Using Dendritic Cell-Based Immunotherapy to Treat HIV: How Can This Strategy be Improved?

Harnessing dendritic cells (DC) to treat HIV infection is considered a key strategy to improve anti-HIV treatment and promote the discovery of functional or sterilizing cures. Although this strategy represents a promising approach, the results of currently published trials suggest that opportunities to optimize its performance still exist. In addition to the genetic and clinical characteristics of patients, the efficacy of DC-based immunotherapy depends on the quality of the vaccine product, which is composed of precursor-derived DC and an antigen for pulsing. Here, we focus on some factors that can interfere with vaccine production and should thus be considered to improve DC-based immunotherapy for HIV infection

SARS-CoV-2 recombinant proteins stimulate distinct cellular and humoral immune response profiles in samples from COVID-19 convalescent patients

OBJECTIVES: In this preliminary study we investigated cellular and humoral immune responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens in blood samples from 14 recovered coronavirus disease 2019 (COVID-19) patients and compared them to those in samples from 12 uninfected/unvaccinated volunteers. METHODS: Cellular immunity was assessed by intracellular detection of IFN-γ in CD3+ T lymphocytes after stimulation with SARS-CoV-2 spike (S1), nucleocapsid (NC), or receptor-binding domain (RBD) recombinant proteins or overlapping peptide pools covering the sequence of SARS-CoV-2 spike, membrane and nucleocapsid regions. The humoral response was examined by ELISAs and/or chemiluminescence assays for the presence of serum IgG antibodies directed to SARS-CoV-2 proteins. RESULTS: We observed differences between humoral and cellular immune profiles in response to stimulation with the same proteins. Assays of IgG antibodies directed to SARS-CoV-2 NC, RBD and S1/S2 recombinant proteins were able to differentiate convalescent from uninfected/unvaccinated groups. Cellular immune responses to SARS-CoV-2 protein stimuli did not exhibit a specific response, as T cells from both individuals with no history of contact with SARS-CoV-2 and from recovered donors were able to produce IFN-γ. CONCLUSIONS: Determination of the cellular immune response to stimulation with a pool of SARS-CoV-2 peptides but not with SARS-CoV-2 proteins is able to distinguish convalescent individuals from unexposed individuals. Regarding the humoral immune response, the screening for serum IgG antibodies directed to SARS-CoV-2 proteins has been shown to be specific for the response of recovered individuals

Uma análise crítica da aplicação do método Janela do Cliente na avaliação da qualidade do Restaurante Universitário

The Costumer Window method has the main function to measure the level of importance and the degree of satisfaction with the products or services provided by the company, being a tool able to identify the weak points and powers of the company. The purpose of this paper is to present a critical analysis of the application of the method the client window in quality evaluation of University Restaurant, from Federal University of Santa Maria – RS (UFSM). The research involved 1,855 users, including students, teachers and administrative staff. The results showed that in the three units of the University Restaurant, most of the items evaluated is in Quadrant A of the Costumer Window, called "competitive force", ie, most of the items is important to the user and are attended of satisfactorily by restaurants. However, some items are in Quadrant D of the Costumer Window, called "competitive vulnerability", these items are important to the user but they are not being attended adequately. The results of this survey were forwarded to the administration of University Restaurant, which can generate subsides for the formulation of proposals to improvements (changes or adaptations) in the assessed attributes.O método da Janela do Cliente tem por principal função medir o nível de importância e o grau de satisfação em relação aos produtos ou serviços prestados pela empresa, sendo uma ferramenta capaz de identificar os pontos falhos e as potências da empresa. O objetivo desse artigo é apresentar uma análise critica da aplicação do método Janela do Cliente na avaliação da qualidade do Restaurante Universitário da Universidade Federal de Santa Maria – RS (UFSM). Participaram da pesquisa, 1.855 usuários, entre alunos, docentes e técnicos administrativos. Os resultados mostraram que, nas três unidades do Restaurante Universitário, a maioria dos itens avaliados encontra-se no Quadrante A da Janela do Cliente, denominado “força competitiva”, ou seja, a maioria dos itens é importante para o usuário e são atendidos de forma satisfatória pelos restaurantes. No entanto, alguns itens encontram-se no Quadrante D da Janela do Cliente, denominado “vulnerabilidade competitiva”, sendo esses itens importantes para o usuário, porém não estão sendo atendidos de forma adequada. Os resultados dessa pesquisa foram encaminhados à administração do Restaurante Universitário, onde poderão gerar subsídios para formulação de propostas que visam melhorias (mudanças ou adaptações) nos atributos avaliados

