Peripheral Mononuclear Cells Of Patients Research Articles

Integrative Single-Cell RNA-Seq and ATAC-Seq Analysis of Peripheral Mononuclear Cells in Patients With Ankylosing Spondylitis.

ObjectiveGenetic studies on ankylosing spondylitis (AS) have identified more than 100 pathogenic genes. Building a bridge between these genes and biologically targeted therapies is the current research hotspot.MethodsWe integrated single-cell assaying transposase-accessible chromatin sequencing (scATAC-seq) and single-cell RNA sequencing (scRNA-seq) to explore the key genes and related mechanisms associated with AS pathogenesis.ResultsWe identified 18 cell types in peripheral mononuclear cells from patients with AS and normal controls and summarized the cell-type-specific abnormal genes by scRNA-seq. Interestingly, we found that the pathogenic gene NFKB involved in AS progression originated from CD8+ T cells. Moreover, we observed an abnormal tumor TNF pathway mediated by abnormal expression of TNF, NFKB, FOS, JUN, and JUNB, and scATAC-seq results confirmed the abnormal accessible binding sites of transcriptional factors FOS, JUN, and JUNB. The final magnetic bead sorting and quantitative real-time PCR(RT-qPCR) confirmed that NFKB, FOS, JUN, and JUNB in CD8+ T cells differed in the AS group.ConclusionsOur results revealed a possible mechanism by which NFKB abnormally regulates FOS, JUN, and JUNB and drives AS progression, providing a novel perspective from a single cell point of view in AS.

Lymphocyte transformation test: History and current approaches

Drug-induced hypersensitivity reactions encompass a variety of different clinical phenotypes ranging from harmless rashes to fatal reactions. They can be classified into allergic (i.e. drug allergy) and non-allergic reactions (i.e. non-allergic hypersensitivity). Drug allergies in turn can either be antibody (e.g. IgE) or T cell-mediated.One of the diagnostic tools for the in vitro detection of drug allergy is the lymphocyte transformation test (LTT) which is based on the activation and expansion of the drug-specific memory T cells following co-incubation of the patient's peripheral mononuclear cells (PMBC) with the suspected drug in vitro. The read-out parameter in the classical LTT is T cell proliferation which can be measured as counts per minute following the addition of radiolabeled thymidine to the cell culture. However, in the course of time different modifications of the classical LTT with regard to the read-out parameters and methods have been proposed. Likewise, variations of the LTT platform itself have been described in the literature.This review article describes the development of the classical LTT and its use in the context of drug allergy detection and summarizes the modifications which have been published over time.

Extracellular vesicles with ubiquitinated adenosine A2A receptor in plasma of patients with coronary artery disease.

Extracellular vesicles (EV) can transfer cellular molecules for specific intercellular communication with potential relevance in pathological conditions. We searched for the presence in plasma from coronary artery disease (CAD) patients of EV containing the adenosine A2A receptor (A2AR), a signalling receptor associated with myocardial ischaemia and whose expression is related to homocysteine (HCy) metabolism. Using protein organic solvent precipitation for plasma EV preparation and Western blotting for protein identification, we found that plasma from CAD patients contained various amounts of EV with ubiquitin bound to A2AR. Interestingly, the presence of ubiquitinated A2AR in EV from patients was dependent on hyperhomocysteinemia, the amount being inversely proportional to A2AR expression in peripheral mononuclear cells in patients with the highest levels of HCy. CEM, a human T cell line, was also found to released EV containing various amounts of ubiquitinated A2AR in stimulated conditions depending on the hypoxic status and HCy level of culture medium. Together, these data show that ubiquitinated A2AR‐containing EV circulate in the plasma of CAD patients and that this presence is related to hyperhomocysteinemia. A2AR in plasma EV could be a useful tool for diagnosis and a promising drug for the treatment of CAD.

HERV-E expression in peripheral mononuclear cells of patients with psoriasis.

