International Immunopharmacology 7 (2007) 701 – 724

www.elsevier.com/locate/intimp

Review
Immunomodulating and anticancer agents in the realm of
macromycetes fungi (macrofungi)
Mohammad-Fata Moradali a , Hossein Mostafavi b,⁎,
Shirin Ghods a , Ghorban-Ali Hedjaroude a
a
Plant Protection Department, Faculty of Agriculture, Tehran University, Karaj, Iran
b
Organic Chemistry and Biochemistry Department, Faculty of Chemistry, Tabriz University, Tabriz, Iran
Received 26 November 2006; received in revised form 7 January 2007; accepted 8 January 2007

Abstract

Nowadays macrofungi are distinguished as important natural resources of immunomodulating and anticancer agents and with
regard to the increase in diseases involving immune dysfunction, cancer, autoimmune conditions in recent years, applying such
immunomodulator agents especially with the natural original is vital. These compounds belong mainly to polysaccharides
especially β -D-glucan derivates, glycopeptide/protein complexes (polysaccharide-peptide/protein complexes), proteoglycans,
proteins and triterpenoids. Among polysaccharides, β (1→3)-D-glucans and their peptide/protein derivates and among proteins,
fungal immunomodulatory proteins (Fips) have more important role in immunomodulating and antitumor activities.
Immunomodulating and antitumor activity of these metabolites related to their effects to act of immune effecter cells such as
hematpoietic stem cells, lymphocytes, macrophages, T cells, dendritic cells (DCs), and natural killer (NK) cells involved in the
innate and adaptive immunity, resulting in the production of biologic response modifiers. In this review we have introduced the
medicinal mushrooms' metabolites with immunomoduling and antitumor activities according to immunological evidences and then
demonstrated their effects on innate and adaptive immunity and also the mechanisms of activation of immune responses and
signaling cascade. In addition, their molecular structure and their relation to these activities have been shown. The important
instances of these metabolites along with their immunomodulating and/or antitumor activities isolated from putative medicinal
mushrooms are also introduced.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Macrofungi; Immunomodulating; Antitumor; Glycopeptide/protein complexes; Fips

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 702
1.1. Macrofungi characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 702
1.2. Macrofungi in medicine as immunomodulating and antitumor agents . . . . . . . . . . . . . . . . . . . . . 702
2. Immunomodulating and antitumor activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 703
3. Effects of macrofungi metabolites on hematopoietic stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 705

⁎ Corresponding author. Tel.: +98 4113393120; fax: +98 4113340191.

E-mail address: hmostafavi1@excite.com (H. Mostafavi).

1567-5769/$ - see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.intimp.2007.01.008
702 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724

4. Effects of macrofungi metabolites on the innate (non-specific) immunity . . . . . . . . . . . . . . . . . . . . . . 705

4.1. Effect on macrophages and APC (Antigen-presenting cell) activation . . . . . . . . . . . . . . . . . . . . . . 705
4.1.1. Effect on dendritic cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 705
4.1.2. Effect on NK cells (cytotoxicity and interferon production) . . . . . . . . . . . . . . . . . . . . . 706
4.1.3. Complement system and macrofungi metabolites. . . . . . . . . . . . . . . . . . . . . . . . . . . 706
5. Effects of macrofungi metabolites on the adaptive (specific) immunity . . . . . . . . . . . . . . . . . . . . . . . 706
5.1. Macrofungi metabolites as antigens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 706
5.2. Biologic response modifiers activation (cytokine production) and macrofungi metabolites . . . . . . . . . . 707
6. Major immunomodulating and antitumor agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 708
6.1. 1-Polysaccharides and glyco-conjugates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 708
6.2. 1-1-Polysaccharides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 708
6.2.1. D- and MD-fraction from G. frondosa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 709
6.2.2. 1-2-Glycoproteins (polysaccharide-protein complexes) . . . . . . . . . . . . . . . . . . . . . . 711
6.2.3. 1-3-Glycopeptides (polysaccharide-peptide complexes) . . . . . . . . . . . . . . . . . . . . . . 712
6.3. 1-4-Proteoglycans . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 712
6.3.1. GLIS from G. lucidum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 712
7. The structure-activity relationship . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 712
8. Chemical and structural modification of polysaccharides and their derivates . . . . . . . . . . . . . . . . . . . . . 713
9. β-D-Glucan receptor(s) and signaling cascade . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 714
10. 2-Terpenoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 715
11. 3-Fungal immunomodulatory proteins (Fips) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 716
11.1. Fips structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 716
11.2. Fips immonumodulating activities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 717
12. Clinical and experimental evidences for anticancer properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 717
13. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 719
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 720

1. Introduction
1.1. Macrofungi characteristics
Fungi are classified in the independent kingdom Fungi
among thorough organisms. They have the cells with true
nucleus (Eucaryotes), and reproduce by spores. They are
C-heterotrophic and obtain the essential nutrients through
breaking down organic substances and absorbing them.
The fungal body (thallus) is either a single cell or a
threadlike structure (hypha). A large number of branching
hypha together constitutes the mycelium which gives
raise to usually multiform spore-producing cells. In most
types of fungi, spore-producing cells (basidia, asci,
conidiophores) form a part of special structure made up
hyphal tissue and called the fruit body (sporocarp).
Although the fruit bodies are given to extraordinary
variations in shape, size and coloring, these do remain
fairly constant in individual fungus groups. The “mush-
room” is a popular term that applied for the fungus has
observable fruit body with the naked eye. Nowadays,
fungi kingdom includes main five phyla including
Chytridiomycota, Zygomycota, Glomeromycota, Asco-
mycota and Basidiomycota. They differ with each other
principally in ontogenesis, phylogeny, sexual and asexual
reproduction, how producing spores, elements constitut-
ing cell wall etc. The mushrooms are also called
“Macromycetes” (a Latin name) in opposite the term
“Micromycetes” (a Latin name for the fungi that are
invisible with the naked eye). Among the Fungi kingdom
the Macromycetes arranged in the phylum Basidiomycota
and a few numbers in the Ascomycota and evolutionary
Basidiomycetes are known as the higher fungi. In human
life fungi are well known and they have important roles in
different fields close to human life as friend or foe.
1.2. Macrofungi in medicine as immunomodulating and
antitumor agents
Applying fungi as medicines dates back to 3000 BC
when macrofungi are used to remedy diseases by
mankind especially in the traditional oriental therapies
and after discovering of Penicillin (1929), fungi were
regarded as rich sources of natural antibiotics and other
bioactive compounds. Macrofungi such as Ganoderma
lucidum, Lentinus edodes, Fomes fomentarius, Fomi-
topsis officinalis and many others have been used to
remedy different diseases for hundreds of years in
China, Japan, Korea and the Slav regions [1]. Now-
adays, constituent molecules of macrofungi organelles
and secondary metabolites are known as bioactive com-
pounds that belong to polysaccharides, glycoproteins,
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Table 1 involved in the innate and adaptive immunity, resulting in

The different groups of bioactive compounds isolated from macrofungi the production of cytokines. The therapeutic effects of
Main compound Example Medical Reference these compounds such as antitumor and antiinfective
group potentiality activity and suppression of autoimmune diseases have
Polysaccharides Grifolan Immunomodulator [61] been associated in many cases with their immunomodu-
Lentinan Antitumor [111] lating effects. Immunomodulators (biological response
Schizophyllan Antiviral [112]
modifiers) can be effective agents in treating and preven-
Antimicrobial [113]
Polysaccharide- PSP Antitumor [35] ting diseases and illnesses and with regard to the increase in
peptide diseases involving immune dysfunction and cancer
PSK Antiviral [114] and along with them infectious diseases in recent years,
Antimicrobial [115] medical researchers and clinicians are interested in
Cytotoxic [116]
Immunotherapy as well as the discovery of novel
Proteins Fips Immunomodulator [87]
Ganoderic acids [117] immune-potentiators and compounds with powerful
Ganoderiol Anti-HIV activity [118] remedy potential without side effects, pathogenic resis-
Ganoderenic acids antitumor [119] tance or affecting normal cell division especially for
Lucidenic acids Cytotoxic [119] anticancer and antiviral agents. In this review article we
Terpenoids Ganolucidic acids Histamine release [120]
have introduced the macrofungi metabolites with immu-
inhibition
Lucidumols Antihypertension [121] nomoduling and antitumor activities and then have
(ACEinhibitor) demonstrated their effects on innate and adaptive immunity
Ganoderols Anti-inflammatory and also mechanisms of activation of immune responses
Applanoxidic acids and signaling cascade. In addition, their molecular structure
Steroids Polyoxygenated Cytotoxic [122]
and their relation to these activities have been shown. The
derivates of
ergosterol important instances of these metabolites along with their
Antitumor [123] immunomodulating and/or antitumor activities isolated
antibacterial [124] from putative medicinal macrofungi are also introduced.
Fatty acids linoleic acid Antimutagenic [125] Hence we have mentioned the scientific names of some
palmitic acid antibacterial [126]
putative medicinal macrofungi that are more studied for
11-octadecanoic
acid finding immunomodulatory and antitumor properties.
Organic Bis-β-carboxyethyl- Antitumor [127]
germanium germanium immunomodulating 2. Immunomodulating and antitumor activity
sesquioxide
Nucleotides Adenosine Platelet aggregate [128]
inhibition
Compounds that are capable of interacting with the
Polyacetylenic Biformyne, As antibiotic [129] immune system to upregulate or downregulate specific
compounds agrocybin Nemotinic aspects of the host response can be classified as immu-
acid, marasmin, nomodulators or biologic response modifiers. Whether
quadrifidins certain compounds enhance or suppress immune res-
ponses can depend on a number of factors, including dose,
proteoglycans, terpenoids, fatty acids, proteins, lectins, route of administration, and timing of administration of
etc that possess certain medicinal properties (Table 1). the compound in question. The type of activity can also
Particularly, they can be added to the diet and used depend on their mechanism of action or the site of activity
orally that they are considered as a safe and useful [2]. In this way these compounds' affect on the different
approach for disease treatment. These compounds are cells types involved hematpoietic stem cells, innate (non-
found in fruiting bodies, mycelia and spores and culture specific) and adaptive (specific) immune systems and also
broth of macrofungi. Several major substances with cytokine networks and signaling pathways. Many com-
immunomodulating and/or antitumor activity have been pounds isolated from macrofungi that are known as
isolated from them. These include mainly polysacchar- immonumodulators that almost related to polysaccharides
ides, glycopeptide/protein complexes, proteoglycans, and their peptide or protein derivates and Fips. Also in
proteins, triterpenoids. some cases triterpenoids have closed to immunomodulat-
Immunomodulating activity of compounds isolated ing activity. These compounds are also worthwhile an-
from medicinal macrofungi, related their effects to act on tiinfective and antitumor agents.
immune effecter cells such as hematpoietic stem cells, A wide range of antitumor activity is proven for these
lymphocytes, macrophages, T cells, DCs, and NK cells fungi. Numerous reports have documented the ability of
704 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724

these compounds especially β-D-glucan derivates to non-
specifically activate cellular and humoral components of
the host immune system so that they increase functional
activity of macrophages, mononuclear cells, and neutro-
phils [3–5]. Although the body's defense against microbial
attack and against spontaneously arising malignant tumor
cells comprises a dynamic orchestrated interplay of innate
and acquired immune responses. Innate immunity (having
macrophages, neutrophils, NK and DCs as gatekeepers), is
regulated by chemical-messengers or cytokines and by
activation of inflammatory and acute phase responses [6].
The mononuclear phagocyte system (e.g., macrophages
and monocytes), DCs and certain lymphocytes (e.g., NK
cells) play a number of important roles including the re-
cognition and destruction of abnormal cells. Stimulated
macrophages and NK cells produce cytokines such as
interferons, interleukins and others that are targeted to-
wards destroying cancer cells. Specific immunity to ab-
normal cells or tissues includes humoral (e.g. generates
antibodies) and cell-mediated immunity (also promotes
inflammatory responses and ultimately kills infected or
abnormal cells). So a fully functional immune response is
critical to the recognition and elimination of tumor cells.
With regard to effects of macrofungi metabolites and ext-
racts on antitumor effector cells, in the recent years, me-
dical researchers have attempted to identify macrofungi
that have compounds that are capable to stimulate com-
ponents of immune system toward discovering more ef-
fective antitumor agents especially with natural origination
and with more safety. In this way the basic strategy under-
lying immunomodulation is to identify aspects of the host
response that can be enhanced or suppressed in such a way
as to augment or complement a desired immune response.
For example lentinan is understood as a pure β-D-glucan
derivate isolated from L. edodes that affects and stimulates
the key immune mechanisms mediate destruction of tumor
cells (Fig. 1). Such mechanisms are also looked upon for
peptide or protein derivates of β-D-glucan [6]. Of course, it
should not be forgotten that there are other pathways and
mechanisms that macrofungi can prevent and suppress
tumor cell growth. In this way, it has proved that different
compounds of G. lucidum act via different pathways