Satisfação dos usuários de um Restaurante Universitário: adaptação de uma escala a partir da Teoria da Resposta ao Item

The client’s opinion is one of the most accurate way to evaluate the performance of an organization. The university restaurants play an important role in public institutions of undergraduate education, representing the democratization of the university space, contributing to dropout rate reduction, and improving living conditions for the clients. Item response theory has been presenting good performance in different management applications, including customer satisfaction assessment. Thus, the goal of this paper is to create a scale to evaluate the user satisfaction level of the Universidade Federal de Santa Maria (UFSM) restaurant considering the two-parameter logistic model (2PL) and item response theory (IRT). The data were obtained based on a form prepared using the Google Docs program, in a total of 1.855 respondents. A descriptive analysis of the considered data is presented and the 2LM model was fitted. The results show at what level of the satisfaction assessment the clients of the UFSM university restaurant are situated and the interpretation of the levels. The 2PL and IRT are identified as suitable tools for the data analysis.A avaliação da satisfação de clientes é uma das formas mais eficientes de analisar o desempenho de uma organização. Os restaurantes universitários desempenham um papel de destaque nas Instituições Públicas de Ensino Superior, representam a democratização do espaço universitário, contribuem para a redução dos índices de evasão escolar e possibilitam melhores condições de vida para os usuários. A Teoria de resposta ao item vem mostrando um bom desempenho em várias aplicações na área da gestão, inclusive na avaliação do nível de satisfação de clientes.  Nesse sentido, o presente trabalho teve como objetivo criar uma escala para mensurar o nível de satisfação dos usuários do Restaurante Universitário da Universidade Federal de Santa Maria, por meio do Modelo Logístico de Dois Parâmetros (ML2) da Teoria da Resposta ao Item (TRI). Os dados foram obtidos por meio de um formulário elaborado utilizando o aplicativo Google Docs, totalizando 1.855 respondentes. Uma análise descritiva dos dados é apresentada e em seguida o ML2 é ajustado.  Foi possível identificar em que nível da escala de satisfação os usuários do RU da UFSM estão situados, assim como a interpretação desses níveis. Verificamos que o ML2 da TRI foi adequado para a análise proposta

Immunogenicity of personalized dendritic-cell therapy in HIV-1 infected individuals under suppressive antiretroviral treatment:interim analysis from a phase II clinical trial

