KIR2DL4 is an important immune modulator expressed in natural killer cells; HLA-G is its main ligand. We have characterized the
KIR2DL4
genetic diversity by considering the promoter, all exons, and ...all introns in a highly admixed Brazilian population sample and by using massively parallel sequencing. We introduce a molecular method to amplify and to sequence the complete
KIR2DL4
gene. To avoid the mapping bias and genotype errors commonly observed in gene families, we have developed and validated a bioinformatic pipeline designed to minimize these errors and applied it to survey the variability of 220 individuals from the State of São Paulo, southeastern Brazil. We have also compared the
KIR2DL4
genetic diversity in the Brazilian cohort with the diversity previously reported by the 1000Genomes consortium.
KIR2DL4
presents high linkage disequilibrium throughout the gene, with coding sequences associated with specific promoters. There are few but divergent promoter haplotypes. We have also detected many new
KIR2DL4
sequences, all bearing nucleotide exchanges in introns and encoding previously described proteins. Exons 3 and 4, which encode the external domains, are the most variable. The ancestry background influences the
KIR2DL4
allele frequencies and must be considered for association studies regarding
KIR2DL4
.
Problem
This study evaluated whether the monocyte inflammatory state in pre‐eclampsia (PE) might be associated with polarization to either M1 classically or M2 alternatively activated monocyte ...subsets.
Method of Study
Eighty‐five women with (PE) and 52 normotensive (NT) pregnant women matched for gestational age were included. Expression of surface receptors characteristic of M1, such as Toll‐like receptor (TLR)2, TLR4, and CD64, or M2, such as CD163 and CD206 monocyte subsets were evaluated in peripheral blood monocytes by flow cytometry. Tumour necrosis factor‐alpha (TNF‐α), interleukin‐(IL)‐12p40, IL‐12p70, and IL‐10 were evaluated in the supernatant of monocyte cultures by ELISA.
Results
Expression of TLR4 and CD64 by monocytes from pre‐eclamptic women was significantly higher, while the expression of CD163 and CD206 expression was significantly lower compared with NT pregnant women. Endogenous production of TNF‐α, IL‐12p40, and IL‐12p70 by monocytes was increased, while synthesis of IL‐10 was lower in women with PE than in NT pregnant women.
Conclusions
Monocytes from women with PE are classically activated, producing higher levels of pro‐inflammatory cytokines, and express surface receptors characteristic of the M1 subset. These results provide evidence that the systemic inflammatory environment in PE may differentiate and polarize these cells to the M1 phenotype.
Pancreatic cancer (PC) has one of the highest mortality rates, with an overall five-year survival rate of only 7%. When diagnosed, PC is limited to the pancreas in only 20% of patients, whereas in ...50% it has already metastasized. This is due to its late diagnosis, which makes the treatments used, such as radiotherapy, difficult, and reduces survival rates. Therefore, the importance of this study in detecting genes that may become possible biomarkers for this type of tumor, especially regarding the human secretome, is highlighted. These genes participate in pathways that are responsible for tumor migration and resistance to therapies, along with other important factors.
To achieve these goals, the following online tools and platforms have been expanded to discover and validate these biomarkers: The Human Protein Atlas database, the Xena Browser platform, Gene Expression Omnibus, the EnrichR platform and the Kaplan-Meier Plotter platform.
Our study adopted a methodology that allows the identification of potential biomarkers related to the effectiveness of radiotherapy in PC. Inflammatory pathways were predominantly enriched, related to the regulation of biological processes, primarily in cytokine-derived proteins, which are responsible for tumor progression and other processes that contribute to the development of the disease.
Radiotherapy treatment demonstrated greater efficacy when used in conjunction with other forms of therapy since it decreased the expression of essential genes involved in several inflammatory pathways linked to tumor progression.
Preeclampsia (PE) is considered the leading cause of maternal and perinatal morbidity and mortality. The placenta seems to play an essential role in this disease, probably due to factors involved in ...its formation and development. The present study aimed to investigate the association between placental lesions, cytokines and angiogenic factors in pregnant women with preeclampsia (PE). We evaluated 20 normotensive pregnant women, 40 with early-onset PE and 80 with late-onset PE. Placental samples were analyzed for histopathology, immunohistochemistry and determination of granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-10 (IL-10), transforming growth factor-beta 1 (TGF-β1), tumor necrosis factor-alpha (TNF-α), placental growth factor (PlGF), vascular endothelial growth factor (VEGF), fms-like tyrosine-kinase-1 (Flt-1) and endoglin (Eng) levels. Higher percentages of increased syncytial knots and increased perivillous fibrin deposits, and greater levels of TNF-α, TGF-β1and Flt-1 were detected in placentas from early-onset PE. Levels of IL-10, VEGF and PlGF were decreased in PE versus normotensive placentas. Both the TNF-α/IL-10 and sFlt-1/PlGF ratios were higher in placental homogenate of early-onset PE than late-onset PE and control groups. The more severe lesions and the imbalance between TNF-α/IL-10 and PlGF/sFlt-1 in placentas from early-onset PE allows differentiation of early and late-onset PE and suggests higher placental impairment in early-onset PE.
