RESUMO
Human immunodeficiency virus 1 (HIV-1) infects blood monocytes that cross the blood-brain barrier to the central nervous system, inducing neuronal damage. This is prompted by the secretion of viral and neurotoxic factors by HIV-infected macrophages, resulting in HIV-associated neurocognitive disorders. One of these neurotoxic factors is cathepsin B (CATB), a lysosomal cysteine protease that plays an important role in neurodegeneration. CATB interacts with the serum amyloid P component (SAPC), contributing to HIV-induced neurotoxicity. However, the neuronal apoptosis pathways triggered by CATB and the SAPC remain unknown. We aimed to elucidate these pathways in neurons exposed to HIV-infected macrophage-conditioned media before and after the inhibition of CATB or the SAPC with antibodies using tandem mass tag proteomics labeling. Based on the significant fold change (FC) ≥ |2| and p-value < 0.05 criteria, a total of 10, 48, and 13 proteins were deregulated after inhibiting CATB, SAPC antibodies, and the CATB inhibitor CA-074, respectively. We found that neurons exposed to the CATB antibody and SAPC antibody modulate similar proteins (TUBA1A and CYPA/PPIA) and unique proteins (LMNA and HSPH1 for the CATB antibody) or (CFL1 and PFN1 for the SAPC antibody). CATB, SAPC, or apoptosis-related proteins could become potential targets against HIV-induced neuronal degeneration.
Assuntos
Catepsina B , Infecções por HIV , Apoptose , Catepsina B/metabolismo , Infecções por HIV/metabolismo , Humanos , Macrófagos/metabolismo , Profilinas/metabolismo , Componente Amiloide P Sérico/metabolismoRESUMO
Human spermatozoa activate neutrophil extracellular traps (NETs) in vitro. NETosis is an efficient mechanism through which polymorphonuclear neutrophils (PMN) capture sperm in vitro. The objective of this study was to establish the role of store-operated Ca+2 entry (SOCE) in human sperm-triggered NETs and its impact on sperm integrity and oocyte binding capacity. PMN isolated from donors were exposed to spermatozoa isolated from normozoospermic donors using the swim-up technique and were divided into the following groups: (1) sperm, (2) PMN, (3) PMN + sperm, (4) PMN (pretreated with 2-APB, SOCE inhibitor) + sperm, (5) (PMN + DNase) + sperm, and (6) (PMN + PMA) + sperm (positive control). NETs were quantified using PicoGreen® and visualised by scanning electron microscopy and immunofluorescence of extracellular DNA and neutrophil elastase. Plasma membrane, acrosome, and DNA integrity were analysed by flow cytometry, and oocyte binding was evaluated using the hemizona pellucida assay. Sperm-triggered NETosis negatively affected the sperm membrane and acrosome integrity and decreased the oocyte binding capacity. These effects were negated by an SOCE inhibitor, thus improving sperm function and achieving high oocyte binding capacity. The SOCE inhibitor significantly reduced NET formation compared with that in control PMN/sperm (P < 0.05). Collectively, these results advance the knowledge about the role of PMN in reproduction and will allow the development of strategies to block NET formation in situations of reduced fertilisation success.
Assuntos
Cálcio/metabolismo , Armadilhas Extracelulares/metabolismo , Neutrófilos/fisiologia , Espermatozoides , Adulto , Compostos de Boro , Proteína C-Reativa/metabolismo , Feminino , Voluntários Saudáveis , Histonas/metabolismo , Humanos , Masculino , Microscopia Eletrônica de Varredura , Componente Amiloide P Sérico/metabolismo , Adulto JovemRESUMO
Pentraxin-3 has been reported as a promising biomarker of pre-eclampsia and its severity; however, available studies have small sample sizes, and analyses are not always adjusted for confounders. The aim of this study is to establish the strength of the association between maternal Pentraxin-3 level and pre-eclampsia or HELLP syndrome. It was a case-control study. Women with pre-eclampsia or HELLP syndrome were defined as cases, and women with healthy pregnancies at term (>37 weeks) were classified as controls. Plasma concentrations of Pentraxin-3 were determined at the time of delivery by quantitative enzyme immunoassay. Associations between Pentraxin-3 and pre-eclampsia and HELLP syndrome were assessed by multinomial logistic regression. Subsidiary analysis for the time of disease onset was also carried out. Odds ratios and 95% confidence intervals are reported. A total of 1024 pregnant women were included (461 controls, 368 pre-eclampsia, 195 HELLP). A positive log-linear relationship was found between the top pentraxin-3 quintile and HELLP syndrome. After adjustment for confounders (maternal age, ethnicity, socioeconomic position, date and place of recruitment, family history of pre-eclampsia, smoking, body mass index at beginning of pregnancy, gestational age and multiple pregnancy), the strength of the association was higher for HELLP syndrome [OR 1.13 (95% CI 1.08; 1.18)] than for pre-eclampsia [OR 1.03 (95% CI 1.03; 1.10)]. No difference according to time of onset or pentraxin-3 level was found. In summary, pentraxin-3 level was associated with pre-eclampsia, but it was more strongly associated with HELLP syndrome. Longitudinal studies with a lower probability of residual confounding are necessary to improve our knowledge about the role of pentraxin-3 in pre-eclampsia.
