Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells
We investigated the effects of teichoic acid (TA) from Staphylococcus aureus Wood 46 on tumor growth and metastasis of the experimental Lewis lung carcinoma (LLC) in mice. Intranasal administration of TA alone aggravated both tumor growth and metastasis, whereas combined administration of TA with a...
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| Cite this: | Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells / V.V. Nikulina, L.V. Garmanchuk, N.V. Senchylo, T.V. Nikolaenko, O.I. Dzhus, L.I. Ostapchenko, N.M. Khranovska // Цитология и генетика. — 2014. — Т. 48, № 6. — С. 56-61. — Бібліогр.: 24 назв. — англ. |
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nasplib_isofts_kiev_ua-123456789-1266752025-02-09T17:57:06Z Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells Комбинированное влияние тейхоевой кислоты из staphylococcus aureus и гетерометаллического Cu/Cd этилен диаминового комплекса на перитонеальные макрофаги и опухолевые клетки Nikulina, V.V. Garmanchuk, L.V. Senchylo, N.V. Nikolaenko, T.V. Dzhus, O.I. Ostapchenko, L.I. Khranovska, N.M. Оригинальные работы We investigated the effects of teichoic acid (TA) from Staphylococcus aureus Wood 46 on tumor growth and metastasis of the experimental Lewis lung carcinoma (LLC) in mice. Intranasal administration of TA alone aggravated both tumor growth and metastasis, whereas combined administration of TA with a synthetic bimetallic (copper: cadmium) ethylene diamine complex PO244 resulted in pronounced antitumor and antimetastatic effects. The group of animals subjected to the combined treatment with TA and PO244 manifested the highest degree of lymphocyte infiltration into the tumor tissue, compared to the control group and those exposed to TA or PO244 alone. Moreover, the combined treatment negatively affected the adhesive properties of peritoneal macrophages in the LLC bearing mice. Co-cultivation of the isolated macrophages with primary LLC cultures revealed significant (p < 0.05) cytotoxic and cytostatic effects, detected as an increased level of apoptosis and a reduced fraction of replicating cells. Исследовали модифицирующее влияние тейхоевой кислоты (ТК) на рост и метастазирование перевиваемой карциномы легких Льюис у мышей. Выявлена гиперактивация роста и метастазирования первичной опухоли при интраназальном введении животным ТК, в то время как при комбинированном влиянии с РО244 наблюдали усиление противо-опухолевого эффекта. Самый высокий уровень инфильтрации опухолевой ткани лимфоцитами зафиксировали в группе животных, которым проводили комбинированную терапию ТК с РО 244. Показано снижение адгезивных свойств перитонеальных макрофагов под влиянием биметаллического комплекса. После сокультивирования макрофагов от животных, которым проводили комбинированную терапию с первичной культурой LLC, выявлено значительное (p < 0.05) цитотоксическое/цитостатическое влияние, что проявлялось в увеличении уровня апоптоза и уменьшении популяции клеток пролиферативного пула. 2014 Article Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells / V.V. Nikulina, L.V. Garmanchuk, N.V. Senchylo, T.V. Nikolaenko, O.I. Dzhus, L.I. Ostapchenko, N.M. Khranovska // Цитология и генетика. — 2014. — Т. 48, № 6. — С. 56-61. — Бібліогр.: 24 назв. — англ. 0564-3783 DOI: 10.3103/S0095452714060085 https://nasplib.isofts.kiev.ua/handle/123456789/126675 616-006.004:618.19:615.373 en Цитология и генетика application/pdf Інститут клітинної біології та генетичної інженерії НАН України |
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Оригинальные работы Оригинальные работы |
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Оригинальные работы Оригинальные работы Nikulina, V.V. Garmanchuk, L.V. Senchylo, N.V. Nikolaenko, T.V. Dzhus, O.I. Ostapchenko, L.I. Khranovska, N.M. Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells Цитология и генетика |
| description |
