This study aims to investigate the associations of hypomethylation and over expression of the TLR9 gene with systemic lupus erythematosus (SLE). Fifteen SLE patients who were diagnosed and not treated, were selected as cases, and 32 healthy subjects were enrolled as controls. DNA and total RNA of peripheral blood mononuclear cells (PBMCs) were extracted. The methylation status of the promoter region CpG motifs of the TLR9 gene was quantitatively measured using bisulfite sequencing PCR, and the mRNA expression of the TLR9 gene was determined using real-time fluorescent quantitative PCR. The methylation level of the 10 TLR9 CpG motifs of gene did not show difference between cases and controls (P>0.05). By contrast, we observed an abnormal increase of TLT9 mRNA expression in patients (P=9.379×10(-8)), which was significantly correlated with SLEDAI (Systemic Lupus Erythematosus Disease Activity Index) (P=9.018×10(-7)). The change of TLR9 gene expression may play an important role in the pathogenesis of SLE.

• MeSH
• dospělí MeSH
• exprese genu MeSH
• leukocyty mononukleární metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA biosyntéza   genetika   MeSH
• metylace DNA fyziologie   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• polymerázová řetězová reakce metody   MeSH
• progrese nemoci * MeSH
• systémový lupus erythematodes krev   diagnóza   genetika   MeSH
• toll-like receptor 9 biosyntéza   genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• messenger RNA MeSH
• TLR9 protein, human MeSH Prohlížeč
• toll-like receptor 9 MeSH

A prelabor rupture of membranes (PROM) and its subtypes, preterm PROM (pPROM) and term PROM (tPROM), are associated with disturbances in the hemostatic system and angiogenesis. This study was designed to demonstrate the role of single nucleotide polymorphisms (SNPs), localized in CSF2 (rs25881), FLT1 (rs722503), TFPI (C-399T) and TLR9 (rs352140) genes, in PROM. A population of 360 women with singleton pregnancy consisted of 180 PROM cases and 180 healthy controls. A single-SNP analysis showed a similar distribution of genotypes in the studied polymorphisms between the PROM or the pPROM women and the healthy controls. Double-SNP TT variants for CSF2 and FLT1 polymorphisms, CC variants for TLR9 and TFPI SNPs, TTC for CSF2, FLT1 and TLR9 polymorphisms, TTT for FLT1, TLR9 and TFPI SNPs and CCCC and TTTC complex variants for all tested SNPs correlated with an increased risk of PROM after adjusting for APTT, PLT parameters and/or pregnancy disorders. The TCT variants for the CSF2, FLT1 and TLR9 SNPs and the CCTC for the CSF2, FLT1, TLR9 and TFPI polymorphisms correlated with a reduced risk of PROM when corrected by PLT and APTT, respectively. We concluded that the polymorphisms of genes, involved in hemostasis and angiogenesis, contributed to PROM.

• Klíčová slova
• angiogenesis, genotyping, hemostasis, pPROM, pregnancy, prelabor rupture of membranes (PROM), restriction fragment length polymorphism (RFLP), single nucleotide polymorphism (SNP), tPROM,
• MeSH
• dospělí MeSH
• faktor stimulující granulocyto-makrofágové kolonie genetika   MeSH
• genetická predispozice k nemoci MeSH
• genetické asociační studie MeSH
• jednonukleotidový polymorfismus * MeSH
• lidé MeSH
• lipoproteiny genetika   MeSH
• mladý dospělý MeSH
• předčasný odtok plodové vody genetika   MeSH
• receptor 1 pro vaskulární endoteliální růstový faktor genetika   MeSH
• studie případů a kontrol MeSH
• těhotenství MeSH
• toll-like receptor 9 genetika   MeSH
• věk matky MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CSF2 protein, human MeSH Prohlížeč
• faktor stimulující granulocyto-makrofágové kolonie MeSH
• FLT1 protein, human MeSH Prohlížeč
• lipoprotein-associated coagulation inhibitor MeSH Prohlížeč
• lipoproteiny MeSH
• receptor 1 pro vaskulární endoteliální růstový faktor MeSH
• TLR9 protein, human MeSH Prohlížeč
• toll-like receptor 9 MeSH

