Laboratory of cell interactions

Department of immunology

Head: Aleksandr Sapozhnikov, D.Sc, professor
amsap@mail.ru+7(495)330-40-11

NamePositionContacts
Aleksandr Sapozhnikov, D.Sc, professordepart. dir.amsap@mail.ru+7(495)330-40-11
Vitaly Fokinl. r. f.
Elena Kovalenko, Ph.D.s. r. f.lenkovalen@mail.ru+7(495)330-40-11
Elena Kirilina, Ph.D.s. r. f.+7(495)330-72-56
Gennadij Lucenko, Ph.D.s. r. f.gvlut@mx.ibch.ru+7(495)330-40-11
Elena Svirshevskaja, Ph.D.s. r. f.esvir@mx.ibch.ru+7(495)330-40-11
Anna Boyko, Ph.D.r. f.boyko_anna@mail.ru+7(495)330-40-11
Marina Shevchenko, Ph.D.r. f.shev@mx.ibch.ru+7(495)330-40-11
Leonid Kanevskiy, Ph.D.r. f.leonid_kanewski@mail.ru+7(495)330-40-11
Natalya Troyanovar. f.troyanatali@gmail.com
Ekaterina Fedotovar. f.
Olga Kotsareva, Ph.D.j. r. f.olga.kotsareva@gmail.com
Ol'ga Shustovaj. r. f.+7(495)330-40-11
Marija Grechihinaj. r. f.marygrec@mail.ru+7(495)330-40-11
Mikhail Efremovj. r. f.mae@mx.ibch.ru+7(495)330-72-56
Anna Klinkovaj. r. f.anna_klinkova@mail.ru+7(495)330-40-11
Ekaterina Servulij. r. f.violet.vulpera@gmail.com
Maria Streltsovaj. r. f.mstreltsova@mail.ru+7(495)3304011
Gul'nar Fattakhovaj. r. f.
Dmitry AronovPhD stud.aronov.mml@gmail.com+7(929)6437489
Dmitriy ChudakovPhD stud.boris-chudakov@yandex.ru+7(916)0761744
Ruslan MirzoevPhD stud.rusfess@mail.ru
Elena KashirinaPhD stud.helen-kas@mail.ru
Sofya ErokhinaPhD stud.sonya.erokhina@gmail.com+7()
Natal'ja Galkinat. q. - lab. as.+7(495)330-40-11
1245, Ph.D.eng.mariya.v.konovalova@gmail.com
Ekaterina Doroninares. eng.

Former members:

Raisa Beljovskaja, Ph.D.r. f.
Nadezhda Lutsan, Ph.D.r. f.
Larisa D'jachkova, Ph.D.j. r. f.
Marija Kuchinaj. r. f.
Olga Tatsyjj. r. f.
Anastasiya Zubarivaj. r. f.zun_88@mail.ru
Maxim KilyachusPhD stud.mskilyachus@gmail.com
Emin Alekperov, Ph.D.res. eng.alekperovea@mail.ru
Anastasija Kuchukovares. eng.
Alexander Prokhorovres. eng.sashapro2006@yandex.ru

Selected publications

  1. Streltsova M.A., Barsov E., Erokhina S.A., Kovalenko E.I. (2017). Retroviral gene transfer into primary human NK cells activated by IL-2 and K562 feeder cells expressing membrane-bound IL-21. J. Immunol. Methods 450, 90–94 [+]

    Natural killer (NK) cells are capable of rapidly recognizing and efficiently killing tumor cells. This makes them a potentially promising agent for cancer immunotherapy. Additional genetic modifications of NK cells may further improve their anti-tumor efficacy. Numerous technical challenges associated with gene delivery into NK cells have significantly tempered this approach. We achieved efficient retroviral vector transduction of primary human NK cells that were stimulated by a combination of IL-2 and engineered K562 cells expressing membrane-bound IL-21. The activated NK cells were in less differentiated state and expressed NK cell activation receptors NKG2D, NKp30, CD16, and were highly HLA-DR-positive. This NK cell population was highly susceptible to the transduction by both GFP- and NGFR-expressing retroviral vectors, with transduction efficiency exceeding 50%. More mature CD57(+) NK cell population was generally resistant to retroviral vector transduction because of poor response to the stimulation. Our findings may facilitate retroviral vector-mediated genetic engineering of human primary NK cells for future immunotherapies.

    ID:1938
  2. Streltsova M.A., Klinkova A.V., Kuchukova A.A., Kadin A.Y., Kanevskiy L.M., Kovalenko E.I. (2016). Ethanol-dependent expression of the NKG2D ligands MICA/B in human cell lines and leukocytes. Biochem. Cell Biol. , 1–9 [+]

    Alcohol consumption affects the human immune system, causing a variety of disorders. However, the mechanisms of development of these changes are not fully understood. We hypothesized that ethanol may influence the expression of MICA and MICB, stress-induced molecules capable of regulating the activity of cytotoxic lymphocytes through the interaction with receptor NKG2D, which substantially affects the functionality of cellular immunity. We analyzed the effects of ethanol on MICA/B expression in tumor cell lines and human leukocytes. In the cell line models, ethanol caused different changes in the surface expression of MICA/B; in particular, it induced the translocation of intracellular proteins MICA/B to the cell surface and shedding of MICA (in soluble and microparticle-associated forms) from the plasma membrane. The observed results are not linked with cell death in cultures, taking place only under higher doses of ethanol. Ethanol at physiologically relevant concentrations (and higher) stimulated expression of MICA/B genes in different cell types. The effect of ethanol was more pronounced in hepatocyte line HepG2 compared with hematopoietic cell lines K562, Jurkat, and THP-1. Among the tested leukocytes, the most sensitive to ethanol action were T cells activated ex vivo with IL-2, in which the increase of MICA/B mRNA expression was registered with the smallest dose of ethanol (0.125%). In human monocytes, ethanol may lead to elevations in surface MICA/B levels. Presumably, changes in MICA/B expression caused by ethanol can affect the functions of NKG2D-positive cytotoxic lymphocytes, modulating immune reactions at excessive alcohol consumption.

    ID:1784
  3. Клинкова А.В., Кузьмина Е.Г., Абакушина Е.В., Каневский Л.М., Неприна Г.С., Павлов В.В., Коваленко Е.И. (2016). Циркулирующий белок MICА у больных злокачественными лимфомами. Медицинская иммунология 18 (2), 151–162 ID:2030
  4. Zubareva A.A., Shcherbinina T.S., Varlamov V.P., Svirshchevskaya E.V. (2015). Intracellular sorting of differently charged chitosan derivatives and chitosan-based nanoparticles. Nanoscale 7 (17), 7942–52 [+]

    Chitosan (Chi) is a biodegradable nontoxic polycation with multiple reactive groups that is easily used to obtain derivatives with a desired charge and hydrophobic properties. The aim of this work was to study the intracellular traffic of positively charged hexanoyl-chitosan (HC) or HC-based nanoparticles (HCNPs) and negatively charged succinoyl-chitosan (SC) and SCNPs in epithelial and macrophage cell lines. By using flow cytometry we demonstrated that positively charged HC adhered to cell membranes quicker and more efficiently than negatively charged SC or NPs. However confocal studies showed that SC and SCNPs penetrated cells much more efficiently than HC while HCNPs did not enter the epithelial cells. Macrophages also phagocyted better negatively charged material but were able to engulf both HC and HCNPs. Upon entering the cells, SC and SCNPs were co-localized with endosomes and lysosomes while HC was found in mitochondria and, to a lesser extent, in lysosomes of epithelial cells. Macrophages, RAW264.7, more efficiently transported all Chi samples to the lysosomal compartment while some positively charged material was still found in mitochondria. Incubation of Chi derivatives and ChiNPs at pH specific to mitochondria (8.0) and lysosomes (4.5) demonstrated the neutralization of Chi charge. We concluded that epithelial cells and, to a lesser extent, macrophages sort charged material to the organelles neutralizing Chi charge.

    ID:1499
  5. Pankova T.M., Sapoznikov A.M., Starostina M.V. (2015). Protective effects of myelopeptides in C-1300 neuroblastoma cell line. Russ. J. Bioorgan. Chem. 41 (3), 333–337 [+]

    Effects of regulatory peptides of the bone marrow, in particular myelopeptides 1–6, were studied in the culture of the mouse C-1300 neuroblastoma. The myelopeptides were shown to stimulate the morphological differentiation of neuroblasts. Neuroprotective properties of the myelopeptides were found in the neuroblastoma culture on models of the morphine toxicity and the oxygen-glucose deprivation.

    ID:1494
  6. Troyanova N.I., Shevchenko M.A., Boyko A.A., Mirzoyev R.R., Pertseva M.A., Kovalenko E.I., Sapozhnikov A.M. (2015). Modulating Effect of Extracellular HSP70 on Generation of Reactive Oxigen Species in Populations of Phagocytes. Russ. J. Bioorgan. Chem. 41 (3), 271–279 [+]

    Reactive oxygen species (ROS) produced by phagocytic cells of the innate immune system play an important role in the first line of defense protecting the host from pathogens. The NADPH oxidase multi-subunit complex is the main source of ROS in all types of the phagocytes. Formation of the membrane-associated enzyme complex and its activity are dependent on many different factors controlling both intensification and suppression of the ROS production rate. However, the evidences are emerging in recent years indicating existence of poorly studied mechanisms of restriction of ROS generation level in phagocytes directed at protection of host tissues in the sites of inflammation from destruction caused by the oxygen free radicals. Our previous data and results of other authors demonstrate that a mechanism of the limitation of ROS production by phagocytes may by connected with immunomodulating activity of extracellular pool. of HSP70. In the present work, we used inhibitors of NADPH oxidase and in vitro cultures of different phagocytes to study a possible relationship between down-regulating effect of exogenous HSP70 on ROS generation and the interaction of the protein with the enzyme subunits. Our results confirmed the literature data concerning the ability of extracellular HSP70 to modulate NADPH oxidase activity and demonstrated for the first time an inhibitory effect of the protein on intracellular ROS generation in phagocytes.