Fig. 1. Mechanisms of antitumor activity of lentinan as a β-D-glucan (according to Chihara [6]). Mac, macrophages; TL(H), T-lymphocyte (helper);
NK, natural killer cells; IL-1, -2 and -13, interleukin-1, -2, -13; CSF, colony-stimulating factor; MAF, macrophage-activating factor; PC-TL, pre-
cytolytic T lymphocyte; CTL, cytolytic (cytotoxic) T lymphocyte; BL, B lymphocyte.
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
recognized as enhancing detoxification of carcinogens,
increased expression and activity of Phase II enzymes,
inhibition of organ exposure of carcinogens due to reduced
absorption or increased excretion, decreased expression
and activity of Phase I enzymes, decreased formation of
toxic metabolites and adduct formation with macromole-
cules, antioxidative and radical-scavenging effect, antipro-
motion effect, antiproliferation, induction of differentiation,
direct cytotoxicity, induction of cell-cycle arrest, antipro-
liferation and modulation of signaling transduction mole-
cules, antiprogression and tumor growth inhibition,
antimetastasis, and anti-angiogenesis [7].
Chihara was one of the first ones that reported the
antitumor properties of the macrofungi. He stated that
lentinan “was found to almost completely regress the solid
type tumors of Sarcoma 180 and several kinds of tumors
including methylchloranthrene-induced in synergic host-
tumor system A” [8]. The antitumor effect of lentinan was
originally confirmed by using Sarcoma 180 transplanted
in CD-1/ICD mice. Later, it showed prominent antitumor
activity not only against allogenic tumors such as Sarcoma
180, but also against various synergic and autochthonous
tumors, and it prevented chemical and viral oncogenesis.
3. Effects of macrofungi metabolites on hematopoietic
stem cells
Many cells involved in the immune response are
derived from undifferentiated hematopoietic stem cells
in the bone marrow and fetal liver. These differentiate
into various cell lineages under the influence of mi-
croenviromental factors such as cell-to-cell interactions
and the presence of soluble or membrane-bound cyto-
kines. In other words hematopoiesis is regulated by
several different cytokine growth factors produced by
bone marrow stromal cells, T cells, and other cell types.
Various metabolites, especially polysaccharides isolated
from medicinal mushrooms that showed to affect bone-
marrow cells (BMCs) and to induce hematopoiesis. It is
proved that these metabolites are able to differentiate the
BMCs to Colony-forming units granulocytes-macro-
phages (CFU-GM) and erythroid burst-forming units
(BFU-E) that will be explained in the follow sections [9].
4. Effects of macrofungi metabolites on the innate
(non-specific) immunity
4.1. Effect on macrophages and APC (Antigen-presenting
cell) activation
Macrophages are involved at all stages of the immune
response. They function in phagocytosis, antigen proces-
705
sing and presentation, secretion of cytokines, and
antibody-dependent cell-mediated cytotoxicity.
The effects of macrofungi extracts and metabolites
on macrophages have been extensively studied in vitro
and in vivo. It has been proven that some macrofungi
extracts can activate macrophages to produce various
mediators. Polysaccharides isolated from these fungi
have a role as APCs and stimulate them to produce
certain cytokines and subsequence of these events is the
activation of other immune cells. Also after contact of
these antigens with macrophages, they mediate process-
es as APCs that involved in the cell-mediate immunity
that will be explain in later sections.
Macrophages have an important role in the initial
response to infection before T- and B-cell enhanced
immunity can act. They act as a rapid protective that can
respond before T-cell-mediated amplification has taken
place. Then they take part in the initiation of T-cell
activation as APCs. APCs leads to T cells respond to
antigen and they release cytokines which activate
macrophages. Following macrophage activation, anti-
microbial, antitumor and production of other cytokines
take place. Macrofungi extracts can mediate these
events. For instance, a glycoprotein, PSPC, isolated
from Tricholoma giganteum could restore and increase
phagocytic function of macrophages of the tumor-
bearing mice. It could also exhibit indirect cytotoxicity
against P815 and L929 by activating macrophages to
release the mediators, such as nitric oxide (NO) and
tumor necrosis factor α (TNF-α) [10].
4.1.1. Effect on dendritic cells
DCs are bone marrow-derived immune accessory
cells that are found in epithelial and lymphoid tissues
that are morphologically characterized by thin membra-
nous projections. They also function as APCs for native
T lymphocytes. Different macrofungi metabolites espe-
cially β-D-glucans and their derivates are capable of
inducing the maturation of bone marrow-derived DCs
and also increase the membrane molecules in them.
Similar to effect of these polysaccharides on macro-
phages as an antigen, DCs also affected by them and
stimulation of cell-mediate immunity occurs. Subse-
quently processes activated in this immunity leads to
activation of antimicrobial and antitumor effector cells
and thus these compounds can be effective to induction
of tumor immunity. Besides activating native T cells,
they can directly activate native and memory B-cells.
DCs at different stages of differentiation can regulate
effectors of innate immunity such as NK cells and NK T
cells. The induction of tumor immunity can be initia-
ted by the effectors of innate immunity and further
706 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
developed by cells of adaptive immunity, with DCs
playing a central regulatory role [9].
4.1.2. Effect on NK cells (cytotoxicity and interferon
production)
NK cells are a subset bone marrow-derived lympho-
cyte that are capable of lysing or killing infected or
tumor cells. Cytotoxicity and cytokines production such
as Interferon-γ (IFN-γ), TNF-α, and granulocyte
macrophage-colony stimulating factor (GM-CSF) that
can modulate natural and specific immune responses are
two relevant functions of NK cells. They osmotically
lyse target cells and induce apoptotic cell death. They
also have an important role in the inflammatory
response so that they can enter sites of inflammation
where they can be stimulated by a cytokine Interleukin-
(IL-) 12 that is produced by activated macrophages.
Some macrofungi metabolites exhibit stimulating
effects on NK cells. They are capable to stimulate
secretion of IL-2 that as known as stimulating of
proliferation of NK cells. Because of NK cells serve as a
rapidly acting first-line defense mechanism that is
involved in the early destructive events following
tumor implantation, stimulation of these cells by these
metabolites is very important in tumor immunity. Not
only IL-2 production induced by these metabolite but
also induction of cytokines and immune factors occurs
[1,2,6]. For instance, oral administration of SSG (a
polysaccharide) isolated from Sclerotinia sclerotiorum
to mice had enhanced the activities of NK cells in spleen
[11].
4.1.3. Complement system and macrofungi metabolites
Complement is a system of serum and cell surface
proteins that interact with one another and with other
molecules of the immune system to generate important
effectors of innate and adaptive immune responses.
Components of the complement system (i.e. activated
components C3a, C3b through to C9) mediate and
amplify immune reactions. Following the release of
chemotactic factors and histamine C3a this induces
considerable inflammation and tissue damage at the
sites of reactions with antibodies. Residual C3b
component bound to the antigen–antibody complexes
attaches to C3b receptors present on macrophages and
thus acts as an opsonin, promoting enhanced phagocy-
tosis. Although there are a few reports concerning the
effects of macrofungi metabolites on complement
system but it is reported that some β-D-glucans (e.g.
lentinan) and glycoproteins and glycopeptides are
capable of activating a complement system and inhibit
tumor growth via alternative pathway [9]. It is eluci-
dated that inhibitory activity of GU-P (a polysaccharide
isolated from Grifola umbellata) and PSK on sarcoma
180 solid tumor growth is due to the C3 activation [12].
Also, an alkali extract isolated from cultured mycelium
of G. lucidum activated classical and alternative path-
ways of a complement system [7].
5. Effects of macrofungi metabolites on the adaptive
(specific) immunity
5.1. Macrofungi metabolites as antigens
Affecting of macrofungi metabolites on the cells,
cytokines and other factors involved in the adaptive
immunity are well known especially for β-glucans and
their derivates and proteoglycans. Concerning these
affecting, it is understood that mentioned metabolites,
act as the antigens, play role in the first step of activation
signaling pathway in the APCs. Subsequent of this
activation and signaling is humoral- and cell-mediate
immunity induction. As mentioned above, in virtue of
appearing of certain different receptors on the APCs and
displaying of Th cells to recognize and interact with
antigen-major histocompatibility complex (MHC) II
molecule complex, various cytokines (IL-12 and IL-4)
are produced and they affect on activation of B cells, Tc
cells, macrophages and other T cells that their results are
as inflammation, antimicrobial and antitumor activities
(Fig. 2). For example, a partially purified polysaccharide
isolated from Antrodia camphorata via promoting a
Th1-dominant state and killer activities and increasing
IL-12 secretion act as an antitumor fungal polysaccha-
ride [13]. Also it is showed that β-glucans have various
effects on the Th1- or Th2-dependent antibody sub-
classes and, in particular, that SSG form S. sclerotiorum
induces the development of Th1 cells via the IL-12
pathway. Stimulating Th1 to produce INF-γ is another
immunomodulating activity that this group of macro-
fungi can cause [14]. Activation Th1 immune response
due to the increased level of IL-12 and -18 by
polysaccharides isolated from Agaricus brasiliensis is
also reported [15].
Thymus-derived regulatory T-cell populations, in-
cluding naturally occurring CD4+CD25+T cells and
inducible IL-10 or TGF-β-producing TR/TH3, develop
in the periphery from TH cells depending on the
tolerance-inducing micro-environment in which these
T cells reside [9]. The downstream immune response
is chosen depending on which subtype of T cell is
activated, which means that the proportion of the
activated sub-types influences phylaxis immunity and
antitumor immunity. This control system is also affected
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724 707

Fig. 2. Schematic representation of the affecting of immunomodulatory macrofungi metabolites on the adaptive immune system leading to activation
of antimicrobial and antitumor pathways. Ag, antigen; PPCs, polysaccharide-peptide/protein complexes; FIPs, fungal immunomodulatory proteins;
APC, antigen-presenting cell; TLR, toll-like receptor; CR1, complement receptor 1; AC, activated complement; NF-kB, nuclear factor kappa B; CD,
cluster designation; MHC II, major histocompatibility complex; TCR, T-cell receptor; NO, nitric oxide; IL, interleukin; IFN-γ, interferon γ; LT,
lymphotoxin; TH, helper T cell; NK cell, natural killer cell; CTL, cytotoxic T lymphocyte.

by the production of IL-1β, IL-12, and IL-18 by APC
[9,16].
5.2. Biologic response modifiers activation (cytokine
production) and macrofungi metabolites
Cytokines are known as biologic response modifiers
that modulates inflammation, immunity and hematopoi-
esis. They are produced during the activation and ef-
fector phases of innate and adaptive immunity and
serves to mediate and regulate immune and inflamma-
tory responses. Many macrofungi metabolites especially
polysaccharides and their derivates and Fips affect on
cytokine production (known as multi-cytokine indu-
cers). Hence these compounds are also known as bio-
logical response modifiers as known as cytokines. These
compounds with affecting cytokine production are cap-
able to induce various changes in immune system. It is
proved that these metabolites can increase messenger
ribonucleic acids (mRNAs) encoding cytokines. For
example, the antitumor activity of GL-B (a polysaccha-
ride from Ganoderma) was derived from promoting
mRNA expression of TNF-α and IFN-γ, resulting in
TNF-α and IFN-γ release [17]. Synthesis of cytokines
happened following recognition of these metabolites
by cell surface receptors. This event leads to the
708 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724

