Recombinant major allergens Phl p 1 from meadow timothy (
Phleum pratense
) and Amb a 1 from ragweed (
Ambrosia artemisiifolia
) were obtained in
E. coli
cells. Allergen purification procedures were ...developed using anion exchange and affinity chromatography, from which we obtain 10 mg Phl p 1 and 7.5 mg Amb a 1 from 1 L of culture medium with an electrophoretic purity of at least 90%. To analyze the ability of IgE antibodies to bind Phl p 1 and Amb a 1, sera obtained from people with IgE to various respiratory allergens, identified by the RIDA method, were used. Part of the sera bound Phl p 1 or Amb a 1, which confirmed the immunoreactivity of the obtained recombinant proteins. Antibody titers to Amb a 1 correlated with RIDA data on sensitivity to a mixture of herbs and rye pollen. Antibody titers to Phl p 1 did not correlate with RIDA data. Of the 20 positive sera selected, 73 and 89% of the sera recognizing Phl p 1 or Amb a 1, respectively, had antibodies to the cross-reactive alder/birch/hazel allergen group.
Type I hypersensitivity is mediated by the production of IgE antibodies in response to normally harmless substances. Debate still continues about the mechanisms underlying allergic reactions. Reduced ...barrier tissue function can be one of the risk factors for allergies. The aim of the present work was to compare the humoral immune response to Epstein-Barr virus in patients allergic to the A. alternata fungus or D. farinae house dust mites and healthy donors. It is known that up to 90% of the world population are infected with EBV. This infection occurs at early age when a child develops allergy. The antibodies were analyzed using immuno-PCR and the recombinant EBV protein rEBNA. We were able to demonstrate that infection occurs at early age in both allergic patients and healthy donors. The proportion of EBP-seropositive individuals was comparable between the groups (75% and 74%). The proportion of patients with high IgG1 titers among patients with allergies was lower (7%) than in healthy donors (18%), suggesting a lower viral load. In patients with allergies (but not in healthy donors) IgG1 titers declined as children grew older (р = 0.037). Besides, IgA1 titers were increased in patients with allergies in comparison with healthy donors, but differed between patients allergic to A. alternata and house dust mites. In allergic individuals, production of IgM against EBV was triggered earlier than in healthy donors. We conclude that IgM production and the IgA1-mediated humoral response occur earlier in patients with allergies, causing a decline in IgG1 titers over time.
Interaction of Lymphocytes with Mesenchymal Stem Cells Svirshchevskaya, E. V.; Poltavtseva, R. A.; Beletskii, I. P. ...
Bulletin of experimental biology and medicine,
08/2016, Letnik:
161, Številka:
4
Journal Article
Recenzirano
We studied the interaction of neural stem cells and dental pulp-derived mesenchymal stem cells with lymphocytes from autologous and heterologous donors. Flow cytometry analysis with the use of ...CFSE-labeled lymphocytes demonstrated an increase in the content of proliferating CD8, CD16 and CD56 cells, but not CD4 cells in cultures of HLA-DR-negative mesenchymal stromal cells from the dental pulp co-cultured with lymphocytes. In neural cultures expressing HLA-DR, all subpopulations of T cells and NK cells were activated. No differences between the autologous and heterologous cultures were revealed.
Successful disease prevention and therapy critically depend on timely diagnosis of infections. Quantitative immuno-PCR (qiPCR) technology improves the sensitivity in the detection of antibodies to ...pathogens. A qiPCR-based assay was developed to determine IgG antibodies to Epstein–Barr virus (EBV) in the human blood serum. EBV nuclear protein 1 fragment (pEBV) was expressed in
Escherichia coli
. A synthetic single-stranded deoxyribonucleotide was conjugated to streptavidin, and the conjugate was used to detect рEBV–IgG1–biotin complexes by qiPCR. The IgG1 titers determined by qiPCR were compared to the results of enzyme-linked immunosorbent assay (ELISA). The sensitivity of qiPCR was one order of magnitude higher than that of ELISA. Thus, a highly sensitive qiPCR-based assay was developed to quantitate antibodies specific to the recombinant EBV antigen.
The article presents the results of international and Russian clinical studies to assess the efficacy and safety of topical antimycotic drugs for the treatment of skin mycoses. The authors ...demonstrate the advantages of Naftifine (compliance and efficacy) as compared to other topical drugs. Taking into consideration a broad range of its action including antifungal, antibacterial and anti-inflammatory effects, Naftifine can be considered as a first-line drug for the treatment of superficial dermatomycoses including complicated with a bacterial infection.