Psoriasis is a common inflammatory skin disease characterized by uncontrolled proliferation of keratinocytes and recruitment of T lymphocytes into the skin. Possible triggers for psoriasis have been attributed to drugs or pathogens such as bacteria and possibly virus. Human endogenous retroviruses (HERVs) might play a role in triggering these antiviral immune responses, since the role of HERVs in the pathogenesis of autoimmune diseases has generated considerable interest. Some studies have also reported an association of HERV-E and psoriasis. None of them investigate the HERV-E expression in peripheral blood of psoriasis. All these considerations have prompted us to perform a survey for HERV-E expression in PBMC from psoriatic patients. Peripheral blood mononuclear cells from 69 psoriatic patients were analyzed. Total RNA was extracted and amplified with reverse transcription polymerase chain reaction. Results were compared with those obtained in a cohort of 20 healthy donors. HERV E was expressed in all samples analyzed but the level of expression was much lower in the psoriasis that in HC P<0.0001. The reasons for the unexpected, low levels of HERV expression in psoriatic patients are unclear and might be in part a consequence of antiviral defense mechanisms.

P6342RANKL expression is increased in peripheral mononuclear cells of patients with severe aortic valve stenosis and promotes pro-calcific differentiation of valve cells

P6342RANKL expression is increased in peripheral mononuclear cells of patients with severe aortic valve stenosis and promotes pro-calcific differentiation of valve cells

Evaluation of tumor necrosis factor (TNF)-α mRNA expression level and the rs1799964 polymorphism of the TNF-α gene in peripheral mononuclear cells of patients with inflammatory bowel diseases.

Crohn's disease (CD) and ulcerative colitis (UC) are types of chronic inflammatory bowel disease (IBD) of which the actual causes remain unknown. Emerging data indicate that alterations in cytokine synthesis may be involved in IBD pathogenesis. The aim of the present study was to determine whether the tumor necrosis factor (TNF)-α mRNA expression level and rs1799964 polymorphism are the genetic susceptibility component of IBD development. The TNF-α mRNA expression level of peripheral blood mononuclear cells (PBMCs) was measured using comparative reverse-transcription quantitative polymerase chain reaction (PCR). Genomic DNA from 201 individuals (CD: n=15; UC: n=86; control subjects: n=100) was analyzed for the presence of the TNF-α-1031 polymorphism by PCR-restriction fragment length polymorphism. An increased TNF-α mRNA expression level was additionally observed in the CC genotype of the -1031 TNF-α gene polymorphism compared with the TC and TT genotypes (P<0.05). Furthermore, the present results revealed that there was no significant difference in the genotype/allele frequencies of the -1031 TNF-α gene polymorphism in Iranian IBD patients. By comparison, the TNF-α mRNA expression level was evaluated in patients with a history of taking medications and demonstrated a significant association in the group that received the 5-ASA + Pred + AZA,5. 5-ASA + Pred + AZA + IFX when compared with the other groups (P<0.05). Thus, these results support the hypothesis that overexpression of the TNF-α gene, which correlated with the CC genotype, may represent a genetic risk factor for Iranian IBD.

Down-regulation of increased TRAF6 expression in the peripheral mononuclear cells of patients with primary Sjögren's syndrome by an EBV-EBER1-specific synthetic single-stranded complementary DNA molecule.