Table 2
Some cytokine-production stimulating or suppressing compounds belong to different groups isolated from macrofungi
Agent Group IL-1 IL-2 IL-3 IL-4 IL-6 IL-8 IL-12 IL-18 LT TNF-α IFN-γ GM-CSF MIF
D-fraction PS SU ST ST ST ST
Grifolan PS ST ST ST
PG101 PS ST SU ST
lentinan PS ST ST ST ST ST ST ST ST
PL PS SU SU ST
SSG PS ST
PSK GP ST ST ST ST ST ST
PSP GP ST ST
LZ-8 Fip ST ST
Fip-vvo Fip ST ST ST
ST, stimulating; SU, suppressing; PS, polysaccharide; PPC, GP, glycopeptide; Fip, fungal immunomodulating protien.

transcription of genes responsible to code cytokines.
Cytokines often influence the action of other cytokines
and act upon many different cell types. For instance, it is
showed that many β-D-glucans induce to produce IL-12.
IL-12 is known as a macrophage-derived activator of
NK cell cytolytic function, although it is appreciated to
be a potent inducer of IFN-γ production by T cell as well
as NK cells. Consequently, macrofungi metabolites are
able to induce to stimulate or suppress cytokines
production (Table 2) that mediates innate immunity
and includes antiviral cytokines (e.g. IL-12 and IL-15),
pro-inflammatory cytokines (e.g. TNF-α, IL-1, IL-6 and
chemokines) and regulatory cytokines (e.g. IL-10).
To evaluate immunomodulating activity of macro-
fungi extracts contain polysaccharides, it is seen that
L. edodes and Grifola frondosa increased TNF-α
production approximately from 500–700 pg/ml and
300–500 pg/ml and NO production 4–7 μM and 5–
7 μM respectively. They finally decreased TNF-α level
to 500 and 260 pg/ml and NO level to 5 and 4 μM
respectively through the growth periods in murine peri-
toneal macrophages [18]. GLP known as a polysaccha-
ride from G. lucidum could induce a marked increase in
the gene expression levels of interleukin IL-lα (2-fold),
IL-lβ (3-fold), TNF-α (2-fold), IL-12 p35 (up to 6-fold),
and IL-12 p40 in the splenocytes. In the macro-
phages, GLP promoted a remarkable increase in the
gene expression levels of IL-lβ (2.5- to 3-fold), TNF-α
(up to 6-fold), and GM-CSF (up to 2-fold) [19].
Studies on the lentinan showed that it increase the
gene expression level of IL-1β and IL-1α cell lines [20].
Also, IL-1 is not produced without this stimulation and
this cytokine is known to have a wide range of biological
activities on many different target tissues, including B
cells, T cells and monocytes. Increasing of production of
IL-1 causes inducing of production of various other
cytokines such as TNF-α and this cytokine induces lysis
of malignant cells and regression of some animal tumor
[20]. It is indicated that the β-glucan-related polysac-
charides of the macrofungi activate macrophages to
release NO which is an important chemical messenger
for the induction of many biological responses [7].
6. Major immunomodulating and antitumor agents
6.1. 1-Polysaccharides and glyco-conjugates
Numerous bioactive polysaccharides, glycoproteins,
glycopeptides, and proteoglycans from macrofungi are
considered as immunomodulators affecting on prolifer-
ation and differentiation of immune cells and cytokines,
interleukins and receptors production due to recognition
these compounds by the certain receptors located on the
leukocytes and other immune cells that lead to enhance
the innate and cell-mediate immune responses. In virtue
of these activities also induction of different types of
antitumor effector cells such as cytotoxic T cells, NK
cells and macrophages occur. In addition to these
activities some of them possess antiviral, antibacterial,
antifungal and antiprotosoal activities [21]. Macrofungi
are rich in such components in their cell wall structures.
These compounds structurally are divided as following:
6.2. 1-1-Polysaccharides
Polysaccharides are carbohydrate polymers that can be
found abundantly in higher fungi cell wall. They involve
different chemical compositions including β-glucans,
hetero-β-glucans, heteroglycans, α-manno-β-glucan com-
plexes that are found in macrofungi and all have
showed immunomodulating and antitumor properties
(Table 3). Among them β-glucans are most important
polysaccharides with immunomodulating and antitumor
activity. β-glucans are glucose polymers that can exist as a
non-branched (1→3)-β-linked backbone or as a (1→3)-β-
linked backbone with (1→6)-β-branches (Fig. 3) and they
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724 709

Table 3 and ribose that may combined with other components.

Some immunomodulating and antitumor polysaccharides isolated Glycans are other polysaccharides that have been found in
from macrofungi
macrofungi. These polysaccharides, in general, contain
Scientific name Bioactive compound Reference units other than glucose in their backbone. They are
Lentinus Galactoglucomannan [130] classified as galactans, fucans, xylans, and mannans by
edodes the individual sugar components in the backbone.
Inonotus Xylogalactoglucan [109]
More than 100 types of polysaccharides with
obliquus
Polyporus Xyloglucan [109] biological activities have been isolated from the fruit
confluens body and mycelia of G. lucidum and most of them have
Flammulina Galactomannoglucan [131] a molecular weight range from 4 × 105 to 1 × 106 in the
velutipes primary structure [22]. Lentinan, schizophyllan (also
Ganoderma Glucogalactan, Arabinoglucan [132]
called SPG, sonifilan, sizofiran), grifolan or GRN, SSG,
tsugae
Ganoderma β-(1→3)-glucuronoglucan, [133] GLPS, D- and MD-fraction, PL, PG101, CA1, SCG are
lucidum Mannogalactoglucan macrofungi polysaccharides are more studied for
Hericium Galactoxyloglucan, Mannoglucoxylan, [109] immunomodulating and/or antitumor activities.
erinaceum Glucoxylan, Xylan
Pleurotus Xyloglucan, Mannogalactoglucan, [134]
6.2.1. D- and MD-fraction from G. frondosa
pulmonarius Mannogalactan, Glucoxylan
Grifola Mannoxyloglucan, Xyloglucan, [132] MD-fraction enhances BMCs growth and differenti-
frondosa β-(1→6); β-(1→3)-glucan, ation into the CFU-GM. This compound that obtained by
Mannogalactofucan further purification of D-fraction has enhanced directly
Agaricus Mannogalactoglucan, β-(1→6); [109] CFU-GM response of BMCs progenitors and has en-
blazei α-(1→3)-glucan, α-(1→4); β-(1→6)-
hanced recovery of the CFU-GM response after doxo-
glucan, α-(1→6); α-(1→4)-glucan,
Riboglucan, Glucomannan, β-(1→2); [133] rubicin induced hematopoietic suppression [23]. Thus,
β-(1→3)-glucomannan, β-(1→6); these results can suggest that this compound can reduce
β-(1→3)-glucan hematopoietic suppression induced by chemotherapy.
In different studies, it is elucidated that antitumor
occur as a primary component in the cell walls of higher activity of D-fraction and MD-fraction relate to their
fungi in the great deals. Heteroglucans side chains contain effects on immune cells and factors that have important
glucuronic acid, xylose, galactose, mannose, arabinose role in the antitumor activity. These compounds are

Fig. 3. Schematic representation of the molecular structure of β-(1→3)-D-glucan (a), α-(1→3)-D-glucan (b), 1,6-Monoglucosyl-branched 1,3-β-D-
glucan (c), polysaccharide peptide/protein complex (d).
710 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
capable of releasing IL-12 from macrophages and DCs
and acts on T lymphocytes and NK cells to stimulate
INF-γ production and cytolytic activity. Monitoring of
levels of NK cell cytotoxic activity in cancer patients
receiving D-fraction has showed that elevated levels of
cytotoxic activity were maintained for one year. To in-
vestigate and elucidate the mechanisms of this activity,
examination in the tumor-bearing mice showed that D-
fraction can markedly suppress tumor growth with
increase TNF-α and INF-γ production from spleen cells
and a significant increase in TNF-α expression in NK
cells [24,25]. Thus clinical study on patient with lung
and breast cancer has showed that D-fraction is able to
enhance and maintain peripheral blood NK cell activity
[24]. Also it is showed their control of the balance
between T lymphocyte subsets Th1 and Th2. This
compound decreases the activation of B cells and
promotes the activation of Th cells, resulting in
enhanced cellular immunity. It also induces the
production of INF-γ, IL-12p70, and IL-18 by whole
spleen cells and lymph node cells, but suppressed that of
IL-4. These results suggest that antitumor activity of D-
fraction arose affecting on IL-12 production from
macrophage and lead to increase NK cells and establish
TH1 dominance which induces cellular immunity [26].
6.2.1.1. Grifolan from G. frondosa. Grifolan is a
macrophage activator which augments cytokine pro-
duction without dependence on endotoxins [27] and it
enhances mRNA level of IL-6, IL-1 and TNF-α of
macrophages [28].
6.2.1.2. PL from Phellinus linteus. PL is an acidic
polysaccharide that is introduced as an immunomodula-
tor, anti-inflammatory and antitumor agent. PL can mo-
dulate circulating cytokine responses in lippolysaccharide
(LPS)-treated mice and administration of this compound
in vivo decreased IL-2 and TNF-α production in
splenocytes and enhance spontaneous cell apoptosis in
macrophages and lymphocytes stimulated with LPS in
vitro [29]. Also this compound can induce maturation of
bone-marrow-derived DCs and readies them for T cell-
mediated immune responses and increase membrane
molecules including MHC class I, II, CD80+, and CD86+,
and IL-12p70 in DCs [30]. The inhibitory effect of PL on
the growth of MCA-120 tumor cells was associated with
its immunoregulating properties including the induction
of IL-12 and IFN-γ production leading to a TH1 domi-
nant state [30]. Also the administration of PL could in-
duce immunomodulating and antitumor activities via
maturation of CD11c+CD8+DCs in tumor-bearing mice.
Also PL selectively activates murine B cells [31].
6.2.1.3. PG101 from Lentinus lepideus. This com-
pound can enhance CFU-GM and BFU-E. These results
that proved in the study on radiation-treated mice suggest
that this compound can recover the radiation-damaged
bone marrow system very efficiently along with it
increases the levels of IL-1β, IL-6, and GM-CSF and also
reduce the level of TNF-α [32]. Such damages including
dysregulation of cytokine expression and increase of
the level TNF-α in the tissue injury are caused by ra-
diation. Thus PG101 can be as a supplement or a major
therapeutic for bone marrow system is damaged.
6.2.1.4. Lentinan from L. edodes. Many researches
have been done on immunomodulating and antitumor
activity of lentinan. This compound is known as a T cell-
oriented adjuvant [33]. The skewing of TH1/TH1
balance to TH1 by lentinan is directed through the
distinctive production of IL-12 versus IL-6, IL10, and
PGE2 by peritoneal macrophages depending on intra-
cellular glutathione redox status [34]. Lentinan induces
non-specific cytotoxicity in macrophages and stimulates
cytokine production and is able to activate the alternate
complement pathway in vitro and it opsonised with C3b
is considered to bind to complement receptor type 1 or 3
(CR1 or CR3) on monocytes. It does not bind to
lymphocytes. Lentinan augments the secretion of IL-1,
colony-stimulating factors (CSF), migration inhibition
factor (MIF), IL-3, IL-6, IFN-γ and generation of
cytotoxic T cells (CTLs) and NK cell activity in the
presence of IL-2 [35].
Study on preoperative intratumor administration of
lentinan for gastric cancer cases showed that lentinan
administered directly into tumors decreased the ratio
of suppressor–inducer T cells and suppressor T cells
tended to increase the ratio of CTLs and IL-2 production
of lymph node lymphocyte [36]. It has been shown to
augment the activities of natural killer cells, lympho-
kine-activated killer cells and CTLs. It can also activate
macrophage differentiation and increases response in
delayed-type hypersensitivity against tumor antigen
[37].
Results obtained of patient orally administered
L. edodes fruit body suggest that tumor growth
inhibition by this macrofungi is at least partly due to
the enhanced production of the superoxide anion by
macrophages or prevention of deterioration of immuno-
phagocytosis on the tumor-bearing state. The cytotoxi-
city is potentiated by oral administration of L. edodes
fruit bodies is that of T-cells, so that it accelerate the
induction of cytotoxic T cells and are very similar to the
mechanisms of lentinan injected intraperitoneally into
mice. Its action is host mediated [38] (Fig. 1).
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724 711