Chitosan, (1-4)-2-amino-2-deoxy-beta-
d-glucan, is a deacetylated form of chitin, an abundant biodegradable, positively charged natural polysaccharide. Chitosan is used for antigen delivery through ...mucosal barrier due to its ability to disrupt tight junctions. Here we produced new water-soluble low-molecular weight chitosan (LMW-Chi) lipid derivatives and compared their ability to stimulate humoral response with the effect of unmodified LMW-Chi or its oligosaccharide derivatives. LMW-Chi effectively penetrated into macrophage-like, lymphoid and epithelial cells. It also stimulated in mice IgG production to model proteins delivered either by subcutaneous or intranasal routes. Adjuvant effect of chitosan derivatives was comparable to or lower than that of unmodified LMW-Chi. Thus, it is possible that adjuvant effect is induced by unmodified glucosamine units of chitosan.
The conjugates of low-molecular weight chitosans with fatty acids and carbohydrates were compared for their ability to stimulate IgG response in mice to model proteins delivered via subcutaneous or mucosal routes.
▪
Reaction of hydroxyl-containing heterocyclic colchicinoids with S-nucleophiles led to the formation of furanoallocolchicinoid sulfides in a high yield.
Proteases play a key role in the physiological processes of the small intestine, supporting its normal physiological functions as a part of the digestive system, in which hydrolysis and assimilation ...of nutrients are implemented. A high concentration of antigens in the intestinal lumen activates immunity and stimulates a chronic weakly expressed inflammatory response in a normal gastrointestinal tract (GIT). Cathepsin
G
, a serine protease controlling the functional state of immune cells, directly participates in the complicated system for the regulation of balance between physiological and pathological inflammations. To determine the role of cathepsin
G
in the small intestine, an immunofluorescent investigation of biopsies from the human duodenal mucosa were investigated using the confocal immunofluorescence microscopy method and human antibodies to cathepsin
G
. It has been shown for the first time that cathepsin
G
, which was regarded conventionally as one of the effectors of the inflammatory process, is a constitutive enzyme of the human duodenum and is constantly present in its normal mucosa. The new cell sources for the cathepsin
G
biosynthesis identified: intraepithelial lymphocytes (IELs), lamina propria lymphocytes,
CD
14-positive intestinal macrophages, and Paneth cells, which are specialized epitheliocytes of intestinal glands. Our data on the cathepsin
G
expression by immunocytes and Paneth cells in the duodenum allow us to attribute cathepsin
G
to the main proteases of intestinal immunity, which indicates the important role of this enzyme in the regulation of human GIT functions.
Along with bright fluorescence in the near-IR range, heptamethine carbocyanine dyes possess affinity to cancer cells. Thus, these dyes could be utilized as fluorescent labels and vectors for drug ...delivery as covalent conjugates with cytotoxic compounds. To test the properties, structure–activity relationship, and scope of such conjugates, we synthesized drug-dye dyads of tricarbocyanine dyes with anthracycline drug daunorubicin. We used hydrophilic zwitterionic and hydrophobic positively charged benzoindoline-benzothiazole-based heptamethine dyes as terminal alkyne derivatives and N-acylated or oxime-linked daunorubicin as azido-derivatives. These two alkynes and two azides were coupled to each other by Cu-catalyzed Huisgen–Meldal–Sharpless cycloaddition (click reaction) to afford four conjugates. Molecules based on hydrophobic dyes possess submicromolar cytotoxicity to HCT116 cells. Cytotoxicity, cell penetration, intracellular distribution, apoptosis induction and the effect of antioxidants on toxicity were evaluated. The results show that the structure of the cyanine–anthracycline conjugate (hydrophilicity/hydrophobicity, charge, linker, attachment site) is important for its biological activity, thus, expansion of the chemical space of such conjugates could provide new molecular research tools for diagnostics and therapy.
Display omitted
•Four novel daunorubicin-heptamethine cyanine (tricarbocyanine) dye were synthesized.•Conjugates with alkyl-substituted dye posessed micromolar cytotoxicity.•Conjugates with sulfo-substituted dye posessed no cytotoxicity.•Tricarbocyanine delivered anthracycline to mitochondria of cancer cells.•ROS generation contributed to cytotoxicity of the conjugates.
PDF
