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Bull Vet Inst Pulawy 54, 181-187, 2010
EFFECT OF BREWERS’ YEAST (SACCHAROMYCES
CEREVISIAE) EXTRACT ON SELECTED PARAMETERS
OF HUMORAL AND CELLULAR IMMUNITY IN LAMBS
ROMAN WÓJCIK
Department of Microbiology and Clinical Immunology, Faculty of Veterinary Medicine,
University of Warmia and Mazury in Olsztyn, 10-957 Olsztyn, Poland
[email protected]
Received for publication September 11, 2009
Abstract
The objective of this study was to determine the stimulating effect of the brewers’ yeast Saccharomyces cerevisiae dietary
supplement on selected parameters of specific and non-specific humoral and cellular immunity in lambs. The study involved 32
lambs aged 30 ±3 d, divided into two equal groups: control and experimental. Animals in the experimental group were fed a C-J
concentrate mixed with a prebiotic, the extract of dried brewers’ yeast containing 10%-15% MOS and 25%-30% β-1,3/1,6-D-glucan
in the amount of 3 g/kg of the concentrate. At the beginning of the experiment (day 0) and on the 15th, 30th, and 60th d of the study,
blood was sampled from the jugular vein to determine γ-globulin levels, lysozyme and ceruloplasmin activity, proliferative response
of blood lymphocytes (MTT) after stimulation with LPS or ConA, metabolic activity (RBA), and potential killing activity (PKA) of
phagocytes. As regards humoral immunity parameters, significantly higher γ-globulin levels and higher lysozyme and ceruloplasmin
activity were noted in the blood serum of experimental lambs supplemented with the yeast extract, in comparison with control lambs
not fed the supplement. No statistically significant differences in serum total protein were found between the control and
experimental groups. The analysis of cellular immunity indicators revealed significantly higher levels of RBA and PKA, and higher
MTT rates after stimulation with LPS or ConA in the experimental group, in comparison with the control group.
Key words: lambs, prebiotics, Saccharomyces cerevisiae, protein content, humoral immunity,
cellular immunity.
Owing
to
its
prebiotic
properties,
Saccharomyces cerevisiae can be widely used as a
natural productivity stimulator added to animals’ feed
(9). The beneficial effects of yeast in the nutrition of
ruminants have been noted in experiments on dairy cows
(8) and suckling lambs (22). The above experiments also
confirmed a stimulating effect of yeast on the health
status of animals. The results of studies involving lambs
(22, 35) indicate that β-1,3/1,6-D-glucan, a structural
component of the cellular wall of Saccharomyces
cerevisiae, plays an important role in this process, in
regard to the indicators of both non-specific humoral
immunity (19) and cellular immunity (35). The
immunomodulating effect of Saccharomyces cerevisiae
yeast is ascribed to mannan-oligosaccharides (MOS),
which build the yeast's cell wall structure (19). By
binding selected pathogens, MOS prevent them from
colonising the host's gastrointestinal system and support
their elimination by specialised immune cells (7, 12, 16).
So far no studies have been conducted concerning the
effect of dried yeast and yeast-derivative prebiotics on
immunity indicators in lambs.
The objective of this study was to determine the
effect of the extract of dried brewers’ yeast
Saccharomyces cerevisiae (Biolex-MB40) with an
increased β-1,3/1,6-D-glucan and MOS content on
selected parameters of humoral and cellular immunity in
lambs.
Material and Methods
Experimental design. Thirty-two suckling
Kamieniec breed lambs from a conservative herd, aged
30 ±3 d, were divided into two equal groups: I – control
and II – experimental. Both groups were identical in
terms of body weight on the second day of life, gender,
birth type, as well as the age of the ewes, to eliminate
possible differences in milk yield. Uniform feeding
standards were applied in both groups in line with lamb
nutrient requirements. Suckling lambs were fed haysilage of grass and legumes, and C-J concentrate. The
quantity of administered feed and leftovers was
monitored throughout the experiment. Experimental
lambs were fed the Biolex–MB40 (Leiber GmbH)
brewer's yeast (Saccharomyces cerevisiae) extract
containing 10%-15% MOS and 25%-30% β-1,3/1,6-Dglucan. The extract was mixed with C-J concentrate in
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the amount of 3 g/kg of the concentrate. C-J concentrate
doses, identical for both groups, were increased every 10
d by 0.05 kg/animal/d, starting from 0.15 kg/animal/d.
