Antiprotozoal potential of Vismia species (Hypericaceae), medicinal plants used to fight cutaneous leishmaniasis.

ETHNOPHARMACOLOGICAL RELEVANCE: Species of Vismia (Hypericaceae), known in Brazil as "lacre", are commonly used in traditional Amazonian medicine for the treatment of skin lesions, including those caused by Leishmania infection. AIM OF THE STUDY: Hexane extracts from the leaves of Vismia cayennensis, V. gracilis, V. sandwithii and V. guianensis, as well as from the fruits of the latter, in addition to the anthraquinones vismiaquinone, physcion and chrysophanol isolated from these species were explored for their anti-promastigote and anti-amastigote activity on Leishmania amazonensis. MATERIALS AND METHODS: Extracts were prepared by static maceration with n-hexane. The compounds, isolated by chromatographic techniques, were identified by spectroscopic methods (1H and 13C NMR). Promastigotes of L.amazonensis were incubated with hexane extracts (1-50 µg/mL) or anthraquinones (1-50 µM) and the parasite survival analyzed. The action of compounds on reactive oxygen species (ROS) production, mitochondrial membrane potential, and membrane integrity of promastigotes were evaluated by flow cytometer, and the cytotoxicity on mammalian cells using MTT assay. Furthermore, the activity of compounds against amastigotes and nitric oxide production were also investigated. RESULTS: Vismiaquinone and physcion were obtained from the leaves of V. guianensis. Physcion, as well as chrysophanol, were isolated from V. sandwithii. Vismia cayennensis and V. gracilis also showed vismiaquinone, compound detected in lower quantity in the fruits of V. guianensis. All extracts were active against the parasite, corroborating the popular use. The greatest activity against promastigotes was achieved with V. guianensis extract (IC50 4.3 µg/mL), precisely the most used Vismia species for treating cutaneous leishmaniasis. Vismiaquinone and physcion exhibited relevant activity with IC50 12.6 and 2.6 µM, respectively. Moreover, all extracts and anthraquinones tested induced ROS production, mitochondrial dysfunction, membrane disruption and were able to kill intracellular amastigote forms, being worthy of further in vivo studies as potential antileishmanial drugs. CONCLUSIONS: The overall data achieved in the current investigation scientifically validate the traditional use of Vismia species, mainly V. guianensis, as an anti-Leishmania agent. Furthermore, the promising results presented here indicate species of Vismia as potentially useful resources of Brazilian flora for the discovery of therapeutic solutions for neglected diseases.

CD300f signalling induces inhibitory human monocytes/macrophages.

The CD300 glycoproteins are a family of related leucocyte surface molecules that regulate the immune response via their paired triggering and inhibitory receptors. Here we studied CD300f, an apoptotic cell receptor, and how it modulates the function of human monocytes and macrophages. We showed that CD300f signalling by crosslinking with anti-CD300f mAb (DCR-2) suppressed monocytes causing upregulation of the inhibitory molecule, CD274 (PD-L1) and their inhibition of T cell proliferation. Furthermore, CD300f signalling drove macrophages preferentially towards M2-type with upregulation of CD274, which was further enhanced by IL-4. CD300f signalling activates the PI3K/Akt pathway in monocytes. Inhibition of PI3K/Akt signalling resulting from CD300f crosslinking leads to downregulation of CD274 expression on monocytes. These findings highlight the potential use of CD300f blockade in cancer immune therapy to target immune suppressive macrophages in the tumour microenvironment, a known resistance mechanism to PD-1/PD-L1 checkpoint inhibitors.

Association between simultaneity of health-risk behaviours and self-rated health in Brazilian adolescents.