Challenges with various TLR ligands (TLRLs)in combination with D-galactosamine (GalN) in rodents may mimic diverse conditions of acute inflammation and organ failure. Here, we report that CpG (ODN1826, TLR9 agonist)/GalN induced a liver-specific injury with modest systemic effects, whereas R848 (resiquimod, TLR7/8 agonist)/GalN exhibited systemic and liver toxicity. We also observed the protective effect of Gr-1+ cells (the population containing neutrophils) against liver injury in both the R848/GalN and CpG/GalN models. In cytokine measurements, the intraperitoneal administration of antibodies showed a non-specific tolerance induction effect, which was more pronounced in the CpG/GalN than in the R848/GalN model. Cytokine analyses also suggested that the TLR9 agonist/GalN induced a limited degree of systemic inflammation compared to TLR7/8 agonist/GalN models. The relevance of this finding to the TLR9-mediated induction of stress tolerance (protective effect) in non-immune cells is discussed.

• MeSH
• galaktosamin toxicita   MeSH
• imidazoly toxicita   MeSH
• lékové postižení jater etiologie   metabolismus   patologie   MeSH
• lipopolysacharidy toxicita   MeSH
• membránové glykoproteiny agonisté   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• stupeň závažnosti nemoci MeSH
• toll-like receptor 7 agonisté   MeSH
• toll-like receptor 8 agonisté   MeSH
• toll-like receptor 9 agonisté   MeSH
• zánět chemicky indukované   metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• galaktosamin MeSH
• imidazoly MeSH
• lipopolysacharidy MeSH
• membránové glykoproteiny MeSH
• resiquimod MeSH Prohlížeč
• Tlr7 protein, mouse MeSH Prohlížeč
• TLR8 protein, mouse MeSH Prohlížeč
• Tlr9 protein, mouse MeSH Prohlížeč
• toll-like receptor 7 MeSH
• toll-like receptor 8 MeSH
• toll-like receptor 9 MeSH

OBJECTIVE: Dendritic cells (DCs) play an important role in pathogenesis of autoimmunity, including type 1 diabetes (T1D). In this study, we investigated DC subpopulations and their responses to TLR stimulation in T1D patients and their relatives. METHODS: We analyzed the frequency of myeloid (mDCs) and plasmacytoid DCs (pDCs) in 97 T1D patients (69 onset, 28 long-term), 67 first-degree relatives, and 64 controls. We additionally tested the IFN-alpha production by pDCs upon stimulation with TLR 7, 8 and 9 agonists. RESULTS: A lower number of mDCs and pDCs were found in T1D patients and their relatives. Of all the tested TLR ligands, only stimulation with CpG 2216 induced IFN-alpha production that was the highest in T1D relatives, except of autoantibody-negative relatives bearing the protective haplotypes. CONCLUSION: Our data demonstrate disturbances in DC number and function expressed most significantly in T1D relatives and point to a potential role of TLR9-induced IFN-alpha production in T1D development.

• Klíčová slova
• Dendritic cell;, Interferon-alpha;, Type 1 diabetes,
• MeSH
• dendritické buňky imunologie   metabolismus   MeSH
• diabetes mellitus 1. typu imunologie   metabolismus   MeSH
• dítě MeSH
• dospělí MeSH
• interferon alfa biosyntéza   krev   MeSH
• leukocyty mononukleární účinky léků   metabolismus   MeSH
• lidé MeSH
• ligandy MeSH
• mladiství MeSH
• mladý dospělý MeSH
• oligodeoxyribonukleotidy farmakologie   MeSH
• počet buněk MeSH
• předškolní dítě MeSH
• rodina MeSH
• toll-like receptor 9 metabolismus   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CpG ODN 2216 MeSH Prohlížeč
• interferon alfa MeSH
• ligandy MeSH
• oligodeoxyribonukleotidy MeSH
• toll-like receptor 9 MeSH

