Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules
The aim: To evaluate anticancer activity of vaccination with chicken embryo proteins (CEP) applied in different schedules. Materials and Methods: C57Bl mice were vaccinated with CEP before (prophylactic schedule) or after (different therapeutic schedules with or without preliminary tumor removal) th...
Saved in:
| Published in: | Experimental Oncology |
|---|---|
| Date: | 2015 |
| Main Authors: | , , , , , , |
| Format: | Article |
| Language: | English |
| Published: |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2015
|
| Subjects: | |
| Online Access: | https://nasplib.isofts.kiev.ua/handle/123456789/145488 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Journal Title: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Cite this: | Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules / T.V. Symchych, N.I. Fedosova, O.M. Karaman, L.M. Yevstratieva, H.S. Lisovenko, I.M. Voyeykova, H.P. Potebnia // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 197-202. — Бібліогр.: 35 назв. — англ. |
Institution
Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1860001263604727808 |
|---|---|
| author | Symchych, T.V. Fedosova, N.I. Karaman, O.M. Yevstratieva, L.M. Lisovenko, H.S. Voyeykova, I.M. Potebnia, H.P. |
| author_facet | Symchych, T.V. Fedosova, N.I. Karaman, O.M. Yevstratieva, L.M. Lisovenko, H.S. Voyeykova, I.M. Potebnia, H.P. |
| citation_txt | Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules / T.V. Symchych, N.I. Fedosova, O.M. Karaman, L.M. Yevstratieva, H.S. Lisovenko, I.M. Voyeykova, H.P. Potebnia // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 197-202. — Бібліогр.: 35 назв. — англ. |
| collection | DSpace DC |
| container_title | Experimental Oncology |
| description | The aim: To evaluate anticancer activity of vaccination with chicken embryo proteins (CEP) applied in different schedules. Materials and Methods: C57Bl mice were vaccinated with CEP before (prophylactic schedule) or after (different therapeutic schedules with or without preliminary tumor removal) the Lewis lung carcinoma cells transplantation. The latent period of tumor development, tumor volume and metastasis rate were evaluated. Results: Potent antimetastatic effect of CEP-based vaccination was seen in case of therapeutic regimen after primary tumor removal. The metastasis inhibition index (MII) reached 96.9 and 97.8% on 18th and 34th day after tumor removal, respectively. When CEP vaccination was performed in the settings of therapeutic regimen without primary tumor removal the anticancer effect was evident only if vaccinations started as early as 24 h after the cancer cells injections. The highest MII achieved in such condition was 77.6%, tumor volume in the group of vaccinated animals was by 53.1–42.1% lower than in the control tumor-bearing mice. CEP vaccination before tumor challenge (prophylactic immunization) led to a statistically significant prolongation of the latent period of tumor development, a reduction of tumor volume (35.8–48.8% compared to control unvaccinated mice) and a marked inhibition of metastasis (MII was 71.1%). Conclusion: Vaccination based on CEP exhibited both prophylactic and therapeutic anticancer effects. The last one is more pronounced when the vaccination starts shortly after the primary tumor resection. Key Words: chicken embryo proteins, anticancer activity, Lewis lung carcinoma.
|
| first_indexed | 2025-12-07T16:36:07Z |
| format | Article |
| fulltext |
Experimental Oncology 37, 197–202, 2015 (September) 197
ANTICANCER EFFECTIVENESS OF VACCINATION BASED
ON XENOGENEIC EMBRYO PROTEINS APPLIED IN DIFFERENT
SCHEDULES
T.V. Symchych*, N.I. Fedosova, O.M. Karaman, L.M. Yevstratieva, H.S. Lisovenko, I.M. Voyeykova, H.P. Potebnia
R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv 03022, Ukraine
The aim: To evaluate anticancer activity of vaccination with chicken embryo proteins (CEP) applied in different schedules. Materials
and Methods: C57Bl mice were vaccinated with CEP before (prophylactic schedule) or after (different therapeutic schedules with
or without preliminary tumor removal) the Lewis lung carcinoma cells transplantation. The latent period of tumor development, tumor
volume and metastasis rate were evaluated. Results: Potent antimetastatic effect of CEP-based vaccination was seen in case of thera-
peutic regimen after primary tumor removal. The metastasis inhibition index (MII) reached 96.9 and 97.8% on 18th and 34th day after
tumor removal, respectively. When CEP vaccination was performed in the settings of therapeutic regimen without primary tumor
removal the anticancer effect was evident only if vaccinations started as early as 24 h after the cancer cells injections. The highest MII
achieved in such condition was 77.6%, tumor volume in the group of vaccinated animals was by 53.1–42.1% lower than in the control
tumor-bearing mice. CEP vaccination before tumor challenge (prophylactic immunization) led to a statistically significant prolonga-
tion of the latent period of tumor development, a reduction of tumor volume (35.8–48.8% compared to control unvaccinated mice)
and a marked inhibition of metastasis (MII was 71.1%). Conclusion: Vaccination based on CEP exhibited both prophylactic and
therapeutic anticancer effects. The last one is more pronounced when the vaccination starts shortly after the primary tumor resection.
