Major depression favors the expansion of Th17-like cells and decrease the proportion of CD39+Treg cell subsets in response to myelin antigen in multiple sclerosis patients.

BACKGROUND: Mood disorders have been associated with risk of clinical relapses in multiple sclerosis (MS), a demyelinating disease mediated by myelin-specific T cells. OBJECTIVES: We aimed to investigate the impact of major depressive disorder (MDD) and cytokine profile of T-cells in relapsing remitting MS patients. METHODS: For our study, plasma and PBMC were obtained from 60 MS patients (30 with lifetime MDD) in remission phase. The PBMC cultures were stimulated with anti-CD3/anti-CD28 beads or myelin basic protein (MBP), and effector and regulatory T cell phenotypes were determined by flow cytometry. The cytokine levels, both in the plasma or in the supernatants collected from PBMC cultures, were quantified by Luminex. In some experiments, the effect of serotonin (5-HT) was investigated. RESULTS: Here, higher Th17-related cytokine levels in response to anti-CD3/anti-CD28 and MBP were quantified in the plasma and PBMC cultures of the MS/MDD group in comparison with MS patients. Further, elevated frequency of CD4+ and CD8+ T cells capable of producing IL-17, IL-22 and GM-CSF was observed in depressed patients. Interestingly, the percentage of myelin-specific IFN-γ+IL-17+ and IFN-γ+GM-CSF+ CD4+ T cells directly correlated with neurological disabilities. In contrast, the occurrence of MDD reduced the proportion of MBP-specific CD39+Tregs subsets. Notably, the severity of both neurological disorder and depressive symptoms inversely correlated with these Tregs. Finally, the addition of 5-HT downregulated the release of Th17-related cytokines in response to anti-CD3/anti-CD28 and myelin antigen. CONCLUSIONS: In summary, our findings suggested that recurrent major depression, by favoring imbalances of effector Th17 and Treg cell subsets, contributes to MS severity.

Metabolites and growth factors produced by airway epithelial cells induce tolerance in macrophages.

Macrophages play a role in preventing inflammation in the respiratory tract. To investigate the mechanisms that lead to tolerance in macrophages, we examined the crosstalk between airway epithelial cells (AECs) and macrophages using a 2D coculture model. Culture of macrophages with AECs led to a significant inhibition of LPS induced pro-inflammatory responses. More importantly, AECs induced the secretion of TGF-ß and IL-10 from macrophages even in the absence of LPS stimulation. In addition, the expression of inhibitory molecule, CD200R was also upregulated on AEC exposed macrophages. Furthermore, the AECs exposed macrophages induced significantly increased level of T regulatory cells. Investigation into the possible mechanisms indicated that a combination of growth factor, G-CSF, and metabolites, Kynurenine and lactic acid produced by AECs is responsible for inducing tolerance in macrophages. Interestingly, all these molecules had differential effect on macrophages with G-CSF inducing TGF-ß, Kynurenine elevating IL-10, and lactic acid upregulating CD200R. Furthermore, a cocktail of these factors/metabolites induced similar changes in macrophages as AEC exposure. Altogether, these data identify factors secreted by AECs that enhance tolerance in the respiratory tract. These mediators thus have the potential to be used for therapeutic purposes to modulate respiratory inflammation following local viral infections and lung diseases.

Neuromyelitis optica is an HLA associated disease different from Multiple Sclerosis: a systematic review with meta-analysis.

Neuromyelitis Optica and Multiple Sclerosis are idiopathic inflammatory demyelinating diseases of the central nervous system that currently are considered distinct autoimmune diseases, so differences in genetic susceptibility would be expected. This study aimed to investigate the HLA association with Neuromyelitis Optica by a systematic review with meta-analysis. The STROBE instrument guided research paper assessments. Thirteen papers published between 2009 and 2020 were eligible. 568 Neuromyelitis Optica patients, 41.4% Asians, 32.4% Latin Americans and 26.2% Europeans were analyzed. Only alleles of the DRB1 locus were genotyped in all studies. Neuromyelitis Optica patients have 2.46 more chances of having the DRB1*03 allelic group than controls. Ethnicity can influence genetic susceptibility. The main HLA association with Neuromyelitis Optica was the DRB1*03:01 allele in Western populations and with the DPB1*05:01 allele in Asia. Differences in the Multiple Sclerosis and Neuromyelitis Optica genetic susceptibility was confirmed in Afro descendants. The DRB1*03 allelic group associated with Neuromyelitis Optica has also been described in other systemic autoimmune diseases.