Following nerve injury, disintegrated axonal mitochondria distal to the injury site release mitochondrial formylated peptides and DNA that can induce activation and inflammatory profiling of Schwann cells via formyl peptide receptor 2 (Fpr2) and toll-like receptor 9 (TLR9), respectively. We studied RT4 schwannoma cells to investigate the regulation of Fpr2 and TLR9 after stimulation with fMLF as a prototypical formylated peptide. RT4 cells were treated with fMLF at various concentrations and times with and without pretreatment with inhibitors (chloroquine for activated TLR9, PBP10 for Fpr2). Western blots of Fpr2, TLR9, p-p38, p-NFκB, and IL-6 were compared in relation to inflammatory profiling of RT4 cells and chemokine receptors (CCR2, CXCR4) as potential co-receptors of Fpr2. fMLF stimulation upregulated Fpr2 in RT4 cells at low concentrations (10 nM and 100 nM) but higher concentrations were required (10 µM and 50 µM) when the cells were pretreated with an activated TLR9 inhibitor. Moreover, the higher concentrations of fMLF could modulate TLR9 and inflammatory markers. Upregulation of Fpr2 triggered by 10 nM and 100 nM fMLF coincided with higher levels of chemokine receptors (CCR2, CXCR4) and PKCβ. Treating RT4 cells with fMLF, as an in vitro model of Schwann cells, uncovered Schwann cells' complex responses to molecular patterns of release from injured axonal mitochondria.

• Klíčová slova
• Wallerian degeneration, chemokines, cytokines, damage-associated molecular patterns, disintegration, mitochondria, receptors,
• MeSH
• chlorochin farmakologie   MeSH
• krysa rodu Rattus MeSH
• N-formylmethionin-leucyl-fenylalanin farmakologie   MeSH
• nádorové buněčné linie MeSH
• neurilemom metabolismus   patologie   MeSH
• receptory CCR2 genetika   metabolismus   MeSH
• receptory CXCR4 genetika   metabolismus   MeSH
• receptory pro formylované peptidy antagonisté a inhibitory   genetika   metabolismus   MeSH
• Schwannovy buňky cytologie   účinky léků   metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• toll-like receptor 9 antagonisté a inhibitory   genetika   metabolismus   MeSH
• upregulace účinky léků   MeSH
• zánět metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chlorochin MeSH
• N-formylmethionin-leucyl-fenylalanin MeSH
• receptory CCR2 MeSH
• receptory CXCR4 MeSH
• receptory pro formylované peptidy MeSH
• toll-like receptor 9 MeSH

Acute lung injury (ALI) caused by lipopolysaccharide (LPS) is a common, severe clinical syndrome. Injury caused by inflammation and oxidative stress in vascular endothelial and alveolar epithelial cells is a vital process in the pathogenesis of ALI. Toll-like receptor 9 (TLR9) is highly expressed in LPS-induced ALI rats. In this study, Beas-2B human pulmonary epithelial cells and A549 alveolar epithelial cells were stimulated by LPS, resulting in the upregulation of TLR9 in a concentrationdependent manner. Furthermore, TLR9 overexpression and interference vectors were transfected before LPS administration to explore the role of TLR9 in LPS-induced ALI in vitro. The findings revealed that inhibition of TLR9 reduced inflammation and oxidative stress while suppressing apoptosis of LPS-induced Beas-2B and A549 cells, whereas TLR9 overexpression aggravated these conditions. Moreover, TLR9 inhibition resulted in downregulated protein expression of myeloid differentiation protein 88 (MyD88) and activator activator protein 1 (AP-1), as well as phosphorylation of nuclear factor-?B (NF-kappaB), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK). The phosphorylation of extracellular-regulated protein kinases 1/2 was upregulated compared to that of cells subjected to only LPS administration, and this was reversed by TLR9 overexpression. These results indicate that inhibition of TLR9 plays a protective role against LPS-induced inflammation and oxidative stress in Beas-2B and A549 cells, possibly via the MyD88/NF-kappaB and MyD88/MAPKs/AP-1 pathways.

• MeSH
• akutní poškození plic * chemicky indukované   metabolismus   prevence a kontrola   MeSH
• epitelové buňky patologie   MeSH
• krysa rodu Rattus MeSH
• lipopolysacharidy * metabolismus   toxicita   MeSH
• NF-kappa B metabolismus   MeSH
• oxidační stres MeSH
• protein MyD88 metabolismus   MeSH
• signální transdukce MeSH
• toll-like receptor 4 metabolismus   MeSH
• toll-like receptor 9 genetika   metabolismus   MeSH
• transkripční faktor AP-1 metabolismus   MeSH
• zánět chemicky indukované   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• lipopolysacharidy * MeSH
• Myd88 protein, rat MeSH Prohlížeč
• NF-kappa B MeSH
• protein MyD88 MeSH
• Tlr9 protein, rat MeSH Prohlížeč
• toll-like receptor 4 MeSH
• toll-like receptor 9 MeSH
• transkripční faktor AP-1 MeSH