INTRODUCTION: This study examined the association between simultaneity of four health-risk behaviours, namely, low levels of moderate-to-vigorous physical activity (insufficient MVPA: <420 min/week), tobacco use, alcohol consumption, and excessive television (TV)-(>2 h/d of TV viewing) and self-rated health (SRH) in Brazilian adolescents. METHODS: We used data of 100,551 adolescents from the National School Health Survey, a national cross-sectional study carried out in 2015. Association between simultaneity of health risk behaviours (i.e. the ratio between observed and expected prevalence rates) and SRH was examined using logistic regression models. RESULTS: The majority of the participants were female (51.9%) and 14 years of age (50.6%), and 26% of the participants' SRH ranged from 'average' to 'extremely poor'. Those who engaged in the following combinations of health-risk behaviours had higher odds of worse SRH than their healthier counterparts: insufficient MVPA and tobacco use (odds ratio-OR: 2.0, 95% confidence interval [CI]: 1.4 to 3.0); insufficient MVPA and alcohol consumption (OR: 1.6, 95%CI: 1.3 to 1.9); insufficient MVPA and >2 h/day of TV viewing (OR: 1.3, 95%CI: 1.1 to 1.6); insufficient MVPA, tobacco use and alcohol consumption (OR: 2.1, 95%CI: 1.7, to 2.7); and insufficient MVPA, alcohol consumption and >2 h/day of TV viewing (OR: 1.6, 95%CI: 1.4 to 2.0). CONCLUSIONS: Insufficient MVPA, alcohol consumption, and other health-risk behaviours were associated with worse SRH in Brazilian adolescents.

Anti-Mouse CD83 Monoclonal Antibody Targeting Mature Dendritic Cells Provides Protection Against Collagen Induced Arthritis.

Antibodies targeting the activation marker CD83 can achieve immune suppression by targeting antigen-presenting mature dendritic cells (DC). This study investigated the immunosuppressive mechanisms of anti-CD83 antibody treatment in mice and tested its efficacy in a model of autoimmune rheumatoid arthritis. A rat anti-mouse CD83 IgG2a monoclonal antibody, DCR-5, was developed and functionally tested in mixed leukocyte reactions, demonstrating depletion of CD83+ conventional (c)DC, induction of regulatory DC (DCreg), and suppression of allogeneic T cell proliferation. DCR-5 injection into mice caused partial splenic cDC depletion for 2-4 days (mostly CD8+ and CD83+ cDC affected) with a concomitant increase in DCreg and regulatory T cells (Treg). Mice with collagen induced arthritis (CIA) treated with 2 or 6 mg/kg DCR-5 at baseline and every three days thereafter until euthanasia at day 36 exhibited significantly reduced arthritic paw scores and joint pathology compared to isotype control or untreated mice. While both doses reduced anti-collagen antibodies, only 6 mg/kg achieved significance. Treatment with 10 mg/kg DCR-5 was ineffective. Immunohistological staining of spleens at the end of CIA model with CD11c, CD83, and FoxP3 showed greater DC depletion and Treg induction in 6 mg/kg compared to 10 mg/kg DCR-5 treated mice. In conclusion, DCR-5 conferred protection from arthritis by targeting CD83, resulting in selective depletion of mature cDC and subsequent increases in DCreg and Treg. This highlights the potential for anti-CD83 antibodies as a targeted therapy for autoimmune diseases.

Psychometric analysis of the Brazilian-version Kidscreen-27 questionnaire.

BACKGROUND: The objective of this study was to verify the reliability, discriminatory power and construct validity of the Kidscreen-27 questionnaire in Brazilian adolescents. METHODS: Adolescents that participated of the pilot study (210 adolescents; 52.9% boys; 13.7 years old) and of the baseline (816 participants; 52.7% girls; 13.1 years old) of the Movimente Project in 2016/2017 composed the sample of the present study. This project was carried out in six public schools in the city of Florianópolis, Santa Catarina, Brazil. Test-retest reproducibility was assessed by the intraclass correlation coefficient and Gwet coefficient; internal consistency through McDonald's Omega; Hankins' Delta G coefficient verified the scale's discriminatory power and; confirmatory factor analysis to assess construct validity. RESULTS: Reproducibility values ranged from 0.71 to 0.78 for the dimensions (ICC), and ranged from 0.60 to 0.83 for the items (Gwet). McDonald's Ômega (0.82-0.91) for internal consistency measures. Discriminatory power ranging from 0.94 for the dimension Social Support and Friends to 0.98 for Psychological Well-Being. The factorial loads were > 0.40, except for item 19 (0.36). The fit quality indicators of the model were adequate (X2[df] = 1022.89 [311], p < 0.001; RMSEA = 0.053 (0.049-0.087); CFI = 0.988; TLI = 0.987), confirming the five-factor structure originally proposed. CONCLUSIONS: The Brazilian-version Kidscreen-27 achieved good levels of reproducibility, internal consistency, discriminatory power and construct validity. Its use is adequate to measure the health-related quality of life of adolescents in the Brazilian context.

Efforts on Changing Lifestyle Behaviors May Not Be Enough to Improve Health-Related Quality of Life Among Adolescents: A Cluster-Randomized Controlled Trial.