Key Words: chicken embryo proteins, anticancer activity, Lewis lung carcinoma.
The development of xenogeneic anticancer vac-
cines (XAV) started at the end of the 1990s, when
it was shown that the use of xenogeneic analogues
of tumor associated antigens enables the body
to overcome immunological toleration for its own
proteins [1]. Now, it is proven that a number of tumor
associated antigens and protein have their counter-
parts of animal or avian origin which can serve as an-
tigens in XAV. These proteins or genes are exploited
in the construction of XAVs, some of which have been
shown to have anticancer effect [2, 3]. Some XAV
successfully passed I–II phases of clinical trials. Their
safety and ability to induce immune response without
autoimmune complications have been proven [4–8].
Among others, genes and proteins of chicken origin
which share homology with human counterparts are
exploited in the XAV construction [9–14].
At the R.E. Kavetsky Institute of Experimental Pathol-
ogy, Oncology and Radiobiology (IEPOR) of the National
Academy of Science of Ukraine XAV based on chicken
embryo proteins (CEP) is under deve lopment.
It is known that anticancer vaccines based on one
or several antigens can lead to an immune edi ting of the
tumor so that it loses antigens targeted by the vac-
cine. Moreover, polyantigenic vaccines potentially can
elicit an immune response to a wider range of cancer
antigens including unidentified ones [15]. That is why
the vaccine which is being constructed is designed
to be polyantigenic and is based on proteins extracted
from the chicken embryo. In preliminary experiments,
it was shown that blood serum of mice bearing different
tumor strains has antibodies which react with CEP [16].
When injected to intact mice CEP caused no side effect
or allergy reactions [17]. The aim of the current work
is to evaluate the anticancer activity of CEP-based vac-
cine administered by different vaccination schedules.
MATERIALS AND METHODS
The study has been carried out on male C57Bl/6 mice
2–2.5 months old weighing 19–20 g, bred in the
IEPOR. The use and care of the experimental animals
have been performed in accordance with the standard
international rules of biologic ethics and was approved
by Institutional Animal Care and Use Committee [18,
19]. The anticancer and antimetastatic efficacy of CEP
was examined when vaccination was applied prior to tu-
mor cells injection (prophylactic schedule), after tumor
transplantation (therapeutic vaccination) or after tumor
removal (post surgery vaccination). Lewis lung carci-
noma (LLC) was used as the model of tumor growth.
CEP was prepared as follows [20]: 7 days chicken
embryos were rinsed two times briefly in cold 0.9%
NaCl solution, homogenized and then extracted with
0.9% NaCl solution containing 0.1% EDTA, for 60 min
at 4 °C by agitation. Following the extraction, chicken em-
bryo tissue was removed by centrifugation at 1.500 g for
30 min. The resulting supernatant was collected and
frozen at −20 °C. Tumor associated antigens of LLC
(LLC-Ag) were prepared by three consecutive cycles
of freezing and melting of cell suspension. Following
the last melting, cell debris was removed by centrifuga-
tion at 1.500 g for 30 min. The resulting supernatants
were collected and frozen at −20 °C. The concentration
of proteins in the extracts was measured by Greenberg
and Craddock assay [21]. The same extracts were used
in all the experiments described in the article.
Submitted: December 29, 2014.
*Correspondence: E-mail: symchychtv@gmail.com
Abbreviations used: CEP — chicken embryo proteins; ITGI — index
of tumor growth inhibition; LLC — Lewis lung carcinoma; LLC-
Ag — Lewis lung carcinoma associated antigens; MII — metastasis
inhibition index; XAV — xenogeneic anticancer vaccine.
Exp Oncol 2015
37, 3, 197–202
198 Experimental Oncology 37, 197–202, 2015 (September)
Irrespective of vaccination schedule, CEP or LLC-
Ag immunizations were performed s.c. with 0.3 ml so-
lution per mouse (protein concentration 0.3 mg/ml).
According to the prophylactic experiment, mice
were immunized with CEP or LLC-Ag (three weekly
injections); LLC cells were transplanted 30 days after
the last immunization.
Therapeutic vaccination has been performed
by three different schemes: on the 1st, 8th, 15th days
(group #1); on the 7th, 14th, 28th days after the tumor
cell transplantation (group #2); in the third group, vac-
cination started when the tumor nodule had become
clearly palpable and was followed with two additional
injections on the 3rd and 10th days after the first vac-
cination (that corresponds to the 12th, 15th and 22nd
days after the tumor transplantation).
Post surgery vaccination started on the 1st, 8th, 15th
days after the tumor removal, which corresponds to the
18th, 24th and 31st days after tumor transplantation.