    ID:1502
  7. Свирщевская Е.В., Матушевская Е.В., Чудаков Д.Б., Матушевская Ю.И. (2015). Роль инфекции в патогенезе аллергодерматозов. Клиническая дерматология и венерология  (2), 4–10 [+]

    This review presents the data about on the role of infection and the disruption of body's barriers integrity in the development of atopic dermatitis.

    ID:1269
  8. Israel L.L., Kovalenko E.I., Boyko A.A., Sapozhnikov A.M., Rosenberger I., Kreuter J., Passoni L., Lellouche J.P. (2015). Towards hybrid biocompatible magnetic rHuman serum albumin-based nanoparticles: use of ultra-small (CeLn)3/4+ cation-doped maghemite nanoparticles as functional shell. Nanotechnology 26 (4), ID:1152
  9. Чудаков Б. (2015). Стимуляция ex vivo экспрессии проаллергических тканевых цитокинов в клетках мышиной трахеи. Российский иммунологический журнал 9 (18), 110–112 [+]

    Тканевые цитокины (интерлейкины-25 и 33 (ИЛ-25 и 33), а также тимический стромальный лимфопоэтин (ТСЛП)) способны индуцировать продукцию цитокинов аллергического воспаления - ИЛ-4, 5 и 13 – в клетках иммунной системы. Целью данной работы было идентифицировать стимулы, вызывающие продукцию данных цитокинов в эпителии трахеи. В качестве стимулов были взяты различные протеазы, липополисахарид (LPS), формил-метионил-лейцил-фенилаланин (fMLP), β-аланин. Стимуляцию проводили ex vivo на эксплантах мышиной трахеи. Экспрессию генов исследовали методом качественной полимеразной цепной реакции ПЦР. Была установлена небольшая спонтанная продукция ИЛ-33, но не ИЛ-25 и ТСЛП, эксплантами трахеи. Ни один из стимулов не индуцировал продукции ИЛ-25. Показано, что протеазы в ряде случаев (особенно коллагеназа) индуцировали продукцию ТСЛП, увеличивали продукцию ИЛ-33. fMLP стимулировал продукцию как ИЛ-33, так и ТСЛП. β-аланин стимулировал продукцию только ИЛ-33. Ни один из стимулов не индуцировал экспрессию цитокинов в тканях пищевода. Таким образом, эпителий трахеи был способен сортировать сигналы в зависимости от типа стимулов. Эпителий пищевода был нечувствителен к стимулам. Для индукции экспрессии ИЛ-33 и ТСЛП в мышиной модели респираторной аллергии могут использоваться коллагеназа и fMLP.

    ID:1408
  10. Чудаков Д.Б., Свирщевская Е.В. (2015). Роль врождённого иммунитета при аллергии. Российский иммунологический журнал 9 (3), 298–306 [+]

    Аллергическая реакция I типа опосредована продукцией В-клетками иммуноглобулинов класса Е (IgE). Переключение В-клеток на синтез IgE вызывают интерлейкины (ИЛ) 4 и 13, которые продуцируются Т-хелперами 2 типа (Тх2). Однако ИЛ-4 и 13 могут продуцировать не только Тх2. Работы последних лет показали, что травма эпителия барьерных органов: желудочно-кишечного тракта, кожи и легких, приводит к продукции молекул, называемых тканевыми цитокинами, к которым относят ИЛ-25 и 33, а также тимический стромальный лимфопоэтин (ТСЛП). Тканевые цитокины, в свою очередь, активируют продукцию цитокинов, характерных для Тх2: ИЛ-4, 5 и 13, клетками, не несущими маркеров Тх2. Эти клетки получили название лимфоидных клеток врожденного иммунитета 2-го типа (ЛКВИ2) из-за экспрессии на их поверхности маркеров лимфоцитов CD44 и CD45 и способности синтезировать Тх2 цитокины. Повышение уровня ИЛ-4, 5 и 13 в крови в результате их синтеза ЛКВИ2 может вызывать переключение В-клеток на синтез IgE без участия классических Тх2, как это наблюдается при паразитарных инфекциях. В обзоре приведены основные характеристики тканевых и лимфоидных цитокинов, ассоциированных с аллергическим ответом, а также описание фенотипа и функций различных ЛКВИ2.

    ID:1409
  11. Semenkov V.F., Michalski A.I., Sapozhnikov A.M. (2015). Heating and ultraviolet light activate anti-stress gene functions in humans. Front Genet 6, 245 [+]

    Different environmental factors (i.e., toxins, heavy metals, ultraviolet (UV) rays, and X-radiation) cause damage to DNA, cell membranes and other organelles and induce oxidative stress, which results in the excessive production of reactive oxygen species (ROS) by phagocytes. All types of cell stress are accompanied by the activation of anti-stress genes that can suppress ROS synthesis. We hypothesized that different environmental factors would affect organisms through the activation of anti-stress genes by autologous serum (AS) proteins, followed by the synthesis of molecules that increase cell resistance to oxidative stress. The goal of this work was to study the influence of AS on ROS production by peripheral blood neutrophils isolated from donors in different age groups. Neutrophils were isolated from 59 donors (38-94 years old). AS was heated at 100°C for 30 s. or irradiated by UV light at 200-280 nm and 8 W for 10 min. Neutrophils were exposed to heat shock at 42°C for 1 min. (short-term heating stress) or 43°C for 10 min., followed by the determination of the chemiluminescence reaction induced by zymosan. AS can increase or decrease ROS production by neutrophils depending on the structure of the proteins in the serum; these structures can be changed by heating or UV treatment and the temperature of their interaction (4 or 37°C). We propose that the effect of environmental factors on AS proteins can cause an adverse increase in oxidative stress levels due to the functional reduction of anti-stress genes. We found a negative correlation between the quantity of intracellular Hsp70 and levels of intracellular ROS production following 10 min of heat shock at 43°C. Short-term heating stress (1 min) at 42°C was followed by a prominent reduction in ROS production. This effect may be a result of the impact of the hormone adrenaline on the functions of anti-stress genes. Indeed, the same effect was observed after treatment of the neutrophils with adrenaline at concentrations of 10(-4) and 10(-5) M. In contrast, dexamethasone from the other stress hormone group did not evoke the same effect at the same concentrations.

    ID:1500
  12. Kovalenko E.I., Boyko A.A., Semenkov V.F., Lutsenko G.V., Grechikhina M.V., Kanevskiy L.M., Azhikina T.L., Telford W.G., Sapozhnikov A.M. (2014). ROS production, intracellular HSP70 levels and their relationship in human neutrophils: effects of age. Oncotarget 5 (23), 11800–12 [+]

    ROS production and intracellular HSP70 levels were measured in human neutrophils for three age groups: young (20-59 years), elders (60-89 years) and nonagenarians (90 years and older). Elders showed higher levels of spontaneous intracellular ROS content compared with young and nonagenarian groups, which had similar intracellular ROS levels. Zymosan-induced (non-spontaneous) extracellular ROS levels were also similar for young and nonagenarians but were lower in elders. However, spontaneous extracellular ROS production increased continuously with age. Correlation analysis revealed positive relationships between HSP70 levels and zymosan-stimulated ROS production in the elder group. This was consistent with a promoting role for HSP70 in ROS-associated neutrophils response to pathogens. No positive correlation between ROS production and intracellular HSP70 levels was found for groups of young people and nonagenarians. In contrast, significant negative correlations of some ROS and HSP70 characteriscics were found for neutrophils from young people and nonagenarians. The observed difference in ROS and HSP70 correlations in elders and nonagenarians might be associated with an increased risk of mortality in older individuals less than 90 years old.

    ID:1140
  13. Kanevskiy L.M., Erokhina S.A., Streltsova M.A., Telford W.G., Sapozhnikov A.M., Kovalenko E.I. (2014). Bacterial lipopolysaccharide activates CD57-negative human NK cells. Biochemistry Mosc. 79 (12), 1339–48 [+]

    NK cells play an important regulatory role in sepsis by induction and augmentation of proinflammatory reactions in early stages of the septic process and by suppression of immune response in later stages of inflammation. The present work was aimed at the effect of bacterial lipopolysaccharide (LPS), the main pathogenic factor of sepsis development, on human NK cells ex vivo. We show that LPS activates immature CD57-negative NK cells, which typically constitute less than half of the normal NK cell population in human peripheral blood. Under conditions of NK cell stimulation with IL-2, addition of LPS provokes an increase in IFN-γ production. However, LPS both increased and inhibited NK cell cytotoxic activity. It is important to note that the activation of NK cells on LPS addition was observed in the absence of TLR4 on the NK cell surface. These results confirm our previous data arguing for a direct interaction of LPS with NK cells and evidence an atypical mechanism of LPS-induced NK cell activation without the involvement of surface TLR4.

    ID:1258
  14. Чудаков Д.Б., Рязанцев Д.Ю., Каширина Е.И., Бержец В.М., Свирщевская Е.В. (2014). Роль дозы аллергена в индукции у мышей IgE антител на белки клещей домашней пыли. Иммунология 35 (6), 321–328 [+]

    Aim: The aim of this work was to study of humoral immune response to different house dust mite protein doses in mice.