We described earlier a simultaneously increased that the increased expression of miRNA-146a/b was accompanied by an increase in the expression of and TRAF6 and a decrease in the expression of IRAK1 genes in the peripheral mononuclear cells (PBMCs) of patients with primary Sjogren's syndrome (pSS) patients. Recently, the expression of EBV encoded. RNA (EBER) was published in the B cells of salivary glands of in pSS. In the present study, we applied an EBV-EBER1 specific synthetic single stranded complementary DNA molecule (EBV-EBER1-cDNA) to test whether any EBER1 related effect exists also in PBMCs of pSS patients. In the PBMCs of pSS patients and healthy controls, we investigated in vitro the effects of a synthetic single stranded EBV-EBER1-cDNA molecule, synthetic double-stranded (ds)RNA polyinosinic-polycytidylic acid [poly (I:C)] and polyadenylic acid potassium salt poly-adenylic acid [poly-(A)] on the expression of TRAF6 gene tested by qRTPCR. The release of interferon -α was detected by ELISA. EBV-EBER1-cDNA resulted in a significant reduction in the expression of TRAF6 in the cells of patients, but in the healthy controls not, whereas the treatments with poly (I:C) and poly-(A) could not reduce the TRAF6 over-expression. No release of EBER1 could be observed in the culture supernatants of patients with pSS. Only the treatment with poly (I:C) resulted in a significant increase of interferon -α release, and only in the heathy controls. No release of EBER1 molecules took place during the culturing of cells. EBV-EBER- cDNA acted functionally on the cells of patients only. These findings give a further evidence of the linkage between EBV and pSS, furthermore, they show the possible role of EBV-EBER1 in the induction of increased TRAF6 expression in the peripheral B cells of Sjögren's patients.

Hepatitis C Virus in Peripheral Mononuclear Cells in Patients on Regular Hemodialysis

Background: Occult hepatitis C virus infection (OCI) was identified as Hepatitis C virus (HCV), characterized by undetectable HCV antibodies and HCV RNA in serum, while HCV RNA is detectable in liver and peripheral blood mononuclear cells (PBMCs) only. Nosocomial transmission in dialysis units maintains a higher prevalence of hepatitis C virus (HCV) infection in patients on maintenance dialysis than in the general population. HCV infection has a detrimental effect on survival in patients on maintenance dialysis and after renal transplantation. The excess risk for death in HCV-positive patients was partially attributed to chronic liver disease with its attendant complications, particularly hepatocellular carcinoma and liver cirrhosis(1). The aim of this study: was to evaluate the hidden infection of hepatitis C virus among regular hemodialysis patients in Bab Al Sharia University Hospital with negative ELISA and PCR by using PCR in mononuclear cells as a marker in the serum of these patients Patients and methods:in this prospective study, 60 patients with end-stage renal disease on regular hemodialysis(for at least 6 months duration)were included. For all patients thorough medical history, clinical examination, kidney function tests, liver function tests, complete blood count, pelvi-abdominal ultrasound, HCVantibodies, hepatitis C viral RNA, quantitative, HbsAg,. HCV PCR done for all patients in serum and mononuclear cells.. Patients with acute or chronic HCV infection as marked by positive hepatitis C antibody,acute or chronic HBV infections marked by hepatitis B surface antigen,other causes of liver dysfunction ( e.g., primary biliary cirrhosis, autoimmune hepatitis, HIV infection) and patient on anti HCV treatment.were excluded. Results: showed detection of HCV-PCR in PBMCs in the absence of HCV-PCR in plasma; was found in three of the 60 patients (3.3%). All patients had negative HIV, HBsAg, HCV Ab and serum HCV PCR. Conclusion: it could be concluded that testing for HCV-RNA in PBMCs is more reliable than hepatitis serological markers in identifying patients with an OCI when a liver biopsy is not available.

Simultaneously increased expression of microRNA-155 and suppressor of cytokine signaling 1 (SOCS1) gene in the peripheral blood mononuclear cells of patients with primary Sjögren's syndrome.

The microRNA-155 (miR-155) is regarded as a central modulator of T-cell responses and could be a potential therapeutic target for certain inflammatory diseases. In our present study we analyzed the expression rate of miR-155 and its functionally linked gene, the suppressor gene of cytokine signaling 1 (SOCS1) in primary Sjögren's syndrome (pSS). We enrolled 23 pSS patients and 10 healthy individuals in the study. The expression of miR-155 and SOCS1 gene were measured by real-time polymerase chain reaction. We observed the over-expression of miR-155 in the peripheral mononuclear cells of patients with pSS. Surprisingly, SOCS1 gene was also over-expressed in pSS patients. This unanticipated phenomenon might be a laboratory characteristic of Sjögren's syndrome, and presumably a consequence of the noteworthy difference in the pSS immune system reacting with Epstein-Barr virus.