In the prophylactic and treatment vaccination experi-
ments cancer cells suspension was injected i.m. into the
right hind leg at a dose of 4 × 105 cells/mouse. Unvac-
cinated mice with the tumor were used as the control.
In the post surgery vaccination experiment, LLC cells
were injected per foot at a dose of 2.5 × 105 cells/mouse.
The tumor removal was performed on the 17th day after
the tumor transplantation. Mice which have undergone
surgical tumor removal but received no vaccination are
referred as the control.
Tumor dimensions were measured with calipers,
and tumor volumes were calculated according to the
formula:
Tumor volume = 2/3 π • width2 • length.
The Index of Tumor Growth Inhibition (ITGI) was
calculated according to the formula:
ITGI = 100%•(Vcontrol mice — Vimmunized mice) / Vcontrol mice,
where Vcontrol mice and Vimmunized mice stand for the
mean tumor volume in control and immunized mice
respectively.
To assess metastasis burden mice were sacrificed
and in each animal lungs were removed; surface lung
metastases were counted and measured. The metas-
tases volume was calculated as following:
V = 4πr3/3,
where r — stands for the metastases radius.
The percentage of mice bearing metastases is referred
as metastases rate. The mean number of metastases
was calculated per all the mice in group and per mice
bearing metastases.
Metastasis Inhibition Index (MII) was calculated
as following:
MII = 100% • (Аc • Вc – Аi • Вi) / (Аc • Вc),
where Аc and Аi stand for the number of mice be-
aring lung metastases in groups of control and immu-
nized mice respectively. Вc and Вi stand for the mean
number of lung metastases in groups of control and
immunized mice respectively [22]. The results were
analyzed for statistical significance by paired t-test
using StatSoft STATISTICA 7.0. Values p < 0.05 were
considered as statistically significant [23, 24]. The
data in figures and tables are presented as M ± SD.
RESULTS
The anticancer activity of CEP applied before
tumor transplantation (prophylactic immuniza-
tion). CEP or LLC-Ag were injected three times with
one-week intervals. Then 30 days after the last immu-
nization, LLC was transplanted into both unvaccinated
animals (the control) and mice vaccinated with CEP
or LLC-Ag. LLC tumor appeared in 81.0% (17 out of 21)
of the control mice (Table 1). In the treatment groups,
77.8% (7 out of 9) and 81.8% (9 out of 11) of mice im-
munized with LLC-Ag or CEP, respectively, developed
LLC tumors. The difference between all the groups was
not significant. The latent period of tumor development
was shorter (p < 0.05) in the group of the control mice
(7.8 days) compared to the mice pre-vaccinated with
LLC-Ag (10.0 days) or CEP (10.9 days).
Table 1. The latent period of tumor development and tumor transplanta-
tion efficacy in the vaccinated and control LLC-bearing mice
Group Tumor transplantation
efficacy, %
Latent period of tumor
development, days
Control 81.0 ± 13.1 7.8 ± 0.4
LLC-Ag 77.8 ± 13.9 10.0 ± 0.7*
CEP 81.8 ± 11.6 10.9 ± 0.6*
Note: *р < 0.05 compared to the control group.
The tumor growth kinetics is shown in Fig. 1. Du-
ring the experiment, the smallest tumor volume was
observed in the group of mice immunized with CEP
(p < 0.05 compared to the control group). In the group
of CEP-immunized mice, the ITGI reached 35.8–48.8%
depending on time after the tumor transplantation.
The tumor volume of mice immunized with LLC-Ag did
not differ significantly compared to both control and CEP-
immunized mice. In the group of LLC-Ag-immunized mice,
the maximal ITGI (28.4%) was observed on the 14th day
after the tumor transplantation (Table 2).
On day 28 after LLC transplantation, all the mice
of the control and treatment groups were euthanized
so the metastases rate to be evaluated. The results
are shown in Table 3.
In the mice vaccinated with CEP, 73.4% reduc-
tion of the mean metastasis volume was registered,
in particular, 51.5 and 72.1% decrease of the metas-
tases number per mouse or per mouse in the group
correspondingly. So, in this group MII reached 59.5%
per metastases-bearing mouse and 71.1% per group.
Contrary to CEP, LLC-Ag vaccination was not effective
against metastases development.