    Results: Specific IgE antibody production was observed only upon multiple administration of low (1ng/injection) recombinant protein doses. The production of IgG and IgA antibodies ware observed only upon administration of relatively high (1-10 microgrammes) allergen doses. IgG and IgA production was also observed during the immunisation with adjuvant upon administration of 0,1 microgramm of proteins. IgE antibodies in our allergic model have low affinity.

    ID:1268
  15. Balabashin D., Kovalenko E., Toporova V., Aliev T., Panina A., Svirshchevskaya E., Dolgikh D., Kirpichnikov M. (2014). Production of anti TNF-α antibodies in eukaryotic cells using different combinations of vectors carrying heavy and light chains. Cytotechnology , ID:1153
  16. Svirshchevskaya E.V., Prokhorov A.V., Zubareva A.A., Berkova N.P. (2014). Tumor immunology and immunotherapy. Recent Research Developments in Cancer 10, ID:1155
  17. Бойко А.А., Ветчинин С.С., Сапожников А.М., Коваленко Е.И. (2014). Изменение уровня белков теплового шока семейства 70 кДа в нейтрофилах человека под действием теплового шока. Биоорг. хим. 40 (5), 528–540 ID:1156
  18. Рязанцев Д.Ю., Дробязина П.Е., Хлгатян С.В., Завриев С.К., Свирщевская Е.В. (2014). Экспрессия аллергенов клещей домашней пыли Der f 1 и Der f 2 в листьях Nicotiana benthamiana. Биоорг. хим. 40 (4), . 468–478 ID:1157
  19. Kuznetsova N.R., Svirshchevskaya E.V., Skripnik I.V., Zarudnaya E.N., Benke A.N., Gaenko G.P., Molotkovskiĭ Yu.G., Vodovozova E.L. (2013). Interaction of liposomes bearing a lipophilic doxorubicin prodrug with tumor cells. Biochemistry (Moscow) Supplement Series A: Membrane and Cell Biology 7 (1), 12–20 [+]

    When used as nanosized carriers, liposomes enable targeted delivery and decrease systemic toxicity of antitumor agents significantly. However, slow unloading of liposomes inside cells diminishes the treatment efficiency. The problem could be overcome by the adoption of lipophilic prodrugs tailored for incorporation into lipid bilayer of liposomes. We prepared liposomes of egg yolk phosphatidylcholine and yeast phosphatidylinositol bearing a diglyceride conjugate of an antitumor antibiotic doxorubicin (a lipophilic prodrug, DOX-DG) in the membrane to study how these formulations interact with tumor cells. We also prepared liposomes of rigid bilayer-forming lipids, such as a mixture of dipalmitoylphosphatidylcholine and cholesterol, bearing DOX in the inner water volume, both pegylated (with polyethylene glycol (PEG) chains exposed to water phase) and non-pegylated. Efficiency of binding of free and liposomal doxorubicin with tumor cells was evaluated in vitro using spectrofluorimetry of cell extracts and flow cytometry. Intracellular traffic of the formulations was investigated by confocal microscopy; co-localization of DOX fluorescence with organelle trackers was estimated. All liposomal formulations of DOX were shown to distribute to organelles retarding its transport to nucleus. Intracellular distribution of liposomal DOX depended on liposome structure and pegylation. We conclude that the most probable mechanism of the lipophilic prodrug penetration into a cell is liposome-mediated endosomal pathway.

    ID:996
  20. Kanevskiy L.M., Telford W.G., Sapozhnikov A.M., Kovalenko E.I. (2013). Lipopolysaccharide induces IFN-γ production in human NK cells. Front Immunol 4, 11 [+]

    Natural killer (NK) cells have been shown to play a regulatory role in sepsis. According to the current view, NK cells become activated via macrophages or dendritic cells primed by lipopolysaccharide (LPS). Recently, TLR4 gene expression was detected in human NK cells suggesting the possibility of a direct action of LPS on NK cells. In this study, effects of LPS on NK cell cytokine production and cytotoxicity were studied using highly purified human NK cells. LPS was shown to induce IFN-γ production in the presence of IL-2 in NK cell populations containing>98% CD56(+) cells. Surprisingly, in the same experiments LPS decreased NK cell degranulation. No significant expression of markers related to blood dendritic cells, monocytes or T or B lymphocytes in the NK cell preparations was observed; the portions of HLA-DR(-bright), CD14(+), CD3(+), and CD20(+) cells amounted to less than 0.1% within the cell populations. No more than 0.2% of NK cells were shown to be slightly positive for surface TLR4 in our experimental system, although intracellular staining revealed moderate amounts of TLR4 inside the NK cell population. These cells were negative for surface CD14, the receptor participating in LPS recognition by TLR4. Incubation of NK cells with IL-2 or/and LPS did not lead to an increase in TLR4 surface expression. TLR4(-)CD56(+) NK cells isolated by cell sorting secreted IFN-γ in response to LPS. Antibody to TLR4 did not block the LPS-induced increase in IFN-γ production. We have also shown that R(e)-form of LPS lacking outer core oligosaccharide and O-antigen induces less cytokine production in NK cells than full-length LPS. We speculate that the polysaccharide fragments of LPS molecule may take part in LPS-induced IFN-γ production by NK cells. Collectively our data suggest the existence of a mechanism of LPS direct action on NK cells distinct from established TLR4-mediated signaling.

    ID:1158
  21. Zubareva A., Ilyina A., Prokhorov A., Kurek D., Efremov M., Varlamov V., Senel S., Ignatyev P., Svirshchevskaya Е. (2013). Characterization of protein and peptide binding to nanogels formed by differently charged chitosan derivatives. Molecules 18 (7), 7848–64 [+]

    Chitosan (Chi) is a natural biodegradable cationic polymer with remarkable potency as a vehicle for drug or vaccine delivery. Chi possesses multiple groups, which can be used both for Chi derivatization and for particle formation. The aim of this work was to produce stable nanosized range Chi gels (nanogels, NGs) with different charge and to study the driving forces of complex formation between Chi NGs and proteins or peptides. Positively charged NGs of 150 nm in diameter were prepared from hexanoyl chitosan (HC) by the ionotropic gelation method while negatively charged NGs of 190 nm were obtained from succinoyl Chi (SC) by a Ca²⁺ coacervation approach. NGs were loaded with a panel of proteins or peptides with different weights and charges. We show that NGs preferentially formed complexes with oppositely charged molecules, especially peptides, as was demonstrated by gel-electrophoresis, confocal microscopy and HPLC. Complex formation was accompanied by a change in zeta-potential and decrease in size. We concluded that complex formation between Chi NGs and peptide/proteins is mediated mostly by electrostatic interactions.

    ID:1164
  22. Кирилина Е.А., Мастернак Т.Б., Ефремов М.А., Камышников О.Ю. (2013). Влияние миелопептида-1 на функциональную активность фагоцитов мышей, обработанных циклофосфаном. Биоорг. хим. 39 (5), 579–585 ID:1165
  23. Луценко Г.В., Гречихина М.В., Дьячкова Л.Г., Сапожников А.М. (2013). Протективное действие аутокринных факторов цитотоксических Т-лимфоцитов в условиях химической гипоксии. Иммунология 34 (5), 251–254 ID:1166
  24. Митрофанов В.С., Свирщевская Е.В. (2013). Аспергиллез легких. Изд. «Фолиант» , 183 ID:1167
  25. Shevchenko M.A., Bolkhovitina E.L., Servuli E.A., Sapozhnikov A.M. (2013). Elimination of Aspergillus fumigatus conidia from the airways of mice with allergic airway inflammation. Respir. Res. , ID:1241
  26. Kovalenko E., Kanevskiy L., Klinkova A., Kuchukova A., Streltsova M., Telford W., Sapozhnikov A. (2012). Stress-Induced Molecules in Regulation of NK Cell Activity. Chapter in a book “Cell Interaction” , 93–120 ID:1168
  27. Svirshchevskaya E.V. (2012). Chronic immune response hypothesis for chronic fatigue syndrome: experimental results and literature overview. Chapter in a book "Immunosuppression" , 147–160 ID:1169
  28. Svirshchevskaya E., Zubkov D., Mouyna I., Berkova N. (2012). Innate immunity and the role of epithelial barrier during Aspergillus fumigatus infection. Chapter in a book: Current Opinion in Immunology , 254–265 ID:1170
  29. Фонина Л.А., Белёвская Р.Г., Трещалина Е.М., Беспалова Ж.Д., Ефремов М.А., Седакова Л.А., Кирилина Е.А. (2012). Синтез и противоопухолевые свойства миелопептида МП-1. Биоорг. хим. 38 (4), 406–412 ID:1171
  30. Сапожников А.М., Зяблицин А.В., Алекперов Э.А., Бойко А.А., Шустова О.А., Луцан Н.И. (2012). Формирование внеклеточного пула БТШ70 в популяциях лимфоидных клеток. Иммунология 33 (1), 20–23 ID:1172
  31. Зяблицин А.В., Алекперов Э.А., Бойко А.А., Клинкова А.В., Троянова Н.И., Сапожников А.М. (2012). Анализ механизмов неклассического пути секреции БТШ70 в популяциях лимфоидных клеток. Иммунология 33 (1), 49–50 ID:1173
  32. Абакушина Е.В., Кузьмина Е.Г., Коваленко Е.И. (2012). Основные свойства и функции NK-клеток человека. Иммунология 33 (4), 220–225 ID:1174
  33. Свирщевская Е.В., Гриневич Р.С., Решетов П.Д., Зубов В.П., Зубарева А.А., Ильина А.В., Варламов В.П. (2012). Наноносители на основе хитозана. Биотехносфера 19 (1), 13–23 ID:1175
  34. Зубарева А.А., Ильина А.В., Курек Д.В., Сизова С.В., Свирщевская Е.В., Варламов В.П. (2012). . Определение физико-химических параметров наночастиц модифицированного хитозана. Российские нанотехнологии 7 (1-2), 46–56 ID:1176
  35. Moiseeva E.V., Kuznetsova N.R., Svirshchevskaya E.V., Bovin N.V., Sitnikov N.S., Shavyrin A.S., Beletskaya I.P., Combes S., Fedorov A.Y.u., Vodovozova E.L. (2011). Liposome formulations of combretastatin A4 and its 4-arylcoumarin analogue prodrugs: The antitumor effect in the mouse model of breast cancer. Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry 5 (3), 276–283 [+]