We investigated the effects of teichoic acid (TA) from Staphylococcus aureus Wood 46 on tumor growth and metastasis of the experimental Lewis lung carcinoma (LLC) in mice. Intranasal administration of TA alone aggravated both tumor growth and metastasis, whereas combined administration of TA with a synthetic bimetallic (copper: cadmium) ethylene diamine complex PO244 resulted in pronounced antitumor and antimetastatic effects. The group of animals subjected to the combined treatment with TA and PO244 manifested the highest degree of lymphocyte infiltration into the tumor tissue, compared to the control group and those exposed to TA or PO244 alone. Moreover, the combined treatment negatively affected the adhesive properties of peritoneal macrophages in the LLC bearing mice. Co-cultivation of the isolated macrophages with primary LLC cultures revealed significant (p < 0.05) cytotoxic and cytostatic effects, detected as an increased level of apoptosis and a reduced fraction of replicating cells. |
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Article |
| author |
Nikulina, V.V. Garmanchuk, L.V. Senchylo, N.V. Nikolaenko, T.V. Dzhus, O.I. Ostapchenko, L.I. Khranovska, N.M. |
| author_facet |
Nikulina, V.V. Garmanchuk, L.V. Senchylo, N.V. Nikolaenko, T.V. Dzhus, O.I. Ostapchenko, L.I. Khranovska, N.M. |
| author_sort |
Nikulina, V.V. |
| title |
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells |
| title_short |
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells |
| title_full |
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells |
| title_fullStr |
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells |
| title_full_unstemmed |
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells |
| title_sort |
combined influence of teichoic acids from staphylococcus aureus and heterometallik cu/cd ethylenediamine complex on peritoneal macrophages and tumor cells |
| publisher |
Інститут клітинної біології та генетичної інженерії НАН України |
| publishDate |
2014 |
| topic_facet |
Оригинальные работы |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/126675 |
| citation_txt |
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Cd ethylenediamine complex on peritoneal macrophages and tumor cells / V.V. Nikulina, L.V. Garmanchuk, N.V. Senchylo, T.V. Nikolaenko, O.I. Dzhus, L.I. Ostapchenko, N.M. Khranovska // Цитология и генетика. — 2014. — Т. 48, № 6. — С. 56-61. — Бібліогр.: 24 назв. — англ. |
| series |
Цитология и генетика |
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ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 656
We investigated the effects of teichoic acid (TA) from
Staphylococcus aureus Wood 46 on tumor growth and
metastasis of the experimental Lewis lung carcinoma (LLC)
in mice. Intranasal administration of TA alone aggravated
both tumor growth and metastasis, whereas combined ad-
ministration of TA with a synthetic bimetallic (copper :
cadmium) ethylene diamine complex PO244 resulted in
pronounced antitumor and antimetastatic effects. The group
of animals subjected to the combined treatment with TA
and PO244 manifested the highest degree of lymphocyte
infiltration into the tumor tissue, compared to the control
group and those exposed to TA or PO244 alone. Moreover,
the combined treatment negatively affected the adhesive
properties of peritoneal macrophages in the LLC bearing
mice. Co-cultivation of the isolated macrophages with pri-
mary LLC cultures revealed significant (p < 0.05) cytotoxic
and cytostatic effects, detected as an increased level of
apoptosis and a reduced fraction of replicating cells.
Key words: teichoic acids, Lewis lung carcinoma, bimetallic
complex, macrophages.
Introduction. In order to overcome the adaptive
mechanisms of tumor cells to form drug resistance
there are a row of modern strategies of antitumor
therapy, one of them is immunotherapy of tumor.
On this base the application of immune modulators
can promote tumor subclone formation. The bacterial
cells components play a significant role as the immu-
nomodulators. They are not antitumor drugs thereby
such components modulate immune response and
favour intensification of antitumor efficiency in com-
bined use with classic chemotherapy [1].
