Total expression of HLA-G and TLR-9 in chronic lymphocytic leukemia patients

Suppressed immune status facilitates immune escape mechanisms that allow chronic lymphocytic leukemia cells to proliferate and expand. The expression of HLA-G could effectively inhibit the immune response. In immune response inhibitory signals follow activation of immune system which might be occur during bacterial or viral infection in CLL patients. In the current study we characterized two components of immune system, inhibitory molecule HLA-G with its receptor – CD85j and Toll-like receptor 9.     

S100A9 promotes human lung fibroblast cells activation through receptor for advanced glycation end‐product‐mediated extracellular‐regulated kinase 1/2, mitogen‐activated protein‐kinase and nuclear factor‐κB‐dependent pathways

S100A9 belongs to the S100 family of calcium‐binding proteins and plays a key role in many inflammatory conditions. Recent studies have found that S100A9 was elevated significantly in the bronchoalveolar lavage fluid of idiopathic pulmonary fibrosis patients, and might be a biomarker for fibrotic interstitial lung diseases. However, the exact function of S100A9 in pulmonary fibrosis needs further studies. We performed this study to investigate the effect of S100A9 on human embryo lung fibroblast (HLF) proliferation and production of cytokines and collagen, providing new insights into the possible mechanism. S100A9 promoted proliferation of fibroblasts and up‐regulated expression of both proinflammatory cytokines interleukin (IL)‐6, IL‐8, IL‐1β and collagen type III. S100A9 also induced HLF cells to produce α‐smooth muscle actin (α‐SMA) and receptor for advanced glycation end‐product (RAGE). In addition, S100A9 caused a significant increase in extracellular‐regulated kinase (ERK)1/2 mitogen‐activated protein kinase (MAPK) phosphorylation, while the status of p38 and c‐Jun N‐terminal kinase (JNK) phosphorylation remained unchanged. Treatment of cells with S100A9 also enhanced nuclear factor kappa B (NF‐κB) activation. RAGE blocking antibody pretreatment inhibited the S100A9‐induced cell proliferation, cytokine production and pathway phosphorylation. S100A9‐mediated cell activation was suppressed significantly by ERK1/2 MAPK inhibitor and NF‐κB inhibitor. In conclusion, S100A9 promoted HLF cell growth and induced cells to secret proinflammatory cytokines and collagen through RAGE signalling and activation of ERK1/2 MAPK and NF‐κB pathways.     

Short DNA sequences and bacterial DNA induce esophageal,gastric, and colorectal cancer cell invasion

Toll‐like receptor 9 (TLR9) recognizes both bacterial and self‐DNA and it is abundantly expressed in the gastrointestinal tract. In this study, we investigated the influences of both bacterial DNA and specific short DNA sequences on TLR9‐mediated gastrointestinal cancer cell invasion. We assessed the effect of various DNA ligands on cellular invasion and on TLR9 and matrix metalloproteinase expression of three gastrointestinal cancer cell lines. DNA‐ligands described in this study include CpG‐ODN M362, 9‐mer (hairpin), human telomeric sequence h‐Tel22 G‐quadruplex, and bacterial DNAs from Escherichia coli and Helicobacter pylori. All of the DNAs studied were demonstrated to induce invasion in the studied cells. The DNA‐induced invasion was inhibited with a broad‐spectrum MMP inhibitor and partly also with chloroquine suggesting that it could be mediated via MMP activation, endosomal signaling, and TLR9. Interestingly, H. pylori DNA was shown to induce a more pronounced invasion in a gastric cancer cell line than in the other cell lines. Our results suggest that bacterial DNA as well as deoxynucleotides having stable secondary structures (i.e. hairpins or G‐quadruplex structures) may serve as endogenous, invasion‐inducing TLR9‐ligands and promote local progression and metastasis of cancers in the alimentary tract.     