    The antimitotic agent combretastatin A-4 (CA-4) has been recently proposed as an antivascular agent for anticancer therapy. In order to reduce systemic toxicity by means of administration in liposome formulations, new lipophilic prodrugs, oleic derivatives of CA-4 and its 4-arylcoumarin analogue (CA4-Ole and ArC-Ole, respectively), have been synthesized in this study. Liposomes with mean diameter of 100 nm prepared on the basis of egg phosphatidylcholine and baker’s yeast phosphatidylinositol quantitatively included up to 15 mol% of CA4-Ole, or 7 mol% of ArC-Ole. To achieve targeting to neovascular endothelium prodrug bearing liposomes decorated with the tetrasaccharide selectin ligand Sialyl Lewis X (SiaLeX) have been also prepared. The antitumor activity was studied in vivo using the model of slow-growing mouse breast cancer. Under the dose used (22 mg/kg) and the administration protocol (four injections, one per a week, starting from the appearance of palpable tumors) cytostatic CA-4 did not reveal any anticancer effect; moreover, it even stimulated tumor growth. The liposome formulations of CA4-Ole did not demonstrate such stimulation. However, to achieve a pronounced antitumor effect, the number of injections of liposomes should be apparently increased. The cytotoxic activity of a novel antimitotic agent ArC was one order of magnitude lower in the human breast carcinoma cell culture in vitro. Nevertheless, in vivo in the mouse model of breast cancer the antitumor effect of this compound corresponded to the double equivalent dose of CA-4. The results demonstrate perspectives of SiaLeX-liposomes loaded with ArC-Ole: the preparation partially inhibited tumor growth already after the second injection. Thus, subsequent optimization of doses and regimens of administration both for ArC and liposomal ArC-Ole formulations are needed.

    ID:671
  36. Balandin T.G., Edelweiss E., Andronova N.V., Treshalina E.M., Sapozhnikov A.M., Deyev S.M. (2011). Antitumor activity and toxicity of anti-HER2 immunoRNase scFv 4D5-dibarnase in mice bearing human breast cancer xenografts. Invest New Drugs 29 (1), 22–32 [+]

    Ribonucleases (RNases) are a non-mutagenic alternative to harmful DNA-damaging anticancer drugs. Targeting of RNases with antibodies to surface antigens that are selectively expressed on tumor cells endows specificity to the cytotoxic actions of RNases. Barnase, a ribonuclease from Bacillus amyloliquefaciens, is a promising candidate for targeted delivery to cancer cells because of its insusceptibility to the ubiquitous cytoplasmic ribonuclease inhibitor, and its high stability and catalytic activity. Here, we characterized in vitro and in vivo an immunoRNase, scFv 4D5-dibarnase, which consists of two barnase molecules that are fused serially to the single-chain variable fragment (scFv) of humanized 4D5 antibody. The latter is directed against the extracellular domain of human epidermal growth factor receptor 2 (HER2), a cancer marker that is overexpressed in many human carcinomas. The scFv 4D5-dibarnase exerted a specific cytotoxic effect on HER2-overexpressing SKBR-3 and BT-474 human breast carcinoma cells (IC(50) = 4.1 and 2.4 nM, respectively) via induction of apoptosis. Ten doses of 0.7 mg/kg scFv 4D5-dibarnase to BALB/c nude mice that bore SKBR-3 human breast cancer xenografts resulted in a 76% reduction in tumor growth. A single injection of scFv 4D5-dibarnase at a total course dose of 7 mg/kg did not cause severe side effects in BALB/c nude or BDF1 mice. The cytotoxicity and selectivity of scFv 4D5-dibarnase merit consideration of this immunoRNase as a potent anticancer agent.

    ID:1177
  37. Kovalenko E.I., Ranjbar S., Jasenosky L.D., Goldfeld A.E., Vorobjev I.A., Barteneva N.S. (2011). The use of HaloTag-based technology in flow and laser scanning cytometry analysis of live and fixed cells. BMC Res Notes 4, 340 [+]

    Combining the technologies of protein tag labeling and optical microscopy allows sensitive analysis of protein function in cells.

    ID:1178
  38. Сапожников А.М., Зяблицин А.В., Алекперов Э.А., Бойко А.А., Клинкова А.В., Луцан Н.И. (2011). Эффекты внеклеточного пула БТШ70 в популяциях лимфоидных клеток. Иммунология 32 (5), 244–248 ID:1179
  39. Константинова Е.В., Хомякова Н.Ф., Константинова Н.А., Подколзина А.В., Сапожников А.М. (2010). Взаимосвязь апоптоза и экспрессии белков теплового шока у лимфоцитов периферической крови больных инфарктом миокарда. Бюллетень экспериментальной биологии и медицины 150 (12), 622–626 ID:1180
  40. Фонина Л.А., Кудрявцева Е.В., Беспалова Ж.Д., Ефремов М.А., Михайлова А.А., Кирилина Е.А. (2010). Синтез и свойства миелопептидов с дифференцировочной активностью. Биоорг. хим. 36 (4), 493–497 ID:1181
  41. Луценко Г.В. (2010). Цитометрический метод регистрации апоптоза с использованием флуорофора 10-N-нонил-акридинового оранжевого. Биологические мембраны 27 (5), 430–439 ID:1182
  42. Glinka E.M., Andryushchenko A.S., Sapozhnikov A.M., Zatsepina O.V. (2009). Construction of the plasmid for expression of ETA-EGFP fusion protein under control of the cytomegalovirus promoter and its effects in HeLa cells. Plasmid 62 (2), 119–27 [+]

    Tumor-targeted vectors encoding toxic protein genes are promising tools for treating malignant tumors. We used the pEGFP-N1 vector to construct a novel plasmid (pCMV-ETA-EGFP) for eukaryotic expression of a truncated Pseudomonas aeruginosa exotoxin A (ETA) that is known to inhibit protein synthesis, and subsequently induce cell death, by inactivation of elongation factor-2. ETA was linked to the enhanced green fluorescent protein (EGFP) gene, and ETA-EGFP gene expression was driven by the cytomegalovirus (CMV) promoter. The time-lapse effects of pCMV-ETA-EGFP expression were examined in transiently transfected HeLa cells. HeLa cells transfected with pCMV-ETA-EGFP or cotransfected with pCMV-ETA-EGFP and small er, Cyrilliccapital IE, CyrillicGFP-N1 showed lower fluorescence intensity than cells transfected with pEGFP-N1 alone. Analysis of the number of dead cells further confirmed the highly toxic effect of the ETA-EGFP fusion protein on cells transfected with pCMV-ETA-EGFP or cotransfected with pCMV-ETA-EGFP and small er, Cyrilliccapital IE, CyrillicGFP-N1. ETA-EGFP fusion protein induced apoptotic cell death through the caspase-3 activation. By using the antibody against a marker nucleolar antigen A3 [Grigoryev, A.A., Bulycheva, T.I., Sheval, E.V., Kalinina, I.A., Zatsepina, O.V., 2008. Cytological indicators of the overall suppression of protein synthesis revealed by staining with new monoclonal antibody. Cell Tissue Biol. 2, 191-199], the distribution of which changes when HeLa cells are treated with known translation inhibitors, we obtained evidence to support the idea that protein synthesis is inhibited in transfected cells in situ. ETA-EGFP fusion protein was identified in lysates of transfected cells using anti-GFP-BL antibodies. Collectively, our results indicate that HeLa cells transfected with pCMV-ETA-EGFP synthesize the ETA-EGFP fusion protein that efficiently inhibits protein synthesis, leading to massive cell death by an apoptosis-mediated pathway with a participation of caspase-3. The constructed vector can be used in suicidal gene therapy of cancer and may also be useful for investigating the general effects of translational downregulation in human cancer cells. We also suggest a novel approach for detecting the activity of new vectors in transfected cells, which is based on the redistribution of nucleolar proteins in transfected cells.

    ID:1183
  43. Svirshchevskaya E.V., Alekseeva L.G., Reshetov P.D., Phomicheva N.N., Parphenyuk S.A., Ilyina A.V., Zueva V.S., Lopatin S.A., Levov A.N., Varlamov V.P. (2009). Mucoadjuvant properties of lipo- and glycoconjugated derivatives of oligochitosans. European journal of medicinal chemistry 44 (5), 2030–7 [+]

    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.

    ID:1185
  44. Moiseeva E.V., Svirshchevskaia E.V., Dozmorov I.M. (2009). [Effect of administration of allogeneic lymphocytes on the survival of sublethally irradiated mice]. Radiobiologiia 28 (2), 235–8 [+]

    The administration of allogenic (CBA----C57B1/6) and semi-allogenic (CBA----F1) lymphocytes to sublethally exposed recipient mice either stimulates or inactivates endogenous colony-formation depending on the dose of lymphocytes administered. The stimulation of endogenous colony-formation correlates with the increased survival rate after radiation doses that decrease the survival rate of the control recipients.