BACKGROUND: We developed a personalized Monocyte-Derived Dendritic-cell Therapy (MDDCT) for HIV-infected individuals on suppressive antiretroviral treatment and evaluated HIV-specific T-cell responses. METHODS: PBMCs were obtained from 10 HIV(+) individuals enrolled in trial NCT02961829. Monocytes were differentiated into DCs using IFN-α and GM-CSF. After sequencing each patient’s HIV-1 Gag and determining HLA profiles, autologous Gag peptides were selected based on the predicted individual immunogenicity and used to pulse MDDCs. Three doses of the MDDCT were administered every 15 days. To assess immunogenicity, patients’ cells were stimulated in vitro with autologous peptides, and intracellular IL-2, TNF, and interferon-gamma (IFN-γ) production were measured in CD4(+) and CD8(+) T-cells. RESULTS: The protocol of ex-vivo treatment with IFN-α and GM-CSF was able to induce maturation of MDDCs, as well as to preserve their viability for reinfusion. MDDCT administration was associated with increased expression of IL-2 in CD4(+) and CD8(+) T-cells at 15 and/or 30 days after the first MDDCT administration. Moreover, intracellular TNF and IFN-γ expression was significantly increased in CD4(+) T-cells. The number of candidates that increased in vitro the cytokine levels in CD4(+) and CD8(+) T cells upon stimulation with Gag peptides from baseline to day 15 and from baseline to day 30 and day 120 after MDDCT was significant as compared to Gag unstimulated response. This was accompanied by an increasing trend in the frequency of polyfunctional T-cells over time, which was visible when considering both cells expressing two and three out of the three cytokines examined. CONCLUSIONS: MDDC had a mature profile, and this MDDCT promoted in-vitro T-cell immune responses in HIV-infected patients undergoing long-term suppressive antiretroviral treatment. Trial registration NCT02961829: (Multi Interventional Study Exploring HIV-1 Residual Replication: a Step Towards HIV-1 Eradication and Sterilizing Cure, https://www.clinicaltrials.gov/ct2/show/NCT02961829, posted November 11th, 2016) SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12981-021-00426-z

TESTE DE ACEITABILIDADE DE BOLOS ELABORADOS COM CASCAS DE BANANAS PRATA EM UMA ESCOLA PÚBLICA DO MUNICÍPIO DE RIO VERDE-GO

This study aimed to evaluate the acceptability of cakes made with silver banana peels in a public school in the municipality of Rio Verde-GO, Brazil. The research was developed by the nutritionists of the Center for Receiving, Storage and Distribution of School Meals (CRADAE) linked to the Municipal Department of Education of Rio Verde-(GO). Acceptability tests were applied to 148 students. The cakes were made at the Municipal Center for Early Childhood Education (CMEI) Ana Pimenta. It was found that in the CMEI Ana Pimenta there is an acceptance of 88% of the students in relation to the cake of the silver banana peel, so because it is a daycare center, the percentage of 90% that the FNDE requires is not credible, but the general acceptance of the students in relation to the food. It is concluded that the high acceptance rate achieved by the cake of the silver banana house at CMEI Ana Pimenta, Rio Verde – GO, indicates its viability as an alternative that can improve the nutritional value and minimize the generation of organic waste through the good use of food.Este estudio tuvo como objetivo evaluar la aceptabilidad de los pasteles hechos con cáscaras de plátano plateado en una escuela pública en el municipio de Rio Verde-GO, Brasil. La investigación fue desarrollada por los nutricionistas del Centro de Recepción, Almacenamiento y Distribución de Comidas Escolares (CRADAE) vinculado al Departamento Municipal de Educación de Río Verde-(GO). Se aplicaron pruebas de aceptabilidad a 148 estudiantes. Los pasteles fueron elaborados en el Centro Municipal de Educación Infantil (CMEI) Ana Pimenta. Se encontró que en el CMEI Ana Pimenta hay una aceptación del 88% de los estudiantes en relación a la torta de la cáscara de plátano plateado, por lo que al ser una guardería, el porcentaje del 90% que exige la FNDE no es creíble, sino la aceptación general de los estudiantes en relación a la alimentación. Se concluye que la alta tasa de aceptación alcanzada por la torta de la casa bananera plateada en CMEI Ana Pimenta, Río Verde – GO, indica su viabilidad como una alternativa que puede mejorar el valor nutricional y minimizar la generación de residuos orgánicos a través del buen uso de los alimentos.Objetivou-se avaliar a aceitabilidade de bolos elaborados com cascas de bananas-prata em uma escola pública do município de Rio Verde-GO, Brasil. A pesquisa foi desenvolvida pelos nutricionistas do Centro de Recebimento, Armazenamento e Distribuição da Alimentação Escolar (CRADAE) vinculada à Secretaria municipal de Educação de Rio Verde-(GO). Foram aplicados testes de aceitabilidade para 148 alunos. Os bolos foram elaborados no Centro Municipal de Educação Infantil (CMEI) Ana Pimenta. Verificou-se que no CMEI Ana Pimenta existe uma aceitação de 88% dos escolares em relação ao bolo da casca de banana prata, sendo assim, por se tratar de uma creche, não é credibilizado a porcentagem de 90% que o FNDE exige, e sim a aceitação geral dos alunos em relação ao alimento. Conclui-se que o alto índice de aceitação alcançado pelo bolo da casa de banana-prata no CMEI Ana Pimenta, Rio Verde – GO, indica sua viabilidade como uma alternativa que pode melhorar o valor nutricional e minimizar a geração de resíduos orgânicos por meio do bom aproveitamento dos alimentos.Objetivou-se avaliar a aceitabilidade de bolos elaborados com cascas de bananas-prata em uma escola pública do município de Rio Verde-GO, Brasil. A pesquisa foi desenvolvida pelos nutricionistas do Centro de Recebimento, Armazenamento e Distribuição da Alimentação Escolar (CRADAE) vinculada à Secretaria municipal de Educação de Rio Verde-(GO). Foram aplicados testes de aceitabilidade para 148 alunos. Os bolos foram elaborados no Centro Municipal de Educação Infantil (CMEI) Ana Pimenta. Verificou-se que no CMEI Ana Pimenta existe uma aceitação de 88% dos escolares em relação ao bolo da casca de banana prata, sendo assim, por se tratar de uma creche, não é credibilizado a porcentagem de 90% que o FNDE exige, e sim a aceitação geral dos alunos em relação ao alimento. Conclui-se que o alto índice de aceitação alcançado pelo bolo da casa de banana-prata no CMEI Ana Pimenta, Rio Verde – GO, indica sua viabilidade como uma alternativa que pode melhorar o valor nutricional e minimizar a geração de resíduos orgânicos por meio do bom aproveitamento dos alimentos