In this study, we hypothesized that the changes localized at angiopoietin-2 (ANGPT2), granulocyte-macrophage colony-stimulating factor (CSF2), fms-related tyrosine kinase 1 (FLT1) and toll-like receptor (TLR) 2, TLR6 and TLR9 genes were associated with spontaneous preterm labor (PTL), as well as with possible genetic alterations on PTL-related coagulation. This case-control genetic association study aimed to identify single nucleotide polymorphisms (SNPs) for the aforementioned genes, which are correlated with genetic risk or protection against PTL in Polish women. The study was conducted in 320 patients treated between 2016 and 2020, including 160 women with PTL and 160 term controls in labor. We found that ANGPT2 rs3020221 AA homozygotes were significantly less common in PTL cases than in controls, especially after adjusting for activated partial thromboplastin time (APTT) and platelet (PLT) parameters. TC heterozygotes for TLR2 rs3804099 were associated with PTL after correcting for anemia, vaginal bleeding, and history of threatened miscarriage or PTL. TC and CC genotypes in TLR9 rs187084 were significantly less common in women with PTL, compared to the controls, after adjusting for bleeding and gestational diabetes. For the first time, it was shown that three polymorphisms-ANGPT2 rs3020221, TLR2 rs3804099 and TLR9 rs187084 -were significantly associated with PTL, adjusted by pregnancy development influencing factors.

• Klíčová slova
• angiogenesis, genotyping, pregnancy, restriction fragment length polymorphism, single nucleotide polymorphism, spontaneous preterm labor,
• Publikační typ
• časopisecké články MeSH

One of the changes brought about by Wallerian degeneration distal to nerve injury is disintegration of axonal mitochondria and consequent leakage of mitochondrial DNA (mtDNA)-the natural ligand for the toll-like receptor 9 (TLR9). RT-PCR and immunohistochemical or Western blot analyses were used to detect TLR9 mRNA and protein respectively in the lumbar (L4-L5) and cervical (C7-C8) dorsal root ganglia (DRG) ipsilateral and contralateral to a sterile unilateral sciatic nerve compression or transection. The unilateral sciatic nerve lesions led to bilateral increases in levels of both TLR9 mRNA and protein not only in the lumbar but also in the remote cervical DRG compared with naive or sham-operated controls. This upregulation of TLR9 was linked to activation of the Nuclear Factor kappa B (NFκB) and nuclear translocation of the Signal Transducer and Activator of Transcription 3 (STAT3), implying innate neuronal immune reaction and a pro-regenerative state in uninjured primary sensory neurons of the cervical DRG. The relationship of TLR9 to the induction of a pro-regenerative state in the cervical DRG neurons was confirmed by the shorter lengths of regenerated axons distal to ulnar nerve crush following a previous sciatic nerve lesion and intrathecal chloroquine injection compared with control rats. The results suggest that a systemic innate immune reaction not only triggers the regenerative state of axotomized DRG neurons but also induces a pro-regenerative state further along the neural axis after unilateral nerve injury.

• Klíčová slova
• axon regeneration, compression, early endosomes, mitochondrial DNA, sciatic nerve, the endoplasmic reticulum, transection,
• MeSH
• krysa rodu Rattus MeSH
• nemoci sedacího nervu imunologie   metabolismus   patologie   terapie   MeSH
• neurony cytologie   imunologie   MeSH
• potkani Wistar MeSH
• přirozená imunita imunologie   MeSH
• spinální ganglia cytologie   MeSH
• toll-like receptor 9 genetika   metabolismus   MeSH
• transkripční faktor STAT3 genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Stat3 protein, rat MeSH Prohlížeč
• Tlr9 protein, rat MeSH Prohlížeč
• toll-like receptor 9 MeSH
• transkripční faktor STAT3 MeSH