    ID:887
  45. Rodin V.V., Voinova I.I., Volkov V.I.a., Volkova L.A., Moiseeva E.V., Dozmorov I.M. (2009). [Study by electron paramagnetic resonance of blood plasma of mice during rotation stress associated with administration of ascorbic acid and GABA]. Kosm Biol Aviakosm Med 24 (5), 57–8 ID:889
  46. Svirshchevskaya E.V., Shevchenko M.A., Huet D., Femenia F., Latgé J.P., Boireau P., Berkova N.P. (2009). Susceptibility of mice to invasive aspergillosis correlates with delayed cell influx into the lungs. Int. J. Immunogenet. 36 (5), 289–299 ID:1184
  47. Нурбаков А.А., Михалева И.И., Сапожников А.М. (2009). Влияние дельта-сониндуцирующего пептида на уровень экспрессии белка теплового шока 70 КД в клетках линии К562. Бюллетень экспериментальной биологии и медицины 147 (1), 40–42 ID:1186
  48. Белёвская Р.Г., Калюжная М.В., Шевченко В.П., Кирилина Е.А. (2009). Миелопептид-5 отменяет вирус-индуцированную иммуносупрессию, восстанавливая функции Т-лимфоцитов. Бюллетень экспериментальной биологии и медицины 147 (1), 62–66 ID:1187
  49. Луценко Г.В., Гречихина М.В., Дьячкова Л.Г. (2009). О влиянии дефицита аутокринных факторов в культуре на протекание апоптоза в цитотоксических Т-клетках. Иммунология 30 (1), 4–7 ID:1188
  50. Mashkina A.P., Tyulina O.V., Solovyova T.I., Kovalenko E.I., Kanevski L.M., Johnson P., Boldyrev A.A. (2009). The excitotoxic effect of NMDA on human lymphocyte immune function. Neurochem. Int. 51 (6-7), 356–60 [+]

    N-Methyl-d-aspartate (NMDA)-activated glutamate receptors are expressed in lymphocytes, but their roles have not yet been defined. We show that incubation of human peripheral blood lymphocytes with NMDA resulted in increased intracellular calcium and reactive oxygen species (ROS) levels through effects on NMDA-activated glutamate receptors. In terms of ROS production, T cells were most affected, followed by NK cells, whereas B cell ROS levels were not increased. In unstimulated T and NK cells, interferon-gamma (IFN-gamma) production was unaffected by NMDA, whereas interleukin-2 stimulation of IFN-gamma production was significantly suppressed by NMDA. Simultaneous incubation of the cells with NMDA and IL-2 resulted in a dramatic increase in the amount of cells expressing the NR1 subunit of the NMDA-activated receptors. We conclude that NMDA-activated glutamate receptor activation, accompanied by the changes in intracellular calcium and ROS levels, may be involved in the modification of immune functions of human T and NK cells.

    ID:1196
  51. Mushenkova N., Moiseeva E., Chaadaeva A., DenOtter W., Svirshchevskaya E. (2009). Antitumor effect of double immunization of mice with mucin 1 and its coding DNA. Anticancer Res. 25 (6B), 3893–8 [+]

    To evaluate the antitumor effect of mouse immunization with human mucin 1 gene (muc 1) DNA plasmids combined with simultaneous injections of human mucin 1 (MUC1) protein.

    ID:1207
  52. Edelweiss E., Balandin T.G., Ivanova J.L., Lutsenko G.V., Leonova O.G., Popenko V.I., Sapozhnikov A.M., Deyev S.M. (2008). Barnase as a new therapeutic agent triggering apoptosis in human cancer cells. PLoS ONE 3 (6), e2434 [+]

    RNases are currently studied as non-mutagenic alternatives to the harmful DNA-damaging anticancer drugs commonly used in clinical practice. Many mammalian RNases are not potent toxins due to the strong inhibition by ribonuclease inhibitor (RI) presented in the cytoplasm of mammalian cells.

    ID:1189
  53. Svirshchevskaya E.V., Mariotti J., Wright M.H., Viskova N.Y., Telford W., Fowler D.H., Varticovski L. (2008). Rapamycin delays growth of Wnt-1 tumors in spite of suppression of host immunity. BMC Cancer 8, 176 [+]

    Rapamycin, an inhibitor of mammalian target of Rapamycin (mTOR), is an immunosuppressive agent that has anti-proliferative effects on some tumors. However, the role of Rapamycin-induced immune suppression on tumor progression has not been examined.

    ID:1190
  54. Svirshchevskaya E.V., Alekseeva L.G., Shevchenko M.A., Kurup V.P. (2008). Pathophysiology and immunotherapy in allergy: new perspectives. Chapter in book: Recent Trends on Aerobiology, Allergy and Immunology , 405–425 ID:1191
  55. Луценко Г.В., Гречихина М.В., Дьячкова Л.Г., Луцан Н.И. (2008). О выживании Т-лимфоцитов в условиях алкалоза и дефицита аутокринных факторов. Биологические мембраны 25 (6), 463–471 ID:1192
  56. Луценко Г.В., Гречихина М.В., Дьячкова Л.Г. (2008). Участие аутокринных факторов в защите клеток цитотоксической линии CTLL-2 от окислительного стресса. Цитология 50 (6), 492–499 ID:1193
  57. Синцов А.В., Коваленко Е.И., Ханин М.А. (2008). Апоптоз, индуцированный гранзимом B. Биоорг. хим. 34 (6), 725–733 ID:1194
  58. Alekseeva L., Nekrasov A., Marchenko A., Shevchenko M., Benevolenskii S., Sapozhnikov A., Kurup V.P., Svirshchevskaya E. (2007). Cryptic B-cell epitope identification through informational analysis of protein sequences. Vaccine 25, 2688–2697 ID:1195
  59. Луценко Г.В., Гречихина М.В., Дьячкова Л.Г., Луцан Н.И. (2007). Влияние дефицита аутокринных факторов в культуре на выживание и энергетический метаболизм клеток линии CTLL-2 в условиях окислительного стресса. Цитология 49 (4), 284–291 ID:1197
  60. Власкин П.А., Каневский Л.М., Стрельникова Ю.И., Сапожников А.М., Коваленко Е.И. (2007). Активирующее действие белка теплового шока 70 кДа на НК-клетки человека. Иммунология 28 (2), 74–79 ID:1198
  61. Svirshchevskaya E., Alekseeva L., Marchenko A., Benevolenskii S., Berzhec V.M., Nekrasov A. (2006). Selection of cryptic B-cell epitopes using informational analysis of protein sequences. J Bioinform Comput Biol 4 (2), 389–402 [+]

    Sub-unit vaccines are synthetic or recombinant peptides representing T- or B-cell epitopes of major protein antigens from a particular pathogen. Epitope selection requires the synthesis of peptides that overlap the protein sequences and screening for the most effective ones. In this study a new method of immunogenic peptide selection based on the analysis of information structure of protein sequences is suggested. The analysis of known B-cell epitope location in the information structure of Aspergillus fumigatus proteins Asp f 2 and Asp f 3 has shown that epitopes are scattered along the sequences of proteins for the exception of sites with Increased Degree Information Coordination (IDIC). Based on these results peptides from different allergens such as Asp f 2, Der p 1, and Fel d 1 were selected and produced in a recombinant form in the context of yeast virus-like particles (VLPs). Immunization of mice with VLPs containing peptides form allergens has induced the production of IgG able to recognize full-length antigens. This result suggests that the analysis of information structure of proteins can be used for the selection of peptides possessing cryptic B-cell epitope activity.

    ID:1200
  62. Glinka E.M., Edelweiss E.F., Sapozhnikov A.M., Deyev S.M. (2006). A new vector for controllable expression of an anti-HER2/neu mini-antibody-barnase fusion protein in HEK 293T cells. Gene 366 (1), 97–103 [+]

    Tumor-targeted vectors with controllable expression of therapeutic genes and specific antitumor antibodies are promising tools for the reduction of malignant tumors. Here we describe a new plasmid for the eukaryotic expression of an anti-HER2/neu mini-antibody-barnase fusion protein (4D5 scFv-barnase-His(5)) with an NH(2)-terminal leader peptide. The 4D5 scFv-barnase-His(5) gene was placed downstream of the tetracycline responsive-element minimal promoter in the vector using the Tet-Off gene-expression system. The Bacillus amyloliquefaciens ribonuclease barnase is toxic for the host cells. To overcome this problem, barstar gene under its own minimal cytomegalovirus promoter was used in designed vector. Barstar inhibits the background level of barnase in the cells in the presence of tetracycline in culture medium. The HEK 293T cells were transfected with the designed vector, and the 4D5 scFv-barnase-His(5) fusion protein was identified by anti-barnase antibodies in cell culture medium and after purification from cell lysates using metal-affinity chromatography. The overexpression of the anti-HER2/neu mini-antibody-barnase fusion protein decreased the intensity of fluorescence of HEK 293T cells co-transfected with the generated plasmid and a plasmid containing the gene of enhanced green fluorescent protein (pEGFP-N1), in comparison with the intensity of fluorescence of HEK 293T cells transfected with pEGFP-N1, in the absence of tetracycline in the medium. The effect of the 4D5 scFv-barnase-His(5) on EGFP fluorescence indicates that the introduced barnase functions as a ribonuclease inside the cells. The anti-HER2/neu mini-antibody could be used to deliver barnase to HER2/neu-positive cells and provide its penetration into the target cells, as HER2/neu is a ligand-internalizing receptor. This expression vector has potential applications to both gene and antibody therapies of cancer.