ATLANTIC EPIPHYTES: a data set of vascular and non-vascular epiphyte plants and lichens from the Atlantic Forest

Epiphytes are hyper-diverse and one of the frequently undervalued life forms in plant surveys and biodiversity inventories. Epiphytes of the Atlantic Forest, one of the most endangered ecosystems in the world, have high endemism and radiated recently in the Pliocene. We aimed to (1) compile an extensive Atlantic Forest data set on vascular, non-vascular plants (including hemiepiphytes), and lichen epiphyte species occurrence and abundance; (2) describe the epiphyte distribution in the Atlantic Forest, in order to indicate future sampling efforts. Our work presents the first epiphyte data set with information on abundance and occurrence of epiphyte phorophyte species. All data compiled here come from three main sources provided by the authors: published sources (comprising peer-reviewed articles, books, and theses), unpublished data, and herbarium data. We compiled a data set composed of 2,095 species, from 89,270 holo/hemiepiphyte records, in the Atlantic Forest of Brazil, Argentina, Paraguay, and Uruguay, recorded from 1824 to early 2018. Most of the records were from qualitative data (occurrence only, 88%), well distributed throughout the Atlantic Forest. For quantitative records, the most common sampling method was individual trees (71%), followed by plot sampling (19%), and transect sampling (10%). Angiosperms (81%) were the most frequently registered group, and Bromeliaceae and Orchidaceae were the families with the greatest number of records (27,272 and 21,945, respectively). Ferns and Lycophytes presented fewer records than Angiosperms, and Polypodiaceae were the most recorded family, and more concentrated in the Southern and Southeastern regions. Data on non-vascular plants and lichens were scarce, with a few disjunct records concentrated in the Northeastern region of the Atlantic Forest. For all non-vascular plant records, Lejeuneaceae, a family of liverworts, was the most recorded family. We hope that our effort to organize scattered epiphyte data help advance the knowledge of epiphyte ecology, as well as our understanding of macroecological and biogeographical patterns in the Atlantic Forest. No copyright restrictions are associated with the data set. Please cite this Ecology Data Paper if the data are used in publication and teaching events. © 2019 The Authors. Ecology © 2019 The Ecological Society of Americ