At the beginning of the experiment (day 0) and on the
15th, 30th, and 60th d of the study, blood was sampled
from the jugular vein to determine selected parameters
of humoral and cellular immunity.
Evaluation
of
non-specific
humoral
immunity parameters. Lysozyme activity was
determined by the turbidimetric method (25) modified
by Siwicki and Anderson (29), ceruloplasmin activity –
by the method developed by Siwicki and Studnicka (31),
total
protein
content
was
determined
by
spectrophotometry as described by Lowry et al. (17) and
modified by Siwicki and Anderson (29), and γ-globulin
level was determined by the precipitation method
modified by Siwicki and Anderson (29).
Lysozyme activity. Whole blood samples were
centrifuged for 5 min at 1,000 g to separate blood cells
from the serum. The serum was diluted 1:1 with
phosphate buffer, and 0.1 ml of the solution was placed
in microplate wells. Next, 0.5 ml of Micrococcus
lysodeikticus suspension (25 mg bacteria/100 ml of
phosphate buffer) (Sigma Chemical Co.) was added.
Absorbance was measured directly after the addition of
bacteria (E0) and after 1, 2, 3, and 30 min (final E). The
final absorbance was subtracted from the initial
absorbance (E0) to determine lysozyme activity with the
use of a standard curve. The standard curve was plotted
based on the optical density values for known lysozyme
concentrations.
Ceruloplasmin activity. Whole blood samples
were centrifuged for 5 min at 1,000 g to separate blood
cells from the serum. The following buffers were
prepared: 1) acetate buffer (pH 5.2, containing
crystalline acetic acid, sodium acetate trihydrate, and 15
mg of EDTA), 2) buffered substrate solution (0.2% pphenyldiamine (PPD) in acetic buffer), 3) sodium azide
solution (0.02% sodium azide solution in deionised
water). 0.5 ml of buffered solution was added to each of
two 16 x 100 mm test tubes immersed in a water bath at
37ºC. One test tube served as an experimental sample,
and the other as control. 50 µl of serum was added to the
experimental sample, which was incubated for 15 min at
37ºC. Next, 2 ml of a sodium azide solution was added
to the experimental and control samples. 50 µl of serum
was added to the control sample, and both samples were
mixed. The absorbance of the experimental sample was
measured at a wavelength of 540 nm, using the control
sample as a blind test. Ceruloplasmin activity was
determined with the use of the standard curve. The
standard curve was plotted based on the optical density
values for known ceruloplasmin concentrations.
Total protein level. Whole blood samples were
centrifuged for 5 min at 1,000 g to separate blood cells
from the serum. 5 µl of serum was placed in the wells,
and 25 µl of reagent A and 200 µl of reagent B were
added (Rio-Rad, Hercules, USA). Well contents were
gently stirred with a pipette. The microplates were
incubated at room temperature for 15 min. Next optical
density was measured in a microplate reader at 620 nm.
Total protein level was determined using a standard
curve as a reference. The standard curve was plotted
based on optical density values for known protein
dilutions.
γ-globulin level. Whole blood samples were
centrifuged for 5 min at 1,000 g to separate blood cells
from the serum. The optical density of total protein was
determined according to following procedure: 0.1 ml of
serum was placed in the microplate wells, and 0.1 ml of
12% polyethylene glycol (10,000 kD) (Sigma Chemical
Co.) suspended in deionised water was added. The
microplates were incubated at room temperature for 2 h,
and well contents were stirred continuously. The
microplates were centrifuged for 10 min at 5,000 g to
separate the γ-globulin fraction bound by polyethylene
glycol (plate sediment) from the remaining total protein
fraction, which constituted the supernatant. The optical
density of supernatant was measured in a microplate
reader at 620 nm. The optical density of supernatant was
subtracted from the optical density of total protein. γglobulin content was determined using a standard curve
(plotted earlier for total protein) as a reference, based on
the ability of γ-globulins to bind with polyethylene
glycol and precipitate.
