Молотковская Ирина Михайловна

Доктор биологических наук


Руководитель подразделения (Группа липидных модуляторов иммунитета)

Тел.: +7 (495) 330-72-56

Эл. почта: immol@ibch.ru

Образование

Период обученияСтрана, городУчебное заведениеДополнительная информация
1973–1978 Москва МГУ им. М. Ломоносова

Избранные публикации

  1. Doronin I.I., Vishnyakova P.A., Kholodenko I.V., Ponomarev E.D., Ryazantsev D.Y., Molotkovskaya I.M., Kholodenko R.V. (2014). Ganglioside GD2 in reception and transduction of cell death signal in tumor cells. BMC Cancer 14, 295 [+]

    Ganglioside GD2 is expressed on plasma membranes of various types of malignant cells. One of the most promising approaches for cancer immunotherapy is the treatment with monoclonal antibodies recognizing tumor-associated markers such as ganglioside GD2. It is considered that major mechanisms of anticancer activity of anti-GD2 antibodies are complement-dependent cytotoxicity and/or antibody-mediated cellular cytotoxicity. At the same time, several studies suggested that anti-GD2 antibodies are capable of direct induction of cell death of number of tumor cell lines, but it has not been investigated in details. In this study we investigated the functional role of ganglioside GD2 in the induction of cell death of multiple tumor cell lines by using GD2-specific monoclonal antibodies.

    ID:1112
  2. Doronin I.I., Kholodenko I.V., Molotkovskaya I.M., Kholodenko R.V. (2013). Preparation of Fab-fragments of GD2-specific antibodies and analysis of their antitumor activity in vitro. Bull. Exp. Biol. Med. 154 (5), 658–63 [+]

    Monoclonal antibodies ME361 specific to ganglioside GD2 were isolated from the conditioned medium of hybridoma HB9326 and mouse ascitic fluid by the method of affinity chromatography; their Fab-fragments were obtained by proteolytic cleavage with papain. Evaluation of Fab-fragment specificity by flow cytometry and dot-blot analysis showed that binding effectiveness of fragments with antigens was close to that for the full-length molecule of antigen. It was shown that Fab-fragments and whole antibodies ME361 dose-dependently inhibit the proliferation of cells of mice T-lymphoma EL-4, and induce apoptosis of these cells 24 h after incubation.

    ID:1121
  3. Astashkin E.I., Bespalova Yu.B., Molotkovskaya I.M., Molotkovsky J.G., Glezer M.G., Grachev S.V. (2009). Effect of sphingosine on Ca(2+)-induced reactions of the HL-60 cells. Dokl. Biol. Sci. 371, 217–20 ID:1116
  4. Kholodenko I.V., Kholodenko R.V., Vodovozova E.L., Oleinikov V.A., Polyakov N.B., Molotkovskaya I.M., Petrov R.V. (2009). Ganglioside GM1-binding sites in interleukin-4: a photoaffinity labeling study. Dokl. Biochem. Biophys. 418, 31–5 ID:1119
  5. Kovalenko E.I., Abakushina E., Telford W., Kapoor V., Korchagina E., Khaidukov S., Molotkovskaya I., Sapozhnikov A., Vlaskin P., Bovin N. (2007). Clustered carbohydrates as a target for natural killer cells: a model system. Histochem. Cell Biol. 127 (3), 313–26 [+]

    Membrane-associated oligosaccharides are known to take part in interactions between natural killer (NK) cells and their targets and modulate NK cell activity. A model system was therefore developed using synthetic glycoconjugates as tools to modify the carbohydrate pattern on NK target cell surfaces. NK cells were then assessed for function in response to synthetic glycoconjugates, using both cytolysis-associated caspase 6 activation measured by flow cytometry and IFN-gamma production. Lipophilic neoglycoconjugates were synthesized to provide their easy incorporation into the target cell membranes and to make carbohydrate residues available for cell-cell interactions. While incorporation was successful based on fluorescence monitoring, glycoconjugate incorporation did not evoke artifactual changes in surface antigen expression, and had no negative effect on cell viability. Glycoconjugates contained Le(x), sulfated Le(x), and Le(y) sharing the common structure motif trisaccharide Le(x) were revealed to enhance cytotoxicity mediated specifically by CD16 +CD56+NK cells. The glycoconjugate effects were dependent on saccharide presentation in a polymeric form. Only polymeric, or clustered, but not monomeric glycoconjugates resulted in alteration of cytotoxicity in our system, suggesting that appropriate presentation is critical for carbohydrate recognition and subsequent biological effects.