6.2.1.5. SSG from S. sclerotiorum. — It is reported Table 4

that SSG can induce the development of TH1 cells via Some structural characteristics of β-D-glucans and glycopeptides and
glycoproteins isolated from macrofungi
the IL-12 pathway [39].
Compound Original Molecular Molecular
structure weight (KDa)
6.2.1.6. GLPS from G. lucidum. GLPS has activated
BALB/c mouse B cells and macrophages, and also could Schizophyllan Schizophyllum 1,6-Monoglucosyl- 350
commune branched 1,3-β-D-
induce IL-1beta production by peritoneal macrophages
glucan
[40]. It could promote the maturation of cultured murine Grifolan Grifola 1,6-Monoglucosyl- 500
bone marrow-derived DCs and the immune response frondosa branched 1,3-β-D-
initiation by DCs [41]. glucan
D-fraction Grifola 1,6-Monoglucosyl- 1000
frondosa branched 1,3-β-D-
6.2.1.7. β-glucan fraction CA1 from Sparassis crispa.
glucan
Oral administration of the CA1 has enhanced the hema- Lentinan Lentinus 1,6-Monoglucosyl- 500
topoietic response in cyclophosphamide (CY)-induced leu- edodes branched 1,3-β-D-
kopenic mice assessed by white blood cell count. The rate glucan
of leukocyte recovery in CY-administered mice was dif- SSG Sclerotinia 1,6-Monoglucosyl- ≥2000
sclerotiorum branched 1,3-β-D-
ferent in each population, such as granulocyte, monocyte,
glucan
NK cell, Â cell, Ò cell, and so on. Administration of CA1 PSK Coriolus 1,3 and 1,6- 100
modulated the recovery rate of each population. In in vitro, versicolor monoglucosyl-
CY-treated spleen cell culture, IL-6 and INF-γ production branched 1,4-β-D-
was enhanced by CA1 treatment. These facts strongly glucan with binding
to aspartic, glutamic
suggested that the enhanced hematopoietic response by
and other acidic
CA1 is due to enhanced cytokine production [42]. amino acids
PSP Coriolus Resemble to PSK 100
6.2.1.8. SCG from S. crispa. It is proved that SCG versicolor structure but is riched
enhances the hematopoietic response. In cyclopho- in glutamic and
aspartic acids
sphamid-(CY)-induced leukopenic mice, SCG recov-
PSPC Tricholoma Consisted of 150
ered monocytes and granulocytes in the peritoneal lobayense galactose, glucose,
cavity, liver, spleen, and bone marrow. The ratio of NK mannose, fucose,
cells and γδT cells in the liver, spleen, and peritoneal arabinose and
cavity was also increased [43]. rhamnose with
binding to aspartic,
glutamic and other
6.2.1.9. Galactomannan from Morchella esculenta. acidic amino acids
This compound enhances macrophage activation. At
3.0 μg/mL the galactomannan increased nuclear factor
kappa B (NF-kB)-directed luciferase expression in THP- molecules such as polypeptide and proteins etc.
1 human monocytic cells to levels of 50% of those Glycoproteins also consist of a protein core that
achieved by maximal activating concentration (10.0 μg/ surrounded with numerous glucan chains (Fig. 3) as
mL) of LPS [44]. these chains bind to protein moiety through O- or N-
glycosidic bones (Table 4). β-glucan-protein, α-glucan-
6.2.1.10. Fucogalactan from Sarcodon aspratus. This protein, heteroglycan-protein complexes are antitumor
carbohydrate at 50 μg/mL could increase the release of glycoproteins that found in macrofungi.
TNF-α and NO in macrophages of mice in vitro Among glycoproteins isolated from macrofungi
approximately 4.3 times than lentinan at 500 μg/mL PSPC and ATOM are well known for immunomodulat-
[45]. It is suggested that fucogalactan can contribute to ing and/or antitumor activities.
antitumor activity in tumor-bearing host as well as
immunomodulating effects. 6.2.2.1. ATOM from Agaricus blazei. It is suggested
that tumor growth inhibitory effect of ATOM is ap-
6.2.2. 1-2-Glycoproteins (polysaccharide-protein parently due to immunological host-mediated mechan-
complexes) isms so that it is caused by increasing the number of
Polysaccharide polymers can reach maximum com- peritoneal macrophages, the phagocytosis of polysty-
plexity when they are covalently attached to other rene latex beads and the proportion of the third
712 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
component of complement (C3)-positive fluorescent
cells in the tumor-bearing mice [46].
6.2.3. 1-3-Glycopeptides (polysaccharide-peptide
complexes)
This group is structurally similar to glycoproteins but
has a smaller chain of amino acids (Fig. 3). PSP and PSK
(Table 4) are well known for immunomodulating and
antitumor activities and they are commercially considered
to develop immunomodulating and anticancer agents.
6.2.3.1. PSK and SPS from Coriolus versicolor. PSK
is classified as biological response modifier that stim-
ulates T-cell activation and induce IFN-γ and IL-2 pro-
duction. The biological activity is characterized by
their ability to increase white blood cell counts, IL-2
production and delayed-type hypersensitivity reactions.
PSK stimulates production of differentiation-inducing
factor (DIF) from macrophages, and activates the function
of polymorphonuclear leukocyte (PMN). PMN plays a
prominent role in the overall course of infectious diseases
and it has a role in the defense against intra- and extra-
cellular pathogens such as bacteria, viruses and some
protozoa. Also, it is clarified PSK induces gene expression
of some cytokines such as TNF-α, IL-1, IL-8 and IL-6 in
vivo and in vitro [47]. It is reported that PSK promotes the
phenotype and functional maturation of DCs derived from
human CD14+ mononuclear cells and can resolve the
immunosuppressive state of a cancer-bearing host and
may be associated with DCs maturation directly [48].
PSK injected directly into human stomach tumors
prior to surgery was taken up specifically by DCs
located in and around the tumors [49]. Aside, this
compound activates killer cells in vivo. Instillation of
PSK into a human gastric tumor mass prior to respective
surgery causes T cells around the site to become tumor-
infiltrating and develops significantly enhanced cyto-
toxic "killer" activity directed at the tumor. Similar
findings have been obtained with a 14-day course of
PSK in bladder cancer patients [50]. PSK also activates
human NK cells in culture at concentrations reached in
the blood by normal oral dosing of 3 g per day [50,51].
Similar to PSK, PSP is classified as biological response
modifier that stimulates T-cell activation and induces IFN-
γ and IL-2 production. In mice with suppression of IL-2
production from cyclophosphamide toxicity, PSP supple-
mentation has restored IL-2 production to normal [52].
Also it is known to stimulate DCs and macrophage or
other immune phagocytic activity in vivo. When mice
were given charcoal intravenously, then fed PSP, both the
phagocytic activity of cells in the blood and the clearance
of the charcoal from the circulation were significantly
accelerated [53]. PSP stimulates lymphokine-activated
killer (LAK) cell proliferation by itself at relatively low
concentrations and in the absence of IL-2 [54].
6.2.3.2. Polysaccharide-polypeptide complexes from
G. lucidum. The fungus G. lucidum (in fruit body
and spore) contains kinds of glycopeptide complexes
with antitumor and immunostimulating activities.
Some of them stimulate the expression of cytokines,
especially IL-1, IL-2 and INF-γ [55,56].
6.3. 1-4-Proteoglycans
This group of glyco-conjugates is proteoglycans that
is known as a special class of glycoproteins that are
heavily glycosylated. They consist of a core protein with
one or more coavalently attached glycosaminoglycan
chain(s).
Glycosaminoglycan molecules are long unbranched
polysaccharides containing a repeating disaccharide
unit. The disaccharide units contain either of two
modified sugars-N-acetylgalactosamine (GalNAc) or
N-acetylglucosamine (GlcNAc) and an uronic acid
such as glucuronate or iduronate. Glycosaminoglycan
molecules are highly negatively charged molecules,
with extended conformation that imparts high viscosity
to the solution. Proteoglycans are remarkable for their
diversity includes different cores, different numbers of
glycosaminoglycans with various lengths and composi-
tions. GLIS is well known as a macrofungi proteoglycan
that has immunomodulating activity. This compound
contains carbohydrates and protein in a ratio of 11.5:1 so
that the carbohydrate portion is formed by seven different
monosaccharides, predominantly D-glucose, D-galactose,
and D-mannose in the molar ratio of 3.0:1:1 [57].
6.3.1. GLIS from G. lucidum
GLIS is proteoglycan that is introduced as a B-cell
stimulating factor. This compound stimulates B lym-
phocyte activation, proliferation, differentiation and
production of immunoglobulin. The activation of B
cells by GLIS may be associated with the expression of
protein kinase C (PKC) α and PKC γ in B cells. This
compound stimulates the proliferation of mouse spleen
lymphocytes, resulting in a three-fold to four-fold
increase in the percentage of B cells [57].
7. The structure-activity relationship
PSs have high capacity for carrying biological infor-
mation because they have great potential for structural
variability. Despite the structural and functional
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
similarities of these glucans, they differ in their
effectiveness against specific tumors and in their ability
to elicit various cellular responses, particularly cytokine
expression and production [58]. The structure of β-D-
glucans has a relationship with the binding characteristics
with receptors that the triple helical solution conforma-
tion, molecular weight, branching ratio, solubility in water
and charge of the glucan polymer are important
determinations in receptor ligand interaction and anti-
tumor activity [59]. For example it is proved that TNF-α
release by macrophage is induced only by β-D-glucans
with high molecular weights and lower branching ratios
[59,60]. Also, it is showed that high molecular weight
glucans appear to be more effective in antitumor activity
than those of low molecular weight [61]. Also bioactive β-
D-glucans present a triple-strand helical structure with
right winding [62]. Triple helix conformation of β-glucans
(Fig. 4) and presence of hydrophilic groups located on the
outside surface of the helix are important for their
biological activities whether immunomodulating or
antitumor activity. Also strong antitumor activities are
found in various hetero-β-D-glucans having a (1→3)-β-D-
glucan chain as the active site such as β-D-glucan,
glucurono-β-D-glucan, arabinoxylo-β-D-glucan, xylo-β-D-
glucan, manno-β-D-glucan and xylomanno-β-D-glucan, as
well as their polypeptide complexes.
Fig. 4. Schematic representation of the triple helix structure of β-(1→3)-D-glucans comparison to other helix structures (a), the higher structure of a β-
(1→3)-D-glucan formed as triple helix structure.
713
8. Chemical and structural modification of
polysaccharides and their derivates
Furthermore, it is proved that increased water
solubility favors enhanced antitumour activity while
the location of substitute groups would also be
important. For example, when the alkali-insoluble,
branched (1→3)-β-D-glucan isolated from Auricularia
auricula-judae was modified by controlled, periodate
oxidation, borohydride reduction, and mild, acid
hydrolysis, resulted in increasing water solubility by
having covalently linked polyhydroxy groups attached
at O-6 of the (1→3)-linked D-glucosyl residues, and
exhibited potent antitumor activity significantly, while
in the native state had no such activity [63].
They cause to improve such polysaccharides to more
effect and their clinical qualities. The main procedures
used for chemical improvement are: Smith degradation
(oxydo–reducto-hydrolysis), formolysis, and carboxy-
methylation [21]. For example, the carboxymethyl
group with low degree of substitution (DS b 0.28) (DS
is a measure of the average number of hydroxyl groups
on each anhydroglucose unit (AGU) which are
derivatized by substituent groups) is introduced as
the best choice on the improvement of immunostimulat-
ing activity of (1→3)-α-D-glucan [64] (Fig. 3) and
714 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
modification of D-glucosyl group of side chains of β-D-
glucans to polyol groups can enhance antitumor activity
[65]. Another finding for carboxymethylation was done
for a polysaccharide with linear structure of (1→3)-α-D-
glucan isolated from Amanita muscaria so that degree
of substitution (DS) of carboxymethyl groups was 0.95
and the substituents were located at O-2, at O-4, at O-6,
at O-2 and O-6, and at O-4 and O-6 on glucose, resulting
potent antitumor activity against sarcoma 180 in mice,
although the native polysaccharide had little effect [66].
Enzymatic reactions also can improve polysacchar-
ides activities. Amylase, cellulose, and protease are
enzymes that used for this purpose. In this way, linear
low molecular α-(1→4)-glucans obtained after enzy-
matic reduction of side chains and protein component
(active hexose correlated compounds) that shows
immunomodulating and anticancer properties [67].
9. β-D-Glucan receptor(s) and signaling cascade
The innate immune system identifies infectious
agents or compounds by using of pattern-recognition
receptors. These receptors recognize the macromole-
cules of pathogens and they called pathogen-associated
molecular patterns. Because of polysaccharide's large
molecular mass, these compounds cannot penetrate cells,
so the first step in the modulation of cellular activity is
binding to immune cell receptors. It is proved that there
are fungal pattern-recognition molecules for the innate
immune system. The mechanism by which the innate
immune system recognizes and responds to fungal cell
wall carbohydrates is a very complex and multifactorial
process [68]. It is clarified that several β-glucan receptors
mediate these activities such as Complement receptor3
(CR3, αM β2-integrin, CD11b/CD18) [69,70], lactosyl-
ceramide, glycosphingolipd [71], scavenger receptors
[72], dectin-1 [73], and tool-like receptors (TLR)-2 and
TLR-4 [74].
CR3 occurs on monocytes, macrophages, neutroph-
lis, NK cells and follicular DCs and is an important
receptor and adhesion molecule. They mediate a variety
of adhesive and cytotoxic functions that are dependent
upon its ability to bind a multiplicity of ligands such as
iC3b, ICAM-1, clotting factory X, certain bacteria and
fibrinogen [69].
It serves as the leukocyte β-glucan receptor [75]
through one or more lectin sites located on the portion of
the α-subunit (CD-11b) known as the CD11b I-domain
that contains the binding sites for iC3b, ICAM, and
fibrinogen [69]. Thus β-glucans mediate cytotoxic and
phagocytic responses by binding to these sites and are
one of the first microbial response modifiers for which
the cellular mechanism of action has been defined at the
specific receptor level. Elsta et al. had found that
opsonized zymosan particles as a derivation of β-
glucans stimulate platelet-activating factor synthesis by
monocytes and they had suggested that CD11b/CD18
facilities binding of the particle and that a β-glucan
receptor transduces the activation signal [76]. platelet-
activating factor has been implicated in the pathogenesis
of septic shock, anaphylaxis, asthma, immune-mediated
tissue injury, and other inflammatory conditions.
Lentinan has been found to bind to monocytes via
CR1, CR3 and may also bind via a beta-glucan receptor.
Lentinan induces non-specific cytotoxicity in macro-
phages and stimulates cytokine production and is able to
activate the alternate complement pathway in vitro and
it opsonised with C3b is considered to bind to CR1 or
CR3 on monocytes. However, lentinan does not bind to
lymphocytes [35].
β-D-glucan can also help override the normal
resistance of iC3b-opsonized tumor cells to the cytotoxic
activation of phagocyte and NK cell CR3, allowing this
important effecter mechanism of the complement system
to function against tumor cells in the same way that it
normally functions against bacteria [70].
Dectin-1 is a type II transmembrane protein and
mammalian cell surface receptor for β-1, 3-D-glucan and
has the typical amino acid sequences of C-type lectins
[77] and is important for recognizing fungal invasion
[78]. It is broadly expressed with highest surface
expression on monocytes, macrophages and neutro-
phils. Also it expressed by dendritic cells and a sub-
population T cells at lower levels [79]. C-type lectins
play important roles in the innate immune response by
recognizing microbial saccharides.
In the study on dectin-1 to recognize the schizo-
phyllan, it is proved that dectin-1 has a carbohydrate
recognition domain consisting of six cysteine residues
that are highly conserved in C-type lectins. C-type
lectins recognize sugar ligands through this carbohy-
drate recognition domain with Ca2+ dependence [78].
This receptor is specific for the (1→3)-β-D-glucosyl
linkage. SPG possess a 1,6-monoglucosyl branch on a
(1→3)-β-D-glucosyl main chain, which forms triple
helix conformation in a physiological solution [80]. The
biologic response transduced by dectin-1 is dependent
upon the TLR pathway. It is determined that dectin-1 in
combination with a β-glucan-enriched zymosan deriv-
ative enhances TLR-2-mediated cell activation [81].
Also it is indicated that for the activation of NF-kB and
induction of TNF-α, dectin-1 needs to cooperate with
TLR-2 (Fig. 5), possibly as part of a heterodimer with
TLR-6 [82]. NF-kB comprises a family of inducible
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
Fig. 5. Schematic representation of the β-glucans recognition by certain receptors on the cell surface and activation of NF-κB leading to transcription
of many genes in both innate and adaptive immune responses.
transcription factors that serve as important regulators of
the host immune and inflammatory response. NF-kB
factors are important in the transcription of many genes
in both innate and adaptive immune responses.
It has been reported that dectin-1 ligation with zymosan
results in tyrosine phosphorylation of the receptor's
immunoreceptor tyrosin-based activating motif (ITAM)-
like signaling motif, generating intracellular signals that
mediate phagocytosis and activation of nicotinamide
adenine dinucleotide phosphate (NADPH) oxidase [83].
In addition to the inflammatory response, dectin-1 binds to
lymphocytes and augments their mitogenic response by
cross-linking T-cell receptors [84].
Recognition of β-glucans by antigen-presenting cells
such as DCs or macrophages can augment pathogen-
specific T-cell activation. Thus β-glucans may be a link
between innate and adaptive immune responses. Despite
the intracellular events that occur what after glucan-
receptor binding has not been fully clarified. However
more studies are required to determine which receptor(s)
715
are essential of the various immunobiological effects
ascribed to β-glucans.
10. 2-Terpenoids
Terpenes are composed from 5-carbon units named
isoprene that they are not with saponification property.
Hitherto, many terpenes are isolated from plants, fungi,
and marine organisms etc. In among terpenes, triterpe-
noids have been found exclusively from macrofungi that
are famous compounds with biological activities and
medicinal properties and they are produced in higher
fungi (Basidiomycetes). Among triterpenoids highly
oxidized lanostane-type triterpenoids are the further type
of triterpenes are isolated from family Polyporaceae and
Ganodermataceae (Fig. 6). They have different biolog-
ical activities especially antiinfectives, cytotoxic and
immunomodulating activities. Ganoderic acids, Gano-
derenic acids, Ganodermic acids, Applanoxidic acids,
Ganoderals, Ganoderols, Lucidone, Ganodermanontriol,
716 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724