Evaluation of non-specific cellular immunity
parameters. The metabolic activity of blood phagocytic
cells was determined based on the intracellular
measurements.
Respiratory burst activity (RBA) after
stimulation with PMA (Phorobol Myristate Acetate,
Sigma), was determined according to the method
described by Chung and Secombes (6) and adapted for
dogs by Siwicki et al. (30). The isolated cells were
resuspended in RPMI-1640 medium (Sigma) at 106
cells/mL. On 96-well U-shaped microplates, 100 µl of
the isolated blood leukocytes was mixed with 100 µl of
a 0.2% nitro blue tetrazolium (NBT, Sigma) solution in
0.2 M phosphate buffer (pH 7.2), and 1 µl of PMA at a
concentration of 1 mg/mL in ethanol was added. After
30 min of incubation at 370C, the supernatant was
removed from each well. The cell pellet was washed
with absolute ethanol, and three times with 70% ethanol,
and then was dried at room temperature. The amount of
extracted reduced NBT after incubation with 2M KOH
and DMSO (dimethylsulfoxide, Sigma) was measured
colorimetrically at 620 nm in a microplate reader (Tecan
Sunrise). All samples were tested in triplicate, and the
results are presented as mean values.
Potential killing activity (PKA) of
mononuclear (MN) phagocytes and polymorphonuclear
(PMN) phagocytes was determined in isolated blood
leukocytes stimulated with killed microorganisms,
according to the method presented by Rook et al. (26)
and adapted for dogs by Siwicki et al. (30). On 96-well
U-shaped microplates, 100 µl of leukocytes was mixed
with 100 µl of 0.2 % NBT in phosphate buffer (pH 7.2),
and 10 µl of killed Staphylococcus aureus strain 209P
(containing 106 bacteria) was added. The mixture was
incubated for 1 h at 370C and the supernatant was
removed. The cell pellet was washed with absolute
ethanol and three times with 70% ethanol, and then was
dried at room temperature. This was followed by the
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differences between groups was verified by the Student's
t-test with the use of GraphPad Prism 5 software.
addition of 2 M KOH and DMSO to each well. The
amount of extracted reduced NBT was measured at 620
nm in a microplate reader (Tecan Sunrise). All samples
were tested in triplicate, and the results are presented as
mean values.
Evaluation of specific cellular immunity
parameters. Lymphocyte proliferation rates after
stimulation with mitogens, concanavalin A (ConA), and
lipopolysaccharide (LPS), were determined by MTT
spectrophotometry (OD 570 nm) using (3-[4,5dimethylthiazol-2yl]-2,5-diphenyltetrazolium bromide –
3-[4,5-dimethyl-2-thiazol]-2,5-diphenyl-2H-tetrazolium
bromide), as described by Mosmann (23).
MTT (Sigma) was dissolved in PBS at a
concentration of 5 mg/ml and filtered. On 96-well
culture plates (Sarstetd, USA), 100 µl of blood
lymphocytes in RPMI - 1640 containing 10% FCS, 2
mM L-glutamine, 0.02 mM 2-mercaptoethanol, 1%
hepes buffer, and penicillin/streptomycin (100 U/100
µg/ml) was mixed with 100 µl of RPMI - 1640
containing mitogens ConA (5 µg/ml), PHA (10µg/ml) or
LPS (20 µg/ml). After 72 h incubation at 370C in a 5%
carbon dioxide atmosphere (Memmert Incubator), 50 µl
of MTT solution was added into each well, and plates
were incubated for 4 h at 370C. After incubation the
plates were centrifuged (1,400 g, 150C, 5 min).