The modern researches reveal anticarcinogenic
influence one of the major bacterial cell wall com-
ponents – teichoic acids (TA). It was shown that
TAs of cell wall of some bacteria are responsible
for enhancement of hypersensitivity reaction and
suppresses antibody synthesis in big concentrations
and is able to activate cell cytotoxicity. It is known
that TA as a component of cell wall of Gram-posi-
tive bacteria as well as lipopolysaccharides of Gram-
negative bacteria can stimulate TNF- production.
Bacterial substances which are responsible for tumor
cell lysis are identified and used for enhancement of
immunogenicity of antitumor vaccines. Indirect toxic
influence of bacterial cells against tumors is realized
by means of recruitment of immune system effect or
sand cross-presentation of tumor antigens. A particu-
lar role in recognizing different bacterial structures and
inducting the antitumor effects is dedicated to Toll-
like receptors (TLRs) [2]. TLRs are known to be used
very often in antitumor therapy [3].
TLR ligands are bacterial cell components,
so-called pathogen-associated molecular patterns
(PAMPs) [4]. In Gram-positive bacteria their role
is fulfilled by teichoic acids.
Teichoic acids of Gram-positive bacteria are po-
tential surface structures, which stimulate TNF-
production and induct IL-12 secretion in CD14-
depended way by monocytes. It results cells the
activation of natural killer cells, which leads to di-
rectly determined IFN- production [5, 6]. More-
over, it has been shown that teichoic acids cause
secretion of Th-1 type cytokines, which activate
T-lymphocytes and natural killer cells, and are ca-
pable to change cytotoxic potential of lymphocytes
with killer activity.
On the other hand, TA stimulates proliferative
activity of both normal and immortalized lymphoid
cells. It can cause progress of lymphoproliferative
diseases [7]. In previous studies we have shown that
TA modifies growth of primary tumor and level of
metastatic dissemination of transplantable lung Lew-
is carcinoma dependently on term of administration.
It is known that motogenic potential of normal
as well as tumor cells is realized by system of adhe-
© V.V. NIKULINA, L.V. GARMANCHUK,
N.V. SENCHYLO, T.V. NIKOLAENKO, O.I. DZHUS,
L.I. OSTAPCHENKO, N.M. KHRANOVSKA, 2014
УДК 616-006.004:618.19:615.373
COMBINED INFLUENCE OF TEICHOIC ACIDS FROM STAPHYLOCOCCUS
AUREUS AND HETEROMETALLIK Cu/Cd ETHYLENEDIAMINE COMPLEX
ON PERITONEAL MACROPHAGES AND TUMOR CELLS
V.V. NIKULINA 1, L.V. GARMANCHUK 1, N.V. SENCHYLO 1,
T.V. NIKOLAENKO 1, O.I. DZHUS 1, L.I. OSTAPCHENKO 1, N.M. KHRANOVSKA 2
1 Educational and scientific centre «Institute of biology» of Taras Shevchenko National University of Kyiv
2 National Cancer Institute, Kyiv
E-mail: liudmylagarmanchuk@rambler.ru
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 6 57
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Ñd
sive molecules. They are responsible for supporting
tissue homeostasis, decreasing adhesive properties
and abnormalities of intercellular contacts, that as-
sist cell migration. Consequently, adhesive poten-
tial is inversely correlated with the ability to migrate.
By our research group it has been synthesized the
complexes of copper and organic ligands in base for
which bactericidal, fungicidal and antitumour activity
have been revealed later. It has been found they permit
to regulate a defensive reaction of organism on molec-
ular and cell levels, promisinge therapeutic advances
in cancer pathogenesis and infection diseases [8]. As
a rule they cause both bactericidal and fungicidal ef-
fective influence, demonstrating antitumor effect and
moderate resistance to microorganisms. Screening of
bimetallic complexes has shown that the complex that
consists of copper ions, cadmium and ethylenedi-
amine – ÐÎ244 ([Cu(en)2][Cd2(CH3COO)6]) dem-
onstrates the highest efficiency [9, 10].
The aim of this study was to investigate a com-
bined influence teichoic acids from Staphylococcus
aureus and heterometallik Cu/Cd ethylenediamine
complex on peritoneal macrophages and tumor
cells in vitro and in vivo.