Table 2. Tumor volume and ITGI in control and vaccinated before tumor transplantation mice bearing LLC
Group Parameter Days after LLC transplantation
10 14 17 21 24 28
Control Tumor volume, mm3 165.0 ± 22.5 493.7 ± 62.2 981.0 ± 120.1 1523.7 ± 85.1 2087.8 ± 114.1 2492.4 ± 268.2
LLC-Ag Tumor volume, mm3 163.5 ± 35.8 353.6 ± 65.0 856.6 ± 125.0 1172.2 ± 190.3 1550.9 ± 291.2 1891.5 ± 332.7
ITGI, % 0.9 27.4 12.7 23.1 25.7 24.1
CEP Tumor volume, mm3 84.6 ± 8.3 301.5 ± 54.6 512.8 ± 73.0 892.3 ± 132.0 1321.1 ± 215.3 1600.8 ± 246.7
ITGI, % 48.8 38.9 47.7 41.4 36.7 35.8
Experimental Oncology 37, 197–202, 2015 (September) 199
0
500
1000
1500
2000
2500
3000
10 14 17 21 24 28
Days after tumor transplantation
Tu
m
or
v
ol
um
e,
m
m
3
Control
LLC-Ag
CEP
Fig. 1. The growth kinetics of LLC in control and pre-vaccinated
animals
Table 3. Metastasis burden in control and vaccinated before tumor trans-
plantation mice bearing LLC
Group Metastases
rate, %
Volume
of metasta-
ses, mm3
Number of metastases
per mouse
bearing me-
tastases
per group
Control 70.0 ± 14.5 38.9 ± 13.9 20.6 ± 6.5 14.4 ± 5.5
LLC-Ag 88.9 ± 9.9 30.4 ± 10.3 22.9 ± 7.1 22.9 ± 7.1
CEP 50.0 ± 20.4 10.4 ± 3.0 11.7 ± 3.5 5.8 ± 3.2
The anticancer activity of the CEP-based vac-
cination applied after the tumor transplantation
(therapeutic vaccination). Therapeutic vaccination
with CEP has been performed according to three dif-
ferent schedules of vaccination (described in details
in the Materials and Methods section). Any of immuni-
zation schedules appeared to be superior in transplan-
tation efficacy and latent period of LLC development,
as far as 85.9–89.6% of the vaccinated and control
mice developed tumors on the 9–11th day after the LLC
cells transplantation.
When it comes to the tumor volume, the most evi-
dent effect on tumor growth was observed in the group
of mice vaccinated according to the schedule #1 (Fig. 2).
Compared to the control group, the difference was sig-
nificant (p < 0.05) till the 20th day after the tumor chal-
lenge. The ITGI reached 53.13% on 14th day after the
LLC transplantation and was decreasing slowly till the
28th day of the experiment. Although the ITGI (42.1%)
observed at this time point of the follow-up period
(the 28th day after the LLC transplantation) was the
lowest for the group #1, it remained to be the highest
among the other groups. The tumor volume of the mice
vaccinated according to the two other schedules did
not differ significantly compared with the control group.
On day 28 after the tumor transplantation, all
the mice were euthanized to assess the metastasis
loading (Table. 4). The results of the group #1 were
out-standing. In this group, the lowest mean metasta-
ses number per group was recorded (0.05 < p < 0.1 com-
pared to the control group). The mean metastases
volume was by 54.4% lower than that in the control
group. So, the MII in group #1 reached 77.6% (per
group) or 66.3% (per mice bearing metastases) and
was the highest among all the treatment and control
groups. In other treatment groups, the results did not
differ significantly from that of the control group.
0
1000
2000
3000
4000
5000
14 18 20 25 28
Days after tumor transplantation
Tu
m
or
v
ol
um
e,
m
m
3
Control
#1
#2
#3
Fig. 2. The tumor growth kinetics in animals vaccinated with
CEP and control mice
It is worth mentioning that only in the group #1 all
the mice were still alive till the end of the experiment
(28 days after tumor transplantation). The worst sur-
vival rate was in the group #3 (here the vaccination
started after the tumor nodule could be palpable),
in which 50% of immunized mice died before the ex-
periment termination.
Table 4. Indexes of metastasis rate in animals vaccinated by different
therapeutic schedules and in control LLC-bearing mice
Group Metastases
rate, %
Volume
of metasta-
ses, mm3
Number of metastases
per mouse
bearing me-
tastases
per group
Control 85.7 ± 12.4 10.1 ± 5.4 10.4 ± 3.4 10.4 ± 3.4
#1 66. 7 ± 19.2 4.6 ± 3.9 4.5 ± 2.0 3.0 ± 1.6*
#2 80.0 ± 19.4 31.3 ± 15.7 17.7 ± 8.4 13.2 ± 7.6
#3 66.7 ± 27.2 13.2 ± 6.2 23.0 ± 8.5 15.3 ± 10.3
Note: *0.05 < р < 0.1 compared to the control group.
The anticancer activity of CEP applied after
the tumor resection (post surgical therapeutic
vaccination). As far as the most prominent anticancer
results were observed in the group of mice immunized
on the 1st, 7th and 14th days after tumor transplantation
(group #1), the same schedule was chosen to be ap-
plied in the study of post surgical therapeutic vaccina-
tion. Mice were transplantated with LLC cells (per foot);
on the 17th day after transplantation the tumor nodule
was removed. All the mice were divided into two groups.
The mice in the CEP group underwent vaccinations with
CEP on the 1st, 7th and 14th days after tumor resection.