Fig. 6. Some lanostane-type triterpenoids isolated from Ganoderma lucidum.

Ganodermanondiol, are of basidiomycetous triterpe- 11. 3-Fungal immunomodulatory proteins (Fips)

noids [85]. Structures of these compounds have a
lanostane skeleton and they are classified into 10 groups
based on their carbon numbers and state of oxidation.
The researches done on G. lucidum have showed that
antitumor activity of such triterpenoids is similar to β-D-
glucans do in some pathways. These compounds, for
instance, could activate NF-kB pathway and modulate
Ras/Erk, c-myc, CREB protein and mitogen-activated
protein kinases [86]. So, these activations can lead to
other immune activations against tumor cells.
In spite of fact that many triterpenoids have been
known of macrofungi, few works have been done to
discovery mode of action of anticancer and immuno-
modulating effects of terpenoids.
11.1. Fips structure
Hitherto, four proteins with relatively similar struc-
ture isolated from macrofungi that have showed immu-
nomodulating activity. These proteins, Fips, are found
merely in the mushrooms including LZ-8, Fip-gts, Fip-
fve and Fip-vvo isolated from G. lucidum, Ganoderma
tsugae, Flammulina velutipes and Volvariella volvacea,
respectively. Fips have a molecular mass of 13 kDa
and share high amino acid sequence homology. Alig-
ment of these proteins revealed 44 % identity and 42%
homology for approximately 110 amino acid residues
(Fig. 7). They are rich in β-structure by secondary

Fig. 7. Comparison of sequences of Fips isolated from macrofungi.

 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
Fig. 8. Amino acid sequence and secondary structure of Fip-gts (this
structure was predicted by Lin et al. [87]).
structure prediction, and contain seven β-strands,
two α-helixes, and one β-turn (Fig. 8). The amphi-
pathic α-helix is a common structural motif, which is
found in a number of functional proteins or peptides
and involved in various functions such as glucagon
binding to its receptor, plasma apolipoproteins solu-
bilization of lipids, antimicrobial peptide disintegra-
tion of bacterial cells, and signal peptide targeting to
mitochondria [87].
11.2. Fips immonumodulating activities
Fips are mitogenic in vitro for human peripheral
blood lymphocytes (hPBLs) and mouse splenocytes,
and induce a bell-shaped dose-response curve similar to
that for lectin mitogens. Activation of hPBLs with Fips
results in the increased production of IL-2, IFN-γ and
TNF-α molecule associated with ICAM-1 expression.
They can also act as immunosuppressive agents. These
proteins could prevent systemic anaphylactic reactions
and significantly decrease foot-pad edema during the
Arthus reactions in vivo [87]. Study on LZ-8 showed
that it is similar to the variable region of immunoglob-
ulin heavy chain both in its sequences and in its
predicted secondary structure by sequencing studies. It
appears to be related to an ancestral protein of the
immunoglobulin superfamily [88,89]. LZ-8 is a potent
T-cell activator, mediating its effects via cytokine
regulation of integrin expression. LZ-8 increases pro-
duction of IFN-γ, TNF-α and IL-1 β [90]. It is proved in
vivo, LZ-8 prevents the production of systemic
anaphylaxis reaction in mice if it has been administered
repeatedly, and reduction of antibody production is
the suggested mechanism. The mechanism of action of
717
LZ-8 on antibody production is unclear but two types of
Fc receptors have been reported; one on the cell surface
of basophiles and mast cells with high affinity for IgE,
and another on the cell surface of T or B lymphocytes
with low affinity. The latter low affinity Fc receptor-
bearing lymphocytes may be important for the regula-
tion of selective IgE production by secreting two IgE
binding factors, one with potentiating and the other
with suppressing activity toward IgE production [88].
Also an immunomodulating protein named Fip-gts has
been purified from G. tsugae that it has the same amino
acid sequence as LZ-8. Using deletion analysis, a
sequence of about 10 amino acids of the N-terminal
amphipathic alpha-helix domain of Fip-gts has been
identified to be responsible for the immunomodu-
latory activity. This region may play an important role
in the formation of homodimers for binding to cell
surface receptors to exert its immunomodulatory
activity [87]. Also it is reported that Fip-vvo induced
most Th1-specific cytokines (IL-2, INF-γ, and lympho-
toxin (LT)) and one Th2-specific cytokine (IL-4)
within 4 hours in mouse spleen cells [91]. These results
suggest that Fip-vvo principally acts on Th1 cells and to
a lesser extent on Th2 cells in the early evet of acti-
vation. Also it is proved Fip-fve selectively stimulates a
Th1 response in hPBMCs and trigger Th1 cytokine
production and oral administration of Fip-fve during
allergen sensitization could induce a Th1-predominant
allergen-specific immune response in mice and protect
the mice from the systemic anaphylaxis-like symptoms
after subsequent oral challenge with same allergen. It
is worth noting that Fip-fve could be administrated
orally and retains its activity, while most protein drugs
cannot [9,92].
12. Clinical and experimental evidences for anticancer
properties
Although, mode of action of many anticancer
components have not been known yet, clinical and
experimental evidences can help us move on these
works. Beyond these evidences, there are mechanisms
act that one of them is immunostimulating effect that
caused by anticancer compounds.
Almost worked studies to investigate anticancer
potentiality of macrofungi metabolites have been done
for Sarcoma 180 and Ehrlich carcinoma in white mice.
But mammalian cell cultures also have been used for
this purpose. In many experiments, study of antitumor
activities on Sarcoma 180 and Ehrlich carcinoma have
showed great antitumor activity to regression tumors
growth even as much as 100% inhibition (Table 5).
718 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724

