scholarly journals POLYMORPHISM OF GENES OF IMMUNOSUPRESSIVE CYTOKINE IL-10 AND TGF-β AT TUBERCULOSIS INFECTION

2014 ◽  
Vol 13 (5) ◽  
pp. 107-113
Author(s):  
Ye. G. Churina ◽  
O. I. Urazova ◽  
V. V. Novitsky ◽  
O. V. Filinyuk
Keyword(s):  
Mononuclear Leukocytes ◽  
Allelic Polymorphism ◽  
Polymorphic Genes ◽  
Allele Specific ◽  
Specific Amplification ◽  
Mononuclear Leucocytes ◽  
Genetically Determined ◽  
Culture Supernatants ◽  
Immunosuppressive Cytokines

The aim of the work was the study of connection of allelic polymorphism of IL10 and TGFВgenes with changes in the basal and BCG-induced production of immunosuppressive cytokines IL-10 and TGF-β by mononuclear leukocytes in vitro in patients with the first diagnosed pulmonary tuberculosis (TB), depending on the clinical form of the disease. The evaluation of the cytokines production was conducted by measuring its concentration in culture supernatants by ELISA. The allele-specific amplification of specific stretches of the genome was used for the study of polymorphic genes of cytokines. The DNA and supernatants of culture suspensions of blood mononuclear leucocytes in healthy volunteers and patients with TB were the material of the research. It was shown in the research conducted that the basal and BCG-induced over-production of IL-10 in vitro occurs in patients with TB, regardless of the genotype of the locus of C-592AIL10 gene. In addition, genotype AA of polymorphism of IL10gene in patients with infiltrative and disseminated TB is associated with the maximum production of IL-10 in vitroand genotype CC – with the minimum production of this cytokine in vitro. Analysis of the production of TGF-β in vitro in patients with TB showed its increase only in case of carriage of allele T (C-509T) of TGFB gene. In patients with disseminated TB and homosygotic genotype TT the increase in both basal and BCG-induced production of TGF-β was determined, and in patients with infiltrative TB – only after induction of cells by BCG-antigen.Thus, the over-production of cytokines with inhibiting activity in patients with TB is genetically determined and promotes the formation of suppressive mode of immune-regulation. The increase in the secretion of cytokines IL-10 and TGF-β in vitro in patients with TB are associated with carriage of allele A and genotype AA (C-592A) of IL10gene and allele T and genotype TT (C-509T) of TGFB gene.

scholarly journals Characterization of proteins from human synovium and mononuclear leucocytes that induce resorption of cartilage proteoglycan in vitro

1983 ◽  
Vol 209 (2) ◽  
pp. 337-344 ◽  
Author(s):  
J Saklatvala ◽  
S J Sarsfield ◽  
L M C Pilsworth
Keyword(s):  
Articular Cartilage ◽  
Synovial Tissue ◽  
Gel Filtration ◽  
Human Articular Cartilage ◽  
Nasal Cartilage ◽  
Cartilage Proteoglycan ◽  
Mononuclear Leucocytes ◽  
Culture Supernatants

Both human synovial tissue in culture and lectin-stimulated mononuclear leucocytes produced a protein that induced proteoglycan resorption in explants of bovine nasal cartilage and human articular cartilage. On gel filtration the protein had Mr 16000-20000 and on isoelectric focusing its pI was 5.2-5.3. The protein corresponded to catabolin, which has previously been identified as a product of cultured porcine synovial tissue and mononuclear leucocytes. The action of partially purified human catabolin was not inhibited by cortisol, although the activity of the leucocyte supernatants from which it had been isolated was inhibited. For this reason it is not possible to be sure that the active factor detected in the bioassay of the crude leucocyte culture supernatants is in fact catabolin.

Simple and Rapid Detection of Factor V Leiden by Allele-specific PCR Amplification

1996 ◽  
Vol 75 (05) ◽  
pp. 757-759 ◽  
Author(s):  
Rainer Blasczyk ◽  
Markus Ritter ◽  
Christian Thiede ◽  
Jenny Wehling ◽  
Günter Hintz ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Activated Protein C ◽  
Factor V Leiden ◽  
Pcr Amplification ◽  
Factor V ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Allele Specific ◽  
Specific Amplification ◽  
Allele Specific Pcr

SummaryResistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506.

scholarly journals LPS Primes Brain Responsiveness to High Mobility Group Box-1 Protein

2021 ◽  
Vol 14 (6) ◽  
pp. 558
Author(s):  
Verena Peek ◽  
Lois M. Harden ◽  
Jelena Damm ◽  
Ferial Aslani ◽  
Stephan Leisengang ◽  
...  
Keyword(s):  
Inflammatory Responses ◽  
Area Postrema ◽  
High Mobility ◽  
High Mobility Group ◽  
Direct Access ◽  
Significant Release ◽  
Factor Α ◽  
Culture Supernatants