Schools have been the main context for physical activity (PA) and sedentary behavior (SB) interventions among adolescents, but there is inconsistent evidence on whether they also improve dimensions of the health-related quality of life (HRQoL). The aim of this study was to evaluate the effects of a school-based active lifestyle intervention on dimensions of HRQoL. A secondary aim was to verify whether sex, age, and HRQoL at baseline were moderators of the intervention effect. A cluster-randomized controlled trial was conducted at three control and three intervention schools in Florianopolis, Brazil. All students from 7th to 9th grade were invited to participate. A school year intervention, designed primarily to increase PA and reduce SB, included strategies focused on (i) teacher training on PA, SB, and nutrition, and availability of teaching materials related to these contents; (ii) environmental improvements (i.e., creation and revitalization of spaces for the practice of PA in school); and (iii) education strategies, with the availability of folders and posters regarding PA, SB, and nutrition. Participants and the research staffs were not blinded to group assignment, but a standardized evaluation protocol was applied at baseline and after the intervention (March and November 2017) using the KIDSCREEN-27 to assess HRQoL across five dimensions. Mixed linear models were performed to evaluate the effect of the Movimente intervention on the five HRQoL dimensions. Of the 921 students who answered the questionnaire at baseline, 300 and 434 completed the study in control and intervention groups, respectively (dropouts: 20%). The results revealed no significant effects of the intervention on any HRQoL dimensions. A reduction of the school environment dimension was observed in both the control (-2.44; 95% CI: -3.41 to -1.48) and intervention groups (-2.09; 95% CI: -2.89 to -1.30). Sensitivity analyses showed that students in the highest baseline tertiles of HRQoL in any dimension had a reduction in their respective scores from pre- to post-intervention in both school groups. In conclusion, our results demonstrated no intervention effect on HRQoL dimensions and those students with the highest levels of HRQoL at baseline on all dimensions reduced from pre to post-intervention. CLINICAL TRIAL REGISTRATION: The trial is registered at the Clinical Trial Registry (Trial ID: NCT02944318; date of registration: October 18, 2016).

Dihydrotestosterone (DHT) Enhances Wound Healing of Major Burn Injury by Accelerating Resolution of Inflammation in Mice.

Androgens have been known to inhibit cutaneous wound healing in men and male mice. However, in children with major burn injuries, a synthetic androgen was reported clinically to improve wound healing. The aim of this study is to investigate the role of dihydrotestosterone (DHT) as a new therapeutic approach in treating major burn injury. In the present study, mice received systemic androgen treatment post major burn injury. Wound healing rate and body weight were monitored over 21 days. The serum level of inflammatory cytokines/chemokines were measured using multiplex immunoassays. In addition, splenocyte enumeration was performed by flow cytometry. Healing phases of inflammation, re-epithelialization, cell proliferation and collagen deposition were also examined. In results, DHT treated mice lost less weight and displayed accelerated wound healing but has no impact on hypermetabolism. Mice, after burn injury, displayed acute systemic inflammatory responses over 21 days. DHT treatment shortened the systemic inflammatory response with reduced splenic weight and monocyte numbers on day 14 and 21. DHT treatment also reduced wound infiltrating macrophage numbers. In conclusion, DHT treatment facilitates local wound healing by accelerating the resolution of inflammation, but not through alterations of post-burn hypermetabolic response.

Protocol paper for the Movimente school-based program: A cluster-randomized controlled trial targeting physical activity and sedentary behavior among Brazilian adolescents.