Table 5 Also it could be effective for the patients with advanced

Tumor growth inhibitory effect of some macrofungi hot water extract or recurrent breast cancer as an agent for supportive
in clinical and experimental studies
therapy. In that, life span prolongation effect of lentinan
Species Against Sarcoma Against Ehrlish had also been observed with statistical significance [98].
180 (%) carcinoma (%)
Schizophyllan has also been shown to increase overall
Auricularia auricula 70–90 60–80 survival of patients with head and neck cancers [99].
A. delicate 42–70 80
Grifolan-D(R) had completed (N 95%) cell death of prostate
A. mesenterica 42–60 60
Tremella sp. 60–100 70–100 cancer cell in vivo conditions with GD ≥ 480 μg/ml [100].
Phlogiotis helvelloides 100 100 PSK and PSP had controlled various carcinomas in
Tremellodon gelationsum 90 90 experimental animals and humans. PSP is active against
Cantharellus sp. 60–100 60–90 Ehrlich ascites carcinoma, P388 leukemia and sarcoma
Craterellus sp. 60–90 60–90
180 [101] and also it enhance the transcription of tumor
Clavaria sp. 60–90 60–100
Clavulinopsis sp. necrosis factor gene in mouse peritoneal macrophages,
Lentaria IL-6 and IL-2 and interferons [102].
Ramaria formosa 60–70 60–70 Also, in an experimental study it is showed that PSP
R. botrytis can be associated with slower deterioration in patient
R. flava
with advanced non-small cell lung cancer. In this ex-
Hydnum sp. 70 90
Polyporus occidentalis 80 100 periment, after 28-day treatment, there was a significant
P. umbellatus 70 improvement in blood leukocyte and neutrophil counts,
Ganoderma applanatum 64.9 serum IgG and IgM, and percent of body fat among the
G. tropicum 70 PSP, but not the control, administrated patients [103].
G. tsugae 77.8
PSP also activates killer cells in situ in the living cancer
Armillariella mellea 70 80
Clitocybe fragrans 70–80 70–80 patient. In the Phase II and III double-blind trials, PSP
Flammulina velutipes 81–100 80 has significantly raised NK cytotoxic activity, IL-2
Lepista nuda 90 100 levels, and significantly improved the CD4 helper/CD8
Mycena pura 60 70 suppressor T-cell ratio. Altogether, these are the primary
Tricholoma gambosum 70–90 70–90
components of anticancer immunity [51]. In Phase II
Buletus edulis 100 90
Agaricus bisporus 90 100 and Phase III trials in China, PSP significantly enhanced
(edible mushroom) immune status in 70 to 97% of patients with cancers of
Coprinus atramentarius 100 100 the stomach, esophagus, lung, ovary, and cervix. In
Pholiota nameko 100 100 these studies, PSK and PSP increased the number of
immune cells and facilitated CDs and cytotoxic T-cell
infiltration of tumors [104].
The purified polysaccharide has been shown in Studies are conducted for antitumor activities of some
animal studies to produce strong tumor regression and macrofungi in the National Cancer Center, Japan. Based
even the disappearance of sarcoma tumors in 5 weeks, on these studies, extracts prepared from Hypsizygus
ascite hepatoma 134, and Ehrlich carcinoma as well as a marmoreus and F. velutipes and contain polysaccharides
number of other experimentally induced cancers in and glycoproteins showed positive effects on the cachexia
allogenic, syngeneic, and autologous hosts [8]. of advanced cancer patients. These extracts had better
In clinical studies for anticancer property, lentinan has effect than methylacetoxyprogestrone in clinical re-
succeeded in prolonging the overall survival of cancer sponse, performance status, and quality of life [105].
patients especially those with gastric and colorectal Ganoderic acids U, V, W, X and Y [85] and lanostane-
carcinomas [93–95]. The study evaluated the ability of type triterpenes isolated from spores of G. ucidum, the
lentinan to modulate Th1 and Th2 responses in patients Ganoderic alcohols Lucidumol A ((24S)-24, 25-dihy-
with digestive cancers. It apparently can cancel Th2- droxylanost-8-ene-3, 7-dione) and Lucidumol B (β,
dominant condition in patients with digestive cancers (24S)-lanosta-7, 9(11)-diene-3β, 24, 25-triol), Ganoder-
and may improve the balance between Th1 and Th2 [96]. manondiol, Ganoderiol F and Ganodermanontriol
In another clinical study, It was effective in metastatic showed cytotoxic effect on Meth-A (sarcoma) and
prostate cancer when incorporated into hormonochem- LLC tumor cells and lucidumol A exhibited the most
otherapy, so that the five-year survival rate of treated potent cytotoxicity (ED50 value 2.3 ìg/ml) against LLC
patients was 43% according to the Kaplan Meier method, tumor cells and Ganodermanondiol (ED50 value 3.4 μg/
while that of control patients was 29% ( p b 0.05) [97]. ml) against Meth-A cells [106].
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
13. Conclusion
The immune system is the body's ultimate defense
against infectious diseases, tumor and cancer growth and it
is complex containing many interacting blood cells, pro-
tein and chemicals. A healthy immune system contains
elements that are in balance with one another and in a
compromised immune system, the components are
unbalanced and unable to protect the body against harmful
agents or processes. The potential immunomodulatory
effect of antimicrobial agents was postulated as early as
the late 19th century with regard to quinine-induced
phagocyte modulation and the emergence of new
pathogen/diseases, microbial resistance to classical anti-
microbial agent, and the increased number of immuno-
compromised individuals makes new therapeutic weapons
increasingly important. Applying immunomodulators
particularly is important when immune system does not
function optimally e.g. in young children, the elderly and
patients especially cancer and AIDS patients (against
opportunistic infections) and surgical procedures. Immu-
nomodulators found in the various organisms that in
among them macrofungi (known as medicinal mush-
rooms) known as important resources of these agents.
Polysaccharides, glycoproteins, glycopeptides, Fips, and
triterpenoids are major immunomodulating agents isolat-
ed from macrofungi. Moreover, antitumor and antimicro-
bial activities are associated with immunomodulating and
immunostimulating activity of these substances. With
regard to that, nowadays, immunotherapy for cancer is
more attended via stimulation of the immune system,
using of such agents and targeting the immune system to
eliminate tumor cells are rapidly developing. In this way,
macrofungi metabolites are well known as antitumor
agents that act with affecting on antitumor effector cells
and factors and immune mechanisms that involved in the
antitumor activity. Polysaccharides are a structurally
diverse class of molecules that are found throughout
nature. They have a considerable capacity for carrying
biological information due to their structural variability.
Their variability arises from the almost endless combina-
tions that the sugar units can join together to form complex
sugars. This makes them very flexible which allows them
to exert the regulatory mechanisms of various cell-to-cell
interactions in higher organisms. It is cleared that
macrofungi are much more important resources of β-D-
glucans and their derivates and other polysaccharide that
are known as immunomodulator and antitumor agents and
using of them in virtue of their safety is important in
biomedical science. Macrofungi polysaccharides and their
peptide/protein derivates are among the emerging new
agents that could directly support or enhance functional
719
autologous hematopoietic stem cells recovery [23].
Overall, the major immunopotentiation effects of these
agents include mitogenicity, stimulation of hematopoietic
stem cells, activation of alternative complement pathway,
and activation of immune cells such as lymphocytes,
macrophages, DCs, NK cells, Th cells, Tc cells and B
cells. On the other hands, their antitumor activity also
associated with such activation and enhancement of
immune system power. In this way, activation of
macrophages due to secretion different cytokines such as
IL-1, IL-6, IL-8, TNF-α, and NO and affecting on cell-
mediate immunity are much important pathways to act
[107]. In preventive medicine, defense against invasion by
foreign bodies is dependent on enhancing the natural
immune system, including activation of macrophages and
NK cells. Because of ability of some macrofungi
immunomodulating agents to enhance NK cells activity
in cancer patients these are also known as biological
responses modifiers. The effects that cause production of
cytokines such as IL-1β IL-6, IL-8, TNF-α and NO lead to
inflammatory responses, but in other cases macrofungi
extracts inhibit the production of NO, IL-1β and TNF-α,
resulting anti-inflammatory effects occur. Thus these
macrofungi might be relevant for clinical use for
inflammatory diseases, including endotoxemia or sepsis
[9]. It is suggested that the immunomodulating action of
these agents is valuable as a mean of prophylaxis, a mild
and noninvasive form of treatment, prevention of
metastatic tumors, and as a co-treatment with chemother-
apy [1]. The enhancement or potentiation of host defense
mechanisms has been recognized as a possible means of
inhibiting tumor growth without harming the host [9]. On
the other hand, Fips as fungal immunomodulatory
proteins, so far, are merely discovered in this group of
fungi. These proteins possess relatively similar structure
and also have shown considerable similarity to the
variable region of immunoglobulin heavy chain both in
their sequences and in their secondary structure [89].
Although their details of mode of action and affecting
mechanisms are still not clearly understood but also it is
clarified that these substances act with affecting on
different cells of the immune system that are responsible
for immune responses in the innate (non-specific) and
adaptive (specific) immunity.
The first step of action of these metabolites is
recognition of them by certain receptors located on
different immune cells and activation of signal trans-
duction pathways [108]. Details of recognition of such
substances whether polysaccharides and their derivates
and Fips is not yet cleared but also some reports and
experiments have introduced some β-glucan receptors.
It is clarified several β-glucan receptors mediate these
720 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
activities such as Complement receptor 3 (CR3, αM β2-
integrin, CD11b/CD18) [69], lactosylceramide, glyco-
sphingolipd [71], scavenger receptors [72], dectin-1
[73], and TLR-2 and TLR-4 [74]. Recognition microbes
and their mediating molecules by macrophages are
mediated by TLRs that are specific for different
components of microbes. NF-kB as inducible transcrip-
tion factor is served as important regulators of the host
immune and inflammatory response after recognition of
microbial products by receptors. The biologic response
transduced by dectin-1 is dependent upon the TLR
pathway. It is determined that dectin-1 in combination
with a β-glucan-enriched zymosan derivative enhances
TLR 2-mediated cell activation [81]. Also it is indicated
that for the activation of NF-kB and induction of TNF-α,
dectin-1 needs to cooperate with TLR-2, possibly as part
of a heterodimer with TLR-6 [82].
The cytoplasmic domain of dectin-1 also has consec-
utive acidic amino acid residues that are a putative
