and the commensal bacteria, is a major driver of intestinal CD4+ T helper (Th) cell differentiation. Dietary compounds can be sensed by nuclear receptors (NRs) that consequently exert pleiotropic effects including immune modulation. Here, we found that under homeostatic conditions the NR Liver X receptor (LXR), a sensor of cholesterol metabolites, regulates RORγt+ CD4 T cells in the intestine draining mesenteric lymph node (MLN). While LXR activation led to a decrease, LXR-deficiency resulted in an increase in MLN Th17 and RORγt+ Tregs. Mechanistically, LXR signaling in CD11c+ myeloid cells was required to control RORγt+ Treg. By contrast, modulation of MLN Th17 was independent of LXR signaling in either immune or epithelial cells. Of note, horizontal transfer of microbiota between LXRα-/- and WT mice was sufficient to only partially increase MLN Th17 in WT mice. Despite LXRα deficiency resulted in an increased abundance of Ruminococcaceae and Lachnospiraceae bacterial families compared to littermate controls, microbiota ablation (including SFB) was not sufficient to dampen LXRα-mediated expansion of MLN Th17. Altogether, our results suggest that LXR modulates RORγt+ Treg and Th17 cells in the MLN through distinct mechanisms.

A committed tissue-resident memory T cell precursor within the circulating CD8+ effector T cell pool.

An increasing body of evidence emphasizes the role of tissue-resident memory T cells (TRM) in the defense against recurring pathogens and malignant neoplasms. However, little is known with regard to the origin of these cells and their kinship to other CD8+ T cell compartments. To address this issue, we followed the antigen-specific progeny of individual naive CD8+ T cells to the T effector (TEFF), T circulating memory (TCIRCM), and TRM pools by lineage-tracing and single-cell transcriptome analysis. We demonstrate that a subset of T cell clones possesses a heightened capacity to form TRM, and that enriched expression of TRM-fate-associated genes is already apparent in the circulating TEFF offspring of such clones. In addition, we demonstrate that the capacity to generate TRM is permanently imprinted at the clonal level, before skin entry. Collectively, these data provide compelling evidence for early stage TRM fate decisions and the existence of committed TRM precursor cells in the circulatory TEFF compartment.

Commensal Bacteria-Specific CD4+ T Cell Responses in Health and Disease.

Over the course of evolution, mammalian body surfaces have adapted their complex immune system to allow a harmless coexistence with the commensal microbiota. The adaptive immune response, in particular CD4+ T cell-mediated, is crucial to maintain intestinal immune homeostasis by discriminating between harmless (e.g., dietary compounds and intestinal microbes) and harmful stimuli (e.g., pathogens). To tolerate food molecules and microbial components, CD4+ T cells establish a finely tuned crosstalk with the environment whereas breakdown of these mechanisms might lead to chronic disease associated with mucosal barriers and beyond. How commensal-specific immune responses are regulated and how these molecular and cellular mechanisms can be manipulated to treat chronic disorders is yet poorly understood. In this review, we discuss current knowledge of the regulation of commensal bacteria-specific CD4+ T cells. We place particular focus on the key role of commensal-specific CD4+ T cells in maintaining tolerance while efficiently eradicating local and systemic infections, with a focus on factors that trigger their aberrant activation.

Multi-level Strategy for Identifying Proteasome-Catalyzed Spliced Epitopes Targeted by CD8+ T Cells during Bacterial Infection.

Proteasome-catalyzed peptide splicing (PCPS) generates peptides that are presented by MHC class I molecules, but because their identification is challenging, the immunological relevance of spliced peptides remains unclear. Here, we developed a reverse immunology-based multi-level approach to identify proteasome-generated spliced epitopes. Applying this strategy to a murine Listeria monocytogenes infection model, we identified two spliced epitopes within the secreted bacterial phospholipase PlcB that primed antigen-specific CD8+ T cells in L. monocytogenes-infected mice. While reacting to the spliced epitopes, these CD8+ T cells failed to recognize the non-spliced peptide parts in the context of their natural flanking sequences. Thus, we here show that PCPS expands the CD8+ T cell response against L. monocytogenes by exposing spliced epitopes on the cell surface. Moreover, our multi-level strategy opens up opportunities to systematically investigate proteins for spliced epitope candidates and thus strategies for immunotherapies or vaccine design.