Supernatants were removed and 100 µl of DMSO
(Sigma) were added into each well and incubated for 15
min at room temperature. After incubation, the
solubilised reduced MTT was measured colorimetrically
at 570 nm in a microplate reader (Tecan Sunrise). All
samples were tested in triplicate, and the results are
presented as mean values. The final results are presented
as the reactivity index (RI).
Statistical analysis. The obtained results were
processed statistically by a one-factorial analysis of
variance in an orthogonal design. The significance of
Results
The obtained results indicate a significant effect
of Biolex-MB40 on the analysed parameters of humoral
and cellular immunity in lambs.
An analysis of humoral immunity indicators–
lysozyme and ceruloplasmin activities (Table 1) –
revealed significantly (P≤0.01) higher values in the
experimental group fed a diet with the addition of dried
brewers’ yeast Saccharomyces cerevisiae than in the
control group administered feed without yeast
supplementation, over the entire experimental period. As
regards the serum levels of γ-globulins (Table 1), their
significant increase in the experimental group was
reported only on the 30th (P≤0.05) and 60th (P≤0.01) d of
the experiment in comparison with control group. In
comparison with day 0 a significant increase in both
ceruloplasmin and lysozyme activities (P≤0.01) was
noted only in the experimental group on the successive
days (15, 30, 60) of the experiment. Serum total protein
content showed no significant differences between the
experimental and control group (Table 1).
In regard to the investigated indicators of nonspecific cellular immunity – RBA and PKA of
phagocytes, and of specific cellular immunity - MTT
stimulated with LPS and ConA (Table 2) – a statistically
significant (P≤0.01 or P≤0.05) increase in their values
was observed in the experimental group in comparison
with the control group. The RBA in the group II was not
marked by a significant increase in comparison with the
group I only on the 60th d of the experiment. The RBA
and MTT stimulated with LPS and ConA (Table 2)
showed a significant increase (P≤0.01) only in the group
I on successive days (15, 30, 60) of the experiment in
comparison with day 0.
Table 1
Effect of Biolex-MB40 on the parameters of non-specific humoral immunity in lambs
Parameter
Group
0
x
Lysozyme activity
(mg/L)
Ceruloplasmin
activity (mg/L)
γ-globulin level (g/L)
Total protein content
(g/L)
I
II
I
II
I
II
I
II
0.79
0.79
31.75
31.20
30.22
34.11
57.18
57.10
15
SD
0.08
0.06
0.61
0.77
5.41
6.81
4.07
2.85
x
B
0.79
1.09 A**
31.28 B
37.18 A**
33.17
36.44
62.28
61.67
Experimental day
30
SD
x
0.09
0.77 B
0.09
1.14 A**
1.15
31.60 B
0.98 37.52 A**
2.83
31.61 b
5.82
35.44 a
3.88
57.30
2.24
56.17
60
SD
0.08
0.06
0.49
0.7
1.27
3.87
2.26
1.53
a, b - P≤0.05; A, B - P≤0.01; SD - standard deviation; I – experimental group; II – control group.
* P≤0.05 in comparison with experimental day 0,
** P≤0.01 in comparison with experimental day 0.
x
B
0.81
1.17 A**
31.22 B
37.48 A**
30.67 B
39.44 A
58.15
56.25
SD
0.06
0.04
0.61
1.38
3.13
3.83
3.22
3.75
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Table 2
Effect of Biolex-MB40 on the parameters of specific and non-specific cellular immunity in lambs
Experimental day
Parameter
0
Group
x
I
RBA (OD 620 nm)
II
I
PKA (OD 620 nm)
II
I
MTT-ConA
(RI)
II
I
MTT-LPS
(RI)
II
0.51
0.51
0.47
0.50
1.15
1.20
1.10
1.12
15
SD
0.06
0.02
0.04
0.04
0.08
0.15
0.05
0.11
x
SD
0.51
0.58
0.56
B
A*
1.30
1.55
b
a**
0.45
b
a**
1.02
1.36
30
B
A**
0.05
0.04
0.02
0.04
0.05
0.21
0.08
0.07
x
0.49
0.57
A**
0.45
0.54
B
A**
0.90
1.32
B
A
1.34
1.61
B
B
A**
60
SD
x
0.05
0.52
0.02
0.03
0.03
0.08
0.19
0.06
0.09
SD
0.02
B
0.02
A
0.03
b
0.12
a**
0.03
B
0.06
A**
0.07
0.56
0.43
0.54
1.35
1.53
1.03
1.33
0.05
**
Symbols as in Table 1.