Identification of pathogenic microorganisms directly from positive blood vials by matrix‐assisted laser desorption/ionization time of flight mass spectrometry

Matrix‐assisted laser desorption/ionization time of flight mass spectrometry (MALDI‐TOF MS) is a promising and fast method for identifying fungi and bacteria directly from positive blood cultures. Various pre‐treatment methods for MALDI‐TOF MS identification have been reported for this purpose. In‐house results for identification of bacterial colonies by MALDI‐TOF MS using a cut‐off score of 1.5 did not reduce the diagnostic accuracy compared with the recommended cut‐off score of 1.8. A 3‐month consecutive study of positive blood cultures was carried out in our laboratory to evaluate whether the Sepsityper? Kit (Bruker Daltonics) with Biotyper 2.0 software could be used as a fast diagnostic tool for bacteria and fungi and whether a 1.5 cut‐off score could improve species identification compared with the recommended score of 1.8. Two hundred and fifty‐six positive blood vials from 210 patients and 19 blood vials spiked with fungi were examined. Using the cut‐off score of 1.8, 81% Gram‐negative bacteria were identified to the species level compared to 84% using a cut‐off score of 1.5. For Gram‐positive bacteria 44% were identified to the species level with a cut‐off of 1.8 compared to 55% with the value of 1.5. The overall identification rate was 63% (cut‐off 1.5) and 54% (cut‐off 1.8). Seventy‐seven per cent of fungal species were identified with both log scores. MALDI‐TOF MS was in this study found to be a powerful tool in fast diagnosis of Gram‐negative bacteria and fungi and to a lesser degree of Gram positives. Using 1.5 as cut‐off score increased the diagnosis for both Gram‐positives and ‐negatives bacteria.     

Molecular characterization of serotype G9 rotaviruses circulating in South Korea between 2005 and 2010

A total of 18 rotavirus G9 strains in South Korea were collected during five rotavirus seasons between 2005 and 2010. The relationship between these strains was examined by analyzing the genetic variation of two major structural genes, VP7 and VP4. All the rotavirus isolates were of the G9P[8] genotype. The VP7 phylogenetic analysis demonstrated that all of the G9 rotaviruses circulating in South Korea belonged to lineage IIId and were within three single clusters. The amino acid comparison of the antigenic regions of the VP7 gene suggests possible common progenitors of these strains. Phylogenetic analysis of P[8] VP4 genotypes indicated three lineages, P[8]‐2, P[8]‐3, and P[8]‐4, with P[8]‐3 being the most common. The results of this study provide information on the genetic relatedness of rotavirus G9 strains circulating in South Korea over recent years and can be utilized for the development of effective vaccines and the identification of reference strains for future efficacy studies. J. Med. Virol. 85:171–178, 2012. © 2012 Wiley Periodicals, Inc.     

Quantification issues of in vivo 1H NMR spectroscopy of the rat brain investigated at 16.4 T

The accuracy and precision of the quantification of metabolite concentrations in in vivo ¹H NMR spectroscopy are affected by linewidth and signal‐to‐noise ratio (SNR). To study the effect of both factors in in vivo ¹H NMR spectra acquired at ultrahigh field, a reference spectrum was generated by summing nine in vivo ¹H NMR spectra obtained in rat brain with a STEAM sequence at 16.4 T. By progressive deterioration of linewidth and SNR, 6400 single spectra were generated. In an accuracy study, the variation in the mean concentrations of five metabolites was mainly dependent on SNR, whereas 11 metabolites were predominantly susceptible to the linewidth. However, the standard deviations of the concentrations obtained were dependent almost exclusively on the SNR. An insignificant correlation was found between most of the heavily overlapping metabolite peaks, indicating independent and reliable quantification. Two different approaches for the consideration of macromolecular signals were evaluated. The use of prior knowledge derived by parameterization of a metabolite‐nulled spectrum demonstrated improved fitting quality, with reduced Cramér–Rao lower bounds, compared to the calculation of a regularized spline baseline. Copyright © 2012 John Wiley & Sons, Ltd.     