Perspectivas das mulheres com câncer de mama sobre a saúde / Health perspectives of women with breast cancer

A compreensão dos aspectos constitutivos da saúde, a partir do olhar das mulheres com câncer de mama, pode colaborar para as tomadas de decisões profissionais e com o plano terapêutico, ampliando os aspectos fundamentais durante o processo do cuidado. O presente estudo objetivou investigar a autoavaliação da saúde e conhecer as percepções de saúde de mulheres participantes do grupo operativo "Guerreiras" e atendidas no Centro Estadual de Atendimento Especializado (CEAE). Foi realizado um estudo transversal, quali-quantitativo, com 15 mulheres que receberam o diagnóstico de câncer de mama. Os dados foram coletados por dois questionários e pelo grupo focal. As participantes possuíam idade mediana de 51 anos, cor parda, sem companheiro(a), aposentada, com ensino médio escolar e renda familiar menores que três salários mínimos e média de dois anos e 6 meses de pós-cirúrgico e três anos e sete meses pós-diagnóstico. A análise categórica dos depoimentos colhidos por meio do grupo focal revelou saúde como: ausência de doença, bem-estar e espiritualidade; e a autoavaliação de saúde mostrou que a maioria dessas mulheres percebem a sua condição de saúde como moderada. Conclui-se que apesar da permanência da visão de saúde como contraposição à doença e sinônimo de bem-estar, as perspectivas sobre saúde do grupo estudado incorporam, ainda, o estado de superação, enfrentamento, valorização da vida, adaptações, crenças, valores e autoconfiança. Dessa forma, amplia-se a visão sobre saúde a partir da percepção da paciente, não focando apenas, no estado em que há a remissão da doença.

The understanding of the constitutive aspects of health, from the perspective of women with breast cancer, can aid professional decision-making and help develop a therapeutic plan, expanding the fundamental aspects during the care process. The present study aimed to investigate the self-assessment of health and to know the health perceptions of women who participated in the "Guerreiras" operating group and attended at the State Center for Specialized Care (CEAE). This cross-sectional, qualitative, and quantitative study was carried out with 15 women who were diagnosed with breast cancer. Data were collected through two questionnaires and the focus group. The participants had a median age of 51 years old, were brown, without a partner, retired, with a high school education, a family income below three minimum wages, and were an average of two years and six months after surgery and three years and seven months post-diagnosis. The categorical analysis of the testimonies collected through the focus group revealed health as: absence of disease, well-being, and spirituality; and the self-rated health showed that most of these women perceive their health condition as moderate. It is concluded that despite the view of health remaining as a counterpoint to the disease and synonymous with well-being, the perspectives on health of the studied group also incorporate the state of overcoming, coping, appreciating life, adaptations, beliefs, values, and self-confidence. Thus, the patient's perception of health has been broadened and not only focuses on the state in which the disease remits.

Characterization of the transport signals that mediate the nucleocytoplasmic traffic of low risk HPV11 E7.

We previously discovered that nuclear import of low risk HPV11 E7 is mediated by its zinc-binding domain via a pathway that is independent of karyopherins/importins (Piccioli et al., 2010. Virology 407, 100-109). In this study we mapped and characterized a leucine-rich nuclear export signal (NES), 76IRQLQDLLL84, within the zinc-binding domain that mediates the nuclear export of HPV11 E7 in a CRM1-dependent manner. We also identified a mostly hydrophobic patch 65VRLVV69 within the zinc-binding domain that mediates nuclear import of HPV11 E7 via hydrophobic interactions with the FG-repeats domain of Nup62. Substitutions of hydrophobic residues to alanine within the 65VRLVV69 sequence disrupt the nuclear localization of 11E7, whereas the R66A mutation has no effect. Overall the data support a model of nuclear entry of HPV11 E7 protein via hydrophobic interactions with FG nucleoporins at the nuclear pore complex.

Farmácias Vivas: caracterização de marcadores químicos ativos para avaliação da qualidade da matéria-prima, intermediário e fitoterápico à base de cidreira (Lippia alba), Quimiotipo II / Live Pharmacies: characterization of active chemical markers for evaluation of the quality of raw material, intermediate and phytotherapeutic based on bushy matgrass (Lippia alba), Chemotype II

A cidreira (Lippia alba (Mill.) N.E. Brown), quimiotipo II, é referidapopularmente como cidreira carmelitana, rica em óleo essencial(limoneno e citral) e foi selecionada pelo Comitê Estadual de Fitoterapiapara integrar o elenco de plantas medicinais de uso nas Farmácias Vivasdo Estado do Ceará, fazendo parte da Relação de Plantas Medicinais-REPLAME/CE (Portaria 275/2012), como terapia complementar àscrises de ansiedade e insônia de leves a moderadas. Vários estudostêm relatado ações farmacológicas do citral - uma mistura de dois isômeros: trans-geranial e cis-neral – especialmente atividade sedativa.O presente trabalho tem como objetivo realizar a caracterização destes marcadores químicos ativos para avaliação da qualidade da matériaprima,intermediário e fitoterápico à base de cidreira (Lippia alba),Quimiotipo II. A metodologia utilizada consistiu em: 1) Caracterização macro e micro morfológica das folhas de L. alba;2) Extração do óleo essencial 3) Preparação do Elixir de L. alba a 8%; 4) Análises por Cromatografia Gasosa acoplada a Espectrômetro de Massas (CG-EM)e Cromatografia em Camada Delgada. Observou-se que a metodologia utilizada pode ser aplicada nos trabalhos de rotina de controle de qualidade de L. alba, quimiotipo II, tanto para caracterização macro e micromorfológica da espécie, como para caracterização do marcadoresquímicos ativos do óleo essencial (citral/neral e geranial). Observouse,ainda, que as técnicas utilizadas para obtenção do extrato fluido edo elixir extraíram estes constituintes ativos do óleo essencial, os quais influenciam na qualidade e atividade destas preparações. O elixir de cidreira pode representar uma alternativa terapêutica simples, segura e de baixo custo para o SUS, em consonância com a Política Nacional de Plantas Medicinais e Fitoterápicos.