Discussion
The conducted study was the first ever attempt
to determine the stimulating effect of a natural
immunostimulator – Biolex-MB40 containing increased
levels of β-1,3/1,6-D-glucan and MOS – on selected
indicators of specific and non-specific humoral and
cellular immunity in lambs.
Glucans are a group of compounds known as
glucose homopolymers. They are isolated from fungi,
yeast, and plants, including oat and barley. Glucans
derived from yeast and fungi have 1-3 bonds, and may
occasionally feature additional 1-6 branches, whose
number varies subject to glucan type. Glucans isolated
from barley and oat are mostly linear compounds
comprising regions with 1-4 bonds that separate smaller
structures with 1-3 bonds (1). The most frequently
described glucans that have been proven to have a
stimulating effect on the immune system are: β-(1,3)(1,6)-glucan extracted from Saccharomyces cerevisiae,
scleroglucan produced by Sclerotium glucanicum,
grifolan (GRN) isolated from Grifola frondosa, SSG
found in Sclerotinia sclerotiorum, and laminarin
extracted from Laminaria digitata (2). The biological
activity of β-glucan is determined by its origin,
occurrence frequency, isolation method, size (molecular
weight), physicochemical properties (such as solubility),
primary structure, shape, degree of branching, and
polymer charge (36). High molecular weight β-glucans
(e.g. zymosan) may directly stimulate leukocytes,
enhancing their phagocytic, cytotoxic, and antibacterial
activity, including the production of reactive oxygen
species and indirect nitrogen compounds. Low and
medium molecular weight β-glucans (e.g. phosphate
glucan) have a weaker effect on immune system cells.
Short β-glucans molecules (e.g. laminarin with
molecular weight of <5,000-10,000) are mostly inactive
(4).
Until recently, MOS were rarely investigated in
regard to their effect on the immune system of animals.
The conducted studies demonstrated that MOS
contributed to a higher production of antibodies (28, 32),
a drop in T cell number in peripheral blood (32, 24), a
decrease in haptoglobin concentrations (5), and had no
effect on the production of IL-6 (5).
In this study, a significant increase in lysozyme
and ceruloplasmin activities, and a rise in γ-globulin
levels were observed in the group of experimental lambs
fed diets with the addition of Biolex-MB40 in
comparison with the control group. No such effect was
noted as regards serum total protein (Table 1). Similar
results were reported by other authors. Kokoshis et al.
(14) demonstrated that after stimulation with β-glucan,
lysozyme
activity
grew
proportionally
with
phagocytosis, which was also enhanced by this
compound. β-glucan binds to the receptors on the cell
surface and activates transcription factors for plasma
proteins. β-glucan and MOS also directly contribute to
higher immunoglobulin levels and support the
proliferation of B cells (7, 12, 16, 28, 32). By
stimulating phagocytes to produce IL-1, IL-6, and TNFα, β-glucan affects the synthesis of acute phase proteins,
including ceruloplasmin. Guzdek and Rokita (10)
demonstrated a stimulating effect of curdlan sulphate
(sulphate derivative of curdlan –1,3-β-glucan) on the
levels of selected blood proteins, including
ceruloplasmin. According to the above authors, the
observed effect is caused mainly by the activation of
transcription factors (mostly NF-κB) inducing protein
synthesis. This observation validates the previous thesis.