骨髓单个核细胞移植后心肌梗死区的血管生成及细胞因子分泌

背景：干细胞移植为治疗心肌梗死带来新的希望,但研究结果不一致,存在争议。细胞移植能否长期持久地改善心脏功能、改善缺血心功能的机制这些问题均不明确。 目的：观察经冠状动脉自体骨髓单个核细胞移植对急性心肌梗死犬心功能、血管生成和细胞因子分泌的影响。 方法：杂种犬分为骨髓单个核细胞组(n=10)和生理盐水组(n=6),前上嵴或髂后上棘穿刺分离得到骨髓单个核细胞,结扎冠状动脉前降支建立急性心肌梗死模型,于心肌梗死后2 h分别经冠状动脉内移植骨髓单个核细胞和生理盐水,心肌梗死后2 h及6周时分别测定超声心动图指标,骨髓单个核细胞移植后6周vWF免疫组化染色检测心肌组织的毛细血管密度,RT-PCR检测血管内皮生长因子(血管内皮生长因子188、血管内皮生长因子164)、碱性成纤维细胞生长因子、基质金属蛋白酶9 mRNA的表达。 结果与结论：经冠状动脉自体骨髓单个核细胞移植后6周,超声心动图显示,骨髓单个核细胞组射血分数和每搏输出量比生理盐水组显著升高;梗死边缘区新生血管数量明显高于生理盐水组。骨髓单个核细胞组梗死区血管内皮生长因子188、血管内皮生长因子164和碱性成纤维细胞生长因子mRNA表达水平显著高于生理盐水组。骨髓单个核细胞组梗死区基质金属蛋白酶9 mRNA 表达水平显著低于生理盐水组。结果说明自体骨髓单个核细胞经冠状动脉内注射移植,改善急性心肌梗死后心功能,促进梗死边缘区血管生成,提高促血管生长因子血管内皮生长因子188、血管内皮生长因子164和碱性成纤维细胞生长因子的mRNA表达水平,减少基质金属蛋白酶9 mRNA表达水平。     

Effects of Interleukin-9 Blockade on Chronic Airway Inflammation in Murine Asthma Models

Purpose

Asthma is a chronic inflammatory disease of the airways associated with structural changes and airway remodeling. Interleukin (IL)-9 has pleiotropic effects on both inflammatory cells and airway structural cells, which are involved in asthma pathogenesis. We evaluated the effects of IL-9 blockade on chronic airway inflammation.

Methods

Acute airway inflammation was induced in Balb/c mice using aerosolized ovalbumin (OVA), whereas chronic asthma was induced by OVA exposure for 5 weeks with anti-IL-9 or isotype-matched antibody (Ab) treatment during the OVA challenge. Inflammatory cells in bronchoalveolar lavage fluid (BALF) were counted and lung tissues were stained to detect cellular infiltration, mucus deposition, and collagen accumulation. The levels of interferon (IFN)-γ, IL-4, IL-5, IL-9, IL-17, and immunoglobulin E (IgE) in BALF were measured using enzyme linked immunosorbent assays, and profiles of inflammatory cells and subsets of T helper (Th) cells were analyzed using flow cytometry.

Results

IL-9, IL-17, and IFN-γ levels were significantly increased in the chronic group compared to the acute asthma group. However, the number of IL-9-positive cells was not affected, with a decrease in Th17 cells in OVA-challenged caspase-1 knockout mice. Numbers of eosinophils, neutrophils, B cells, mast cells, and Th17 cells decreased after administration of anti-IL-9 Ab. Total IgE, IL-5, IL-9, and IL-17 levels were also lower in the anti-IL-9 group.

Conclusions

Our results suggest that anti-IL-9 Ab treatment inhibits pulmonary infiltration of inflammatory cells and cytokine production, especially IL-17. These results provide a basis for the use of an anti-IL-9 Ab to combat IL-17-mediated airway inflammation.