internalizing signal sequence for the lysosomal endosome
and it is also has a putative ITAM-like region consisting of
an YXXL amino acid sequence [77]. This ITAM can be
phosphorylated by stimulation with particulate β-glucans.
It has been reported that this phosphorylation can be
involved in superoxide production by macrophages [81].
There for dectin-1 may contribute not only to phagocy-
tosis of fungal cells but also to induction of fungicidal
effector molecules [78].
Overall such findings suggest that using of macro-
fungi as immunomodulator is important in the field
infectious and immunodeficiency diseases especially
when immune system does not function optimally. Also
such compounds can mobilize the immune system to
ward off viral, bacterial, fungal and protozoan infec-
tions resistant to current antibiotics. Much attention is
now being paid to the use of immune modulators,
colony stimulating factors and cytokines as adjunctive
therapy in immunocompromised and infected patients.
However, to discover interaction between antitumor
agents and the immune system can cause to select more
effective drugs and, ideally, to combine antitumor
efficacy and “biological response modifiers” activity.
Consequently, macrofungi can be important resources
of immunomodulating and antitumor agents as well as
they known as antiviral, antimicrobial, antimutagenic,
antihypertension, antiinflammatory, antiallergic etc.
Macrofungi growth wildly in forests and rangelands
and nowadays some of them are cultivated by human.
They are more regarded as natural products and dietary
supplements and they are produced in various formula-
tions in world. Nowadays, different formulations of
medicinal macrofungi are produced with the intention
of certain aims. For example PSK under the name
Krestin was produced in Japan with the intention of
prevention and remedy of a number of cancers. In 1993,
Krestin comprised 25% of the anticancer drug market in
Japan and sales totaled US $350 million [109]. Also an
analogous product under the name PSP was developed
in China from this fungus (but from other strain) that in
a period of 10 years was soled totaled US $75 million or
10 billion yen. The market values of G. lucidum-based
natural healthcare products in 1995 were estimated as
US $215 million in Taiwan, US $350 million in China,
US $600 million in Korea and US $350 million in Japan
[110]. G. lucidum dietary supplement are valued for
their immunomodulating, anticancer and antiviral
properties. The market value of macrofungi dietary
supplement products worldwide is estimated at US
$6 billion per year. The market value of G. lucidum
mushroom-based dietary supplement alone in 1995 was
estimate at more than US $1.628 billion [110]. At
present, L. edodes is one of the five most cultivated
edible mushrooms in the world. Its production (2 mil-
lion tones) is second only to button mushroom Agari-
cus bisporus [8]. These macrofungi almost are applied
as multipurpose medicines. There is an increasing
interest in finding natural biological response modifiers
and immunoenhancers or immunomodulators from
them. In 2003, the value of world macrofungi
production and medicinal mushroom products was
estimated to be worth approximately 21 billion US
dollars. However, in this discipline, there are the
powerful sources to discover novel pharmaceutical
products.
References
[1] Wasser SP. Medicinal mushrooms as a source of antitumor and
immunomodulating polysaccharides. Appl Microbiol Biotech-
nol 2002;60:258–74.
[2] Tzianabos AO. Polysaccharide immunomodulators as thera-
peutic agents: structural aspects and biologic function. Clin
Microbiol Rev 2000;13:523–33.
[3] Liang J, Melican D, Cafro L, Palace G, Fisette L, et al. Enhanced
clearance of a multiple antibiotic resistant Staphylococcus aureus
in rats treated with PGG-glucan is associated with increased
leukocyte counts and increased neutrophil oxidative burst activity.
Int J Immunopharmacol 1998;20:595–614.
[4] Wakshull E, Brunke-Reese D, Lindermuth J, Fisette L, Nathans
RS, et al. PGG-glucan, a soluble beta-(1,3)-glucan, enhances the
oxidative burst response, microbicidal activity, and activates an
NF-kappa B-like factor in human PMN: evidence for a glyco-
sphingolipid beta-(1,3)-glucan receptor. Immunopharmacology
1999;41:89–107.
[5] Burgaleta C, Territo MC, Quan SG, Golde DW. Glucan-
activated macrophages: functional characteristics and surface
morphology. J Reticuloendothel Soc 1978;23:195–204.
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
[6] Chihara G. Immunopharmacology of Lentinan, a polysaccha-
ride isolated from Lentinus edodes: its applications as a host
defense potentiator. Int J Orient Med 1992;17:57–77.
[7] Wasser SP. Reishi or Ling Zhi (Ganoderma lucidum). Encyclop
Diet Suppl 2005:603–22.
[8] Wasser SP. Shiitake (Lentinus edodes). Encyclop Diet Suppl
2005:653–64.
[9] Lull C, Wichers HJ, Savelkoul HFJ. Antiinflammatory and immu-
nomodulating properties of fungal metabolites. Mediat Inflamm
2005;2:63–80.
[10] Ooi VE. Pharmacological studies on certain mushrooms from
China. Int J Med Mushroom 2001;4.
[11] Suzuki I, Hashimoto K, Ohno N, Tanaka H, Yadomae T.
Immunomodulation by orally administred beta-glucan in mice.
Int J Immunopharmacol 1989;11:761–9.
[12] Ito H. Effects of the antitumor agents from various natural
sources on drug-metabolizing system, phagocytic activity and
complement system in Sarcoma 180-bearing mice. Jpn J
Pharmacol 1986;40:435–43.
[13] Liu JJ, Huang TS, Hsu ML, Chen CC, Lin WS, et al. Antitumor
effects of the partially purified polysaccharides from Antrodia
camphorata and the mechanism of its action. Toxicol Appl
Pharmacol 2004;201:186–93.
[14] Ohno N, Jinushi T, Yadomae T. Th1-Oriented Immunomodulat-
ing Activity of Gel-Forming Fungal (1®3)-Beta-Glucans. Int J
Med Mushroom 2002;2.
[15] Mizuno M, Kawakami S, Fujitake N. Macrophages stimulated
by polysaccharide purified from Agaricus brasiliensis S.Wasser
et al. (Agaricomycetideae) Enhance mRNA expression of Th1
cytokine including IL-12 and 18. Int J Med Mushroom 2003;5.
[16] Leung MY, Fung KP, Choy YM. The isolation and character-
ization of an immunomodulatory and anti-tumor polysaccharide
preparation from Flammulina velutipes. Immunopharmacology
1997;35:255–63.
[17] Zhang Q, Lin Z. Study on antitumor activity and mechanism of
Ganoderma polysaccharides B. Zhongguo Zhong Xi Yi Jie He
Za Zhi 1999;19:544–7.
[18] Minato KI, Mizuno M, Kawakami S, Tatsuoka S, Denpo Y, et al.
Changes in immunomodulating activities and content of
antitumor polysaccharides during the growth of two medicinal
mushrooms, Lentinus edodes (Berk.) Sing, and Grifola frondosa
(Dicks.: Fr.) S. F. Gray. Int J Med Mushroom 2001;3.
[19] Ooi LSM, Ooi VEC, Fung MC. Induction of gene expression of
immunomodulatory cytokines in the mouse by a polysaccharide
from Ganoderma lucidum (Curt.: Fr.) P. Karst. (Aphyllophor-
omycetideae). Int J Med Mushroom 2002;4.
[20] Liu F, Ooi VEC, Fung MC. Analysis of immunomodulating
cytokine mRNAs in the mouse induced by mushroom poly-
saccharides. Life Sci 1999;64:1005–11.
[21] Wasser SP, Weis AL. Therapeutic effects of substances
occurring in higher basidiomycetes mushrooms: a modern
perspective. Crit Rev Immunol 1999;19:65–96.
[22] Su C, Sun C, Juan SW, Hu C, Ke W, Sheu M. Fungal mycelia as
the source of chitin and polysaccharides and their applications
as skin substitutes. Biomaterials 1997;18:1169–74.
[23] Lin H, She YH, Cassileth BR, Sirotnak F, Cunningham
Rundles S. Maitake beta-glucan MD-fraction enhances bone
marrow colony formation and reduces doxorubicin toxicity in
vitro. Int Immunopharmacol 2004;4:91–9.
[24] Kodama N, Komuta K, Nanba H. Effect of Maitake (Grifola
frondosa) D-fraction on the activation of NK cells in cancer
patients. J Med Food 2003;6:371–7.
721
[25] Kodama N, Komuta K, Sakai N, Nanba H. Effects of D-
fraction, a polysaccharide from Grifola frondosa on tumor
growth involves activation of NK cells. Biol Pharm Bull
2002;25:1647–50.
[26] Inoue A, Kodama N, Nanba H. Effect of Maitake (Grifola
frondosa) D-fraction on the control of the T lymph node Th-1/
Th-2 proportion. Biol Pharm Bull 2002;25:536–40.
[27] Adachi Y, Okazaki M, Ohno N, Yadomae T. Enhancement of
cytokine production by macrophages stimulated with (1→3)-
Beta-D-glucan, Grifolan (GRN), isolated from Grifola frondosa.
Biol Pharm Bull 1994;17:1554–60.
[28] Ooi VEC, Liu F. A review of pharmacological activities of mush-
room polysaccharides. Int J Med Mushroom 1999;1:195–206.
[29] Kim GY, Roh SI, Park SK, et al. Alleviation of experimental
septic shock in mice by acidic polysaccharide isolated from
the medicinal mushroom Phellinus linteus. Biol Pharm Bull
2003;26:1418–23.
[30] Park SK, Kim GY, Lim JY, et al. Acidic polysaccharides
isolated from Phellinus linteus induces phenotypic and
functional maturation of murine dendritic cells. Biochem
Biophys Res Commun 2003;312:449–58.
[31] Kim GY, Park SK, Lee MK, et al. Proteoglycan isolated from
Phellinus linteus activates murine B lymphocytes via protein
kinase C and protein tyrosine kinase. Int Immunopharmacol
2003;3:1281–92.
[32] Jin M, Jeon H, Jung HJ, et al. Enhancement of repopulation and
hematopoiesis of bone marrow cells in irradiated mice by oral
administration of PG101, a water-soluble extract from Lentinus
lepideus. Exp Biol Med (Maywood) 2003;228:759–66.
[33] Chihara G, Hamuro J, Maeda YY, et al. Antitumor and
metastasis-inhibitory activities of lentinan as an immunomod-
ulator: an overview. Cancer Detect Prev Suppl 1987;1:423–43.
[34] Murata Y, Shimamura T, Tagami T, Takatsuki F, Hamuro J.
The skewing to Th1 induced by lentinan is directed through
the distinctive cytokine production by macrophages with ele-
vated intracellular glutathione content. Int Immunopharmacol
2002;2:673–89.
[35] Maeda YY, Chihara G. Immunomodulator agents from plants.
In: Wagner H, editor. 1999.
[36] Watanabe T, Ogawa K. Study on preoperative intratumor
administration of lentinan for gastric cancer cases.II Antitumor
immune response of regional lymph nodes. J Jpn Soc Cancer
Ther 1995;30:948–55.
[37] Nakano H, Namatama K, Nemoto H, Motohashi H, Nishiyama K,
Kumada K. A multi-institutional prospective study of lentinan in
advanced gastric cancer patients with unresectable and recurrent
diseases: effect on prolongation of survival and improvement of
quality of life. Hepato-Gastroenterol 1999;46:2662–8.
[38] Nanba H, Kuroda H. Antitumor mechanism of orally adminis-
tered Shiitake fruit bodies. Chem Pharm Bull 1987;35:2459–64.
[39] Suzuki Y, Adachi Y, Ohno N, Yadomae T. Th1/Th2-balancing
immunomodulating activity of gel-forming (1→3)- β-glucans
from fungi. Biol Pharm Bull 2001;24:811–9.
[40] Shao BM, Dai H, Xu W, Lin ZB, Gao XM. Immune receptors
for polysaccharides from Ganoderma lucidum. Biochem
Biophys Res Commun 2004;323:133–41.
[41] Cao LZ, Lin ZB. Regulation on maturation and function of
dendritic cells by Ganoderma lucidum polysaccharides. Immu-
nol Lett 2002;83:163–9.
[42] Ohno N, Harada T, Masuzawa S, Miura NN, et al. Antitumor
activity and hematopoietic response of a β-glucan extracted
from an edible and medicinal mushroom Sparassis crispa
722 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
Wulf.: Fr. (Aphyllophoromycetideae). Int J Med Mushroom
2002;4.
[43] Harada T, Miura N, Adachi Y, Nakajima M, Yadomae T, Ohn N.