Phenotypic analysis of T follicular helper and T follicular regulatory cells in primary selective IgM deficiency.

Selective IgM deficiency (SIgMD) is a rare immunodeficiency characterized by serum IgM below two standard of mean, and normal IgG and IgA levels. Both in human and mice with selective IgM deficiency, germinal centers cells are decreased. The development of germinal center and humoral immunity are regulated in part by follicular helper T (TFH) and follicular regulatory T (TFR) cells. However, the analysis of circulating TFH (cTFH) and TFR (cTFR) cells in the pathogenesis of SIgMD has not been explored. We observed lower percentage of cTFR cells in SIgMD patients than in control group. However, we did not observe any significant difference in the percentage of cTFH cells and their subsets between both experimental groups. When data were analyzed according to specific antibody response to pneumococcal polysaccharide, we observed a higher percentage of cTFH cells in SIgMD patients with specific antibody deficiency than in SIgMD patients with normal specific antibody response. Our results suggest that cTFH cells and their subsets are preserved in SIgMD patients. However, the role of lower percentage of cTFR cells in the pathogenesis of this immunodeficiency is not clear.

Phenotypic and Functional Analysis of T Follicular Cells in Common Variable Immunodeficiency.

INTRODUCTION: One of the most frequent abnormalities of B cells in common variable immunodeficiency (CVID) is reduced number of class-switched memory B cells, suggesting an impaired germinal center response. Therefore, due to its pivotal role in regulating the development of humoral immunity, the objective of this study was to evaluate the role of circulating T follicular helper (cTFH) and circulating T follicular regulatory (cTFR) cells in the pathogenesis of CVID. METHODS: cTFH and cTFR cells from CVID patients and healthy subjects were phenotypically characterized by flow cytometry. cTFH and memory B cells from CVID patients and healthy subjects were isolated and cocultured. RESULTS: Our results showed a reduced proportion of cTFH17 cells in patients with CVID and an increased ratio of cTFH/cTFR cells in CVID patients with autoimmune diseases. Furthermore, the proportion of IL-21-producing cTFH cells was directly related to the proportion of CD27+ IgD- B cells. Interestingly, coculture assay showed that CVID-derived cTFH cells are able to help memory B cells from healthy controls to produce immunoglobulins. CONCLUSIONS: The proportions of cTFH17 and cTFR cells are altered in CVID patients; however, the cTFH function in assisting B cells to produce antibodies in vitro is preserved.

Is Asian type MS an MS phenotype, an NMO spectrum disorder, or a MOG-IgG related disease?