On the 35th and 50th days after the tumor transplanta-
tion (the 18th and 34th days after the tumor removal,
respectively) the mice of both (control and treatment)
groups were euthanized to assess the metastases
burden (Table 5).
Table 5. Metastasis indexes in mice vaccinated with CEP after surgical removal of LLC
Group
The day 18 after tumor removal The day 34 after tumor removal
Metastases
rate, %
Volume of me-
tastases, mm3
Number of metastases Metastases
rate, %
Volume of me-
tastases, mm3
Number of metastases
per mouse be-
aring metastases per group per mouse be-
aring metastases per group
Control 72.7 ± 13.4 69.3 ± 25.8 16.5 ± 3.6 12.0 ± 3.5 66.7 ± 13.6 133.1 ± 98.5 4.2 ± 2.02 2.4 ± 1.12
CEP 27.3 ± 13.41 1.2 ± 0.91 3.7 ± 1.11 1.0 ± 0.61 15.4 ± 10.01 0.8 ± 0.4 1.5 ± 0.7 0.2 ± 0.2
Note: 1р < 0.05 compared to the control; 2р < 0.05 compared to the 18th day after tumor removal.
200 Experimental Oncology 37, 197–202, 2015 (September)
In this experimental setting, CEP showed evident
and long-lasting antimetastatic effect. Independently
on observation time, CEP immunization led to reduc-
tion of the metastases rate, metastases number and
volume. For example, on the 18th day after the tumor
removal, only 27.3% of the immunized mice had me-
tastases, while in the control group this index reached
72.7% (the difference was statistically significant).
The metastases volume in the group of vaccinated
mice was by 98.3% lower (p < 0.05) when compared
to the control mice. The mean number of metastases
per metastases-bearing mouse or per group in total
was statistically significantly lower in the group of CEP
vaccinated mice. So, MII reached 91.7% per mouse
and 96.9% per group.
On the day 34th after the tumor removal, mice in the
control group showed disease progression. For example,
the metastases volume increased by 1.92 times, com-
pared with the 18th day after the tumor removal. The mean
number of metastases slightly decreased possibly due
to the merging of small metastases. The metastasis rate
did not change significantly (66.7 ± 13.6 and 72.7 ± 13.4%
of control mice had metastases on the 34th and 18th days
after the tumor removal respectively).
On the other hand, mice immunized with CEP
showed stabilization of metastatic process. In particu-
lar, the metastases volume was 0.8 ± 0.4 mm3 (to com-
pare, it was 1.2 ± 0.9 mm3 on the 18th day after the
tumor removal); the metastases number per mouse
bearing it was 1.5 ± 0.7 (3.7 ± 1.1 on the day 18th
of examination). In control mice, metastasis rate in the
group of vaccinated mice did not differ significantly
from the previous point of observation.
So, we can assume that on the 18th day after the
tumor removal almost all the mice (of both control and
treatment groups) which were prone to develop me-
tastases developed them, as long as the metastasis
rate did not differ significantly on the 18th and 34th days
of observation. But the metastasis rate was statistically
lower in the CEP vaccinated group during all the experi-
ment (i.e. on the 18th and 34th days after tumor removal)
compared to the control. What is important, the vac-
cinated mice showed inhibition of metastases growth,
whereas in the control group the mean metastases vo-
lume increased by almost 2 times. As a result, the mean
metastases volume in the group of immunized mice was
by 94.4% lower than that in the control group. In the
CEP group, the MII calculated per group was equivalent
to 97.8%. So, the antimetastatic effect of CEP-based
vaccination was observed for a prolonged period of time
even after the termination of the vaccination.
DISCUSSION
So, as it was shown in the model of LLC, vaccination
with CEP appears to have anticancer and antimeta-
static effects. In the previous experiments it has been
shown that there were CEP-specific antibodies in the
blood serum of mice bearing different tumor strains
(LLC, sarcoma 37, Ehrlich carcinoma, melanoma
B-16) [16]. The presence of CEP-specific antibodies
in the blood serum of unimmunized tumor-bearing
mice may be explained by at least two reasons: poly-
specific antibody circulation [25, 26] and the pre sence
of some homologous proteins in CEP. It is known
that some proteins of chicken origin share homology
with mammals proteins, including that of human and
mice [9, 10, 12, 27–29]. The anticancer effect of CEP
seems to be based on the last assumption. Especially,
it looks possible when we consider the antitumor and
antimetastatic effects of CEP applied before the tu-
mor challenge — in so called prophylactic settings.
According to the prophylactic schedule which was
applied in the experiment, the tumor cell injection was
performed on the 30th day after the last immunization.
Till the 30th day after the CEP injection, the immune
response induced by the immunization was expected
to terminate [30], but immune memory cells had been
already established [31]. The immune memory is ca-
pable of mounting a rapid response to subsequent
antigen stimulation [32]. In the experiment, LLC cell
suspension in the dose sufficient to induce tumors
was used instead of the antigen re-challenge. Since
a statistically significant prolongation of the latent pe-
riod of tumor development in the groups of immunized
mice was observed, it points to the generation of the
rapid immune response to the cancer cells injection.