BACKGROUND: A better understanding of how multicomponent school-based interventions work and their effects on health and education outcomes are needed. This paper described the methods of the Movimente Program, a school-based intervention that aims to increase physical activity (PA) and decrease sedentary behavior (SB) among Brazilian students. METHODS: This is a cluster randomized controlled trial with adolescents from 7th to 9th grade in public schools from Florianopolis, Southern Brazil. After agreement, 6 schools were randomly selected to intervention or control groups (3 schools each), and all eligible students were invited to the study. The Movimente intervention program was performed during a school year and included 3 main components: Teacher training (including face-to-face meeting, social media platform, and handbook with lesson plans); improvements in the PA environment in school; and educational strategies. Control schools continued with their traditional schedule. Baseline (March/April 2017), postintervention (November/December 2017), and maintenance (June/July 2018) evaluations included PA and SB as primary outcomes (assessed by self-report and accelerometry). Secondary outcomes included psychosocial factors related to PA and SB (e.g., social support and self-efficacy), as well as health (e.g., quality of life and nutritional status) and education (e.g., academic achievement) outcomes. A program evaluation was performed based on the RE-AIM framework. Participants, intervention staffs, and evaluators were not blinded to group assignment, but a standardized evaluation protocol was applied independently of the trial allocation. RESULTS: Statistical analyses will include a multilevel approach for repeated measurements and mediation analysis. Any side effects of the intervention will be recorded. The sample size close to that expected (n = 1090) was reached (n = 999). The results of this trial will involve valuable information about the effect and the evaluation of a multicomponent intervention carried out in a middle-income country. CONCLUSION: By creating opportunities for adolescents to be active at school using multicomponent strategies, the Movimente program has the potential to enhance students health and academic performance which may encourage the school community (e.g., teachers, principals) to adopt the program. Also, this trial will provide evidence for practitioners, policy makers, and researchers on how multicomponent program may be implemented in a school setting. TRIAL REGISTRATION: The trial is registered at the Clinical Trial Registry (Trial ID: NCT02944318; date of registration: 18 October 2016).

Targeting CD83 in mantle cell lymphoma with anti-human CD83 antibody.

OBJECTIVES: Effective antibody-drug conjugates (ADCs) provide potent targeted cancer therapies. CD83 is expressed on activated immune cells including B cells and is a therapeutic target for Hodgkin lymphoma. Our objective was to determine CD83 expression on non-Hodgkin lymphoma (NHL) and its therapeutic potential to treat mantle cell lymphoma (MCL) which is currently an incurable NHL. METHODS: We analysed CD83 expression on MCL cell lines and the lymph node/bone marrow biopsies of MCL patients. We tested the killing effect of CD83 ADC in vitro and in an in vivo xenograft MCL mouse model. RESULTS: CD83 is expressed on MCL, and its upregulation is correlated with the nuclear factor κB (NF-κB) activation. CD83 ADC kills MCL in vitro and in vivo. Doxorubicin and cyclophosphamide (CP), which are included in the current treatment regimen for MCL, enhance the NF-κB activity and increase CD83 expression on MCL cell lines. The combination of CD83 ADC with doxorubicin and CP has synergistic killing effect of MCL. CONCLUSION: This study provides evidence that a novel immunotherapeutic agent CD83 ADC, in combination with chemotherapy, has the potential to enhance the efficacy of current treatments for MCL.

Targeting CD300f to enhance hematopoietic stem cell transplantation in acute myeloid leukemia.

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) significantly reduces the rate of relapse in acute myeloid leukemia (AML) but comes at the cost of significant treatment-related mortality. Despite the reduction in relapse overall, it remains common, especially in high-risk groups. The outcomes for patients who relapse after transplant remains very poor. A large proportion of the morbidity that prevents most patients from accessing allo-HSCT is due to toxic nonspecific conditioning agents that are required to remove recipient hematopoietic stem and progenitor cells (HSPCs), allowing for successful donor engraftment. CD300f is expressed evenly across HSPC subtypes. CD300f has transcription and protein expression equivalent to CD33 on AML. We have developed an anti-CD300f antibody that efficiently internalizes into target cells. We have generated a highly potent anti-CD300f antibody-drug conjugate (ADC) with a pyrrolobenzodiazepine warhead that selectively depletes AML cell lines and colony forming units in vitro. The ADC synergizes with fludarabine, making it a natural combination to use in a minimal toxicity conditioning regimen. Our ADC prolongs the survival of mice engrafted with human cell lines and depletes primary human AML engrafted with a single injection. In a humanized mouse model, a single injection of the ADC depletes CD34+ HSPCs and CD34+CD38-CD90+ hematopoietic stem cells. This work establishes an anti-CD300f ADC as an attractive potential therapeutic that, if validated in transplant models using a larger cohort of primary AML samples, will reduce relapse rate and toxicity for patients with AML undergoing allo-HSCT.

Is Hematopoietic Stem Cell Transplantation Required to Unleash the Full Potential of Immunotherapy in Acute Myeloid Leukemia?