Materials and methods. The investigation was
carried out in C57Bl/6 female mice weighing 20–
25 g aged 2 to 3 months from the vivarium of the
Educational and scientific centre «Institute of bio-
logy» of Taras Shevchenko National University, in
two groups of experiments (Table 1). All researches
on animals were carried out according to Guide
for the Care and Use of Laboratory Animals [11].
The strain of metastatic Lewis lung carcinoma
(LLC) was kindly given by National Bank of Cell
Lines and Tumor Strains of R.E. Kavetsky Institute
of experimental pathology, oncology and radiobiol-
ogy. The cell suspension of LLC (0.2 ml of 20 %
cell suspension) was inoculated subcutaneously into
the site of sacrum.
Administration of PO244 to the animals was car-
ried out intraperitoneally at the total dose 0.8 mg/
kg of body weight, whereas TA from Staphylococ-
cus aureus Wood 46 was administrated intranasally
in concentration 2 ng/kg of animal weight. Both
TA and PO244 were administrated on 8th day after
tumor cell inoculation. Antimetastatic and antitu-
mor effects were defined according to the inhibi-
tion index of tumor weight, volume and number of
metastases in lung as described [12].
The primary culture was obtained from trans-
plantable Lewis lung carcinoma after 2–3 times
trypsinization of tumor tissue in trypsin-EDTA so-
lution with pH 7.0. Cell cultivation was conducted
under standard and serum free conditions at 37 °C,
100 % humidity and 5 % ÑÎ2.
Mononuclear phagocyte fraction of peritoneal
exudate of mice was obtained by standard procedure
of Pietrangeli [13]. Macrophages were incubated
under standard conditions at 37 °C, 100 % humidity
and 5 % ÑÎ2 for 4 h and then their cultural medium
was added to LLC culture.
The lymphocyte infiltration degree of tumor
(1000 cells/1 g tumor tissue) was estimated after
their fractionation in phycoll-verografin gradient
with density 1,077 g/ml by centrifugation 40 min
at 1.500 rpm. The adhesive capacity of macrophage
and tumor cells was determined by the percent of
cells which were attached to the substrate, after
Table 1. The influence of heterobimetallic complex and teichoic acids on the growth and metastasis
of transplantable Lewis lung carcinoma (27 days after tumor cell inoculation)
* ð < 0,05. **3 animals died before 27th day.
Animal group
Control
(n1 = 8; n2 = 6)
TA **
(n1 = 7; n2 = 6)
PO244
(n1 = 6; n2 = 7)
TA + PO244
(n1 = 8; n2 = 6)
Number of LLC bearing mice (%)
(nt/n1 + n2)
Tumor weight (g)
WI
Number of animals with metastases (%)
(nm/n1 + n2)
Number of metastases in lungs
Metastatic index Nk-N(exp)/Nk · 100 (%)
85,7 % (12/14)
5,3 ± 1,2
78,6 % (11/14)
21,7 ± 3,2
100 %(14/14) *
7,8 ± 1,4 *
>47,2 %
100 % (14/14) *
37,3 ± 5,4 *
>71,9,2 %
69,2 % (9/13) *
4,1 ± 0,7 *
<22,6 %
63 % (8/13) *
15,2 ± 3,6
64,3 % (8/14) *
2,5 ± 0,6 *
<52,8 %
57,2 % (8/14) *
9,3 ± 2,1 *
<57,3 %
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 658
V.V. Nikulina, L.V. Garmanchuk, N.V. Senchylo et al.
crystal violet staining by measuring absorbance at
570 nm using a multiwell spectrophotometer, Syn-
ergie Biotec (USA).