That is, the mice immunized with CEP or LLC-Ag in the
prophylactic mode mounted a rapid immune response
to cancer cells as if it was an antigen re-challenge.
Subsequently, the observed results indicate with
high probability that CEP contains some proteins which
share homology with LLC antigens and immunization
with CEP leads to immune memory formation. More-
over, in terms of its antimetastatic activity, vaccination
with CEP was much more effective than application
of LLC-Ag. This finding can be considered as an ad-
ditional demonstration that xenogeneic homologous
proteins are useful for breaking immune tolerance
towards autologous cancer antigens.
In case of therapeutic immunization, the anticancer
effect of CEP was evident only when applied at the very
early stage of tumor formation (24 h after tumor cells in-
jection, group #1), when tumor burden is minimal. When
vaccination was postponed to only 7 days (group #2)
the anticancer effect could hardly be observed. Fur-
thermore, vaccination with CEP has no anticancer ef-
fect when applied to mice with the already established
tumor (group #3). So, it can be concluded that without
prior tumor removal the application of anticancer vac-
cine based on CEP will have a minimal anticancer ef-
fect in clinical settings. On the other hand, it confirms
a generally acknowledged statement that benefit
of an anticancer vaccine is most evident when it is ap-
plied in earlier and less aggressive disease settings,
that is in settings of minimal residual disease [33, 34].
Owning to this, the third experiment — application
of CEP after the surgical resection of the tumor —
was conducted. In this case, CEP application had
a pronounced and long-lasting antimetastatic effect.
The number of metastases bearing mice and the mean
Experimental Oncology 37, 197–202, 2015 (September) 201
metastases volume were significantly reduced in the
group of treated mice. These effects were evident till
the 34th day after the tumor removal — the day of the
experiment termination. It should be mentioned that
the mean metastases volume in the CEP group was
60 and 168.5 times smaller than that of the control
group on the 18th and 34th days after the tumor re-
section respectively. MII was very high and reached
96.9 and 97.8% per group in total on the 18th and 34th
day respectively. It can be assumed that this vaccine
when applied after the surgical removal of a tumor may
dramatically improve therapeutic efficacy of cancer
treatment, as long as metastatic spread of a tumor
is the main death cause of cancer patients [35].
It has been shown that some genes or proteins
of chicken origin, when used as a xenogeneic vaccine,
can elicit anticancer effect or tumor specific immune
response. For example, xenogeneic vaccines based
on chicken HSP70 [11], MMP-2 [10, 14], Tie-2 [9]
or FGFR [12, 13] were effective in case of LLC [10, 14],
fibrosarcoma Meth A [13, 10], hepatoma H22 [9, 10],
melanoma B-16 [9], CT26 colon adenocarcinoma [14],
canine cancer [11]. Anticancer effects of CEP are com-
parable with these of the vaccines mentioned above.
Whether CEP contains some of abovementioned pro-
teins or its anticancer effect is based on other antigens
it remains to be elucidated.
CONCLUSION
Vaccination based on CEP exhibited both prophy-
lactic and therapeutic anticancer effects. The last one
is more pronounced when the vaccination starts shortly
after the primary tumor resection. In this case, the MII
reaches 91.7%. So, CEP are suitable to be used in xe-
nogeneic cancer vaccine construction.
REFERENCES
1. Naftzger C, Takechi Y, Kohda H, et al. Immune
response to a differentiation antigen induced by altered anti-
gen: A study of tumor rejection and autoimmunity. Proc Natl
Acad Sci USA 1996; 93: 14809–14.
2. Potebnya GP, Symchych TV, Lisovenko GS. Xenogenic
cancer vaccines. Exp Oncol 2010; 32: 61–5.
3. Sioud M, Sоrensen D. Generation of an effective anti-
tumor immunity after immunization with xenogeneic antigens.
Eur J Immunol 2003; 33: 38–45.
4. Seledtsov VI, Shishkov AA, Surovtseva MA, et al. Xeno-
vaccinotherapy for melanoma. Eur J Dermatol 2006; 6: 655–61.
5. Seledtsov VI, Niza NA, Felde MA, et al. Xenovaccino-
therapy for colorectal cancer. Biomed Pharmacother 2007;
61: 125–30.
6. Yuan J, Ku GY, Gallardo HF, et al. Safety and immunoge-
nicity of a human and mouse gp100 DNA vaccine in a phase I trial
of patients with melanoma. Cancer Immunity 2009; 9. 5 p.
7. Ginsberg BA, Gallardo HF, Rasalan TS, et al. Immu-
nologic response to xenogeneic gp100 DNA in melanoma pa-
tients: comparison of particle-mediated epidermal delivery with
intramuscular injection. Clin Cancer Res 2010; 16: 4057–65.