Phenotypic abnormalities of peripheral blood mononuclear cells in patients with Behçet’s disease and association with HLA-B51 expression

The aim of this study was to investigate the subclasses and the immunophenotypic profile of peripheral mononuclear cells in patients with Behçet’s disease (BD) and to assess associations between the expression of HLA-B51 antigen and that of other cell markers. Thirty healthy volunteer blood donors and forty patients with BD were enrolled into this study. Phenotyping was performed using two color flow cytometry. HLA-B51 typing was performed using the complement dependent microlymphocytotoxicity assay. Unlike controls, patients with BD presented a modified immunophenotypic profile of lymphocytes. Compared to those in the remission phase, patients with active BD showed an increased mean of MFI ratio of CD56 on CD16+CD56+ cells (32.47 ± 14.26 versus 23.87 ± 10.3; p = 0.032), increased absolute numbers of CD4−CD8bright and CD4+CD8+ cells (657.1 ± 463.6 cells/µL versus 319.24 ± 116.4 cells/µL; p = 0.017 and 40.77 ± 36.41 cells/µL versus 10.77 ± 9.78 cells/µL; p < 0.0001, respectively) and an elevated mean of MFI ratio of CD19 on B cells (252.3 ± 56.7 versus 205.67 ± 32.3; p = 0.021). However, expression of HLA-B51 was not associated with any specific immunophenotypic profile. In conclusion, abnormal immunophenotypic profile of peripheral lymphocytes was found in patients with BD, especially in active phase, reflecting an immune dysregulation. Moreover, HLA-B51 expression was not found to be related to the expression of other cell markers.

Vitiligo-Inducing Phenols Activate the Unfolded Protein Response in Melanocytes Resulting in Upregulation of IL6 and IL8

Vitiligo is characterized by depigmented skin patches due to loss of epidermal melanocytes. Oxidative stress may play a role in vitiligo onset, while autoimmunity contributes to disease progression. In this study we sought to identify mechanisms that link disease triggers and spreading of lesions. A hallmark of melanocytes at the periphery of vitiligo lesions is dilation of the endoplasmic reticulum (ER). We hypothesized that oxidative stress results in redox disruptions that extend to the ER, causing accumulation of misfolded peptides, which activates the unfolded protein response (UPR). We used 4-tertiary butyl phenol (4-TBP) and monobenzyl ether of hydroquinone (MBEH), known triggers of vitiligo. We show that expression of key UPR components, including the transcription factor X-box binding protein 1 (XBP1), are increased following exposure of melanocytes to phenols. XBP1 activation increases production of immune mediators interleukin-6 (IL6) and IL8. Co-treatment with XBP1 inhibitors reduced IL6 and IL8 production induced by phenols, while over-expression of XBP1 alone increased their expression. Thus, melanocytes themselves produce cytokines associated with activation of an immune response following exposure to chemical triggers of vitiligo. These results expand our understanding of the mechanisms underlying melanocyte loss in vitiligo and pathways linking environmental stressors and autoimmunity.

Increased microRNA-146a/b, TRAF6 gene and decreased IRAK1 gene expressions in the peripheral mononuclear cells of patients with Sjögren's syndrome

MicroRNA-146a (miR-146a) is a microRNA supposed to regulate innate immune, inflammatory response and antiviral pathway negatively. Recently, its potential use as a biomarker for disease diagnosis, prevention and treatment has become widely investigated. In the current study, we measured the expression of miR-146a/b, and their target genes, IRAK1, IRAK4, TRAF6 in the peripheral mononuclear cells of patients with Sjögren's syndrome ( n = 21) and healthy controls ( n = 10) by quantitative reverse transcription polymerase chain reaction. We found that both miR-146a and miR-146b, furthermore, the gene of TRAF6 were significantly overexpressed in the Sjögren's patients, whereas the expression of IRAK1 gene was significantly decreased. The expression of IRAK4 did not differ significantly. These results suggest that in the peripheral mononuclear cells of Sjögren's patients, the transcriptional repression of IRAK1 is taking place, whereas the other NF-κB pathway regulating gene, TRAF6 is overexpressed. As IRAK1 has been regarded a crucial gene in the pathogenesis of systemic lupus erythematosus, TRAF6 can be a Sjögren's syndrome specific biomarker, confirming and partly explaining the existance of different pathogenic pathways in the two diseases. These observations, however, need still wider confirmations.