BACKGROUND: A specific particularity of neurological diseases in Asia is the relative commonality of neuromyelitis optica (NMO) and Asian type MS (OSMS). Both conditions also occur in South American patients. The Brazilian population differs from the European and the Asian populations due to the mixture of ancestralities between European colonizers and African slaves. To better know the clinical characteristics of Brazilian patients with Asian type MS this study aimed to analyze the clinical, radiological and serological data that would help to distinguish between OSMS and NMO and clarify, in a Non-Asian population, if OSMS is an MS phenotype, an NMO spectrum disorder by 2015 classification, or a complement activating antibody to myelin oligodendrocyte glycoprotein (MOG-IgG) antibody-related disease. METHODS: We selected cases retrospectively with NMO and OSMS in the medical registry of patients with idiopathic inflammatory demyelinating diseases under follow-up since 1997 in Federal Hospital da Lagoa, the principal reference center for MS treatment in Rio de Janeiro, Brazil. OSMS has selective involvement of the optic nerve and spinal cord with no cerebral or cerebellar symptoms associated with small spinal cord lesions and negativity for the aquaporin-4 antibody (AQP4-IgG). NMO full-filled the revised criteria (2006) associated with longitudinally extensive transverse myelitis (LETM). We recorded the following data: ethnicity/skin color, neurologic impairment "at nadir" and "at recovery" of the index events (optic neuritis and transverse myelitis), long term disability, mortality, health quality of life scores by the SF-36 questionnaire, CSF IgG oligoclonal bands and serological AQP4-IgG and MOG-IgG antibodies tested by Cell-based assay. The last brain MRIs were classified as either satisfying or not satisfying MAGNIMS radiologic criteria for MS or typical or not typical for NMOSD. The new classification of NMO spectrum disorders (2015) was applied. RESULTS: Forty-one OSMS and 122 NMO cases were analyzed. OSMS affected mainly young white women, causing unilateral optic neuritis and partial myelitis with excellent recovery. After a mean disease duration of 20 years, 90% of the patients had free ambulation, and 70% had a mild disability or no disability. Only 7.2% presented a secondary progressive course, and no deaths occurred. All cases had negativity to AQP4-IgG and MOG-IgG biomarkers. 95% had resonance criteria for MS. OSMS differed from NMO by ethnicity, morbidity, and mortality: most were African descendants, with severe motor and visual dysfunction, and one third died. Only NMO cases full-filled the new NMOSD classification (52 AQP4-IgG positive, 29 AQP4-IgG negative, and 41 AQP4-IgG unknown). CONCLUSION: In Brazilian patients, OSMS and NMO are different immune-mediated diseases. OSMS is a milder MS phenotype.

Reactivation of latent HIV-1 in vitro using an ethanolic extract from Euphorbia umbellata (Euphorbiaceae) latex.

Euphorbia umbellata (E. umbellata) belongs to Euphorbiaceae family, popularly known as Janauba, and its latex contains a combination of phorbol esters with biological activities described to different cellular protein kinase C (PKC) isoforms. Here, we identified deoxi-phorbol esters present in E. umbellata latex alcoholic extract that are able to increase HIV transcription and reactivate virus from latency models. This activity is probably mediated by NF-kB activation followed by nuclear translocation and binding to the HIV LTR promoter. In addition, E. umbellata latex extract induced the production of pro inflammatory cytokines in vitro in human PBMC cultures. This latex extract also activates latent virus in human PBMCs isolated from HIV positive patients as well as latent SIV in non-human primate primary CD4+ T lymphocytes. Together, these results indicate that the phorbol esters present in E. umbellata latex are promising candidate compounds for future clinical trials for shock and kill therapies to promote HIV cure and eradication.

Cognitive changes in nurses working in intensive care units / Alteraciones cognitivas en enfermeros actuantes en unidades de terapia intensiva / Alterações cognitivas em enfermeiros que atuam em unidades de terapia intensiva

ABSTRACT Objective: To measure the levels of stress, anxiety, and depression of nurses working in ICUs, relating them to levels of attention before and after 24 hours. Method: An observational, quantitative, analytical study with 18 nurses undergoing an inventory of stress, anxiety, and depression, as well as assessment of attention levels and psychomotor functioning. Results: Sixty-one percent showed positive for stress. Depression was observed in 33%; and anxiety in 99.9%. A strong correlation between stress and depression (ρ = 0.564 with p <0.05) and anxiety (ρ = 1 with p <0.05) was observed. There was a weak correlation between stress and task execution time in M2 (ρ = 0.055) for TMT A, a fact that did not occur in M0 (ρ = -0.249). Conclusion: The study shows that the workload of the nurses working in 24-hour shifts in the ICU is correlated with high levels of stress, decreases in the attention process, and psychomotor decline.

