Interleukin-32 is a novel inflammatory mediator that has been described to be important in the immunopathogenesis and control of infections caused by Leishmania parasites. By performing experiments with primary human cells in vitro, we demonstrate that the expression of IL-32 isoforms is dependent on the time exposed to L. amazonensis and L. braziliensis antigens. Moreover, for the first time we show the functional consequences of three different genetic variations in the IL32 (rs4786370, rs4349147, rs1555001) modulating IL-32γ expression, influencing innate and adaptive cytokine production after Leishmania exposure. Using a Brazilian cohort of 107 American Tegumentary Leishmaniasis patients and a control cohort of 245 healthy individuals, the IL32 rs4786370 genetic variant was associated with protection against ATL, whereas the IL32 rs4349147 was associated with susceptibility to the development of localized cutaneous and mucosal leishmaniasis. These novel insights may help improve therapeutic strategies and lead to benefits for patients suffering from Leishmania infections.

This study was designed to assess in vitro metacyclogenesis of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis clinical field isolates obtained from patient lesions (L. braziliensis IMG3 and PPS6m; L. amazonensis MAB6). Metacyclogenesis was evaluated by different criteria, namely, promastigote size (morphometric analysis and flow cytometry), surface modifications (loss of lectin or monoclonal antibody (mAb) binding, complement resistance), and infectivity to human macrophages. Growth curves were similar for all parasites evaluated. The various features analyzed were expressed in a high percentage of promastigotes at 6th and 10th days of culture and a low percentage at the 2nd day. However, in most isolates, these features, considered as markers of metacyclogenesis, seemed to develop with different time courses, since the percentages of metacyclic forms detected with each technique were usually different. Parasites from 6th or 10th day and those negatively selected with lectin or mAb similarly infected human macrophages. From all isolates analyzed, L. amazonensis PH8 and MAB6 showed the highest and the lowest levels of susceptibility, respectively, to leishmanicidal activity of IFN-γ/LPS-activated macrophages. Our results showed that by using different techniques to evaluate different aspects of metacyclogenesis (morphological and biochemical modifications) different percentages of metacyclic promastigotes can be detected in each isolate culture.

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Phenotypic and functional aspects of monocytes from Localized Cutaneous Leishmaniasis (LCL) patients were evaluated. The frequencies of monocyte subsets and TLR2/TLR4 expression were evaluated in fresh peripheral blood whereas cytokine production was evaluated in whole blood cell cultures stimulated with TLR agonists or Leishmania braziliensis antigen (Ag). CD16+ monocytes frequency was increased in patients compared with controls. A TLR4 agonist (LPS) induced expression of TNF and IL‐10 in monocyte subsets of patients and controls. The CD14+CD16+ monocytes expressed higher levels of these cytokines than CD14+CD16− cells. The levels of secreted TNF were higher in whole blood cell cultures from patients than controls after LPS/TLR4 or Ag stimulation. Whereas in controls there was a positive correlation between TNF and IL‐10 levels, this was not observed in stimulated cell cultures from patients. The high levels of LPS‐induced TNF were associated with the number of lesions and the percentages of CD14hiCD16+ monocytes. The levels of TLR2‐induced TNF were also associated with number of lesions. All monocyte subsets from patients expressed higher levels of TLR2 and TLR4 than controls. Data suggest that systemically activated monocytes contribute for an imbalance in pro‐ and anti‐inflammatory cytokine production during LCL, participating in the immunopathogenesis of the disease.

Este estudo teve por objetivo avaliar o número de linfócitos T CD4 + e carga viral em pacientes infectados com HIV, atendidos em laboratório de referência em Goiânia, Goiás. Trata-se de pesquisa  observacional, longitudinal e retrospectiva. Foram avaliados prontuários eletrônicos de pacientes submetidos ao exame de contagem de linfócitos T CD4 + e carga viral, no período de janeiro de 2015 a dezembro de 2017. Dos 14 pacientes avaliados, 35,7% (n=5/14) obtiveram, no primeiro exame, resultados abaixo de 200 células/mm³ e a quantificação da carga víral foi expressiva. Na quantificação de linfócitos T CD4+, 85,7% (n=12/14) dos pacientes possuíam valores abaixo do valor de referência (VR) mínimo, quando fizeram o primeiro exame, após o diagnóstico, e todos estes tiveram a carga de RNA viral acima do valor mínimo quantificável (VR: 560 células/mm³ a 2700 células/mm³). Foi constatado que, dos 12 dos pacientes que no início apresentaram valores abaixo do normal, 4 deles (33,3%) conseguiram atingir valores normais de linfócitos T CD4+, após o tratamento, e 50,0% (n=6/12) dos pacientes alcançaram níveis indetectáveis de carga viral. O monitoramento imune, feito pela citometria de fluxo, aliado a quantificação da carga viral de RNA, ganha notoriedade, pois, por meio destas ferramentas, pode-se determinar o grau de comprometimento imunológico dos pacientes e a eficácia do tratamento com antirretroviral.