Quantitative gene expression of transcription factors RORγτ and FoxP3 in patients with liver cystic echinococcosis Turgun.

Objective To investigate the mRNA expression and significance of transcription factors RORγτ and FoxP3 in peripheral mononuclear cells in patients with liver cystic echinococcosis, nethotis Ffity-four subjects were enrolled and derided into three groups： ealthy donor （HD, n = 20）, liver cystic echinococcosis （CE,n = 20）, ecuured cystic echinococcosis （RCE,n = g）. The expression of transcrition factors RORγτ and FoxP3 mRNA in peripheral mononuclear cells of patients with liver cystic echinococcosis was detected by real-time polymerase chain reaction （PCR）. Results The amplification efficiency of quantification of RORγτ and FoxP3 with real-time PCR was 90% -100%. The Ct in intra- and interassay was 〈 0. 20. As compared with HD group, the expression of RORγτ（ RORγτ copies/GAPDH copies） was decreased in CE and RCE groups （ t = 2. 91, t = 2. 79, P 〈 0.05 ） , but there was no statistically significant difference between CE and RCE groups （ t = 0. 56, P 〉 0.05 ）. Oppositely, the expression of FoxP3 （FoxP3 copies/GAPDH copies） was increased in CE and RCE groups as compared with HD group, but there was statistically significant difference （ t = 0.49, t = 0. 02, P 〉 0. 05 ）. The ratio of RORγτ and FoxP3 was obviously decreased in CE and RCE groups as compared with HD group （ t = 0. 33, t = 0. 35, P 〈0. 01 ）, but there was no significant difference between CE and RCE groups （ t = 0. 48, P 〉 0. 05 ）. Conclusion The real-time PCR assay for quantification of RORγτ and FoxP3 is efficient, special and productive. The increased expression of FoxP3 and decreased expression of RORγτ and ROR3,＇r/FoxP3 are related to special immunity during the pathogenesis of liver cystic echinococcosis, The decreased expression of RORγτ may play a major role and the RORγτ/FoxP3 expression ratio may be used as a more objective marker than RORγτ and FoxP3 alone for estimation of special immunovasion during the infection. Key words: Liver echinococcosis; T cells; Transcription factor; Immune evasion

Rs5848 Variant Influences GRN mRNA Levels in Brain and Peripheral Mononuclear Cells in Patients with Alzheimer's Disease

Mutations in the progranulin gene (GRN), causative for Frontotemporal Lobar Degeneration with ubiquitin-immunoreactive neuronal inclusions (FTLD-U), could also be associated with Alzheimer's disease (AD). The influence of GRN genetic variability on susceptibility to AD and on expression levels in a series of neuropathologically-confirmed AD patients as well as in peripheral mononuclear cells (PBMC) and in cells isolated from cerebrospinal fluid (CSF) was investigated. An association study of rs9897526 and rs5848 was carried out in an Italian population and in a replication population of European American patients and controls. None of the variants tested act as unequivocal susceptibility factor in both populations although rs9897526 anticipated the onset of the disease in the Italian population. GRN expression in the parietal lobe of AD cases showed a 0.76-fold decrease compared with controls (1.31 +/- 0.07 versus 1.73 +/- 0.12, P = 0.0025). Patients carrying the rs5848 TT genotype had the lowest GRN expression levels (0.96 +/- 0.12, P = 0.014). Despite no significant differences were found in the relative PBMC and CSF GRN expression in patients compared to controls, stratifying patients according to the presence of rs5848 T allele, a 0.57-fold decrease in GRN mRNA levels over C carriers was found in PBMC (1.22 +/- 0.23 versus 0.70 +/- 0.12, P = 0.04). Similarly to data obtained in brain samples, patients carrying the TT genotype showed the lowest GRN mRNA levels (TT = 0.46 +/- 0.14, CC = 1.22 +/- 0.23; P = 0.013). These data argue against a direct role of GRN as a susceptibility factor for sporadic AD but support a role of GRN as a disease-modifying gene, possibly contributing to the failure of neuronal survival.