Cytotoxic/cytostatic effect, necrotic/apoptotic in-
dex and proliferative activity after the influence of
ÐÎ244 ([Cu(en)2][Cd2(CH3COO)6]) and TA with
respect to cells of primary culture of LLC were as-
sessed by cytofluorimetry, counting cells stained with
trypan blue and MTT-test assay, cells were incubated
with TA and PO244 for 24 and 48 h under normal
conditions in 96 well plates. The initial cell concentra-
tion was approximately 5·104 cells/ml in the sample
volume of 100 l. It has been used a culture media
RPMI («Sigma», USA) with 10 % FBS («Sigma»,
USA), 2 mM L-glutamine, and 40 g/ml gentamicin.
Different concentrations (1 · 10–6–1,25 · 10–4 M) of
PO244 and TA (1 ng/ml–1 g/ml) were added to cell
cultures in 100 l of media after the period of cell
adaptation under standard conditions (5 % CO2, 100
humidity, 37 ºC) during 4 h. The number of living
cells was determined in wells using MTT-colorimetric
assay and cell counting were performed using a tripan
blue dye after 48 h incubation with test-agents [14].
The cytotoxic effect was evaluated as a percent of
live cells relative to control and characterized by IC50
index [15] and mitogenic effect was evaluated as per-
cent increase number alive cells relative to control.
The isolated peritoneal macrophages from tu-
mor bearing mice after TA and PO244 treatment
have been co-cultivated with primary LLC culture
during 48 h. Apoptotic level and distribution of cells
in phases of cell cycle were assessed by cytofluorim-
etry [16]. For this purpose the samples were stained
with PI, which selectively joins with intercalating
places in DNA. Cytofluorymetry was carried out
on the instrument FACS Ñalibur («Becton Dick-
inson», USA). Special mathematical program Mod
Fit LT 2.0 (BDIS, USA) for Macintosh computers
was used for acquisition and data analysis. Narrow-
band filter 585/42 nm was used in order to measure
the fluorescence of PI. It has been performed the
histological mounts of tumor samples after their
exposure to TA, ÐO244, and their combination by
standard methods. Coloring of Lewis carcinoma tu-
mor sections was performed by the standard method
of hematoxylin-eosin staining [17].
Results and conclusions. For the first time, it has
been shown antitumor and antimetastatic activity of
heterometallic coordinational composition on the
model of transplantable Lewis lung carcinoma. Un-
der the condition of ÐÎ244 administration tumors
were found out in 69,2 % of experimental animals
(Table 1), the administration of TA to LLC-bearing
mice resulted in 100 % tumor outcome, in compari-
son with 85,7 % in control group, whereas com-
bined application of ÐÎ244 with teichoic acids led
to 64,3 % tumor outcome
The tumors in animals with certain therapy dif-
fered in the rate of growth and weight from each
other. The administration of TA increased the tu-
mor weight by 42,7 % compared to control, whilst
PO244 injection resulted in the decrease of tumor
weight by 22,6 %, whereas LLC-bearing mice that
received PO244 + TA combined treatment had
52,8 % smaller tumor weight. Moreover, this pat-
tern remained in determining the tumor weight at
different stages of growth. From the dynamics of
tumor growth the growth rate of the tumor was seen
to be reduced in the application of TA with PO244
compared to the control group (Fig. 1).
Number of animals with metastases in control
group was 78,6 %, where as number of metastases in
lungs was 21,7 ± 3,2, TA treatment resulted in the
increase metastatic level to 100 % and 37,3 ± 5,4
correspondently, PO244 utilization led to decrea-
se metastasis to 63 % animals with metastases and
15,2 ± 3,6 metastases in lungs, after combined use
PO244 and TA antimetastatic effect was the high-
est: 52,7 % of animals with metastases had 9,3 ±
± 2,1 metastases in lungs.
On experimental model of metastatic Lewis lung
carcinoma hyperactivation of primary tumor growth
simultaneously with metastasis was as certained in
intranasal administration of TA on the stage of
primary node formation. During the application
Fig.1. The influence of teichoic acids and PO244 (mono
and combined therapy) on growth of primary tumors.