    ID:1199
  63. Гурьянов С.А., Кирилина Е.А., Хайдуков С.И., Суворов Н.И., Молотковская И.М., Михайлова А.А. (2006). Специфическое связывание и проникновение внутрь клетки-мишени флуоресцентно меченного миелопептида-4, обладающего дифференцировочной активностью. Биоорг. хим. 32 (6), 574–578 ID:1201
  64. Коваленко Е.И., Власкин П.А., Каневский Л.М., Стрельникова Ю.И., Сапожников А.М. (2006). Влияние белков теплового шока 70 кДа на продукцию γ-интерферона натуральными киллерами человека. ДАН 406 (1), 1–4 ID:1202
  65. Мельникова В.И., Афанасьева М.А., Сапожников А.М., Захарова Л.А. (2006). Динамика апоптоза и пролиферации в тимусе и селезенки крыс в перинатальном онтогенезе. Онтогенез 37 (4), 1–6 ID:1203
  66. Свирщевская Е.В., Алексеева Л.Г., Марченко А.М., Бержец В.М., Шевченко М.А., Андронова Т.М., Беневоленский С.В. (2006). Снижение продукции IgE рекомбинантными пептидами из основных белков аллергенов. Изд. «Фолиант»  (2), 91–97 ID:1205
  67. Мушенкова Н.В., Моисеева Е.В., Чаадаева А.М., Свирщевская Е.В. (2006). Индукция гуморального и клеточного ответа на муцин-1 с использованием ДНК иммунизации. Иммунология 27 (4), 216–221 ID:1210
  68. Novoselova T.V., Margulis B.A., Novoselov S.S., Sapozhnikov A.M., vanderSpuy J., Cheetham M.E., Guzhova I.V. (2005). Treatment with extracellular HSP70/HSC70 protein can reduce polyglutamine toxicity and aggregation. J. Neurochem. 94 (3), 597–606 [+]

    The accumulation of insoluble protein aggregates is a feature of neurodegenerative disease. Overexpression of Heat Shock Protein 70 (HSP70) can protect cells with protein aggregates from apoptosis. Another trait of HSP70 is its ability to cross the plasma membrane. Therefore, we purified a preparation of HSP70/HSC70 from bovine muscle and used it in a model of Huntington's disease. Human neuroblastoma SK-N-SH cells were transfected with huntington exon 1 with short (25) or long (103) CAG trinucleotide repeats coupled to green flourescent protein (GFP). Cells transfected with the long polyCAG repeat had insoluble protein aggregates and died through apoptosis. Biotinylated HSP70/HSC70 incorporated into the culture medium appeared inside the cells within 3-6 h of incubation. This incorporation correlated with a reduction in apoptotic cells by 40-50%. Confocal microscopy revealed that labelled internalized HSP70/HSC70 co-localized with the polyglutamine inclusions. The measurement of the number and size of inclusions showed that HSP70/HSC70 was able to reduce both these parameters. A filter trap assay and immunoblotting demonstrated that the introduction of HSP70/HSC70 also decreased protein aggregation. Together with earlier data on the effects of exogenously administered HSP70/HSC70 on cultured cells and on animals, these data show that preparations based on HSP70 may have some potential as therapies for a variety of neurodegenerative pathologies.

    ID:1208
  69. Svirshchevskaya E.V., Kurup V.P. (2005). Pathogenesis of allergy and specific features of allergic aspergillosis. In book: Mold Allergy, Biology and Pathogenesis , 257–274 ID:1206
  70. Пономарёв А.Д., Семенков В.Ф., Сапожников А.М. (2005). Влияние белков теплового шока на продукцию активных форм кислорода нейтрофилами человека. Иммунология 26, 72–75 ID:1209
  71. Свирщевская Е.В., Митрофанов В.С., Шендорова М.И., Чужова Н.М. (2005). Иммунитет при туберкулезе и аспергиллезе. Проблемы медицинской микологии 7 (1), 3–13 ID:1211
  72. Свирщевская Е.В., Шевченко М.А., Вискова Н.Ю., Шеховцова Е.Л. (2005). Повышение аффинности антител к основным белкам гриба Aspergullus fumigatus зависит от дозы антигена и формирования герминальных центров. Успехи медицинской микологии 5, 108–111 ID:1212
  73. Луценко Г.В., Гречихина М.В., Дьячкова Л.Г. (2005). Регуляция уровня АТФ в нормальных и трансформированных Т-клетках аутокринными факторами. Иммунология 26, 91–95 ID:1213
  74. Kalinichenko V.V., Gusarova G.A., Kim I.M., Shin B., Yoder H.M., Clark J., Sapozhnikov A.M., Whitsett J.A., Costa R.H. (2004). Foxf1 haploinsufficiency reduces Notch-2 signaling during mouse lung development. Am. J. Physiol. Lung Cell Mol. Physiol. 286 (3), L521–30 [+]

    The forkhead box (Fox) f1 transcription factor is expressed in the mouse splanchnic (visceral) mesoderm, which contributes to development of the liver, gallbladder, lung, and intestinal tract. Pulmonary hemorrhage and peripheral microvascular defects were found in approximately half of the newborn Foxf1(+/-) mice, which expressed low levels of lung Foxf1 mRNA [low-Foxf1(+/-) mice]. Microvascular development was normal in the surviving newborn high-Foxf1(+/-) mice, which compensated for pulmonary Foxf1 haploinsufficiency and expressed wild-type Foxf1 levels. To identify expression of genes regulated by Foxf1, we used Affymetrix microarrays to determine embryonic lung RNAs influenced by Foxf1 haploinsufficiency. Embryonic Foxf1(+/-) lungs exhibited diminished expression of hepatocyte growth factor receptor c-Met, myosin VI, the transcription factors SP-3, BMI-1, ATF-2, and glucocorticoid receptor, and cell cycle inhibitors p53, p21(Cip1), retinoblastoma, and p107. Furthermore, Notch-2 signaling was decreased in embryonic Foxf1(+/-) lungs, as evidenced by significantly reduced levels of the Notch-2 receptor and the Notch-2 downstream target hairy enhancer of split-1. The severity of the Notch-2-signaling defect in 18-day postcoitus Foxf1(+/-) lungs correlated with Foxf1 mRNA levels. Disruption of pulmonary Notch-2 signaling continued in newborn low-Foxf1(+/-) mice, which died of lung hemorrhage and failed to compensate for Foxf1 haploinsufficiency. In contrast, in newborn high-Foxf1(+/-) lungs, Notch-2 signaling was restored to the level found in wild-type mice, which was associated with normal microvascular formation and survival. Foxf1 haploinsufficiency disrupted pulmonary expression of genes in the Notch-2-signaling pathway and resulted in abnormal development of lung microvasculature.

    ID:1214
  75. Svirshchevskaya E.V., Viskova N., Shevchenko M., Alekseeva L., Marchenko A., Benevolensky S., Kurup V.P. (2004). High-affinity IgG to a major A. fumigatus allergen, Asp f 2, retards allergic response. Med Sci Monit 10, 371–380 ID:1215
  76. Коваленко Е.И., Хирова Е.В., Молотковская И.М., Овчинникова Т.В., Саблина М.А., Сапожников А.М., Хайдуков С.В., Бовин Н.В. (2004). . Модификация поверхности клеток с помощью липофильных гликоконъюгатов и взаимодействие модифицированных клеток с натуральными киллерами. Биоорг. хим. 30 (3), 281–292 ID:1216
  77. Шевченко М.А., Вискова Н.Ю., Свирщевская Е.В. (2004). Аллергенность и иммуногенность протеолитически деградированных белков из гриба Aspergillus fumigatus. Иммунология 1, 23–28 ID:1217
  78. Свирщевская Е.В., Попова И.С., Матушевская Е.В., Коцарева О.Д., Эртнеева И.Я. (2004). Плацебо-контролируемый эффект антигистаминного препарата кларотадин на продукцию ИЛ-13 при атопическом дерматите. Вестник дерматологии и венерологии 5, 27–32 ID:1218
  79. Сапожников А.М., Тарасенко Т.Н., Пономарёв А.Д., Гусарова Г.А., Мурашко Д.А. (2004). Анализ изменения экспрессии белков теплового шока 70 кДа в процессе апоптоза тимоцитов. Иммунология 3, 135–142 ID:1219
  80. Svirshchevskaya E.V., Kurup V.P. (2003). Immunotherapy of allergic bronchopulmonary aspergillosis: a clinical and experimental approach. Front. Biosci. 8, s92–101 [+]

    Allergic bronchopulmonary aspergillosis (ABPA) is a severe allergic pulmonary complication caused by the saprophytic fungus Aspergillus fumigatus. The present review examines the pathogenesis of this disease describing in detail the role of innate and acquired immunity in the induction of sensitivity to A.fumigatus. Different approaches in developing specific immunotherapeutic treatments such as induction of anergy, regulatory cells, a switch from Th2 to Th1 type of immune response, CpG and genetic immunization and the usage of altered peptides or modified allergens are critically examined.

    ID:1220
  81. Marchenko A.N., Kozlov D.G., Svirshchevskaya E.V., Viskova N.Y., Benevolensky S.V. (2003). The p1 protein of the yeast transposon Ty1 can be used for the construction of bi-functional virus-like particles. J. Mol. Microbiol. Biotechnol. 5 (2), 97–104 [+]

    Virus-like particles (VLPs) containing heterologous proteins are often used as vaccines. Two approaches for the construction of bi-functional VLPs using hybrid protein pl-380 of the TY1 transposon of Saccharomyces yeast are described. We have shown that both C- and N-termini of p1-380 can be used for the expression of heterologous peptides. Peptides from A. Fumigatus Asp f 2, expressed at the C- and/or N-termini of p1-380, did not interfere with VLP self-assembling, were accessible for antibodies and hence were exposed at the VLP surface. Another way to obtain bivalent VLPs is the formation of mixed particles, which co-express two hybrid pl proteins with different heterologous protein fragments at the C-terminus. To do it the yeast cells were transfected with a mixture of two recombinant DNA coding Asp f 2 peptide and green fluorescent protein (Gfp). We have shown that both Asp f 2 peptide and Gfp are expressed within the same particle. To evaluate biological activity of bi-functional VLP a construction containing peptides representing dominant T- and B-cell epitopes of Asp f 2 was produced. Bi-functional particles were more potent in stimulating memory immune responses. These results demonstrate new possibilities of pl-380 based expression system to produce multifunctional VLPs.

