Determination of specificity of the MAbs to Der p 21
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Laimis Silimavicius
PROVIDER: E-MTAB-13625 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:For determination of reactivity with rDer p 21 and Der p 21 in extracts, a serially diluted rDer p 21 from 100 to 0.00305 µg/mL and 6 different HDM extracts diluted 16 times from their stock concentrations were printed. Human serum albumin, rMBP, HRP and PBS were printed as negative controls. Each allergen and control were printed as a single droplet (~450 pL/drop) in three replicates onto 2D-Epoxy glass slides (PolyAn GmbH, Germany) using sciFLEXARRAYER SX microarray printer (SCIENION GmbH, Germany). The slides were blocked with PBS-T containing 2% BSA (blocking buffer) for 30 min at RT. After that, the slides were incubated for 2 h with MAbs 0.2 µg/mL, diluted with blocking buffer, 80 µL/well). Then the slides were incubated for 30 min with the goat anti-mouse IgG Fc Alexa Fluor® 647 (SouthernBiotech, USA) (1 μg/mL, diluted with blocking buffer, 80 μL/well). The slides were scanned using InnoScan 710 AL microarray scanner (Innopsys, France). The images were analyzed with MAPIX software (Innopsys, France). The average mean ± standart deviation value of each allergen and control was calculated using spots’ median fluorescence intensity (MFI) values
2023-12-24 | E-MTAB-13590 | biostudies-arrayexpress
Project description:goal was to determine antigenic properties of rDer p 21 and 6 different HDM using sera samples
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2013-11-01 | E-MTAB-2026 | biostudies-arrayexpress
Project description:Rationale: During excessive pressure or overload, cardiac cells are subjected to increased mechanical stress. We investigate how the stress response of cardiac cells to mechanical stress can be compared to genotoxic stresses induced by DNA damaging agents. Methods: Cultures enriched for cardiomyocytes and cultures of fibroblasts were derived from ventricles of neonatal rat hearts. In three independent experiments, both cell cultures were subjected to mechanical stress (cyclic stretch at 60 cycles/min), UV irradiation (10 J/m2), or X-rays (8.5 Gy). Cells were harvested 24 h. after (the onset of) treatment. Total RNA was isolated using an RNeasy® kit (Qiagen). Single stranded cDNA was synthesized from RNA from myocyte-enriched populations (10-14 microgram) and populations of fibroblasts (7.5-14 microgram), using Superscript II reverse transcriptase and T7-oligo(dT)24 primers at 42ºC for 1 h. Double stranded cDNA was obtained by using DNA ligase, DNA polymerase I and RNAse H at 16ºC for 2 h, followed by T4 DNA polymerase at 16ºC for 5 min. cRNA was synthesized from cDNA using the BioArray HighYield® RNA transcript labeling kit (Enzo) in the presence of biotin-labeled CTP and UTP. cRNA was fragmented in a buffer containing 40 mM Tris-acetate (pH 8.1), 100 mM KAc and 30 mM MgAc, at 94ºC for 35 min. Fragmented cRNA was hybridized to GeneChip RG-U34A arrays (Affymetrix) at 45ºC for 16 h. Arrays were washed and incubated with a streptavidin-phycoerythrin complex. Fluorescent signals were determined with a GeneChip scanner.
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Project description:Multiple sclerosis (MS) is a chronic, inflammatory, and demyelinating disease of the central nervous system (CNS). Ursolic acid (UA) can be used in the MS treatment with anti-inflammatory and neuroprotective activities. However, UA is insoluble in water, which may affect its medication effectiveness. In this study, we evaluated the pharmacological effects of UAOS-Na, a water-soluble UA derivative, on experimental autoimmune encephalomyelitis (EAE) mouse, explored its underlying mechanism, and verified the mechanism by in vitro and in vivo experiments. As we expected, UAOS-Na (30 mg/kg/d) delayed the onset time of EAE from 11.78 days post immunization (dpi) to 14.33 dpi, reduced the incidence from 90.0% to 42.9%, and was more effective than UA. UAOS-Na (60 mg/kg/d) significantly decreased the serum levels of IFN-γ, IL-17A, TNF-α and IL-6, reduced the mononuclear cell infiltration of spinal cord, and inhibited the overexpression of key transcription factors T-bet and ROR-γt of EAE mouse spinal cord and spleen. In addition, UAOS-Na attenuated demyelination and astrogliosis in the CNS of EAE and Cuprizone-induced mice. Mechanically, proteomics showed that 217 differential expression proteins (DEPs) were enriched and 215 were upregulated in EAE mice. After UAOS-Na treatment, 52 DEPs were enriched and 49 were downregulated, and these DEPs were markedly enriched in inositol phosphate metabolism, calcium, sphingolipid, cAMP, and antigen processing and presentation (APP) signaling pathways. Among them, there were few studies on APP signaling pathway related with MS. Therefore, we further investigated the effect of UAOS-Na on APP signaling pathway and found that UAOS-Na downregulated the protein levels of Tapbp and H2-T23 in MHC-I antigen presentation pathway and decreased the proliferation of splenic CD8 T cells, thereby inhibiting the CNS infiltration of CD8 T cells. Together, our findings demonstrated that UAOS-Na have both direct anti-demyelination and anti-inflammation effects. And it could reduce the inflammation of MS by downregulating the expression of Tapbp and H2-T23 in the MHC-I antigen presentation pathway.
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Project description:Inducing the long-term protection via effector memory CD8+ T cells residing in the peripheral tissues is the ultimate goal of T cell-based vaccination strategies. CD8+ T cell reacting against a particular set of antigenic peptides show propensity to generate stable pools of memory inflating cells with profound protective capacity. While the epitopes that induce memory inflation have been thoroughly characterized and used as excellent constituents of vaccines such as recombinant adenoviruses, the identity of the tissue factors responsible for maintaining memory inflating CD8+ T cells remains elusive. Here, we have used a Cre recombinase-dependent, -galactosidase (gal)–recombinant adenovirus vector that allowed for cell-specific expression of gal epitopes and identification of cell types involved in generation of inflationary responses. While targeting antigen expression to classical hematopoietic antigen presenting cells was insufficient to activate gal-specific CD8+ T cells, expressing the antigen exclusively in CCL19-producing fibroblastic stromal cells (FSCs) induced robust anti-gal CD8+ T cell response. Using bone marrow chimera mice to abolish the expression of major histocompatibility complex I (MHC-I) and Basic Leucine Zipper ATF-Like Transcription Factor 3 (BATF3) in hematopoietic cells we demonstrated that CCL19-expressing FCS function as antigen libraries, gradually releasing the antigen to CD103+ DCs to maintain gal-specific memory inflating population. Moreover, targeted ablation of CCL19-producing cells revealed that pulmonary fibroblastic stromal cells act as the principal organizer of the nurturing niches that support the metabolic conversion in CD8+ T cells necessary for the generation of long-lasting protective inflationary responses. Overall, our data reveal a hitherto unknown function of CCL19-expressiong fibroblastic stromal in supporting the metabolic fitness of CD8+ T cells, thereby promoting the generation of tissue-specific and long-lasting protective CD8+ T cell responses.
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