Here and Fig. 3, 4 *P < 0.05 vs control
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 6 59
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Ñd
in combined therapy TA with synthetic bimetallic
complex (ÐÎ244), which demonstrated both an-
titumor and antimetastatic influence, intensifying
of antitumor effect of PO244 was shown (Fig. 1).
Our results argue that considerable depression of
tumor growth occurs to all phases of tumor forma-
tion with combined use PO244 and TA. It is known
that recognition of bacterial structures including LTA
and TA passes with involvement of toll-like receptors
[18]. This receptors (including TLR2 and TLR4,
there ligands are respectively TA and LTA) are ex-
pressed on surface of different cell types including
different types of tumors [19]. The interaction TA
and LTA with specific receptors results in activation
of cytokine production. Moreover, in depending on
ligand doze the activation both pro-inflammatory
and anti-inflammatory cytokines are possible [20].
For the purpose to define the mechanisms of
combined TA and PO244 influence, cytotoxic/cy-
tostatic screening of these substances was carried
out on primary LLC culture. However, the inten-
sification of cytotoxic effect of PO244 and TA on
tumor cells of primary culture was not detected in
such conditions (Table 2). IC50 index was 2.20 ±
± 0.04·10–5 Ì under the influence PO244, addition
of TA didn’t impact on the index 3.03 ± 0.06·10–5 Ì,
whereas monotherapy by TA stimulated of primary
LLC culture proliferation insignificantly.
The PO244 substance showed distinct cytotoxic
effect whereas TA in certain concentrations activat-
ed cell proliferation, as we can see from mentioned
data. The survival curve under combine influence
of these substances practically doesn’t differ from
the curve under the influence of ÐÎ244.
The modern researchers consider the degree of
lymphocytes infiltration of tumor as diagnostic and
prognostic marker of clinical course [20, 21].
Fig. 2. Slide mount of mouse lung demonstrates level
of lymphocyte infiltration in control (a) and under the
influence of teichoic acid and PO244 (infiltrative lym-
phocyte is marked) (b)
Table 2. The influence of ÐÎ244 and TA on LLC cells
with their separated and combined application in vitro.
Incubation of cells with above-mentioned agents was
carried out 48 h
*Apoptotic level, viability and mitogenic index were as-
sessed after cells incubation with TA (concentration of
TA was 0.25 ng/ml).
Cells
PO244 TA+PO244
M·10–5
IC50
IC50/10
Viability (IC50/10,%)
Apoptotic level
(IC50/10,%)
2.20 ± 0.04
0.22 ± 0.00
71.6 ± 7.3
24.4 ± 4.3
3.03 ± 0.06
0.30 ± 0.01
79.4 ± 3.1
18.1 ± 0.4
Cells Control TA*
Mitogenic index
Viability (%)
Apoptotic level (%)
92.4 ± 3.7
11.7 ± 2.7
1.12
94.3 ± 5.6
9.1 ± 0.5
Fig. 3. The influence of agents (TA, ÐÎ244 and ÒA+
+ ÐÎ244) degree of infiltrative lymphocytes in tumor
tissue (a). The adhesion parameter of peritoneal mac-
rophages of LLC bearing (b)
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 660
V.V. Nikulina, L.V. Garmanchuk, N.V. Senchylo et al.
In order to determine possible mechanism of TA
impact on the tumor through the immune cells we
studied macrophages that were received from differ-
ent groups of animals at the last stage of carcinogen-
esis. Macrophages from animals after therapy and
control group in vivo were added and cocultivated
with primary LLC culture during 48 h; we founded
cytotoxic/cytostatic influence which was expressed
as increasing of apoptotic level and decreasing of
cell population of proliferative pool (Fig. 4). There-
by, the screening of potential antitumor agent PO
244 (mono and combined application with TA) was
carried out. The results of the study of influence
of teichoic acids on transplantable Lewis lung car-
cinoma results strongly suggest about implication
this bacteria cell wall component of Staphylococcus
aureus Wood 46 in antitumor and antimetastatic
response in vivo and modification influence in vitro.