Desempenho de smmentes de crambe expostas à tratamentos pré-germinativos / Performance of Seeds Crambe Exposed to Pre-germination Treatments / Comportamiento de semillas de Crambe sometidas a tratamientos pre-germinativoss

Com o estímulo à produção e uso de biodiesel, sementes de crambe constituem hoje uma das melhores opçõ es para o fornecimento de matéria-prima, sendo também excelente alternativa para cultivo de outono-inverno visando rotação de culturas. O objeto deste estudo foi avaliar o efeito da combinação de tratamentos de pré-germinação sobre o comportamento de sementes de crambe. A partir de um lote de sementes da cultivar FMS Brilhante, foi proposto o delineamento experimental inteiramente casualizado, em esquema fatorial 2 x 5. O delineamento foi baseado na combinação de duas condiçõ es estruturais: sementes com e sem pericarpo, e ambas submetidas a tratamentos pré-germinativos com e sem ácido giberélico, sendo: testemunha (sem tratamento); sementes pré-embebidas em água destilada sem adição de giberelina por 24 horas como controle; e sementes préembebidas em solução de ácido giberélico a 4 %, em diferentes concentraçõ es (400; 500 e 600 mg.L-1). Determinou-se o teor de água, primeira contagem de germinação, porcentagem de germinação, emergência de plântulas e índice de velocidade de emergência. Diante dos resultados conclui-se que a retirada do pericarpo em sementes de crambe, cultivar FMS Brilhante, acelera a velocidade de germinação, porém, diminui a sua porcentagem final. A pré-embebição em ácido giberélico (400, 500 e 600 mg.L-1) por 24 horas proporcionou incrementos na germinação e no vigor das sementes de crambe com pericarpo.

Encouraging the production and use of biodiesel, seeds of crambe today constitute one of the best options for the supply of raw material, is also an excellent alternative for autumn-winter crop rotation order. The aim of this study was to evaluate the efficiency of combined pre-germination treatments on the seed behavior of crambe. From a seed sample of FMS Brilhante cultivar, an experimental design completely randomized with a 2 x 5 factorial arrangement was performed. It was formed from combination of two structural conditions, seeds with or without pericarp, and treatments with or without giberelic acid, being: control (no treatment); seeds pre-soaked in distilled water for 24 hours as control; and seeds pre-soaked in gibberellic acid at 4 % at different concentrations (400, 500 and 600 mg.L-1). Water content, first count germination, germination, seedling emergence and emergence rate index were determined. From these results it is concluded that removal of the pericarp in seed of crambe, cultivar FMS Brilhante, accelerated the germination rate, however, decreased your final percentage. The pre-soaking in gibberellic acid (400, 500 and 600 mg L-1) for 24 hours, increased the germination and seed vigor crambe with pericarp.

Como un estímulo a la producción y uso del biodiesel, las semillas de crambe constituyen hoy una de las mejores opciones para el suministro de materia prima, también son una excelente alternativa en la rotación de cultivos de otoã±oinvierno. El objetivo de este estudio fue evaluar la eficiencia de la combinación de tratamientos pre-germinativos sobre el comportamiento de semillas de crambe. A partir de una muestra de semillas del cultivar FMS Brilhante, se propuso un diseã±o experimental completamente aleatorizado, con un esquema factorial 2 x 5. El diseã±o se basó en la combinación de dos condiciones estructurales: semillas con y sin pericarpio, y ambos sometidos a tratamentos pre-germinativos con y sin ácido giberélico, siendo: control (sin tratamiento); semillas pre-embebidas en agua destilada sin giberelina por 24 horas como control; y semillas pre-embebidas en solución de ácido giberélico a 4 %, en diferentes concentraciones (400; 500 e 600 mg.L-1). Se determinó el contenido de agua, primer contaje de germinación y porcentaje de germinación, emergencia de plántulas e índice de velocidad de emergencia. A partir de estos resultados se concluye que la remoción del pericarpio en semillas de crambe, cultivar FMS Brilhante, aceleró la velocidad de germinación, pero, disminuyó su porcentaje final. La pre-imbibición en ácido giberélico (400, 500 e 600 mg.L-1) por 24 horas proporcionó incrementos en la germinación y en el vigor de las semillas de crambe con pericarpio.