RESUMEN Objetivo: Medir los niveles de estrés, ansiedad y depresión en enfermeros actuantes en UTI, relacionándolos con los niveles de atención anteriores y posteriores a jornada de 24 horas. Método: Estudio observacional, analítico, de abordaje cuantitativo, realizado con 18 enfermeros que completaron inventario de estrés, ansiedad y depresión; evaluación de niveles de atención y funcionamiento psicomotor. Resultados: 61% sufre estrés positivo. Verificada depresión en 33%; ansiedad en 99,9%. Fuerte correlación entre estrés y depresión (ρ=0,564 con p<0,05) y ansiedad (ρ=1 con p<0,05). Correlación débil entre estrés y tiempo de ejecución de la tarea en M2 (ρ = 0,055) para el TMT A, hecho que no replicado en M0 (ρ=-0,249). Conclusión: El estudio muestra que la carga laboral de los enfermeros actuantes en UTI en turnos de 24 horas está correlacionada con la elevación de los niveles de estrés, disminución del proceso de atención y declinación psicomotora.

RESUMO Objetivo: Medir os níveis de estresse, ansiedade, depressão dos enfermeiros que atuam em UTI, relacionando-os com os níveis de atenção do antes e após jornada de 24 horas. Método: Estudo observacional analítico, de abordagem quantitativa, realizado com 18 enfermeiros submetidos a um inventário de estresse, ansiedade e depressão, avaliação dos níveis de atenção e funcionamento psicomotor. Resultados: 61% possuem estresse positivo. Depressão foi verificada em 33%; de ansiedade, em 99,9%. Forte correlação entre estresse e depressão (ρ=0,564 com p<0,05) e ansiedade (ρ=1 com p<0,05). Correlação fraca entre estresse e o tempo de execução da tarefa em M2 (ρ = 0,055) para o TMT A, fato que não ocorreu em M0 (ρ=-0,249). Conclusão: O estudo mostra que a carga trabalho dos enfermeiros que atuam em UTI, em turnos de 24 horas, está correlacionada com a elevação dos níveis de estresse, diminuição do processo de atenção e declínio psicomotor.

Cognitive changes in nurses working in intensive care units.

OBJECTIVE: To measure the levels of stress, anxiety, and depression of nurses working in ICUs, relating them to levels of attention before and after 24 hours. METHOD: An observational, quantitative, analytical study with 18 nurses undergoing an inventory of stress, anxiety, and depression, as well as assessment of attention levels and psychomotor functioning. RESULTS: Sixty-one percent showed positive for stress. Depression was observed in 33%; and anxiety in 99.9%. A strong correlation between stress and depression (ρ = 0.564 with p <0.05) and anxiety (ρ = 1 with p <0.05) was observed. There was a weak correlation between stress and task execution time in M2 (ρ = 0.055) for TMT A, a fact that did not occur in M0 (ρ = -0.249). CONCLUSION: The study shows that the workload of the nurses working in 24-hour shifts in the ICU is correlated with high levels of stress, decreases in the attention process, and psychomotor decline.

Enhanced Th17 phenotype in uninfected neonates born from viremic HIV-1-infected pregnant women.

Our objective was to evaluate the in vitro functional profile of T cells from uninfected neonates born from HIV-1-infected pregnant women who controlled (G1) or not (G2) the virus replication. We demonstrated that the lymphoproliferation of T cell to polyclonal activators was higher in the G2 as compared with G1. Nevertheless, no detectable proliferative response was observed in response to HIV-1 antigens in both neonate groups. Cytokine dosage in the supernatants of these polyclonally activated T cell cultures demonstrated that, while IL-10 was the dominant cytokine produced in G1, Th17-related cytokines were significantly higher in G2 neonates. The higher Th17 phenotype tendency in G2 was related to high production of IL-23 by lipopolysaccharide-activated monocyte-derived dendritic cells from these neonates. Our results demonstrated immunological disorders in uninfected neonates born from viremic HIV-1-infected mothers that can help to explain why some of these children have elevated risk of clinical morbidity and mortality due to pathological hypersensitivity.