    ID:1221
  82. Луценко Г.В., Дьячкова Л.Г. (2003). Роль энергетического метаболизма клеток цитотоксической линии CTLL-2 в механизме контроля их выживания аутокринными факторами. Биологические мембраны 20 (5), 401–408 ID:1222
  83. Сапожников А.М., Тарасенко Т.Н., Пономарёв А.Д., Гусарова Г.А., Мурашко Д.А., Петров Р.В. (2003). Адреналин-опосредованная активация экспрессии белков теплового шока 70 кДа в популяции тимоцитов. ДАН 392 (2), 277–279 ID:1223
  84. Вискова Н.В., Шевченко М.А., Свирщевская Е.В. (2003). Индукция синтеза IgE при первичном ответе и ответе памяти на растворимые белки. Иммунология 24 (4), 227–231 ID:1224
  85. Свирщевская Е.В. (2003). Клинические и экспериментальные аспекты патогенеза аллергических реакций на условно патогенный грибок Aspergillus fumigatus. Иммунология  (3), 188–192 ID:1225
  86. Свирщевская Е.В., Матушевская Е.В., Коцарева О.Д., Багаева Л.В., Дзуцева И.Р., Лысенко А.А. (2003). Блокада акантолиза эпидермиса при вульгарной пузырчатке протективными антителами. Вестник дерматологии и венерологии  (4), 4–7 ID:1226
  87. Sapozhnikov A.M., Gusarova G.A., Ponomarev E.D., Telford W.G. (2002). Translocation of cytoplasmic HSP70 onto the surface of EL-4 cells during apoptosis. Cell Prolif. 35 (4), 193–206 [+]

    Heat shock proteins (HSPs) are involved in a variety of intracellular processes and can have both pro- and anti-apoptotic action. However, little is known about the role of HSPs in the progression of apoptosis. Translocation of HSPs to the surface of apoptotic cells is a previously observed phenomenon demonstrating participation of these proteins in execution of the terminal stages of apoptosis. In a previous study we showed that development of EL-4 lymphoma cell apoptosis in vitro is accompanied by elevation of surface HSP expression. In this study we used this model to analyse the relationship of HSP70 expression and its translocation to the cell surface during apoptosis with some key intracellular events. Our data demonstrate a synchronization of surface and intracellular HSP70 expression with bcl-2 expression, intracellular reactive oxygen species (ROS) concentration and caspase-3 activity. A maximum level of surface and intracellular HSP70 expression was observed at an irreversible phase of EL-4 cell apoptosis after mitochondrial potential depolarization. In addition, an enhancement of the relative level of cytoplasmic HSP70 translocation to the cell surface was concomitant with EL-4 cell apoptosis. However, the size of surface and intracellular pools of HSP70, increasing for initial and intermediate stages of cell death, decreased at the terminal phase of apoptosis. Western blot analysis of HSP70 in conditioned supernatant obtained from EL-4 cell tissue showed that the observed decrease of HSP70 cell content might be due to surface HSP70 shedding into the intercellular space.

    ID:1229
  88. Svirshchevskaya E.V., Alekseeva L., Marchenko A., Viskova N., Andronova T.M., Benevolenskii S.V., Kurup V.P. (2002). Immune response modulation by recombinant peptides expressed in virus-like particles. Clin. Exp. Immunol. 127 (2), 199–205 [+]

    Aspergillus fumigatus, a ubiquitous fungus, is implicated in the pathogenesis of a number of clinically different allergic diseases in man, including allergic bronchopulmonary aspergillosis. Peptide-based immunotherapy may offer an alternative treatment strategy for the management of allergic disease. The objective of this study was to alter the allergen-specific immune response using dominant T cell epitopes of a major A. fumigatus allergen, Asp f2, expressed in yeast as virus-like particles (VLP). The T cell epitopes of Asp f2, recognized in mice with an H-2d background, were determined by producing T-cell hybridomas. Two dominant T cell epitopes, aa60--71 and aa235--249, were identified and expressed in a yeast VLP system. To induce tolerance VLP-peptides were injected subcutaneously into mice previously immunized with recombinant Asp f2. The T cell immune response was abrogated totally in 3 weeks following a single injection of VLP but was restored 2 months later following intranasal antigen exposure. T-cell depletion resulted in the reduction of 20-30% of all antigen-specific immunoglobulin classes. Thus, recombinant peptides expressed in the VLP system can be used successfully in the modulation of Asp f2-induced immune response in mice, although a single administration is not sufficient to maintain a state of tolerance for a long period of time.

    ID:1228
  89. Svirshchevskaya E.V., Kurup V.P. (2002). IgE regulation and pathogenesis in ABPA. In book: Recent Res. Devel. Immunol. , 103–114 ID:1227
  90. Баев Д.В., Гусарова Г.А., Сапожников А.М. (2002). Взаимосвязь протективного эффекта ингибиторов хлорных каналов с экспрессией белков теплового шока 70 в моделях апоптоза и осмотического стресса клеток лимфомы EL-4. Биологические мембраны 19 (4), 275–282 ID:1230
  91. Сапожников А.М., Баев Д.В., Гусарова Г.А. (2002). Вовлеченность белков теплового шока в феномен защиты клеток от апоптоза ингибиторами хлорных каналов плазматических мембран. ДАН 384 (5), 712–714 ID:1231
  92. Свирщевская Е.В., Вискова Н.Ю., Чудновская Т.Н., Гурина О.Н., Матушевская Е.В. (2002). Участие Т-хелперов 2 типа в патогенезе аутоиммунной пузырчатки. Иммунология  (2), 112–114 ID:1232
  93. Пономарёв А.Д., Гусарова Г.А., Моисеева Е.В., Сапожников А.М. (2002). Разработка нового подхода к созданию противоопухолевых вакцин на основе белков теплового шока. Аллергия, астма и клиническая иммунология  (3), 3–8 ID:1234
  94. Свирщевская Е.В., Вискова Н.Ю., Шевченко М.А. (2002). Механизмы патогенеза аллергического аспергиллеза. Аллергия, астма и клиническая иммунология  (4), 6–14 ID:1235
  95. Svirshchevskaya E.V., Alekseeva L.G., Andronova T.M., Kurup V.P. (2001). Do T helpers 1 and 2 recognize different class II MHC molecules? Humoral and cellular immune responses to soluble allergen from Aspergillus fumigatus Asp f2. Clin. Immunol. 100 (3), 349–54 [+]

    Cellular signals leading to T helper (Th)1/Th2 shift are not well known. Here we demonstrate that Th1 possibly recognizes peptides presented by the IE molecule of MHC class II while Th2 is activated by the recognition of peptides presented by the IA molecule. BALB/c mice immunized with Asp f2 developed stable IA-restricted Th2 immune response to the 12th day after immunization, as analyzed by IL-2 production. On the contrary, early Th0 cells did not secrete IL-2 upon Asp f2 stimulation but did produce a high level of IL-2 if stimulated in the presence of anti-IA Abs. This effect of anti-IA Abs on early Th0 cells was both MHC IE and CD4(+) cell restricted. In vivo blocking of Asp f2 peptide presentation by the IA molecule led to the formation of antigen-specific cytotoxicity as demonstrated using immune splenocytes as effector cells and Asp f2 loaded P815 cells as targets.

    ID:1236
  96. Ponomarev E.D., Tarasenko T.N., Sapozhnikov A.M. (2000). Splenic cytotoxic cells recognize surface HSP70 on culture-adapted EL-4 mouse lymphoma cells. Immunol. Lett. 74 (2), 133–9 [+]

    Heat shock proteins (HSPs) are intracellular proteins which function as molecular chaperones. At the same time, translocation of HSPs to the cell surface has been observed in stressed, infected and transformed cells. It seems plausible that surface HSPs may represent molecular targets for recognition and elimination of 'altered' cells by cytotoxic lymphocytes. Previously we demonstrated that EL-4 mouse lymphoma cells growing in vitro express HSPs on their plasma membrane. In this study, we tested the hypothesis that surface HSPs present on EL-4 cells may mediate their recognition and killing by cytotoxic lymphocytes. We have found that susceptibility of culture-adapted EL-4 cells to in vitro lysis by syngeneic and allogeneic splenocytes correlated with the expression of HSP70 on EL-4 cells. Moreover, cytotoxicity was blocked by pretreatment of EL-4 target cells with anti-HSP70 antibody, whereas antibodies to MHC class I molecules and Thy1 did not have such effect. Cytotoxicity against EL-4 lymphoma was not MHC class I-restricted, and was not decreased after depletion of CD8(+) cells from the effector cell population. We conclude that in vitro killing of EL-4 cells is mediated, at least in part, by NK cells via recognition of HSPs present on the surface of tumor cells. Thus, cytotoxic response against EL-4 lymphoma should serve as a good model to study the role of HSPs in anti-tumor immunity.