8. Eriksson F, Totterman T, Maltais AK, et al. DNA vac-
cine coding for the rhesus prostate specific antigen delivered
by intradermal electroporation in patients with relapsed pros-
tate cancer. Vaccine 2013; 31: 3843–8.
9. Luo Y, Wen YJ, Ding ZhY, et al. Іmmunotherapy
of tumors with protein vaccine based on chicken homologous
Tie-2. Clin Cancer Res 2006; 12: 1813–9.
10. Su J-M, Wei Y-Q, Tian L, et al. Active immunogene
therapy of cancer with vaccine on the basis of chicken homolo-
gous matrix metalloproteinase-2. Cancer Res 2003; 63: 600–7.
11. Yu W-Y, Chuang T-F, Guichard C, et al. Chicken
HSP70 DNA vaccine inhibits tumor growth in a canine cancer
model. Vaccine 2011; 29: 3489–500.
12. Shaojiang Zh, Fengying H, Shaoping Zh, et al. Vac-
cination with a recombinant chicken FGFR-1 bypasses immu-
nological tolerance against self-FGFR-1 in mice. J Huazhong
Univ Sci Technolog Med Sci 2006; 26: 389–91.
13. Shaoping Zh, Junzhi Zh, Shaojiang Zh, et al. Anti-
angiogeneic target therapy for cancer with vaccine based
on the recombinant chicken FGFR-1 in tumor-bearing mice.
J Huazhong Univ Sci Technolog Med Sci 2007; 27: 120–3.
14. Yi T, Wei Y-Q, Tian L, et al. Humoral and cellular
immunity induced by tumor cell vaccine based on the chicken
xenogeneic homologous matrix metalloproteinase-2. Cancer
Gene Therapy 2007; 14: 158–64.
15. Srinivasan R, Wolchok JD. Tumor antigens for cancer
immunotherapy: therapeutic potential of xenogeneic DNA
vaccines. J Transl Med 2004; 2. 12 p.
16. Symchych TV, Karaman OM, Voyeykova IM, et al.
Experimental approaches to elaboration on the diagnostic test
based on embryo proteins. Naukovi Zapysky NAUKMA 2010;
106: 29–32 (in Ukrainian).
17. Symchych TV, Karaman OM, Yudina OY, et al. Toxic
and immunomodulating effects evaluation of chicken embry-
onic proteins on Balb/c mice. Naukovi Zapysky NAUKMA
2009; 93: 31–6 (in Ukrainian).
18. Kozhemyakin YM, Kchromov OS, Filonenko
MA, et al. Scientific-practical recommendations on manage-
ment of laboratory animals and work with them. Kyiv, 2002.
179 p. (in Ukrainian).
19. Council Directive 2010/63/EU of 22 september
2010 on the protection of animals used for scientific purposes.
Official J Eur Commun 2010; 276: 33–79.
20. Isokawa K, Rezaee M, Wunsch A, et al. Identification
of transferrin as one of multiple EDTA-extractable extracellular
proteins involved in early chick heart morphogenesis. J Cell
Biochem 1994; 54: 207–18.
21. Greenberg CS, Craddock P. Rapid single-step mem-
brane protein assay. Clin Chem 1982; 28: 1725–6.
22. Cherdinceva NV, Kokorev OV, Konovalova NP,
Kagiya VT. Enhancement of cytotoxic and cytostatic activi-
ties of spleen cells and macrophages by radiosensitizing drug
АК-2123 in mice bearing Lewis lung carcinoma and treated
with cyclophosphan. Exp Oncol 1997; 19: 333–7 (in Russian).
23. Lakin GF. Biometry. Moskow: Vyschaya Shkola, 1980.
290 p. (in Russian).
24. Sydenko AV, Vyshnyakov VV, Isaev SM. Theory of statis-
tics. Manual. Moskow: MAKS-Press, 2011. 343 p. (in Russian).
25. Wing MG. The molecular basis for a polyspecific an-
tibody. Editorial review. Clin Exp Immunol 1995; 99: 313–5.
26. Dimitrov JD, Pashov AD, Vassilev TL. Antibody
polyspecificity: what does it matter? In: Lutz HU, eds.
Naturally occurring antibodies (NAbs). New York: Springer
Science+Business Media, 2012: 213–26.
27. Bassuk JA, Iruela-Arispe ML, Lane TF, et al. Mo-
lecular analysis of chicken embryo SPARC (osteonectin). Eur
J Biochem 1993; 218: 117–27.
28. Schneider J, Linares R, Martinez-Arribas F, et al.
Developing chick embryos express a protein which shares ho-
202 Experimental Oncology 37, 197–202, 2015 (September)
mology with the nuclear pore complex protein Nup88 present
in human tumors. Int J Dev Biol 2004; 48: 339–42.