Characterization of dendritic cells used in an anti-HIV therapeutic vaccine phase I/II clinical trial

INTRODUÇÃO: A imunoterapia baseada em células dendríticas derivadas de monócitos (MoDCs) constitui uma estratégia promissora para o tratamento de indivíduos infectados pelo HIV. Devido à sua notória plasticidade, populações heterogêneas de MoDCs podem ser obtidas in vitro, dependendo das condições da cultura. Consequentemente, a capacidade dessas células em secretar citocinas e expressar moléculas que participam do processo de apresentação antigênica (MHC, moléculas de adesão e coestimuladoras) é variável, podendo interferir no perfil e eficácia da resposta imune induzida pela terapia. Em nosso laboratório foi desenvolvido um protocolo clínico de vacinação terapêutica baseada em MoDCs e HIV autólogo inativado para o tratamento de indivíduos cronicamente infectados pelo HIV, não expostos à terapia antirretroviral. Deste modo tornou-se oportuna uma investigação in vitro mais aprofundada sobre a produção viral e as características das MoDCs utilizadas como produto vacinal. OBJETIVOS: Caracterizar o produto vacinal constituído por vírus autólogo e MoDCs de indivíduos infectados pelo HIV utilizados em imunoterapia, com relação a aspectos fenotípicos e funcionais. MÉTODOS: Foram incluídos no estudo 17 indivíduos cronicamente infectados pelo HIV, participantes de um estudo clínico de fase I/II de imunoterapia com MoDCs. Células mononucleares do sangue periférico (PBMCs) foram obtidas a partir de leucaférese e parte do material foi utilizada para isolamento e expansão de HIV em sistema de cultura autólogo ou alogênico. Outra parte das PBMCs foi utilizada como fonte de monócitos para diferenciação em MoDCs imaturas que foram pulsadas ou não com o HIV quimicamente inativado pelo aldrithiol-2 (HIV-AT-2), denominadas respectivamente MoDCs HIV-AT-2 e MoDCs maduras, e posteriormente ativadas com citocinas pró-inflamatórias. MoDCs foram avaliadas fenotípica e funcionalmente quanto à expressão de moléculas de superfície, capacidade fagocítica, potencial migratório, produção de citocinas e habilidade em gerar resposta celular in vitro, avaliada por meio da capacidade em induzir proliferação, produção de citocinas e atividade citotóxica em linfócitos T autólogos. RESULTADOS: O rendimento de partículas virais foi mais elevado quando a expansão do HIV foi realizada em sistema alogênico em comparação ao sistema autólogo. Após estímulo para maturação, tanto MoDCs maduras quanto MoDCs HIV-AT-2 apresentaram aumento na expressão de moléculas de coestimulação, ativação e migração, comparado às MoDCs imaturas. Com relação à caracterização funcional, observamos que MoDCs foram capazes de fagocitar partículas de dextran-FITC, exibiram baixo potencial migratório e baixa produção de citocina polarizante para Th1. Ainda, observamos reduzida atividade citotóxica induzida tanto por MoDCs HIV-AT-2 quanto por MoDCs maduras. Por outro lado, MoDCs HIV-AT-2 promoveram proliferação de linfócitos T autólogos e maior polifuncionalidade em células TCD4+ e TCD8+ em comparação às MoDCs maduras. CONCLUSÃO: A produção de vírus autólogo através de sistema alogênico resulta em maior rendimento viral e potencial imunogênico. O produto vacinal composto por MoDCs HIV-AT-2 é capaz de induzir resposta polifuncional antígeno especifica in vitroINTRODUCTION: Immunotherapy based on monocyte-derived dendritic cells (MDDCs) is a promising strategy for the treatment of HIV-infected individuals. Due their plasticity, using different combinations of cytokines cocktail in vitro it is possible to obtain a heterogeneous MDDCs population. Consequently the capacity of these cells to secrete cytokines and express molecules that participate in antigen presentation varies (MHC, adhesion and costimulatory molecules) and can interfere in the profile and efficacy of the immune response induced by this therapy. A clinical trial was conducted in our laboratory to evaluate a immunotherapy based on dendritic cells sensitized with autologous inactivated HIV for the treatment of antiretroviral naive chronically HIV-infected individuals. Therefore, it was a good opportunity to study deeply the virus production and expansion in vitro and to characterize MDDCs used as a vaccine. OBJECTIVE. To characterize MDDCs in context of their phenotype and function as well as investigating viral production and expansion in autologous and allogenic systems. METHODS: 17 patients underwent apheresis before vaccination and their peripheral blood mononuclear cells (PBMCs) were used for autologous virus production and expansion of the virus was carried out in both autologous and allogenic systems. Monocytes were differentiated into immature MDDCs that were pulsed/or not with autologous chemically (aldrithiol-2) inactivated HIV particles (HIV-AT-2). These pulsed (HIV-AT-2 MDDCs) and non-pulsed (mature MDDCs) cells were then activated by proinflammatory cytokines. Phenotypic (cell surface marker) and functional analysis (phagocytosis, transmigration and cytokines production) of MDDCs and their priming and stimulation of lymphocyte (proliferation, polyfunctionality and cytotoxicity) was performed using flow cytometry. RESULTS. Viral yield was higher when expanded in allogenic compared to autologous system. After stimulation with proinflammatory cytokines, both HIV-AT-2 MDDCs and mature MDDCs presented increased costimulation expression, activation and migratory molecules compared to immature MDDCs. Regarding to functional characterization, we observed that MDDCs were able to phagocytize FITC-Dextran and exhibitted a low migratory potential and low production of Th1 polarizing response cytokines. Moreover we observed reduced cytotoxic activity induced by HIV-AT-2 MDDCs and mature MDDCs. On the other hand we also observed that HIV-AT-2 MDDCs were capable of inducing proliferation and polyfunctionality of autologous CD4+ and CD8+ T-lymphocytes compared to mature MDDCs. CONCLUSION. Allogenic system was found to be more efficient in increased viral yield in relation to autologous system. Besides, virus expanded in allogenic system showed a more immunogenic profile. Vaccine product (HIV-AT-2 MDDCs) was able to induce antigen specific polyfunctional respons

Phenotypic and functional profile of IFN-α-differentiated dendritic cells (IFN-DCs) from HIV-infected individuals

Dendritic cell (DC)-based immunotherapy is a promising strategy for the treatment of HIV-infected individuals. Different from the conventional protocol for DC differentiation based on the cytokine IL-4 (IL4-DCs), several studies have suggested obtaining DCs by culturing monocytes with type I IFN (IFN-α) to yield IFN-DCs, as performed in cancer therapy. To evaluate the phenotypic and functional characteristics, monocytes from HIV-infected subjects were differentiated into IFN-DCs or IL4-DCs, pulsed with chemically inactivated HIV and stimulated with pro-inflammatory cytokines. A comparative analysis between both types of monocyte-derived DCs (MoDCs) showed that immature IFN-DCs were phenotypically distinct from immature IL4-DCs at the baseline of differentiation, presenting a pre-activated profile. From the functional profile, we determined that IFN-DCs were capable of producing the cytokine IL-12 p70 and of inducing the production of IFN-γ by CD4 + T lymphocytes but not by TCD8+ lymphocytes. Our results suggest that IFN-DCs derived from HIV-infected individuals are able to recognize and present viral antigens to induce TCD4+ cellular immunity to HIV