ÊÎÌÁÈÍÈÐÎÂÀÍÍÎÅ ÂËÈßÍÈÅ ÒÅÉÕÎÅÂÎÉ
ÊÈÑËÎÒÛ ÈÇ STAPHYLOCOCCUS AUREUS
È ÃÅÒÅÐÎÌÅÒÀËËÈ×ÅÑÊÎÃÎ Cu/Cd
ÝÒÈËÅÍ ÄÈÀÌÈÍÎÂÎÃÎ ÊÎÌÏËÅÊÑÀ
ÍÀ ÏÅÐÈÒÎÍÅÀËÜÍÛÅ ÌÀÊÐÎÔÀÃÈ
È ÎÏÓÕÎËÅÂÛÅ ÊËÅÒÊÈ
Â.Â. Íèêóëèíà, Ë.Â. Ãàðìàí÷óê,
Í.Â. Ñåí÷èëî, Ò.Â. Íèêîëàåíêî, Å.È. Äæóñ,
Ë.È. Îñòàï÷åíêî, Í.Í. Õðàíîâñêàÿ
Èññëåäîâàëè ìîäèôèöèðóþùåå âëèÿíèå òåéõîåâîé
êèñëîòû (ÒÊ) íà ðîñò è ìåòàñòàçèðîâàíèå ïåðåâè-
âàåìîé êàðöèíîìû ëåãêèõ Ëüþèñ ó ìûøåé. Âûÿâ-
ëåíà ãèïåðàêòèâàöèÿ ðîñòà è ìåòàñòàçèðîâàíèÿ ïåð-
âè÷íîé îïóõîëè ïðè èíòðàíàçàëüíîì ââåäåíèè æè-
âîòíûì ÒÊ, â òî âðåìÿ êàê ïðè êîìáèíèðîâàííîì
âëèÿíèè ñ ÐÎ244 íàáëþäàëè óñèëåíèå ïðîòèâî-
îïóõîëåâîãî ýôôåêòà. Ñàìûé âûñîêèé óðîâåíü èí-
ôèëüòðàöèè îïóõîëåâîé òêàíè ëèìôîöèòàìè çàôèê-
ñèðîâàëè â ãðóïïå æèâîòíûõ, êîòîðûì ïðîâîäèëè
êîìáèíèðîâàííóþ òåðàïèþ ÒÊ ñ ÐÎ244. Ïîêàçàíî
ñíèæåíèå àäãåçèâíûõ ñâîéñòâ ïåðèòîíåàëüíûõ ìàêðî-
ôàãîâ ïîä âëèÿíèåì áèìåòàëëè÷åñêîãî êîìïëåêñà.
During the experiment of primary tumor therapy
we registered the increase of lymphocyte infiltra-
tion in animal groups with application of bimetallic
complex PO244 or TA (Fig. 2, a, b).
The highest degree of lymphocyte infiltration
in tumor was detected in animal group which was
treated by mixture of TA and PO244. It should be
noted that the increase of rate of lymphocyte in-
filtration in tumor tissue after mono or combined
therapy is one probable mechanism of inhibition of
primary tumor (Fig. 3, a) [22–24].
The TLR-mediated effect can be modifying of
cell-cell interaction and cell interaction with mole-
cules of extracellular matrix. We have tested the TA
influence on adhesive potential of peritoneal mac-
rophages of LLC bearing mice or intact animals.
It was shown that application of synthetic
bimetallic complex PO244 in therapeutic con-
centration (1.1 ± 0.01 · 10–5Ì/g weigh animal)
didn’t result in a cytotoxic impact on peritoneal
macrophages, whereas the modifying effect on
adhesive behavior of peritoneal macrophages was
observed (Fig. 3, b).
Since, adhesive potential inversely correlates
with cell ability to migrating, the in vitro findings
indicate that migration and tumor infiltration can
be activate when tumor growing in vivo. We have
shown it in combined therapeutic scheme applica-
tion of TA and PO244 on transplantable Lewis lung
carcinoma (Fig. 1). Monotherapy by TA stimulates
tumor infiltration by lymphocytes insignificantly,
whereas in combined therapy with PO244 this pa-
rameter is increased 2.4 times (p < 0.05).