Genomic (epigenetic) DNA methylation in patients with open‐angle glaucoma

Abstract Purpose DNA methylation occurs by transfer of a methyl group to cytosine residues in the CpG1 islands. The extent of DNA methylation positively correlates with the extent of gene inactivation. The present study was performed to investigate whether there is an altered global DNA methylation in patients with open‐angle glaucoma. Methods This prospectice case control study included 59 patients with primary open‐angle glaucoma (POAG, age: 68 (SD 8) years), 54 patients with secondary open‐angle glaucoma due to pseudoexfoliation syndrome (PEXG, age: 72 (SD 8) years), and 53 patients with cataract as controls (age: 69 (SD 11) years). Total DNA was extracted from frozen EDTA‐blood using QIAmp DNA Blood Mini Kit (Qiagen). Global methylation status [DNA methylation in %, 1‐(HpaII/MspI)] was measured with a modified non‐radioactive assay. Statistics: Mann‐Whitney‐Test (2‐tailed), the results are presented as means (SD), significance level p&lt;0.05. Results There was a significantly elevated genomic DNA methylation (in %) 1‐(HpaII/MspI) in peripheral mononuclear cells in patients with POAG (68%, SD 18; U=868, Z = ‐2.79, p=0.005), but not in PEXG (55%, SD 17; U=1049, Z=‐0.57, p=0.57) when compared with healthy controls (55%, SD 24). Thus, the difference between POAG and PEXG was exactly 13% of methylated HpaII/MspI restriction sites. Conclusion Since methylation of DNA is an important epigenetic factor in regulation of gene expression these findings may have implications for a possible subsequent derangement of epigenetic control in patients with POAG. Further studies including gene‐specific analyses are needed to clarify the differences between POAG and PEXG.

The expression of sialic acid-binding immunoglobulin-like lectin 1 on peripheral mononuclear cells in patients with coronary heart disease and its clinical significance

Objective To investigate the expression of sialic acid-binding immunoglobulin-like lectin-one (Siglec-1, also called CD169) in lymphocytes, monocytes and neutrophils in peripheral blood in patients with coronary heart disease(CHD), and explore the relationship between Siglec-1 expression and atheresclerosis. Methods CD145 CD169 positive cell proportion and CD169 mRNA levels were respectively measured by flow cytometry and real-time quantitative reverse transcription-polymerase chain reaction (FQ-RT-PCR) in 57 CHD patients and 38 healthy controls. And the levels of serum hpids were determined by automatic biochemistry analyzer. Results The flow cytometry analysis showed that CD169 protein was not found in lymphocytes and neutrophils in both CHD patients and healthy controls. The rate of CD14 CD169 double positive ceils in monocytes in CHD group was significandy higher than that in healthy controls [(12.7±2.4)% vs (1.0±0.3)% ,t =23.2,P 0.05] nor mRNA levels [3.64 fold vs 2.79 fold when compared with healthy controls,t =0. 98, P > 0. 05] were found between CHD patients with normal and abnormal levels of serum Lipids. Conclusions CD169 is mainly expressed in human tissue-resident macrophages but not expressed in peripheral blood monecytes. And when the monocytes is stimulated by inflammation, the expression of CD169 is increased. In patients with CHD, the increased expression of CD169 protein and mRNA level has demonstrated the activation of monocytes in peripheral blood. CD169 and CD169-mediated monocytes activation may play an important role in the development and progression of atherosclerosis. Key words: Coronary disease; Leukocytes, mononuclear; Membrane glycoproteins; Receptors,immunologic; Flow cytometry