High mobility group box (HMGB)1 action contributes to late phases of sepsis, but the effects of increased endogenous plasma HMGB1 levels on brain cells during inflammation are unclear. Here, we aimed to further investigate the role of HMGB1 in the brain during septic-like lipopolysaccharide-induced inflammation in rats (LPS, 10 mg/kg, i.p.). HMGB-1 mRNA expression and release were measured in the periphery/brain by RT-PCR, immunohistochemistry and ELISA. In vitro experiments with disulfide-HMGB1 in primary neuro-glial cell cultures of the area postrema (AP), a circumventricular organ with a leaky blood–brain barrier and direct access to circulating mediators like HMGB1 and LPS, were performed to determine the direct influence of HMGB1 on this pivotal brain structure for immune-to-brain communication. Indeed, HMGB1 plasma levels stayed elevated after LPS injection. Immunohistochemistry of brains and AP cultures confirmed LPS-stimulated cytoplasmatic translocation of HMGB1 indicative of local HMGB1 release. Moreover, disulfide-HMGB1 stimulation induced nuclear factor (NF)-κB activation and a significant release of interleukin-6, but not tumor necrosis factor α, into AP culture supernatants. However, only a few AP cells directly responded to HMGB1 with increased intracellular calcium concentration. Interestingly, priming with LPS induced a seven-fold higher percentage of responsive cells to HMGB1. We conclude that, as a humoral and local mediator, HMGB1 enhances brain inflammatory responses, after LPS priming, linked to sustained sepsis symptoms.

scholarly journals Identification of NAD+ Synthetase from Streptococcus sobrinus as a B-Cell-Stimulatory Protein

2004 ◽  
Vol 186 (2) ◽  
pp. 419-426 ◽  
Author(s):  
Isabel Veiga-Malta ◽  
Margarida Duarte ◽  
Márcia Dinis ◽  
Pedro Madureira ◽  
Paula Ferreira ◽  
...  
Keyword(s):  
B Cell ◽  
Active Form ◽  
Lymphocyte Activation ◽  
Amino Acid Residues ◽  
Streptococcus Sobrinus ◽  
Reading Frame ◽  
High Sequence Homology ◽  
Culture Supernatants ◽  
Nad Synthetase

ABSTRACT Streptococcus sobrinus, one agent of dental caries, secretes a protein that induces lymphocyte polyclonal activation of the host as a mechanism of immune evasion. We have isolated from culture supernatants of this bacterium a protein with murine B-cell-stimulatory properties and subsequently cloned the relevant gene. It contains an open reading frame of 825 bp encoding a polypeptide with 275 amino acid residues and a molecular mass of 30 kDa. The protein displays high sequence homology with NAD+ synthetases from several organisms, including a conserved fingerprint sequence (SGGXD) characteristic of ATP pyrophosphatases. The polypeptide was expressed in Escherichia coli as a hexahistidine-tagged protein and purified in an enzymatically active form. The recombinant NAD+ synthetase stimulates murine B cells after in vitro treatment of spleen cell cultures, as demonstrated by its ability to induce up-regulation of the expression of CD69, an early marker of lymphocyte activation. Stimulation with the recombinant NAD+ synthetase was also observed with other B-cell markers, such as CD19+, B220+, and CD21+. Cell proliferation follows the activation induced by the recombinant NAD+ synthetase.

Typing forHLA-DPB1 * 03 andHLA-DPB1 * 06 using allele-specific DNA in vitro amplification and allele-specific oligonucleotide probes. Detection of “new” DPB1*06 variants

1989 ◽  
Vol 30 (3) ◽  
pp. 208-213 ◽  
Author(s):  
L. Fugger ◽  
N. Morling ◽  
L. P. Ryder ◽  
N. Ødum ◽  
J. Georgsen ◽  
...  
Keyword(s):  
Oligonucleotide Probes ◽  
Allele Specific

scholarly journals Metagenomics, gene discovery and the ideal biocatalyst

2004 ◽  
Vol 32 (2) ◽  
pp. 298-302 ◽  
Author(s):  
D.A. Cowan ◽  
A. Arslanoglu ◽  
S.G. Burton ◽  
G.C. Baker ◽  
R.A. Cameron ◽  
...  
Keyword(s):  
New Technologies ◽  
Rapid Development ◽  
Gene Discovery ◽  
Metagenomic Library ◽  
Metagenomic Sequencing ◽  
Optimum Conditions ◽  
Enzyme Screening ◽  
Specific Amplification ◽  
The Ideal

With the rapid development of powerful protein evolution and enzyme-screening technologies, there is a growing belief that optimum conditions for biotransformation processes can be established without the constraints of the properties of the biocatalyst. These technologies can then be applied to find the ‘ideal biocatalyst’ for the process. In identifying the ideal biocatalyst, the processes of gene discovery and enzyme evolution play major roles. However, in order to expand the pool genes for in vitro evolution, new technologies, which circumvent the limitations of microbial culturability, must be applied. These technologies, which currently include metagenomic library screening, gene-specific amplification methods and even full metagenomic sequencing, provide access to a volume of ‘sequence space’ that is not addressed by traditional screening.

Arachidonic acid and LTB4 enhance aggregation of psoriatic peripheral blood mononuclear leukocytes in vitro

1987 ◽  
Vol 279 (5) ◽  
pp. 347-350 ◽  
Author(s):  
P. D. Pigatto ◽  
M. M. Polengni ◽  
G. F. Altomare ◽  
G. L. Tadini ◽  
S. Villa
Keyword(s):  
Arachidonic Acid ◽  
Peripheral Blood ◽  
Mononuclear Leukocytes ◽  
Peripheral Blood Mononuclear
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