From monoclonal antibodies (mAbs) to Chimeric Antigen Receptor (CAR) T cells, immunotherapies have enhanced the efficacy of treatments against B cell malignancies. The same has not been true for Acute Myeloid Leukemia (AML). Hematologic toxicity has limited the potential of modern immunotherapies for AML at preclinical and clinical levels. Gemtuzumab Ozogamicin has demonstrated hematologic toxicity, but the challenge of preserving normal hematopoiesis has become more apparent with the development of increasingly potent immunotherapies. To date, no single surface molecule has been identified that is able to differentiate AML from Hematopoietic Stem and Progenitor Cells (HSPC). Attempts have been made to spare hematopoiesis by targeting molecules expressed only on later myeloid progenitors as well as AML or using toxins that selectively kill AML over HSPC. Other strategies include targeting aberrantly expressed lymphoid molecules or only targeting monocyte-associated proteins in AML with monocytic differentiation. Recently, some groups have accepted that stem cell transplantation is required to access potent AML immunotherapy and envision it as a rescue to avoid severe hematologic toxicity. Whether it will ever be possible to differentiate AML from HSPC using surface molecules is unclear. Unless true specific AML surface targets are discovered, stem cell transplantation could be required to harness the true potential of immunotherapy in AML.

[Fruit and vegetable consumption over a decade among adolescents in the State of Santa Catarina, Brazil]. / Consumo de frutas e verduras por adolescentes catarinenses ao longo de uma década.

Understanding the changes in the trends of fruit and vegetable consumption among adolescents is important in order to implement public health actions. The scope of this article is to investigate the changes over a ten-year period in the daily fruit and vegetable consumption among adolescents from Santa Catarina, according to sex, age and housing area. It is a secondary analysis of a panel survey entitled "Lifestyle and Risk Behavior of Adolescents in the State of Santa Catarina, Brazil (ComPAC)." Adolescents (15-19 years of age) of state schools in 2001 (n=5.028) and 2011 (n=6.529) answered a questionnaire about fruit and vegetable consumption as well as about sociodemographic aspects. Descriptive statistics and logistic regression were applied, according to sex, age and housing area. There was a decrease in daily fruit (39.1% and 16.6%) and vegetable (40.1% and 20.6%) consumption. Different prevalence levels were observed when analyzing subgroups, mainly among girls. From 2001 to 2011, daily vegetable consumption among boys and adolescents in rural areas remained the same. The decrease in daily consumption of fruit and vegetables among adolescents aged 15 to 19 in Santa Catarina highlights the need for the development of strategies to reverse this scenario.

Compreender mudanças no consumo de frutas e verduras por adolescentes é importante para criação de ações de saúde pública. O objetivo deste artigo é investigar mudanças, em dez anos, no consumo diário de frutas e verduras por adolescentes, de acordo com sexo, idade e área de moradia. Estudo de painel (análise secundária) da pesquisa "Estilo de vida e comportamentos de risco de jovens catarinenses". Adolescentes (15-19 anos) de escolas estaduais, em 2001 (n = 5.028) e 2011 (n = 6.529), responderam questionário sobre consumo de frutas e verduras e aspectos sociodemográficos. Estatística descritiva e regressão logística (2001 vs 2011), estratificada para sexo, idade e área de moradia. Houve diminuição no consumo diário de frutas (39,1% e 16,6%) e de verduras (40,1% e 20,6%). Diferentes prevalências são observadas de acordo com subgrupos, principalmente entre moças. A chance para consumo diário de verduras entre rapazes e adolescentes de área rural continuou a mesma. A diminuição na prevalência do consumo diário de frutas e verduras por adolescentes catarinenses de 15 a 19 anos aponta necessidade de criação de estratégias para reversão deste cenário.

Consumo de frutas e verduras por adolescentes catarinenses ao longo de uma década / Fruit and vegetable consumption over a decade among adolescents in the State of Santa Catarina, Brazil

Resumo Compreender mudanças no consumo de frutas e verduras por adolescentes é importante para criação de ações de saúde pública. O objetivo deste artigo é investigar mudanças, em dez anos, no consumo diário de frutas e verduras por adolescentes, de acordo com sexo, idade e área de moradia. Estudo de painel (análise secundária) da pesquisa "Estilo de vida e comportamentos de risco de jovens catarinenses". Adolescentes (15-19 anos) de escolas estaduais, em 2001 (n = 5.028) e 2011 (n = 6.529), responderam questionário sobre consumo de frutas e verduras e aspectos sociodemográficos. Estatística descritiva e regressão logística (2001 vs 2011), estratificada para sexo, idade e área de moradia. Houve diminuição no consumo diário de frutas (39,1% e 16,6%) e de verduras (40,1% e 20,6%). Diferentes prevalências são observadas de acordo com subgrupos, principalmente entre moças. A chance para consumo diário de verduras entre rapazes e adolescentes de área rural continuou a mesma. A diminuição na prevalência do consumo diário de frutas e verduras por adolescentes catarinenses de 15 a 19 anos aponta necessidade de criação de estratégias para reversão deste cenário.

