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1.
Mol Biol (Mosk) ; 51(5): 787-796, 2017.
Artigo em Russo | MEDLINE | ID: mdl-29116065

RESUMO

Therapeutic monoclonal antibodies and recombinant proteins including cytokines are commonly used in the treatment of cancer and inflammatory diseases. In most cases, these protein-based drugs exhibit a high therapeutic efficacy, which is unfortunately frequently associated with a variety of side effects. We have investigated the in vitro and in vivo immunogenicity of recombinant antitumor protein lactaptin (RL2). Based on the qRT-PCR analysis, we have shown that, in MDA-MB-231 human breast adenocarcinoma cells, RL2 suppresses the NF-kB signaling cascade that regulates the reactions of innate immunity. RL2 inhibits the expression of the CXCL1 protein and apoptosis inhibitor A20 and enhances expression of IkB, NF-kB repressor. The ELISA method has been used to evaluate the antibody titer in the blood of mice, which received single and triple intravenous or intraperitoneal injections of RL2. The multiplex immunoassay of 23 cytokines in the mice blood has shown that the RL2 injections lead to a slight increase in the levels of systemic pro-inflammatory cytokine interleukin-5 (IL-5) and keratinocyte chemoattractant (KC), a homologue of human macrophage inflammatory protein-1 (MIP-1). These observations indicate the low immunogenicity of the recombinant lactaptin analog, which can be considered to be a potential molecular drug candidate for further clinical development.


Assuntos
Antineoplásicos , Caseínas , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/imunologia , Adenocarcinoma/patologia , Antineoplásicos/imunologia , Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Caseínas/genética , Caseínas/imunologia , Caseínas/farmacologia , Citocinas/imunologia , Humanos , Células MCF-7 , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto
2.
Ross Fiziol Zh Im I M Sechenova ; 100(5): 613-23, 2014 May.
Artigo em Russo | MEDLINE | ID: mdl-25669100

RESUMO

The aim of the main research was investigated of interaction of neural, endocrine and immune systems under experimental postviral fatigue, behavioral reactions, level of corticosterone, changes of IL-3 and IL-10 gene expression in rats' hypothalamus and INF-α in hypothalamus and spleen were analyzed. It has been shown the decrease of physical activity, increasing of corticosterone's level and gene expression of cytokines after injection of Poly I:C as a model of postviral fatigue. After remedication of Poly I:C increasing of physical activity was shown.


Assuntos
Síndrome de Fadiga Crônica/fisiopatologia , Hipotálamo/fisiopatologia , Atividade Motora/imunologia , Baço/fisiopatologia , Animais , Corticosterona/sangue , Modelos Animais de Doenças , Síndrome de Fadiga Crônica/induzido quimicamente , Síndrome de Fadiga Crônica/genética , Síndrome de Fadiga Crônica/imunologia , Expressão Gênica , Hipotálamo/imunologia , Imunidade Inata , Interferon-alfa/genética , Interferon-alfa/imunologia , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-3/genética , Interleucina-3/imunologia , Masculino , Atividade Motora/genética , Poli I-C , Ratos , Ratos Wistar , Baço/imunologia , Natação
3.
J Mol Recognit ; 26(8): 368-75, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23784993

RESUMO

A convenient and simple approach for the preparation of molecularly imprinted polymers (MIPs) based on polyamide (nylon-6) was developed. The polymer matrix formation occurred during the transition of nylon from dissolved to solid state in the presence of template molecules in the initial solution. 2,2,2-Trifluoroethanol (TFE) was chosen as a main solvent for the polyamide. It provides a high solubility of nylon and does not significantly change the structure of biopolymers. The alteration of the polymer matrix structure after the addition of different types of porogens in the nylon/TFE solution was investigated. The structured polymers in the form of films and microparticles were prepared in the chosen optimal conditions. Different model biomolecular templates (of low- and high-molecular weight) were used for the preparation of MIPs, which were shown to specifically recognize these molecules upon binding experiments. The binding of the template molecules to MIPs was monitored using spectrophotometric, radioisotopic, or fluorometric detection. The selectivity coefficients of the MIPs were estimated to be 1.4-4.6 depending on the type of templates and conditions of the polymer matrix formation.


Assuntos
Caprolactama/análogos & derivados , Impressão Molecular/métodos , Polímeros/química , Trifosfato de Adenosina/química , Adsorção , Animais , Caprolactama/química , Bovinos , Dicroísmo Circular , Elétrons , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Soroalbumina Bovina/metabolismo , Solventes/química
4.
Bioorg Khim ; 36(5): 700-13, 2010.
Artigo em Russo | MEDLINE | ID: mdl-21063457

RESUMO

The features of UV-induced immobilization of oligonucleotides on a nylon membranes and the effectiveness of enzymatic labeling of immobilized probes at heterophase detection of nucleic acids are studied. Short terminal oligothymidilate (up to 10 nt) sequences are suggested to attach to the probe via a flexible ethylene glycol based linker. The presence of such fragment enhances the intensity of immobilization and reduces UV-dependent degradation of the targeted (sequence-specific) part of the probe by reducing the dose needed for the immobilization of DNA. The optimum dose of UV-irradiation is determined to be ~0.4 J/cm(2) at the wavelength 254 nm. This dose provides high level of hybridization signal for immobilized probes with various nucleotide composition of the sequence specific moiety. The amide groups of the polyamide are shown to play the key role in the photoinduced immobilization of nucleic acids, whereas the primary amino groups in the structure of PA is not the center responsible for the covalent binding of DNA by UV-irradiation, as previously believed. Various additives in the soaking solution during the membrane of UV-dependent immobilization of probes are shown to influence its effectiveness. The use of alternative to UV-irradiation system of radical generation are shown to provide the immobilization of oligonucleotides onto the nylon membrane.


Assuntos
Membranas Artificiais , Hibridização de Ácido Nucleico/métodos , Nylons , Sondas de Oligonucleotídeos/química , Oligonucleotídeos/química , Bacteriófago T4/genética , Sequência de Bases , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos/instrumentação , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Sondas de Oligonucleotídeos/genética , Oligonucleotídeos/genética , Raios Ultravioleta
5.
Bioorg Khim ; 36(6): 802-14, 2010.
Artigo em Russo | MEDLINE | ID: mdl-21317947

RESUMO

The isothermal amplification of reporter signal via limited probe extension (minisequencing) upon hybridization of nucleic acids has been studied. The intensity of reporter signal has been shown to increase due to enzymatic labeling of multiple probes upon consecutive hybridization with one DNA template both in homophase and heterophase assays using various kinds of detection signal: radioisotope label, fluorescent label, and enzyme-linked assay. The kinetic scheme of the process has been proposed and kinetic parameters for each step have been determined. The signal intensity has been shown to correlate with physicochemical characteristics of both complexes: probe/DNA and product/DNA. The maximum intensity has been observed at minimal difference between the thermodynamic stability of these complexes, provided the reaction temperature has been adjusted near their melting temperature values; rising or lowering the reaction temperature reduces the amount of reporting product. The signal intensity has been shown to decrease significantly upon hybridization with the DNA template containing single-nucleotide mismatches. Limited probe extension assay is useful not only for detection of DNA template but also for its quantitative characterization.


Assuntos
DNA/análise , Oligonucleotídeos/química , Análise de Sequência de DNA/métodos , DNA/química , Hibridização de Ácido Nucleico , Termodinâmica
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