29. Yamaguchi S, Iwata K, Shibuya M. Soluble Flt-1 (so-
luble VEGFR-1), a potent natural antiangiogenic molecule
in mammals, is phylogenetically conserved in avians. Biochem
Biophys Res Commun 2002; 291: 554–9.
30. Fahey JL, Sell S. The immunoglobulins of mice: the
metabolic (catabolic) properties of five immunoglobulin
classes. J Exp Med 1965; 122: 41–58.
31. Inamine A, Takahashi Y, Baba N, et al. Two waves
of memory B-cell generation in the primary immune response.
Int Immunol 2005; 17: 581–9.
32. Cellular and molecular immunology. In: Abbas AK,
Lichtman AH, Pillai Sh, eds. Saunders Elsevier; 6th ed,
2007: 215–42.
33. Bergman PJ. Anticancer vaccines. Vet Clin Small Anim
2007; 37: 1111–9.
34. Hale DF, Clifton GT, Sears AK, et al. Cancer vac-
cines: should we be targeting patients with less aggressive
disease? Expert Rev Vaccines 2012; 11: 721–31.
35. Hart I. The spread of tumors. In: Knowles MA,
Selby PJ, eds. Introduction to the cellular and molecular
bio logy of cancer. 4th ed. New York: Oxford University Press
Inc, 2005: 278–88.
Copyright © Experimental Oncology, 2015
|
| id | nasplib_isofts_kiev_ua-123456789-145488 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-12-07T16:36:07Z |
| publishDate | 2015 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | Symchych, T.V. Fedosova, N.I. Karaman, O.M. Yevstratieva, L.M. Lisovenko, H.S. Voyeykova, I.M. Potebnia, H.P. 2019-01-22T12:58:10Z 2019-01-22T12:58:10Z 2015 Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules / T.V. Symchych, N.I. Fedosova, O.M. Karaman, L.M. Yevstratieva, H.S. Lisovenko, I.M. Voyeykova, H.P. Potebnia // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 197-202. — Бібліогр.: 35 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/145488 The aim: To evaluate anticancer activity of vaccination with chicken embryo proteins (CEP) applied in different schedules. Materials and Methods: C57Bl mice were vaccinated with CEP before (prophylactic schedule) or after (different therapeutic schedules with or without preliminary tumor removal) the Lewis lung carcinoma cells transplantation. The latent period of tumor development, tumor volume and metastasis rate were evaluated. Results: Potent antimetastatic effect of CEP-based vaccination was seen in case of therapeutic regimen after primary tumor removal. The metastasis inhibition index (MII) reached 96.9 and 97.8% on 18th and 34th day after tumor removal, respectively. When CEP vaccination was performed in the settings of therapeutic regimen without primary tumor removal the anticancer effect was evident only if vaccinations started as early as 24 h after the cancer cells injections. The highest MII achieved in such condition was 77.6%, tumor volume in the group of vaccinated animals was by 53.1–42.1% lower than in the control tumor-bearing mice. CEP vaccination before tumor challenge (prophylactic immunization) led to a statistically significant prolongation of the latent period of tumor development, a reduction of tumor volume (35.8–48.8% compared to control unvaccinated mice) and a marked inhibition of metastasis (MII was 71.1%). Conclusion: Vaccination based on CEP exhibited both prophylactic and therapeutic anticancer effects. The last one is more pronounced when the vaccination starts shortly after the primary tumor resection. Key Words: chicken embryo proteins, anticancer activity, Lewis lung carcinoma. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules Article published earlier |
| spellingShingle | Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules Symchych, T.V. Fedosova, N.I. Karaman, O.M. Yevstratieva, L.M. Lisovenko, H.S. Voyeykova, I.M. Potebnia, H.P. Original contributions |
| title | Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules |
| title_full | Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules |
| title_fullStr | Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules |
| title_full_unstemmed | Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules |
| title_short | Anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules |
| title_sort | anticancer effectiveness of vaccination based on xenogeneic embryo proteins applied in different schedules |
| topic | Original contributions |
| topic_facet | Original contributions |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/145488 |
| work_keys_str_mv | AT symchychtv anticancereffectivenessofvaccinationbasedonxenogeneicembryoproteinsappliedindifferentschedules AT fedosovani anticancereffectivenessofvaccinationbasedonxenogeneicembryoproteinsappliedindifferentschedules AT karamanom anticancereffectivenessofvaccinationbasedonxenogeneicembryoproteinsappliedindifferentschedules AT yevstratievalm anticancereffectivenessofvaccinationbasedonxenogeneicembryoproteinsappliedindifferentschedules AT lisovenkohs anticancereffectivenessofvaccinationbasedonxenogeneicembryoproteinsappliedindifferentschedules AT voyeykovaim anticancereffectivenessofvaccinationbasedonxenogeneicembryoproteinsappliedindifferentschedules AT potebniahp anticancereffectivenessofvaccinationbasedonxenogeneicembryoproteinsappliedindifferentschedules |