    ID:1238
  97. Svirshchevskaya E., Frolova E., Alekseeva L., Kotzareva O., Kurup V.P. (2000). Intravenous injection of major and cryptic peptide epitopes of ribotoxin, Asp f 1 inhibits T cell response induced by crude Aspergillus fumigatus antigens in mice. Peptides 21 (1), 1–8 [+]

    Aspergillus fumigatus, a ubiquitous fungus, is implicated in the pathogenesis of a number of clinically different allergic diseases in man. Peptide-based immunotherapy may offer an alternative in patient care and management. The purpose of this study was to evaluate the role of T cell epitopes of A. fumigatus ribotoxin, Asp f 1 in inducing tolerance in mice exposed to A. fumigatus antigen. The epitope analysis in BALB/c mice using synthetic peptides of Asp f 1 demonstrated both cryptic and dominant epitopes detected from 42 through 54 and 155 through 167 aa, accordingly. Intravenous injection of these peptides markedly inhibited the response induced by the exposure to crude A. fumigatus extract in mice as evidenced by the in vitro interleukin-2 (IL-2) production and proliferation of T-lymphocytes. Cytokine transcription studies indicate that, when stimulated with the peptides in immunogenic conditions, the major peptide (aa 155-167) specific T cell clone produced only IFN-gamma, but not IL-4. The ability of both dominant and cryptic peptide epitopes of a single molecule to induce tolerance against the immune response to a multi-molecular allergen complex has significant implication for peptide-based immunotherapy.

    ID:1237
  98. Сапожников А.М., Пономарёв Е.Д., Гусарова Г.А. (2000). О взаимосвязи апоптоза клеток лимфомы EL-4 с экспрессией белков теплового шока. ДАН 375 (4), 576–579 ID:1239
  99. Sapozhnikov A.M., Ponomarev E.D., Tarasenko T.N., Telford W.G. (1999). Spontaneous apoptosis and expression of cell surface heat-shock proteins in cultured EL-4 lymphoma cells. Cell Prolif. 32 (6), 363–78 [+]

    The expression of heat-shock proteins (HSPs) is enhanced in stressed cells and can protect cells from stress-induced injury. However, existing data about the relationship between apoptosis and HSP expression is contradictory. In this paper, a mouse lymphoma cell death model system is used to detect simultaneously both the process of apoptosis and the level of HSP expression. The model was established after discovering that spontaneous apoptosis and spontaneous cell surface HSP expression occurs in EL-4 mouse lymphoma cells during normal optimal culture conditions. The data show that apoptotic EL-4 cells had higher levels of hsp25, hsp60, hsp70 and hsp90 exposed on the plasma membrane surface than viable cells. The level of surface HSPs was found to increase through several stages of early and late apoptotic death as measured by flow cytometry, with the highest levels observed during the loss of cell membrane phospholipid asymmetry. Heat shock and actinomycin D significantly increased the proportion of apoptotic cells in culture. However, hyperthermia only stimulated a weak and temporary increase in surface HSP expression, whereas actinomycin D strongly elevated the level of surface and intracellular HSPs, particularly in live cells. These results show an associative relationship between apoptosis and HSP expression. The relationship between the progression of cell death and HSP expression suggests a role for membrane HSP expression in programmed cell death.

    ID:1240

Aleksandr Sapozhnikov

  • Russia, Moscow, Ul. Miklukho-Maklaya 16/10 — On the map
  • IBCh RAS, build. 52, office. 457
  • Phone: +7(495)330-40-11
  • E-mail: amsap@mail.ru

Retroviral gene transfer into primary human NK cells activated by IL-2 and K562 feeder cells expressing membrane-bound IL-21 (2017-12-13)

Natural killer (NK) cells are capable of rapidly recognizing and efficiently killing tumor cells. This makes them a potentially promising agent for cancer immunotherapy. Additional genetic modifications of NK cells may further improve their anti-tumor efficacy. Numerous technical challenges associated with gene delivery into NK cells have significantly tempered this approach. We achieved efficient retroviral vector transduction of primary human NK cells that were stimulated by a combination of IL-2 and engineered K562 cells expressing membrane-bound IL-21. The activated NK cells were in less differentiated state and expressed NK cell activation receptors NKG2D, NKp30, CD16, and were highly HLA-DR-positive. This NK cell population was highly susceptible to the transduction by both GFP- and NGFR-expressing retroviral vectors. More mature CD57+ NK cell population was generally resistant to retroviral vector transduction because of poor response to the stimulation. Our findings may facilitate retroviral vector-mediated genetic engineering of human primary NK cells for future immunotherapies.

Publications

  1. Streltsova M.A., Barsov E., Erokhina S.A., Kovalenko E.I. (2017). Retroviral gene transfer into primary human NK cells activated by IL-2 and K562 feeder cells expressing membrane-bound IL-21. J. Immunol. Methods 450, 90–94 [+]

    Natural killer (NK) cells are capable of rapidly recognizing and efficiently killing tumor cells. This makes them a potentially promising agent for cancer immunotherapy. Additional genetic modifications of NK cells may further improve their anti-tumor efficacy. Numerous technical challenges associated with gene delivery into NK cells have significantly tempered this approach. We achieved efficient retroviral vector transduction of primary human NK cells that were stimulated by a combination of IL-2 and engineered K562 cells expressing membrane-bound IL-21. The activated NK cells were in less differentiated state and expressed NK cell activation receptors NKG2D, NKp30, CD16, and were highly HLA-DR-positive. This NK cell population was highly susceptible to the transduction by both GFP- and NGFR-expressing retroviral vectors, with transduction efficiency exceeding 50%. More mature CD57(+) NK cell population was generally resistant to retroviral vector transduction because of poor response to the stimulation. Our findings may facilitate retroviral vector-mediated genetic engineering of human primary NK cells for future immunotherapies.

    ID:1938

Ethanol-dependent expression of the NKG2D ligands MICA/B in human cell lines and leukocytes (2017-12-13)

Stress-induced molecules MICA and MICB are capable to regulate activity of cytotoxic lymphocytes through the interaction with receptor NKG2D, which substantially affects the functionality of cellular immunity. In cell line models ethanol caused different changes in surface expression of MICA/B, particularly it induced the translocation of intracellular proteins MICA/B to the cell surface and shedding of MICA (in soluble and microparticle-associated forms) from the plasma membrane. The observed results are not linked with cell death in cultures, taking place only under higher doses of ethanol. Ethanol at physiologically relevant concentrations (and higher) stimulated expression of MICA/B genes in different cell types. Presumably, changes in MICA/B expression, caused by ethanol, can affect functions of NKG2D-positive cytotoxic lymphocytes, modulating immune reactions at excessive alcohol consumption.

Publications

  1. Streltsova M.A., Klinkova A.V., Kuchukova A.A., Kadin A.Y., Kanevskiy L.M., Kovalenko E.I. (2016). Ethanol-dependent expression of the NKG2D ligands MICA/B in human cell lines and leukocytes. Biochem. Cell Biol. , 1–9 [+]

    Alcohol consumption affects the human immune system, causing a variety of disorders. However, the mechanisms of development of these changes are not fully understood. We hypothesized that ethanol may influence the expression of MICA and MICB, stress-induced molecules capable of regulating the activity of cytotoxic lymphocytes through the interaction with receptor NKG2D, which substantially affects the functionality of cellular immunity. We analyzed the effects of ethanol on MICA/B expression in tumor cell lines and human leukocytes. In the cell line models, ethanol caused different changes in the surface expression of MICA/B; in particular, it induced the translocation of intracellular proteins MICA/B to the cell surface and shedding of MICA (in soluble and microparticle-associated forms) from the plasma membrane. The observed results are not linked with cell death in cultures, taking place only under higher doses of ethanol. Ethanol at physiologically relevant concentrations (and higher) stimulated expression of MICA/B genes in different cell types. The effect of ethanol was more pronounced in hepatocyte line HepG2 compared with hematopoietic cell lines K562, Jurkat, and THP-1. Among the tested leukocytes, the most sensitive to ethanol action were T cells activated ex vivo with IL-2, in which the increase of MICA/B mRNA expression was registered with the smallest dose of ethanol (0.125%). In human monocytes, ethanol may lead to elevations in surface MICA/B levels. Presumably, changes in MICA/B expression caused by ethanol can affect the functions of NKG2D-positive cytotoxic lymphocytes, modulating immune reactions at excessive alcohol consumption.

    ID:1784
  2. Клинкова А.В., Кузьмина Е.Г., Абакушина Е.В., Каневский Л.М., Неприна Г.С., Павлов В.В., Коваленко Е.И. (2016). Циркулирующий белок MICА у больных злокачественными лимфомами. Медицинская иммунология 18 (2), 151–162 ID:2030

The localization of positively and negatively charged chitosan nanoparticles in tumor cells has been characterized (2016-03-30)

In collaboration with Centre of Bioengineering RAS, Moscow

 

Using in vitro culture of RAW264 cells and nanoparticles fabricated from positive and negative chitosan derivations we demonstrated penetration of the particles into tumor cells and their intracellular localization: in mitochondria for positively charged nanoparticles and in lysosomes for negatively charged nanoparticles. Accumulation of both types of the particles resulted in reducing mitochondrial membrane potential and in exocytosis of mitochondria and lysosomes from live tumor cells. Our results suggest that positively charged chitosan derivates may be used for development of nanopreparations for treatment of the diseases with mitochondrial dysfunctions, and negatively charged chitosan derivates – for anticancer preparations.

Zubareva A.A., Shcherbinina T.S., Varlamov V.P., Svirshchevskaya E.V. Intracellular Sorting of Differently Charged Chitosan Derivatives and Chitosan-Based Nanoparticles. NanoScale. 2015 Nanoscale, 2015, 7, 7942 - 7952.