Upregulation of the immunoproteasome in peripheral blood mononuclear cells of patients with IgA nephropathy

In order to present an antigen to T-cells, the antigen must first be degraded by proteasomes. Following exposure to interferons, some proteasome subunits (ss1,ss2,ss5) are replaced by others (LMP2, LMP7, MECL-1) that have more optimal catalytic properties for peptide presentation; this more efficient organelle is termed the immuno-proteasome. Here we measured gene expression of various subunits in peripheral mononuclear cells of patients with IgA nephropathy, a disease with features of immune dysregulation. We used quantitative PCR to measure the expression of proteasomal subunit mRNA in mononuclear cells from IgA nephropathy patients, a group of proteinuric control patients with idiopathic nephrotic syndromes, and healthy controls. A significant switch in the expression of trypsin- and chymotrypsin-like proteasome subunits to corresponding immuno-proteasome subunits was found in patients as compared to healthy controls. Further, we found that nuclear translocation of NF-kappaB p50 and p65 was significantly greater in the IgA nephropathy patients, but this did not correlate with the switch to the immuno-proteasome phenotype. Patients with proteinuria greater than 0.5 g/1.73 m(2)/day had a significant switch of the chymotryptic-like beta5 protease to the LMP7 subunit, but this did not occur in patients with idiopathic nephrotic syndrome. The switch to an immuno-proteasome in peripheral blood mononuclear cells of patients with IgA nephropathy suggests an increased efficiency of antigen processing and presentation. This switch appears to be independent of a coincidental activation of the NF-kappaB pathway but is associated with high levels of proteinuria, a well known risk factor for progression of IgA nephropathy.

Toll-like receptor pathway signaling is differently regulated in neutrophils and peripheral mononuclear cells of patients with sepsis, severe sepsis, and septic shock*

Toll-like receptor pathway signaling is differently regulated in neutrophils and peripheral mononuclear cells of patients with sepsis, severe sepsis, and septic shock*

Toll-like receptor pathway signaling is differently regulated in neutrophils and peripheral mononuclear cells of patients with sepsis, severe sepsis, and septic shock*

Up- and down-regulation of inflammatory response was described in blood cells from septic patients, according to the stage of sepsis and the cells evaluated. This study aimed to evaluate the Toll-like receptor (TLR) signaling pathway gene expression in peripheral blood mononuclear cells (PBMC) and neutrophils in patients throughout the different stages of sepsis. Prospective, observational study. Two emergency rooms and two intensive care units in one university and one teaching hospital. A total of 15 septic patients, five with sepsis, five with severe sepsis, and five with septic shock, in addition to five healthy volunteers were enrolled. None. The Human-TLR Signaling Pathway, which comprises 84 genes related to TLR-mediated signal transduction, was evaluated by real time polymerase chain reaction in PBMC and neutrophils obtained from patients and controls. The fold change for each gene (2(-Delta DeltaCt)) was compared between the groups. Genes with fold changes greater than 2 and significant changes in DeltaCT are reported as differently expressed. The fold change ratios in PBMC gene expression between septic patients and healthy controls revealed a dynamic process according to the stage of sepsis, tending toward down-regulation of the TLR signaling pathway in PBMC in the more severe forms of the disease. However, the differential gene expression was restricted to five down-regulated genes in septic shock patients, which are found in the effector and downstream pathways. Neutrophils showed a different pattern of adaptation. Patients with sepsis, severe sepsis, and septic shock presented a broad gene up-regulation, which included all functional groups evaluated and persisted throughout the stages of the disease. TLR-signaling pathway genes are differently regulated in PBMC and neutrophils of septic patients, and are dynamically modulated throughout the different stages of sepsis.

Toll-like receptor pathway signaling is differently regulated in neurophils and peripheral mononuclear cells of patients with sepsis, severe sepsis and septic shock

Toll-like receptor pathway signaling is differently regulated in neurophils and peripheral mononuclear cells of patients with sepsis, severe sepsis and septic shock