Abstract Understanding the changes in the trends of fruit and vegetable consumption among adolescents is important in order to implement public health actions. The scope of this article is to investigate the changes over a ten-year period in the daily fruit and vegetable consumption among adolescents from Santa Catarina, according to sex, age and housing area. It is a secondary analysis of a panel survey entitled "Lifestyle and Risk Behavior of Adolescents in the State of Santa Catarina, Brazil (ComPAC)." Adolescents (15-19 years of age) of state schools in 2001 (n=5.028) and 2011 (n=6.529) answered a questionnaire about fruit and vegetable consumption as well as about sociodemographic aspects. Descriptive statistics and logistic regression were applied, according to sex, age and housing area. There was a decrease in daily fruit (39.1% and 16.6%) and vegetable (40.1% and 20.6%) consumption. Different prevalence levels were observed when analyzing subgroups, mainly among girls. From 2001 to 2011, daily vegetable consumption among boys and adolescents in rural areas remained the same. The decrease in daily consumption of fruit and vegetables among adolescents aged 15 to 19 in Santa Catarina highlights the need for the development of strategies to reverse this scenario.

Smoking among industrial workers in Brazil: association with sociodemographic factors, alcohol consumption, and stress levels.

OBJECTIVE: To determine the prevalence of smoking, as well as its association with sociodemographic factors, alcohol consumption, and stress levels, among industrial workers in Brazil. METHODS: This was a nationwide survey, conducted in 24 capitals in Brazil through the application of a pre-tested questionnaire. The response to the question "What is your smoking status?" was the outcome variable. To determine the associations, we performed Poisson regression analyses in which the inputs were blocks of variables: block 1 (age and marital status); block 2 (level of education and gross family income); block 3 (geographic region); and block 4 (alcohol consumption and stress level). All analyses were stratified by gender. RESULTS: The sample consisted of 47,328 workers ≥ 18 years of age, of whom 14,577 (30.8%) were women. The prevalence of smoking was 13.0% (15.2% in men and 7.9% in women). Advancing age, alcohol consumption, and a high stress level were positively associated with smoking. A lower risk of smoking was associated with being married, having a higher level of education, and living in the northeastern region of the country (versus the southern region). CONCLUSIONS: The prevalence of smoking was greater in men than in women. Alcohol consumption and high stress levels appear to promote smoking.

Distinguishing human peripheral blood CD16+ myeloid cells based on phenotypic characteristics.

Myeloid lineage cells present in human peripheral blood include dendritic cells (DC) and monocytes. The DC are identified phenotypically as HLA-DR+ cells that lack major cell surface lineage markers for T cells (CD3), B cells (CD19, CD20), NK cells (CD56), red blood cells (CD235a), hematopoietic stem cells (CD34), and Mo that express CD14. Both DC and Mo can be phenotypically divided into subsets. DC are divided into plasmacytoid DC, which are CD11c- , CD304+ , CD85g+ , and myeloid DC that are CD11c+ . The CD11c+ DC are readily classified as CD1c+ DC and CD141+ DC. Monocytes are broadly divided into the CD14+ CD16- (classical) and CD14dim CD16+ subsets (nonclassical). A population of myeloid-derived cells that have DC characteristics, that is, HLA-DR+ and lacking lineage markers including CD14, but express CD16 are generally clustered with CD14dim CD16+ monocytes. We used high-dimensional clustering analyses of fluorescence and mass cytometry data, to delineate CD14+ monocytes, CD14dim CD16+ monocytes (CD16+ Mo), and CD14- CD16+ DC (CD16+ DC). We sought to identify the functional and kinetic relationship of CD16+ DC to CD16+ Mo. We demonstrate that differentiation of CD16+ DC and CD16+ Mo during activation with IFNγ in vitro and as a result of an allo-hematopoietic cell transplant (HCT) in vivo resulted in distinct populations. Recovery of blood CD16+ DC in both auto- and allo-(HCT) patients after myeloablative conditioning showed similar reconstitution and activation kinetics to CD16+ Mo. Finally, we show that expression of the cell surface markers CD300c, CCR5, and CLEC5a can distinguish the cell populations phenotypically paving the way for functional differentiation as new reagents become available.