The results of other studies (22) also point to an
increase in γ-globulin levels after the supplementation of
lamb diets with β-1,3/1,6-D-glucan. Krakowski et al.
(15) studied the immunostimulating effect of β-1,3/1,6D-glucan on pregnant mares and did not observe
significant differences between the total protein content
and γ-globulin levels in the offspring of mares fed
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glucan. However, an increase in the total protein content
and selected immunoglobulin levels in mare colostrum
was observed.
In this study, Biolex-MB40 also stimulated the
proliferative activity of T and B cells. β-glucan and
MOS bind with the C3R receptor on the surface of B
and T cells to induce a cascade reaction and activate the
NF-κB transcription factor. This factor induces the
expression of cytokines, mostly IL-2 and IL-4, which
stimulate the proliferation of B and T cells. NF-κB also
exerts an indirect effect by activating phagocytes that
remain in a mutual relationship with lymphocytes. The
results of this study, relating to the stimulating effect of
β-glucan and MOS on the proliferative activity of
lymphocytes (Table 2), are consistent with the findings
of other authors. Suzuki et al. (33) reported higher levels
of proliferative activity of B cells (stimulated with LPS)
and T cells (stimulated with ConA) in mice orally
administered SSG (β-1,3-glucan isolated from
Sclerotinia sclerotorium) for 5 and 10 consecutive days
in the amount of 40 and 80 mg/kg/d. Similar results
were reported by Szymańska-Czerwińska et al. (34) in a
study of calves orally administered Alphamune (MOS
and β-glucan) in daily doses of 14 g per animal. Zhao et
al. (37) noted increased activity of lymphocytes
stimulated with LPS and ConA following the
administration of a different glucan type, FPS-1 (1,6-βglucan found in Aconitum carmichaeli). Mao et al. (20)
studied the effect of β-glucan extracted from Astragalus
membranaceus on lymphocyte proliferation in pigs
twice immunised with LPS and in non-immunised pigs.
The lymphocytes of both immunised and nonimmunised pigs receiving β-glucan were marked by
higher proliferative activity stimulated by ConA. The
above authors also demonstrated β-glucan's contribution
to higher levels of IL-2 activity, thus confirming the
previous observations that β-glucan activates
lymphocytes through the stimulation of cytokine
synthesis.
In this study, the supplementation of lamb diets
with Biolex-MB40 enhanced the activity of monocytes
and granulocytes, leading to an increase in the RBA and
PKA of phagocytes (Table 2). RBA and PKA tests
evaluate changes in the oxygen metabolism of
neutrophil granulocytes, known as the respiratory burst,
and they are an indirect measure of the phagocytic
activity of neutrophils. Nevertheless, the above tests are
characterised by a different mechanism of neutrophil
activation. In the RBA test, the switching of cell
metabolism to glucose oxidation in the pentose cycle
and the activation of the membrane enzyme complex –
NADPH oxidase (nicotinamide adenine dinucleotide
phosphate-oxidase) takes place with the involvement of
phorbol ester (PMA) by the extra-receptor pathway,
with the omission of signal transduction at the receptorNADPH oxidase level, and through direct stimulation
(phosphorylation) of protein kinease C (PKC). Active
NADPH oxidase continues to catalyse oxygen reduction
to the superoxide anion radical O2-. As a substrate in
various biochemical reactions, this radical supports the
formation of other reactive oxygen species, including
hydrogen peroxide, hydroxyl radical, singlet oxygen,
and hyperchlorous acid. They have one or more lone
electron pairs, which make them highly reactive. Found
in phagolysosomes, they are toxic for bacteria, fungi,
parasites, and neoplastic cells, and they determine the
oxygen-dependent mechanism of microbial killing by
phagocytes. Hyperchlorous acid and chloramine are the
most potent antimicrobial oxidising agents produced by
neutrophils. In the PKA test, the activation of the
respiratory burst, as a result of the phosphorylation of
cytoplasmic components of NADPH oxidase, starts from
signal transmission at the ligand-receptor level, where
the phagocyted strain of Staphylococcus aureus binds to
the TLR2/TLR6 receptors of neutrophil granulocytes via
the ligand modulin (PAMP – pathogen associated
molecular patterns). Following the formation of
superoxide radicals, the added NBT is reduced to
insoluble formazan in the neutrophil cytoplasm.