An apoptotic index increase and reduction of pro-
liferative cells quantity in non-contact coculture with
primary culture LLC were observed as a result of
adding the peritoneal macrophages from intact ani-
mals, the animals bearing the tumors with and without
combined influence of TA and PO244 (Fig. 4, a, b).
Fig. 4. Apoptotic level, % (a) of LLC
cells under condition of contactless
cocultivation with macrophages, which
were obtained from LLC bearing mice,
LLC bearing mice after effective com-
bine therapy by teichoic acid and
PO244 and distribution of LLC cells
in phases of cell cycle, % (b); 1 – LLC
(control), 2 – LLC(Mph_tumor), 3 –
LLC(Mph_tumor+TA+PO244)
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2014. Ò. 48. ¹ 6 61
Combined influence of teichoic acids from Staphylococcus aureus and heterometallik Cu/Ñd
Ïîñëå ñîêóëüòèâèðîâàíèÿ ìàêðîôàãîâ îò æèâîòíûõ,
êîòîðûì ïðîâîäèëè êîìáèíèðîâàííóþ òåðàïèþ ñ
ïåðâè÷íîé êóëüòóðîé LLC, âûÿâëåíî çíà÷èòåëüíîå
(p < 0.05) öèòîòîêñè÷åñêîå/öèòîñòàòè÷åñêîå âëèÿ-
íèå, ÷òî ïðîÿâëÿëîñü â óâåëè÷åíèè óðîâíÿ àïîïòî-
çà è óìåíüøåíèè ïîïóëÿöèè êëåòîê ïðîëèôåðàòèâ-
íîãî ïóëà.
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Received 28.12.12
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/NLD (Gebruik deze instellingen om Adobe PDF-documenten te maken die zijn geoptimaliseerd voor prepress-afdrukken van hoge kwaliteit. De gemaakte PDF-documenten kunnen worden geopend met Acrobat en Adobe Reader 5.0 en hoger.)
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/TUR <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>
/UKR <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>
/ENU (Use these settings to create Adobe PDF documents best suited for high-quality prepress printing. Created PDF documents can be opened with Acrobat and Adobe Reader 5.0 and later.)
>>
/Namespace [
(Adobe)
(Common)
(1.0)
]
/OtherNamespaces [
<<
/AsReaderSpreads false
/CropImagesToFrames true
/ErrorControl /WarnAndContinue
/FlattenerIgnoreSpreadOverrides false
/IncludeGuidesGrids false
/IncludeNonPrinting false
/IncludeSlug false
/Namespace [
(Adobe)
(InDesign)
(4.0)
]
/OmitPlacedBitmaps false
/OmitPlacedEPS false
/OmitPlacedPDF false
/SimulateOverprint /Legacy
>>
<<
/AddBleedMarks false
/AddColorBars false
/AddCropMarks false
/AddPageInfo false
/AddRegMarks false
/ConvertColors /ConvertToCMYK
/DestinationProfileName ()
/DestinationProfileSelector /DocumentCMYK
/Downsample16BitImages true
/FlattenerPreset <<
/PresetSelector /MediumResolution
>>
/FormElements false
/GenerateStructure false
/IncludeBookmarks false
/IncludeHyperlinks false
/IncludeInteractive false
/IncludeLayers false
/IncludeProfiles false
/MultimediaHandling /UseObjectSettings
/Namespace [
(Adobe)
(CreativeSuite)
(2.0)
]
/PDFXOutputIntentProfileSelector /DocumentCMYK
/PreserveEditing true
/UntaggedCMYKHandling /LeaveUntagged
/UntaggedRGBHandling /UseDocumentProfile
/UseDocumentBleed false
>>
]
>> setdistillerparams
<<
/HWResolution [2400 2400]
/PageSize [612.000 792.000]
>> setpagedevice
|