The effect of an intervention on physical activity of moderate-and-vigorous intensity, and sedentary behavior during adolescents' time at school. / Efeito de uma intervenção sobre atividade física moderada a vigorosa e comportamento sedentário no tempo escolar de adolescentes.

INTRODUCTION: This study evaluated the effect of an intervention on the engagement in physical activity (PA) and sedentary behavior (SB) of sixth to ninth grade students during school-time, physical education (PE) classes, and recesses at two public schools in Florianopolis, SC, Brazil. METHOD: Schools were divided into control and experimental groups. Participants wore accelerometers during school-time, and PA and SB were estimated for school-time, PE classes and recesses at the baseline and after the intervention. The intervention was composed of four components: changes were made in the PE classes, including giving instruction to teachers; sports equipment was made available for use during recesses; educational sessions on the format of classes were conducted; folders and posters were distributed. Data was analyzed using an Analysis of Covariance for repeated measures comparing baseline data with post intervention data, and for independent samples when comparing control and intervention groups. RESULTS: A low proportion of engagement in PA and a large engagement in SB was observed on the baseline. PA decreased in the intervention group during PE classes, while it increased in the control group with regard to school-time, PE classes, and recess. The intervention group accumulated more SB during school-time and PE classes after the intervention, while a decrease in the control group's SB during school-time was observed. CONCLUSION: The intervention was not effective in increasing PA or decreasing SB. Environmental and school's organizational factors impact how interventions are conducted, and should be considered beforehand.

INTRODUÇÃO: Este estudo objetivou avaliar o efeito de uma intervenção sobre o engajamento em atividade física (AF) e comportamento sedentário (CS) no período escolar, em aulas de educação física (EF) e nos recreios em estudantes do sexto ao nono ano de escolas públicas de Florianópolis, Santa Catarina. MÉTODOS: Duas escolas foram alocadas em grupo experimental e controle. Os participantes utilizaram acelerômetros no período escolar e o tempo em AF e CS foram estimados no período escolar, aulas de EF e recreios antes e após a intervenção. A intervenção foi composta de quatro componentes: mudanças nas aulas de EF, com formação dos professores; disponibilização de materiais esportivos no recreio; sessões educativas no formato de aulas; e distribuição de folders e cartazes com informações sobre os desfechos da intervenção. Foram empregadas análises de covariância para medidas repetidas comparando a linha de base e pós-intervenção e para amostras independentes, comparando o grupo controle com o grupo intervenção. RESULTADOS: Observou-se uma baixa proporção de AF na escola na linha de base e um elevado volume de CS. O grupo intervenção diminuiu a AF em aulas de EF, enquanto o grupo controle aumentou em todos os períodos. O grupo intervenção também acumulou mais o CS no período escolar e em aulas de EF após a intervenção, enquanto o controle diminuiu o CS no período escolar. CONCLUSÃO: A intervenção proposta não foi efetiva em aumentar a AF e diminuir o CS. Fatores ambientais e de organização escolar podem ter impactado os resultados e devem ser considerados no planejamento de intervenções.

CD300f epitopes are specific targets for acute myeloid leukemia with monocytic differentiation.

Antibody-based therapy in acute myeloid leukemia (AML) has been marred by significant hematologic toxicity due to targeting of both hematopoietic stem and progenitor cells (HSPCs). Achieving greater success with therapeutic antibodies requires careful characterization of the potential target molecules on AML. One potential target is CD300f, which is an immunoregulatory molecule expressed predominantly on myeloid lineage cells. To confirm the value of CD300f as a leukemic target, we showed that CD300f antibodies bind to AML from 85% of patient samples. While one CD300f monoclonal antibody (mAb) reportedly did not bind healthy hematopoietic stem cells, transcriptomic analysis found that CD300f transcripts are expressed by healthy HSPC. Several CD300f protein isoforms exist as a result of alternative splicing. Importantly for antibody targeting, the extracellular region of CD300f can be present with or without the exon 4-encoded sequence. This results in CD300f isoforms that are differentially bound by CD300f-specific antibodies. Furthermore, binding of one mAb, DCR-2, to CD300f exposes a structural epitope recognized by a second CD300f mAb, UP-D2. Detailed analysis of publicly available transcriptomic data indicated that CD34+ HSPC expressed fewer CD300f transcripts that lacked exon 4 compared to AML with monocytic differentiation. Analysis of a small cohort of AML cells revealed that the UP-D2 conformational binding site could be induced in cells from AML patients with monocytic differentiation but not those from other AML or HSPC. This provides the opportunity to develop an antibody-based strategy to target AMLs with monocytic differentiation but not healthy CD34+ HSPCs. This would be a major step forward in developing effective anti-AML therapeutic antibodies with reduced hematologic toxicity.