    ID:1120
  6. Molotkovskaya I.M., Kholodenko R.V., Molotkovsky J.G. (2002). Influence of gangliosides on the IL-2- and IL-4-dependent cell proliferation. Neurochem. Res. 27 (7-8), 761–70 [+]

    Ganglioside-induced apoptosis in the cells of IL-2-dependent cytotoxic murine cell line CTLL-2 was shown to be caspase dependent: GM1-, GM2-, and GD3-induced suppression of cell proliferation was cancelled by a general caspase inhibitor Z-VAD-FMK. Ganglioside-induced apoptosis pathways are different for different individual glycolipids; the differences exist both at the initiation and effector stages of the caspase cascade. Only for GM1-induced process, molecular mechanisms of signal transduction coincide with the ones for CD95 and TNFalpha: the participation of both the main initiation caspases 8, 1, and 4, and caspases 3 and 9 as well, has been shown. Caspase 3 participates in the pathway induced by GM3, GD1a, GD1b, and GT1b, but not by GM2. As morphological features show, tumor-associated ganglioside GM2 is also a stimulus of programmed cell death (PCD) for CTLL-2 cell line: addition of GM2 into cell culture has resulted in appearance of annexin V-positive cells and in accumulation of DNA breaks (shown by the TUNEL direct dyeing of the open ends). But a caspase 3 inhibitor Z-DEVD-FMK did not restore the cell proliferation suppressed by GM2, and addition of a fluorescent substrate of caspase 3 Ac-DEVD-AFC did not result in the fluorescence development. So caspase 3 does not participate in downstream pathways of GM2-induced cell apoptosis, and a PCD-effector system other than the apoptosome-mediated one is involved here.

    ID:1118
  7. Molotkovskaya I.M., Kholodenko R.V., Zelenova N.A., Sapozhnikov A.M., Mikhalev I.I., Molotkovsky J.G. (2000). Gangliosides induce cell apoptosis in the cytotoxic line CTLL-2, but not in the promyelocyte leukemia cell line HL-60. Membr Cell Biol 13 (6), 811–22 [+]

    Gangliosides induce apoptosis in the cells of the IL-2-dependent cytotoxic mouse line CTLL-2. Upon incubation with gangliosides for 24 h, their effect resulting in appearance of apoptotic cells, falls in a series GM2 > GM3 > GM1 > GD1a > GD1b > GT1b. In the presence of rIL-2, apoptosis induced by GM1 is suppressed, whereas that induced by GM2 is enhanced (the effect of intracellular agent C2-Cer is independent of this cytokine). The GM1-induced apoptosis is cancelled by the caspase I inhibitor. The gangliosides under study are not able to induce apoptosis in the promyelocyte leukemia cell line HL-60. Physiological aspects of the phenomenon found are discussed.

    ID:1117
  8. Molotkovskaya I.M., Malyukova I.V., Zakharova L.A. (1999). Opioid agonist modulation of cytoplasmic free Ca2+ level in concanavalin A-stimulated mouse lymphocytes. Biochemistry Mosc. 64 (5), 546–51 [+]