Tumor Necrosis Factor Receptor 2 (TNFR2) expression is increasingly being linked to tolerogenic immune reactions and cells with suppressor function including a subset of T-regulatory cells. B-regulatory cells play an important role in control of T-cell responses and inflammation. Recently, we described TNFR2 as a marker for IL-10-producing B cells, a hallmark of this cell subset. Here, we demonstrate that proliferation of T cells is reduced in the presence of TNFR2 positive human memory B cells generated with TLR9 ligand, while TNFR2- and TNFR2+CD27- B cells display costimulatory activity. Our data further reveal that IL-10 secretion is characteristic of IgM+ naïve and memory B cells but suppressive activity is not restricted to IL-10: (i) the inhibitory effect of TNFR2+ switched memory B cells was comparable to that exerted by TNFR2+ IgM+ memory B cells although IL-10 secretion levels in the cocultures were lower; (ii) supernatants from TNFR2+ memory B cells failed to suppress T-cell proliferation. Based on our findings, we propose that formation of Breg is a specific characteristic of human memory B cells undergoing terminal differentiation. Our data further corroborate that TNFR2 represents a viable marker for identification of memory B cells with regulatory function.

• Klíčová slova
• B cells, Breg, IL-10, TLR9, TNFR2,
• MeSH
• aktivace lymfocytů genetika   imunologie   MeSH
• B-lymfocyty imunologie   metabolismus   MeSH
• běžná variabilní imunodeficience etiologie   metabolismus   MeSH
• buněčná diferenciace imunologie   MeSH
• cytokiny metabolismus   MeSH
• imunologická paměť * MeSH
• imunomodulace genetika   MeSH
• interleukin-10 metabolismus   MeSH
• lidé MeSH
• mezibuněčná komunikace imunologie   MeSH
• receptory TNF - typ II genetika   metabolismus   MeSH
• regulace genové exprese * MeSH
• regulační B-lymfocyty imunologie   metabolismus   MeSH
• studie případů a kontrol MeSH
• T-lymfocyty imunologie   metabolismus   MeSH
• toll-like receptor 9 metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH
• interleukin-10 MeSH
• receptory TNF - typ II MeSH
• TLR9 protein, human MeSH Prohlížeč
• toll-like receptor 9 MeSH

The elevated plasma cell-free DNA (cfDNA) concentrations were repeatedly reported in association with the process of inflammation. The qualitative and quantitative characteristics of plasma cfDNA in active (newly diagnosed) celiac disease patients (CD) have not yet been studied despite the fact that cfDNA of healthy individuals is able to regulate immune response. We determined the total cfDNA concentration and relative content of telomeric sequences in plasma cfDNA in CD (n = 10) and healthy age- and sex-matched controls (HC, n = 10) by quantitative PCR. To obtain the evidence that the observed biological effects are caused solely by cfDNA molecules, we applied the treatment of paired plasma samples with DNase. Using paired samples of plasma (non-treated/native and treated by DNase), we analyzed the contribution of cfDNA to the activation of TLR9 and TNF-α mRNA expression in THP1 monocytic cell line. There were no significant differences in the quantities of plasma cfDNA and relative contents of telomeric sequences in their pools. When we compared the levels of TNF-α mRNA expression in THP1 cells achieved after stimulation with native CD and HC plasma samples, we found significantly (p = .031) higher expression after stimulation with CD samples. We documented also the ability of cfDNA contained in CD plasma samples to stimulate the production of TLR9 mRNA. The TLR9 mRNA expression levels were significantly (p = .014) lowered after cfDNA removal from CD plasma samples. The design of our experiments allowed us to study the effects of cfDNA without its isolation from plasma. cfDNA contained in CD plasma samples differs significantly in its immunoregulatory capacity from cfDNA in HC plasma. The differences are caused neither by different concentrations of cfDNA in plasma samples nor by different relative abundance of telomeric sequences. Further studies are needed to elucidate the role of plasma cfDNA in celiac disease pathogenesis.

• Klíčová slova
• Celiac disease, TNF-α, cell-free DNA, telomeric sequences, toll-like receptor 9,
• MeSH
• celiakie krev   MeSH
• dospělí MeSH
• imunologické faktory * krev   imunologie   farmakologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• pilotní projekty MeSH
• regulace genové exprese * účinky léků   imunologie   MeSH
• THP-1 buňky MeSH
• TNF-alfa * biosyntéza   imunologie   MeSH
• toll-like receptor 9 * biosyntéza   imunologie   MeSH
• volné cirkulující nukleové kyseliny * krev   imunologie   farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• imunologické faktory * MeSH
• TLR9 protein, human MeSH Prohlížeč
• TNF-alfa * MeSH
• toll-like receptor 9 * MeSH
• volné cirkulující nukleové kyseliny * MeSH