Peripheral inflammation, particularly mediated by monocytes, can cause neuroinflammation in Parkinson's disease (PD). We investigated the mechanism of TLR2-induced cytokine impairment in peripheral monocytes from PD patients and the association between the presence of CD14 + TLR10 + monocytes and PD severity. Peripheral blood mononuclear cells from PD patients and healthy individuals were evaluated for TLR expression on monocyte subsets (CD14 and CD16 expression) using flow cytometry. Moreover, cytokines were evaluated using flow cytometry after stimulation with Pam 3 Cys (TLR2/TLR1 agonist) in the absence or presence of neutralizing antibodies to TLR10. The severity of PD was assessed using the unified PD rating scale (UPDRS) and motor activity, anxiety (BAI), depression (BDI), and fatigue (PD Fatigue Scale-16) scales. The frequency of CD14 + TLR10 + monocytes and expression intensity of TLR2 and TLR10 were higher in patients with PD than healthy individuals. The frequency of intermediate monocytes (CD14 ++ CD16 + ) was not significantly increased in patients with PD, but was the main monocyte subset expressing TLR10.The TLR2/TLR1-impaired cytokine production TNFα, in PD patients was reversed by neutralizing TLR10. The high frequency of total CD14 + TLR10 + monocytes was associated with a reduction in the severity of PD according to the evaluation of motor and nonmotor symptoms. Peripheral monocytes from patients with PD showed phenotypic and functional alterations. The expression of TLR10 on monocytes can protect against PD by controlling TLR2-induced cytokine production. Furthermore, data suggested that a low frequency of CD14 + TLR10 + monocytes indicates the severity of PD. The results identified new opportunities for the development of novel PD neuroprotective therapies.

Objetivo: Ressaltar o monitoramento da HPN por imunofenotipagem por citometria de fluxo (CMF) e avaliar a eficácia do tratamento com o anticorpo monoclonal eculizumab. Metodologia: O trabalho é uma análise retrospectiva de dados de prontuários dos pacientes encaminhados a um Instituto de referência em doenças hematológicas no Estado de Goiás para diagnóstico e monitoramento da HPN no período de janeiro de 2010 a dezembro de 2017. Resultados: Dos 31 pacientes diagnosticados com HPN, 5 foram selecionados com dados completos nos prontuários. No paciente 1, houve uma melhora significativa com o tratamento, com um aumento do número relativo para os marcadores dos eritrócitos (CD55 e CD59) de 22,7% para 71,7%. Para o paciente 3, a variação média de marcadores foi de 0,5% para 5,2%. No paciente 5 a variação média ficou entre 12,6% e 36,9%. O eculizumab reduziu o quadro hemolítico, melhorando os sintomas e estabilizando os resultados dos testes de hemólise. Conclusão: A imunofenotipagem por CMF é padrão ouro na investigação de HPN por avaliar a expressão de proteínas ancoradas pela GPI com alta sensibilidade e especificidade. Este estudo é o primeiro a demonstrar a eficácia do eculizumab, sem a progressão da doença.

Os meningiomas são tumores benignos intracranianos de evolução lenta que se originam nas meninges. Por serem assintomáticos, a maioria desses tumores, geralmente, são de difícil identificação. Entretanto, se não diagnosticados, podem comprimir vasos sanguíneos cerebrais levando ao óbito. A Ressonância Magnética (RM) é um método de imagem vital para o diagnóstico e caracterização desses tumores, particularmente importante para se estabelecer um plano de tratamento oncológico. O objetivo deste estudo foi descrever a importância da RM no diagnóstico e acompanhamento evolutivo dos meningiomas e na caracterização tissular, correlacionando os achados imagenológicos com análises histológicas do tumor. Este estudo constitui uma revisão bibliográfica narrativa. Os meningiomas são, geralmente, isointensos em T1 e T2, com variações ligadas à textura mais ou menos compacta ou hidratada do tecido, associados à presença de cistos, calcificações ou metaplasia adiposa. Podem provocar hiperostose, levando ao espessamento do osso afetado. Em imagens ponderadas em T1, pós contraste, os meningiomas, geralmente, aparecem com hipersinal homogêneo. A RM contribui, de forma promissora, para a detecção e caracterização dos meningiomas fornecendo dados prognósticos para o seu tratamento. Um diagnóstico preciso auxilia no acompanhamento evolutivo e pós cirúrgico. Em um período de 24 horas pós cirúrgico, a RM é capaz de identificar resíduos tumorais e recorrências tardias, comuns entre 10 a 20 anos depois da ressecção total do tumor.