    In this study the influence of mu-, delta-, and kappa-selective opioid agonists (DAMGO, DSLET, and dynorphin A (1-13)) on cytoplasmic free Ca2+ ([Ca2+]i) level in normal and concanavalin-A (Con A)-activated mouse lymphocytes was investigated. [Ca2+]i was measured using the fluorescent dye FURA-2AM. The opioid peptides at 10-12-10-7 M induced some increase in [Ca2+]i in non-activated lymphocytes. However, DAMGO and DSLET (10-13-10-7 M) considerably inhibited a Con A-induced increase in [Ca2+]i. The inhibiting effect of both peptides was higher after 20-min preincubation compare to 2-h preincubation. The effect of the kappa-agonist dynorphin A (1-13) was significantly different depending on the duration of cell pretreatment and the concentration of the peptide used. After preincubation for 20 min at low concentrations (10-12-10-11 M) it slightly stimulated, while at higher (10-10-10-7 M) concentrations it inhibited lymphocyte response to Con A. After preincubation for 2 h, pronounced stimulation of mitogen-induced Ca2+ flux was observed at peptide concentration 10-9 M. The effects of opioids were antagonized by naloxone. These data indicate that functionally active opioid receptors expressed on lymphocytes could be involved in early stages of mitogen activation.

    ID:1114
  9. Molotkovskaya I.M., Zelenova N.A., Lutsenko G.V., Sapozhnikov A.M., Mikhalyov I.I., Molotkovsky J.G. (1999). Immunosuppressive activity of glycosphingolipids. Influence of serum factors on ganglioside inhibition of IL-4-dependent cell proliferation. Membr Cell Biol 12 (6), 783–91 [+]

    Gangliosides have been shown to inhibit proliferation of the interleukin-4 (IL-4) responsive cell line CT.4R. Kinetic analysis has revealed that ganglioside GT1b is a competitive inhibitor of proliferation, while GM and GM3 show a mixed pattern of inhibition, i.e., exhibit more than one inhibition type. Contribution of the competitive cell inhibition for GM1 and GM3 depends on serum factors added: the higher is the percentage of FCS, the larger is the contribution of competitive inhibition. The pattern of proliferation inhibition shown for GT1b does not depend on the FCS content. We have also studied the interaction of the recombinant IL-4 with fluorescent (anthrylvinyl-labelled) gangliosides GM1 and GM3 and lactosylceramide incorporated into liposomes. Dissociation constants of the IL-4-ganglioside complexes have been determined; lactosylceramide does not interact with rIL-4. The K(d) values for the lymphokine complexes with gangliosides support the conclusion based on the kinetic analysis that IL-4 has a higher affinity for GM3 (K(d) = 5 nM) than for GM1 (K(d) = 0.28 microM).

    ID:1115
  10. Molotkovsky J.G., Manevich Y.M., Gerasimova E.N., Molotkovskaya I.M., Polessky V.A., Bergelson L.D. (1982). Differential study of phosphatidylcholine and sphingomyelin in human high-density lipoproteins with lipid-specific fluorescent probes. Eur. J. Biochem. 122 (3), 573–9 [+]

    Modified phosphatidylcholine and sphingomyelin containing an anthryl end group attached to one of the fatty acyl chains were used as fluorescent probes in an investigation of the molecular organization of human high-density lipoproteins (HDL). Monolayer experiments and NMR measurements showed the anthryl-labeled lipids to mimic closely the corresponding host phospholipids, the fluorophores being located near to the terminal CH3 groups of the fatty acid residues. The above fluorescent phospholipid probes made it possible for the first time to study differentially the behaviour of phosphatidylcholine and sphingomyelin in HDL. The probes were shown to interact in a different way with the apoprotein tryptophans and to be non-randomly distributed at the surface of the globules. The probable sphingomyelin binding site of apolipoprotein A-I was defined. Evidence was obtained suggesting the existence in high-density lipoproteins of two slowly exchanging phospholipid pools: one strongly bound to apoproteins, and the other free or loosely bound. Fluorescence parameters characterizing the fluidity of HDL phospholipids and their interaction with the apoprotein tryptophans were found to correlate with the HDL cholesterol level. The possible significance of the obtained results for a better understanding of the relation of high-density lipoproteins to coronary heart diseases is discussed.

    ID:1113