Imunomodulação materna no ciclo gestatório normal e patológico / The Maternal immunomodulation in normal and pathological pregnancy

O conhecimento sobre as mudanças imunoendócrinas às quais as gestantes são condicionadas ajuda a entender muito da relação materno-fetal envolvida na aceitação e rejeição de um corpo semi-estranho. Este trabalho teve por objetivo trazer à comunidade científica a discussão sobre a imunobiologia materna no ciclo gestatório normal e patológico. Nesse contexto, uma rede de citocinas participa de todas as etapas da gestação. Enquanto as citocinas inflamatórias, como TNF-a, IL-1b e o IFN-y, estão envolvidas na fase de implantação e no pré-parto, a produção de citocinas antiinflamatória é responsável pela sobrevivência do feto na cavidade uterina. Muitos pesquisadores acreditam que níveis elevados de estrogênio e, principalmente, de progesterona induzem uma resposta imune materna mediada por células T reguladoras dos tipos 1 e 3, que garante a não responsividade às estruturas fetais. Estes linfócitos T reguladores, quando ativados, produzem IL-10 e TGF-b que contra-regulam os efeitos embriotóxicos das citocinas inflamatórias na interface decídua-trofoblasto. A presença de cortisol em tempos mais tardios da gestação auxilia na manutenção sistêmica desse fenótipo por, dentre outras funções, amplificar a secreção da IL-10 no sangue periférico materno. Esse equilíbrio é tênue e pode ser quebrado por algumas intercorrências, como na pré-eclâmpsia, infecções e nas doenças auto-imunes

Produção de interleucina-10 na gestação reduz a taxa de replicação do HIV-1 em culturas de linfócitos maternos / Interleukin-10 production during pregnancy reduces HIV-1 replicaction in cultures of maternal lymphocytes

OBJETIVO: avaliar a proliferação de células T e a produção de citocinas em gestantes infectadas pelo HIV-1 e seu impacto na replicação viral in vitro. MÉTODOS: sangue periférico de 12 gestantes infectadas pelo HIV-1 e de seus neonatos, bem como de 10 gestantes HIV-1 negativas, foi colhido e a quantidade de linfócitos TCD4+ e TCD8+ periféricos foi avaliada por citometria de fluxo. Para obter plasma ou células mononucleares periféricas (PBMC), as amostras foram centrifugadas na ausência ou presença de um gradiente de Ficoll-Hypaque, respectivamente. As PBMC foram mantidas em cultura por sete dias na presença de fito-hemaglutinina mais IL-2 recombinante e a resposta linfoproliferativa de células T foi analisada pelo método de exclusão em azul de Trypan. Em alguns experimentos, as culturas foram mantidas na presença adicional de anticorpo anti-IL-10. Os plasmas e sobrenadantes das culturas de PBMC ativadas foram submetidos à análise da produção de citocinas, pelo método ELISA indireto, e a carga viral, detectada pelo RT-PCR. RESULTADOS: independente da carga viral plasmática, a resposta linfoproliferativa em culturas de células obtidas de gestantes infectadas pelo HIV foi inferior às amostras normais [4,2±0,37 vs 2,4±0,56 (x 10(6)) células/mL; p<0.005]. Tanto em gestantes normais quanto em pacientes com cargas virais baixas, os níveis de IL-10 foram mais elevados que os das gestantes com elevada replicação viral (9.790±3.224 vs 1.256±350 pg/mL; p=0.002). Níveis elevados de TNF-alfa no soro (7.200±2.440 vs 510±476 pg/mL) e nas culturas de células (21.350±15.230 vs 1.256±350 pg/mL) correlacionaram-se à carga viral plasmática elevada e a infecção neonatal. O bloqueio da IL-10 endógena com anticorpos anti-IL-10 aumentou a replicação do HIV-1 em cultura de células. CONCLUSAO: nossos resultados sugerem que a IL-10 exerce influência negativa na replicação viral, o que provavelmente contribui para reduzir o risco de infecção vertical.