The exact mechanism of β-glucan's and MOS's
impact on phagocyte functions has not been fully
elucidated. β-glucans and MOS bind with the C3R
complement receptor to activate phagocytes. By
becoming attached to specific receptors on the cell
surface, including the C3R complement receptor,
lactosylceramide (CDw17), Dectin-1, and selected
scavenger receptors (1, 3), β-glucan induces a reaction
cascade, which activates the nuclear factor κB (NF-κB).
NF-κB becomes attached to the promoter region of
cytokines, such as IL-1, IL-6, and TNF-α, to induce their
synthesis. It may also stimulate reactions, enhance the
expression of iNOS (nitrogen oxide ligase), and other
enzymes that participate in the processes of active
elimination of microorganisms, such as the production
of reactive oxygen species and lysozyme synthesis.
The results noted by other authors concerning a
stimulating effect of β-glucans on phagocyte activity in
various animal species are consistent with the findings
of this study involving the Biolex-MB40 supplement.
Luhm et al. (18) demonstrated that NFκB induced
higher levels of IL-8 and IL-1 synthesis in persons
supplemented with 1,3-β-glucan. Similar results were
noted by Kataoka et al. (13), who observed a strongly
activating influence of linear glucan (curdlan) on NFκB. Linear β-glucans bind with phagocytes not only
through the above receptors, but also through the TLR
receptor (Toll-like receptors) and the TIR domain
(similar to the domain in IL-1 receptors). Other studies
(21) point to a stimulating effect of soluble glucan PGG
(Betafectin®, β-1,3/1,6-D-glucan) on TNF-α and IL-1β
synthesis with the involvement of monocytes and
macrophages. An increase in superoxide anion
production was also noted. The above authors found that
PGG was able to stimulate the synthesis of the above
compounds only when immobilised on the carrier. They
argued that the above is explained by the fact that
macrophage activation requires the cross-coupling of
ligands with the receptor, as observed in molecular
(insoluble) β-glucans, such as β-1,3/1,6-D-glucan
investigated in this study. Sakurai et al. (27)
demonstrated a stimulating effect of another soluble β1,3/1,6-D-glucan (SSG) on the production of TNF-α,
nitrogen oxide, IL-1, and IL-6 by macrophages. The best
results were noted during co-stimulation with INF-γ,
186
which testifies to a weaker effect of soluble glucans.
Several years earlier, Hamuro et al. (11) confirmed the
positive effects of eight types of 1,3-β-glucans (lentinan
isolated from Lentinus edodes, pachyman extracted from
Poria cocos and its chemically modified derivatives) on
the cytotoxicity of macrophages. The above authors also
suggested another possible mechanism of phagocyte
activation. They argued that the studied β-glucans were
capable of activating the complement receptor in an
alternative manner, i.e. by activating its components, in
particular C3b. The activated complement receptor
stimulates the potential killing activity of phagocytes,
increasing their ability to eliminate pathogens. Dietary
supplementation of β-1,3/1,6-D-glucan (Biolex-Beta
HP) (35) stimulated phagocyte metabolism and the
intercellular elimination of pathogens in lambs.
The results of this study indicate that BiolexMB40 stimulates humoral and cellular immunity. These
findings offer a valuable incentive for further research
concerning the use of this supplement in the treatment of
impaired immunity in animals and humans, especially in
periods of increased susceptibility to bacterial and viral
infections. Biolex-MB40 can be safely used as an
effective immunostimulator without the risk of toxicity.
12.
13.
14.
15.
16.
17.
18.
References
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