REVIEWS
Interleukin 33 (IL-33) is a member of the IL-1 family, which is widely expressed on all types of cells. IL-33 was identified as a functional ligand for the plasma membrane receptor complex, which is a heterodimer consisting of a membrane bound ST2 receptor (growth stimulating factor). IL-33 is involved in the development of immune response with predominant release of pro-inflammatory T helper type 2 cytokines. IL-33 is widely expressed on various structure-forming cells, such as epithelial, endothelial and smooth muscle cells. Increased expression of IL-33 is observed during necrosis of these cells (after tissue or cell damage), and it is released into extracellular space, and acts as an endogenous danger signal, sending a sort of warnings to neighboring cells and tissues. Recently, many studies have shown that IL-33 can participate in development and progression of fibrosis in various organs. However, it exerts anti-inflammatory effects upon development of other diseases. This review will discuss biological characteristics of IL-33 and a role of the IL-33/ST2 signaling pathway in the development of fibrosis.
ORIGINAL ARTICLES
Authenticity evaluation of proteins obtained with recombinant DNA technology is an important step in confirming efficacy and safety of the drugs based on them. One of the main ways to assess the authenticity is to compare molecular structure of the test and standard samples using the peptide mapping method with chromatographic separation of the products obtained by enzymatic degradation. Proper selection of a standard reference sample is essential in order to achieve reliable results. A standard sample of Interferon (CRS, Chemical Reference Substances) recommended by the European Agency for the Quality of Medicines for interferon alpha-2b substances containing N-terminal methionine is inappropriate, since the Interferon CRS sample doesn’t contain methionine. We present the results of development of qualification procedure for methionine form of Interferon alfa-2b industrial standard sample (ISS). The range of use for this ISS is authenticity confirmation for the methionine form of interferon alpha-2b substance using peptide mapping method with reverse-phase high-performance liquid chromatography (reverse-phase HPLC). The quality assessment was performed for all the parameters specified by the manufacturer of this candidate substance at the initial stage of qualification procedure, due to changed application area, and changed package size. Further, 30 peptide cards of the ISS candidate substance were obtained after pre-trypsinolysis of the protein followed by validated HPLC method with proven repeatability.
It was shown that the hydrolysis conditions, i.e., the choice of trypsin preparations, may significantly affect the peptide map profile. Therefore, a reference to specific manufacturer and the catalog number of the product should be provided in description of application conditions for the ISS proposed.
A set of eight reference peaks (peaks of comparison) has been justified, as based on evaluation of peptide maps and results of high-resolution mass spectrometry. The peak with maximally stable yield and intensity was selected as the main peak with an established absolute retention time. Two peaks with relative retention times were chosen as essential peaks for evaluation, i.e., the 1st peak containing N-terminal methionine, and the 2nd peak of highest molecular weight with an established amino acid sequence covering 11% of the studied interferon molecule.
We have also qualified ISS parameters expressed as absolute (minimum for one reference peak), and relative (for the remaining reference peaks) retention time periods. Authenticity of the ISS candidate was further confirmed by the peptide mapping method, as compared with interferon CRS reference standard. Their peak patterns proved to be near-similar, except of a peak with eluted peptide containing N-terminal methionine as revealed by high-resolution mass spectrometry
Studies of probable significance of different immunological mediators for the development of chronic allergic inflammation in patients with allergic bronchopulmonary aspergillosis (ABPA) are necessary in order to specify potential targets for therapeutic intervention and timely diagnosis of the disease. The purpose of present study was to determine the features of immune response regulation, and to identify diagnostic markers associated with development of ABPA in patients with bronchial asthma, and to evaluate clinical and immunological efficacy of specific antimycotic therapy.
The study involved 13 patients with ABPA, 14 patients with bronchial asthma with fungal sensitization (BAFS), 17 patients with bronchial asthma (BA) and 12 apparently healthy individuals. Levels of thymic stromal lymphopoietin (TSLP), thymus and activation-regulated chemokine (TARC), IL-8, as well as levels of total IgE and specific IgE to Aspergillus fumigatus (A. fumigatus) were measured in blood serum by enzyme immunoassay; blood eosinophil counts were also made. Monitoring of these immunological markers in the course of antimycotic therapy was carried out.
Significantly higher numbers of eosinophils, increased levels of total IgE and sIgE for A. fumigatus, as well as TARC and IL-8 in serum were revealed in patients with ABPA when compared to the patients with BA. No significant differences in TSLP content were found between the examined groups of patients. A positive correlation between the levels of sIgE to A. fumigatus and contents of TARC and IL-8, numbers of eosinophils, and total IgE levels confirms the important diagnostic value of proinflammatory cytokines in ABPA patients. In the course of itraconazole medication, a positive clinical and immunological dynamics in ABPA patients was revealed. After 12 weeks of therapy, a significant increase in AST, FEV1 and Tiffno respiratory indexes, along with decreased number of eosinophils, total IgE levels, and a trend towards a decrease in TARC and IL-8 levels were documented. This dynamics confirms clinical efficiency of antifungal drugs when treating chronic allergic inflammation in ABPA patients.
Implementation of modern immunological biomarkers, alongside with traditional indicators, will allow to differentially evaluate a probability for ABLA development in patients with bronchial asthma, to present additional evidence for discerning early stages of the disease, and to conclude about the efficiency of the therapy applied.
The aim of present study was to evaluate safety and clinical efficacy of inhalatory immunotherapy based on intranasal delivery of bioactive factors produced by M2 macrophages applied for treatment of patients with organic brain syndrome (OBS).
Materials and methods. The study under the NCT02957123 protocol (www.ClinicalTrails.gov) included thirty patients with OBS of various genesis (10 men and 20 women aged 18 to 81; Me, 62.5 years). Neurological assessment and the levels of 32 cytokines in the blood serum of patients were evaluated before and 2-3 days after completion of inhalation immunotherapy.
Intranasal inhalations of cell-free culture medium of M2 macrophages (2 mL, once a day for 28-30 days) were safe and well tolerated. None of 30 treated patients had severe adverse events and serious treatmentrelated side reactions. One month after starting the inhalations, a positive dynamics in neurological status was noted in all the patients. A marked clinical response was documented in twenty out of thirty patients (67%), which manifested as improvement, according to all scales and questionnaires. The neurological improvement was not reversed over 6 months of follow-up period. In other ten patients (33%), a moderate clinical response was shown as improvement of individual scores. The positive changes were as follows: 1) a 43% decrease in anxiety and depression scores (according to HADS scale, pU = 0.0008); 2) an increase of total motor activity (stability and gait) by 25%, pU = 0.0001); 3) correction of cognitive functions (MoCa test, pU = 0.007); 4) reduced number and intensity of the disease symptoms by 52% (pU = 0.0001). This marked clinical response to immunotherapy is shown to be associated with correction/normalization of serum hepatocyte growth factor (HGF) level.
Conclusion. Inhalation immunotherapy based on intranasal delivery of bioactive factors produced by M2 macrophages can improve neurological and functional recovery in patients with organic brain syndrome.
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ISSN 2313-741X (Online)