Assuntos
Proteína C-Reativa/metabolismo , Síndrome HELLP/sangue , Pré-Eclâmpsia/sangue , Componente Amiloide P Sérico/metabolismo , Adolescente , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Adulto JovemAssuntos
Proteína C-Reativa/metabolismo , Infecções/diagnóstico , Lúpus Eritematoso Sistêmico/complicações , Componente Amiloide P Sérico/metabolismo , Adulto , Biomarcadores/sangue , Feminino , Humanos , Infecções/sangue , Infecções/complicações , Lúpus Eritematoso Sistêmico/sangue , Masculino , Pessoa de Meia-Idade , Exacerbação dos Sintomas , Adulto JovemRESUMO
Reperfusion of an ischemic tissue is the treatment of choice for several diseases, including myocardial infarction and stroke. However, reperfusion of an ischemic tissue causes injury, known as Ischemia and Reperfusion Injury (IRI), that limits the benefit of blood flow restoration. IRI also occurs during solid organ transplantation. During IRI, there is activation of the innate immune system, especially neutrophils, which contributes to the degree of injury. It has been shown that PTX3 can regulate multiple aspects of innate immunity and tissue inflammation during sterile injury, as observed during IRI. In humans, levels of PTX3 increase in blood and elevated levels associate with extent of IRI. In mice, there is also enhanced expression of PTX3 in tissues and plasma after IRI. In general, absence of PTX3, as seen in PTX3-deficient mice, results in worse outcome after IRI. On the contrary, increased expression of PTX3, as seen in PTX3 transgenic mice and after PTX3 administration, is associated with better outcome after IRI. The exception is the gut where PTX3 seems to have a clear deleterious role. Here, we discuss mechanisms by which PTX3 contributes to IRI and the potential of taming this system for the treatment of injuries associated with reperfusion of solid organs.
Assuntos
Proteína C-Reativa/metabolismo , Neutrófilos/imunologia , Traumatismo por Reperfusão/metabolismo , Componente Amiloide P Sérico/metabolismo , Animais , Proteína C-Reativa/genética , Regulação da Expressão Gênica , Humanos , Imunidade Inata , Camundongos , Camundongos Knockout , Especificidade de Órgãos , Componente Amiloide P Sérico/genéticaRESUMO
The purpose of this study was to investigate the role of pentraxin 3 (PTX3), a pivotal component of the innate immune system, in gout. Levels of PTX3 and IL-1ß in human samples were evaluated by ELISA. Development of murine gout was evaluated through the levels of cytokines (PTX3, CXCL1, and IL-1ß) and neutrophil recruitment into the joint cavity. Phagocytosis of monosodium urate (MSU) crystals and caspase-1 activation were determined by flow cytometer. Acute gout patients showed elevated concentration of PTX3 in plasma and synovial fluid as compared with healthy and osteoarthritic subjects. Moreover, there was a positive correlation between intra-articular PTX3 and IL-1ß levels. PTX3 was induced in the periarticular tissue of mice postinjection of MSU crystals. Importantly, Ptx3-deficient mice showed reduced inflammation in response to MSU crystal injection compared with wild-type mice, including reduction of neutrophil recruitment into the joint cavity and IL-1ß and CXCL1 production. Interestingly, addition of PTX3 in vitro enhanced MSU crystal phagocytosis by monocytes and resulted in higher production of IL-1ß by macrophages. This contribution of PTX3 to the phagocytosis of MSU crystals and consequent production of IL-1ß occurred through a mechanism mainly dependent on FcγRIII. Thus, our results suggest that PTX3 acts as a humoral pattern recognition molecule in gout facilitating MSU crystal phagocytosis and contributing to the pathogenesis of gouty arthritis.