Abstract
As whole-genome sequencing (WGS) becomes the gold standard tool for studying population genomics and medical applications, data on diverse non-European and admixed individuals are still ...scarce. Here, we present a high-coverage WGS dataset of 1,171 highly admixed elderly Brazilians from a census-based cohort, providing over 76 million variants, of which ~2 million are absent from large public databases. WGS enables identification of ~2,000 previously undescribed mobile element insertions without previous description, nearly 5 Mb of genomic segments absent from the human genome reference, and over 140 alleles from HLA genes absent from public resources. We reclassify and curate pathogenicity assertions for nearly four hundred variants in genes associated with dominantly-inherited Mendelian disorders and calculate the incidence for selected recessive disorders, demonstrating the clinical usefulness of the present study. Finally, we observe that whole-genome and HLA imputation could be significantly improved compared to available datasets since rare variation represents the largest proportion of input from WGS. These results demonstrate that even smaller sample sizes of underrepresented populations bring relevant data for genomic studies, especially when exploring analyses allowed only by WGS.
Abstract Preeclampsia (PE) is a complication of human pregnancy associated with an intense inflammatory response involving leukocyte activation, as well as elevated production of pro-inflammatory ...cytokines. The nuclear transcription factor-kappa B (NF-κB) is present in cells of the immune system and is responsible for transcription of genes coding for pro-inflammatory proteins. Silibinin is the main component of silymarin, a polyphenolic extract obtained from fruits and seeds of Silybum marianum with potent hepatoprotective and anti-inflammatory activities. In this study, we assessed whether silibinin modulated NF-κB activity and the production of pro-inflammatory cytokines by peripheral blood mononuclear cells (PBMCs) from preeclamptic patients. PBMC from women with PE, normotensive (NT) pregnant women, and nonpregnant (NP) women were cultured with or without silibinin (5 μM and 50 μM) and 1 μg/mL lipopolysaccharide (LPS) for 18 h. The supernatants were assayed for tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) by ELISA. Cells were cultured for 30 min to evaluate NF-κB activity. There was increased endogenous activation of NF-κB as well as TNF-α and IL-1β release by PBMC in the PE group compared with the NT and NP groups. A positive correlation between NF-κB activity and cytokine production was also observed in the PE group. Silibinin was capable of reducing, at least in part, the levels of NF-κB and cytokines TNF-α and IL-1β in preeclamptic women. We conclude that silibinin exhibits potent anti-inflammatory activity on PBMC from preeclamptic women by downmodulation of NF-κB activation and inflammatory cytokine production.
Abstract Preeclampsia (PE), a specific syndrome of pregnancy, can be classified into early and late onset, depending on whether clinical manifestations occur before or after 34 weeks’ gestation. We ...determined whether plasma concentrations of Hsp60 and Hsp70 were related to circulating cytokine levels, as well as kidney and liver functions, in early- and late-onset PE. Two hundred and thirty-seven preeclamptic women (95 with early- and 142 with late-onset PE) were evaluated. Plasma levels of Hsp60, Hsp70, and their specific antibodies, tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1, IL-10, IL-12, and soluble TNF-α-receptor I (sTNFRI) concentrations, were determined by enzyme-linked immunosorbent assay (ELISA). Concentrations of Hsp70, TNF-α, IL-1β, IL-12, and sTNFRI were significantly elevated in patients with early-onset PE compared with women with late-onset PE; IL-10 levels were significantly lower in the early-onset PE group. Concentrations of urea, uric acid, proteinuria, glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), and lactate dehydrogenase (LDH) were also significantly higher in early-onset PE. The percentage of infants with intrauterine growth restriction was also significantly higher in women with early-onset PE. There were positive correlations between Hsp70 levels and TNF-α, TNFRI, IL-1β, IL-12, GOT, GPT, LDH, and uric acid concentrations in early-onset PE group. Thus, early-onset PE was associated with greater maternal and fetal impairment. There are differences in pathophysiology between early- and late-onset PE, highlighting by the difference in Hsp70 levels.
Highlights • ADA activity and hyperuricemia were highly correlated in preeclampsia. • ADA activity may be involved in inflammatory activation of peripheral blood mononuclear cells in preeclampsia. • ...ADA activity may play a role in systemic inflammatory response in women with preeclampsia.
•ADA activity and hyperuricemia were highly correlated in preeclampsia.•ADA activity may be involved in inflammatory activation of peripheral blood mononuclear cells in preeclampsia.•ADA activity may ...play a role in systemic inflammatory response in women with preeclampsia.
Preeclampsia is a specific disorder of human pregnancy that is associated with hyperuricemia and higher levels of pro-inflammatory cytokines. Adenosine deaminase (ADA) is an enzyme present in all human tissues, and is considered an indicator of cellular inflammation. In the present study we assess whether adenosine deaminase (ADA) activity is altered in women with preeclampsia (PE) and contributes to elevated levels of uric acid and pro-inflammatory cytokine production.
The population studied consisted of 60 women with PE, 30 normotensive pregnant women (NT) and 20 non-pregnant women (NP). Uric acid concentration and ADA activity were determined in the serum. Peripheral blood mononuclear cells (PBMCs) were isolated and evaluated for intracellular nuclear transcription factor kappa B (NF-κB) levels and for endogenous tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) production. The data were evaluated with parametric or non-parametric tests with significance set at P<0.05.
ADA levels were higher in the PE group compared with the NT and NP groups (P<0.001). A positive correlation between ADA and uric acid levels was identified in women with PE (P<0.001). Endogenous production of IL-1β and TNF-α, as well as intracellular NF-κB levels, were higher in PBMCs from the PE group than from NT and NP women (P<0.01) and correlated with the ADA concentration in preeclamptic women (P<0.01).
An elevation in ADA activity in women with PE may contribute to their increased levels of uric acid and pro-inflammatory immune activity.
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