CD83: Activation Marker for Antigen Presenting Cells and Its Therapeutic Potential.

CD83 is a member of the immunoglobulin (Ig) superfamily and is expressed in membrane bound or soluble forms. Membrane CD83 (mCD83) can be detected on a variety of activated immune cells, although it is most highly and stably expressed by mature dendritic cells (DC). mCD83 regulates maturation, activation and homeostasis. Soluble CD83 (sCD83), which is elevated in the serum of patients with autoimmune disease and some hematological malignancies is reported to have an immune suppressive function. While CD83 is emerging as a promising immune modulator with therapeutic potential, some important aspects such as its ligand/s, intracellular signaling pathways and modulators of its expression are unclear. In this review we discuss the recent biological findings and the potential clinical value of CD83 based therapeutics in various conditions including autoimmune disease, graft-vs.-host disease, transplantation and hematological malignancies.

Examination of CD302 as a potential therapeutic target for acute myeloid leukemia.

Acute myeloid leukemia (AML) is the most common form of adult acute leukemia with ~20,000 new cases yearly. The disease develops in people of all ages, but is more prominent in the elderly, who due to limited treatment options, have poor overall survival rates. Monoclonal antibodies (mAb) targeting specific cell surface molecules have proven to be safe and effective in different haematological malignancies. However, AML target molecules are currently limited so discovery of new targets would be highly beneficial to patients. We examined the C-type lectin receptor CD302 as a potential therapeutic target for AML due to its selective expression in myeloid immune populations. In a cohort of 33 AML patients with varied morphological and karyotypic classifications, 88% were found to express CD302 on the surface of blasts and 80% on the surface of CD34+ CD38- population enriched with leukemic stem cells. A mAb targeting human CD302 was effective in mediating antibody dependent cell cytotoxicity and was internalised, making it amenable to toxin conjugation. Targeting CD302 with antibody limited in vivo engraftment of the leukemic cell line HL-60 in NOD/SCID mice. While CD302 was expressed in a hepatic cell line, HepG2, this molecule was not detected on the surface of HepG2, nor could HepG2 be killed using a CD302 antibody-drug conjugate. Expression was however found on the surface of haematopoietic stem cells suggesting that targeting CD302 would be most effective prior to haematopoietic transplantation. These studies provide the foundation for examining CD302 as a potential therapeutic target for AML.

On the Other Side: Manipulating the Immune Checkpoint Landscape of Dendritic Cells to Enhance Cancer Immunotherapy.

Monoclonal antibodies targeting co-inhibitory immune checkpoint molecules have been successful in clinical trials of both solid and hematological malignancies as acknowledged by the 2018 Nobel Prize in Medicine, however improving clinical response rates is now key to expanding their efficacy in areas of unmet medical need. Antibodies to checkpoint inhibitors target molecules on either T cells or tumor cells to stimulate T cells or remove tumor mediated immunosuppression, respectively. However, many of the well-characterized T cell immune checkpoint receptors have their ligands on antigen presenting cells or exert direct effects on those cells. Dendritic cells are the most powerful antigen presenting cells; they possess the ability to elicit antigen-specific responses and have important roles in regulation of immune tolerance. Despite their theoretical benefits in cancer immunotherapy, the translation of DC therapies into the clinic is yet to be fully realized and combining DC-based immunotherapy with immune checkpoint inhibitors is an attractive strategy. This combination takes advantage of the antigen presenting capability of DC to maximize specific immune responses to tumor antigens whilst removing tumor-associated immune inhibitory mechanisms with immune checkpoint inhibition. Here we review the expression and functional effects of immune checkpoint molecules on DC and identify rational combinations for DC vaccination to enhance antigen-specific T cell responses, cytokine production, and promotion of long-lasting immunological memory.