Assuntos
Artrite Gotosa/imunologia , Proteína C-Reativa/imunologia , Interleucina-1beta/imunologia , Fagocitose/imunologia , Componente Amiloide P Sérico/imunologia , Ácido Úrico/imunologia , Animais , Artrite Gotosa/metabolismo , Artrite Gotosa/patologia , Proteína C-Reativa/metabolismo , Humanos , Interleucina-1beta/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Componente Amiloide P Sérico/metabolismo , Ácido Úrico/metabolismoRESUMO
Hepatitis C virus (HCV) is the main cause of chronic liver disease, cirrhosis and hepatocellular carcinoma (HCC) worldwide. The risk to develop HCC increases with the severity of liver inflammation and fibrosis. Long pentraxin 3 (PTX3) is a soluble pattern-recognition receptor produced by phagocytes and nonimmune cells at sites of inflammation or injury. The aim of the present study was to determine the association of PTX3 polymorphisms and its plasma levels with HCC occurrence among patients with HCV. Samples from 524 patients with chronic hepatitis C were evaluated in this study. Two polymorphisms (rs1840680 and rs2305619) in the PTX3 gene were determined by real-time PCR. PTX3 plasma levels were measured by Enzyme-linked Immunosorbent Assay (ELISA). Our data show a significant association between PTX3 polymorphisms and HCC occurrence in univariate and multivariate analysis (P = 0.024). Patients with HCC had higher PTX3 plasma levels compared to individuals with mild or severe fibrosis (P < 0.0001 and P = 0.002, respectively). In addition, PTX3 rs2305619 polymorphism and plasma levels were correlated with Child-Pugh scores B and C in HCC individuals. PTX3 seems to be a risk factor for HCC occurrence in chronic hepatitis C. This is the first study that evaluates PTX3 in the context of hepatitis C.
Assuntos
Proteína C-Reativa/genética , Proteína C-Reativa/metabolismo , Carcinoma Hepatocelular/sangue , Hepatite C Crônica/virologia , Neoplasias Hepáticas/sangue , Componente Amiloide P Sérico/genética , Componente Amiloide P Sérico/metabolismo , Carcinoma Hepatocelular/virologia , Ensaio de Imunoadsorção Enzimática , Feminino , Hepacivirus , Humanos , Cirrose Hepática/virologia , Neoplasias Hepáticas/virologia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Reação em Cadeia da Polimerase em Tempo Real , Fatores de RiscoAssuntos
Proteína C-Reativa/metabolismo , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/diagnóstico , Doença de Chagas/sangue , Doença de Chagas/diagnóstico , Componente Amiloide P Sérico/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Doenças Cardiovasculares/epidemiologia , Doença de Chagas/epidemiologia , Doença Crônica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de RiscoRESUMO
OBJECTIVE: To evaluate the concentration of PTX3 in amniotic fluid (AF) during the final weeks of normal pregnancies and in pregnancies complicated by preterm delivery (PTD). STUDY DESIGN: A cross-sectional study was conducted with 95 pregnant women followed to term and 25 who presented with PTD. Samples of AF from all patients were obtained during cesarean section and the PTX3 concentration was determined by enzyme immunoassay (ELISA). Maternal characteristics were compared by ANOVA and the Kruskal-Wallis and Chi square tests. Comparison between PTX3 concentrations in the "PTD in labor" and "PTD not in labor" groups were performed using the Mann-Whitney test. A p value <0.05 was considered statistically significant. RESULTS: Regarding term pregnancies, PTX3 concentrations were not statistically different across the period studied (37 weeks to 40 weeks). Among preterm pregnancies, those in preterm labor (PTL) presented higher PTX3 levels than those not in labor (p=0.001) and the risk of occurrence of PTL increased by 1% with a rise of 1pg/mL in PTX3. CONCLUSION: PTX3 is a physiological constituent of the AF, and its concentration is elevated in the presence of spontaneous PTL, reinforcing the theory that PTX3 plays a role in the innate immune response during gestational complications associated with infectious/inflammatory conditions.
Assuntos
Líquido Amniótico/química , Proteína C-Reativa/metabolismo , Trabalho de Parto Prematuro/metabolismo , Componente Amiloide P Sérico/metabolismo , Adolescente , Adulto , Estudos Transversais , Feminino , Idade Gestacional , Humanos , Gravidez , Complicações Infecciosas na Gravidez/imunologia , Nascimento PrematuroRESUMO
Mechanical ventilation is the major cause of iatrogenic lung damage in intensive care units. Although inflammation is known to be involved in ventilator-induced lung injury (VILI), several aspects of this process are still unknown. Pentraxin 3 (PTX3) is an acute phase protein with important regulatory functions in inflammation which has been found elevated in patients with acute respiratory distress syndrome. This study aimed at investigating the direct effect of PTX3 production in the pathogenesis of VILI. Genetically modified mice deficient and that over express murine Ptx3 gene were subjected to high tidal volume ventilation (V(T)=45 mL/kg, PEEP(zero)). Morphological changes and time required for 50% increase in respiratory system elastance were evaluated. Gene expression profile in the lungs was also investigated in earlier times in Ptx3-overexpressing mice. Ptx3 knockout and wild-type mice developed same lung injury degree in similar times (156±42 min and 148±41 min, respectively; p=0.8173). However, Ptx3 over-expression led to a faster development of VILI in Ptx3-overexpressing mice (77±29 min vs 118±41 min, p=0.0225) which also displayed a faster kinetics of Il1b expression and elevated Ptx3, Cxcl1 and Ccl2 transcripts levels in comparison with wild-type mice assessed by quantitative real-time polymerase chain reaction. Ptx3 deficiency did not impacted the time for VILI induced by high tidal volume ventilation but Ptx3-overexpression increased inflammatory response and reflected in a faster VILI development.