Effect of SCG, 1,3-β-D-glucan from Sparassis crispa on the
hematopoietic response in cyclophosphamide induced leuko-
penic mice. Biol Pharm Bull 2002;25:931–9.
[44] Duncan CJ, Pugh N, Pasco DS, Ross SA. Isolation of a galac-
tomannan that enhances macrophage activation from the edible
fungus Morchella esculenta. J Agric Food Chem 2002;50:5683–5.
[45] Mizuno M, Shiomi Y, Minato K, Kawakami S, Ashida H,
Tsuchida H. Fucogalactan isolated from Sarcodon aspratus
elicits release of tumor necrosis factor-α and nitric oxide from
murine macrophages. Immunopharmacology 2000;46:113–21.
[46] Ito H, Shimura K, Itoh H, Kawade M. Antitumor effects of a new
polysaccharide-protein complex (ATOM) prepared from Agaricus
blazei (Iwade strain 101) ʽʽHimematsutake" and its mechanisms in
tumor-bearing mice. Anticancer Res 1997;17:277–84.
[47] Kato M, et al. Induction of gene expression for immunomodulat-
ing cytokines in peripheral blood mononuclear cells in response to
orally administrated PSK, an immunomodulating protein-bound
polysaccharide. Cancer Immunol Immunother 1995;40:152–6.
[48] Kanazawa M, Mori Y, Yoshihara K, et al. Effect of PSK on the
maturation of dendritic cells derived from human peripheral
blood monocytes. Immunol Lett 2004;91:229–38.
[49] Tomochika H, Gouchi A, Okanobu K, et al. The effect and
distribution of a protein-bound polysaccharide preparation, PSK
(Krestin), intratumorally injected prior to surgery into gastric
cancer patients. Acta Med Okayama 1989;43:289–97.
[50] Mizutani Y, Yoshida O. Activation by the protein-bound
polysaccharide PSK (Krestin) of cytotoxic lymphocytes that
act on fresh autologous tumor cells and T24 human urinary
bladder transitional carcinoma cell line in patients with urinary
bladder cancer. J Urol 1991;145:1082–7.
[51] Kariya Y, Inoue N, Kihara T, et al. Activation of human natural
killer cells by the protein-bound polysaccharide PSK independent-
ly of interferon and interleukin 2. Immunol Lett 1992;31:241–6.
[52] Yang Q. Yun Zhi polysaccharopeptide (PSP) and the general
aspects of its research. In: Yang Q, editor. Advanced Research
in PSP, 1999. Hong Kong: Hong Kong Association for Health
Care Ltd; 1999. p. 29–38.
[53] Yang Q, Hu Y, Li X, et al. A new biological response modifier
substance PSP. In: Yang Q, Kwok C, editors. PSP International
Symposium 1993. Hong Kong: Fudan University Press; 1993.
p. 56–72.
[54] Jian Z, Zhang W, Li S. The effect of PSP on LAK cell functions.
In: Yang Q, Kwok C, editors. PSP International Symposium
1993. Hong Kong: Fudan University Press; 1993. p. 143–50.
[55] Wang YY, Khoo KH, Chen ST, Lin CC, Wong CH, Lin CH.
Studies on the immuno-modulating and antitumor activities of
Ganoderma lucidum (Reishi) polysaccharides: functional and
proteomic analyses of a fucose-containing glycoprotein fraction
responsible for the activities. Bioorg Med Chem 2002;10:1057–62.
[56] Won SJ, Lee SS, Ke YH, Lin MT. Enhancement of splenic NK
cytotoxic activity by extracts of Ganoderma lucidum mycelium
in mice. J Biomed Lab Sci 1989;55:201–13.
[57] Zhang J, Tang Q, Zimmerman-Kordmann M, Reutter W, Fan H.
Activation of B lymphocytes by GLIS, a bioactive proteoglycan
from Ganoderma lucidum. Life Sci 2002;71:623–38.
[58] Borchers AT, Stern JS, Hackman RM, Keen CL, Gershwin ME.
Proc Soc Exp Biol Med 1999;221:281–93.
[59] Okazaki M, Adachi Y, Ohno N, Yadomae T. Structure-activity
relationship of (1→3)-beta-D-glucans in the induction of
cytokine production from macrophages, in vitro. Biol Pharm
Bull 1995;18:1320–7.
[60] Maeda YY, Watanabe ST, Chihara C, Rokutanda M. Denatur-
ation and renaturation of a β-1,6: 1,3-glucan, lentinan associated
with expression of T-cell mediated responses. Cancer Res
1988;48:671–5.
[61] Bohn JA, BeMiller JN. (1-3)-β-D-glucans as biological response
modifiers: a review of structure-functional activity relation-
ships. Carbohydr Polym 1995;28:3–14.
[62] Bluhm TL, Sarco A. The triple helical structure of lentinan, a β-
(1-3)-D-glucan. Can J Chem 1977;55:293–9.
[63] Misaki A, Kakuta M, Sasaki T, Tanaka M, Miyaji H. Studies on
interrelation of structure and antitumor effects of polysacchar-
ides: antitumor action of periodate-modified, branched (1 goes
to 3)-beta-D-glucan of Auricularia auricula-judae, and other
polysaccharides containing (1 goes to 3)-glycosidic linkages.
Carbohydr Res 1981;92:115–29.
[64] Bao X, Duan J, Fang X, Fang J. Chemical modifications of the
(1-(3)-alpha-D-glucan from spores of Ganoderma lucidum and
investigation of their physicochemical properties and immuno-
logical activity. Carbohydr Res 2001;336:127–40.
[65] Eo SK, Kim YS, Lee CK, Han SS. Antiviral activity of various
water and methanol soluble substances isolated from Gano-
derma lucidum. J Ethnopharmacol 1999;68:129–36.
[66] Kiho T, Yoshida I, Katsuragawa M, Sakushima M, et al.
Polysaccharides in Fungi. XXXIV. A polysaccharide from the
fruiting bodies of Amanita muscaria and the antitumor acti-
vity of its carboxymethylated product. Biol Pharm Bull
1994;17:1460–2.
[67] Kidd PM. The use of mushroom glucans and proteoglycans in
cancer treatment. Altern Med Rev 2000;5:4–27.
[68] Lowe E, Rice P, Ha T, et al. A (1→3)-β-D-linked heptasacchar-
ide is the unit ligand for glucan pattern recognition receptors on
human monocytes. Microbes Infect 2001;3:789–97.
[69] Thornton BP, Větvička V, Pitman M, Goldman RC, Ross GD.
Analysis of the sugar specificity and molecular location of the
β-glucan-binding lectin site of complement receptor type 3
(CD11b/CD18). J Immunol 1996;156:1235–46.
[70] Ross GD, Cain JA, Myones BL, Newman SL, Lachmann PJ.
Specificity of membrane complement receptor type three (CR3)
for beta-glucans. Complement 1987;4:61–74.
[71] Zimmerman JW, Lindermuth J, Fish PA, Palace GP, Stevenson
TT, DeMong DE. A novel carbohydrate-glycosphingolipid
interaction between a β-(1-3)-glucan immunomodulator, PGG-
glucan, and lactosylceramide of human leukocytes. J Biol Chem
1998;273:22014–20.
[72] Rice PJ, Kelley JL, Kogan G, et al. Human monocyte scavenger
receptors are pattern recognition receptors for (1→3)-β-D-
glucans. J Leukoc Biol 2002;72:140–6.
[73] Brown GD, Gordon S. Immune recognition. A new receptor for
beta-glucans. Nature 2001;413:36–7.
[74] Shao BM, Dai H, Xu W, Lin ZB, Gao XM. Immune receptors
for polysaccharides from Ganoderma lucidum. Biochem
Biophys Res Commun 2004;323:133–41.
[75] Ross GD, Cain JA, Lachmann PJ. Memberan complement
receptor type three (CR3) has lectin-like properties analogous to
abovine conglutinin and functions as a receptor for Zymosan and
rabbit erythrocytes as well as a receptor for iC3b1. J Immunol
1985;134:3307.
[76] Elstad MR, et al. CD11b/CD18 integrin and a β-glucan receptor
act in concert to induce the synthesis of platelet-activating factor
by monocytes. J Immunol 1994;152:220.
 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
[77] Ariizumi K, Shen GL, Shikano S, Xu S, Ritter R, Kumamoto T,
et al. Identification of a novel, dendritic cell-associated mol-
ecule, dectin-1, by subtractive cDNA cloning. J Biol Chem
2000;275:20157–67.
[78] Adachi Y, Ishii T, Ikeda Y, Hoshino A, Tamura H, et al.
Characterization of β-glucan recognition site on C-Type lectin,
dectin 1. Infect Immun 2004;72:4159–71.
[79] Herre J, Gordon S, Brown GD. Dectin-1 and its role in the
recognition of β-glucans by macrophages. Mol Immunol
2004;40:869–76.
[80] Young SH, Dong WJ, Jacobs RR. Observation of a partially
opened triple-helix conformation in 1→3-β-glucan by fluores-
cence resonance energy transfer spectroscopy. J Biol Chem
2000;275:11874–9.
[81] Gantner BN, Simmons RM, Canavera SJ, Akira S, Underhill
DM. Collaborative induction of inflammatory responses by
dectin-1 and Toll-like receptor 2. J Exp Med 2003;197:1107–17.
[82] Borchers AT, Keen CL, Gershwin ME. Mushrooms, tumors,
and immunity: an update. Exp Biol Med 2004;229:393–406.
[83] Pinheiro F, Faria RR, de Camargo JLV, Spinardi-Barbisan ALT,
da Eira AF, Barbisan LF. Chemoprevention of preneoplastic
liver foci by dietary mushroom Agaricus blazei Murill in the rat.
Food Chem Toxicol 2003;41:1543–50.
[84] Travnikova IG, Shutov VN, Bruk GY, Balonov MI, Skuterud L,
Strand P, et al. Assessment of current exposure levels in dif-
ferent population groups of the Kola Peninsula. J Environ
Radioact 2002;60:235–48.
[85] Wasser SP, Weis AL, Nevo E. Medicinal Mushrooms: Gano-
derma lucidum (Curtis: Fr.) P. Karst. Reishi Mushroom. Haifa,
Israel: Pedelfus Publishing House; 1997.
[86] Gao Y, Zhou SH, Chen G, Dai X, Ye J. A phase I/II study of a
Ganoderma lucidum (Curt.: Fr.) P. Karst. extract (ganopoly)
in patients with advanced cancer. Int J Med Mushroom 2003;4:
207–14.
[87] Lin WH, Hung CH, Hsu CI, Lin JY. Dimerzation of the N-
terminal amphipathic alpha-helix domain of the fungal
immunomodulatory protein from Ganoderma tsugae (Fip-gts)
defined by a yeast two-hybride system and site-directed
mutagenesis. J Biol Chem 1997;272:20044–8.
[88] Kino K, et al. Isolation and characterization of a new immu-
nomodulatory protein, Ling Zhi-8 (LZ-8), from Ganoderma
lucidum. J Biol Chem 1989;264:472–8.
[89] Tanaka S, et al. Complete amino acid sequence of an
immunomodulatory protein, Ling Zhi-8 (LZ-8). An immuno-
modulator from a fungus, Ganoderma lucidum, having similarity
to immunoglobulin variable regions. J Biol Chem 1989;264:
16372–7.
[90] Haak-Frendscho M, Kino K, Sone T, Jardieu P. Ling Zhi-8: a
novel T cell mitogen induces cytokine production and upregula-
tion of ICAM-1 expression. Cell Immunol 1993;150:101–13.
[91] Hsu HC, Hsu CI, Lin RH, Kao CL, Lin JY. Fip-vvo, a new
fungal immunomodulatory protein isolated from Volvariella
volvacea. Biochem J 1997;323:557–65.
[92] Hsieh KY, Hsu CI, Lin JY, Tsai CC, Lin RH. Oral
administration of an edible-mushroom-derived protein inhibits
the development of food-allergic reactions in mice. Clin Exp
Allergy 2003;33:1595–602.
[93] Furue H, Kitoh I. Phase 111-study on Lentinan. Jpn J Cancer
Chemother 1981;8:944–60.
[94] Taguchi T, Furue H, Kimura T, Kondo T, Hattori T, Itoh T, et al.
End point results of phase III study of lentinan. Jpn J Cancer
Chemother 1985;12:366–80.
723
[95] Taguchi T, Furue H, Kimura T, Kondo T, Hattori T, Itoh T, et al.
End point result of a randomized controlled study of the treatment
of gastrointestinal cancer with a combination of lentinan and
chemotherapeutic agents. Excerpta Medica 1985:151–65.
[96] Yashino S, et al. Immunoregulatory effects of the antitumor
polysaccharide lentinan onTh1/Th2 balances in patients with
digestive cancers. Anticancer Res 2000;20:4707–11.
[97] Tari K, et al. Effect of lentinan for advanced prostate carcinoma.
Hinyokika Kiyo 1994;40:119–23.
[98] Taguchi T. Effects of lentinan in advanced or recurrent cases of
gastric, colorectal, and breast cancer. Gan To Kagaku Ryoho
1983;10:387–93.
[99] Kimura Y, Mizuno H, Satake K, Tahara H, Tsukuda M. Clinical
evaluation of Sizofiran an assistant immunotherapy in treatment
of head and neck cancer. Acta Otolaryngol 1994;511:192–5.
[100] Fullerton SA, Samadi AA, Tortorelis DG, Choudhury MS,
Mallouh C, Tazaki H, et al. Mol Urol 2000;4:7–13.
[101] Yang QY, Jong S, Li XY, Zhou JX, Chen RT, Xu LZ. Antitumor
and immunomodulating activities of the polysaccharide-
peptide (PSP) of Coriolus versicolor. J Immunol Immunophar-
macol 1992;12:29–34.
[102] Yang QY, Yi J, Xy LI. A new biological response modifier-PSP.
PSP International Symposium; 1993. p. 56–72.
[103] Tsang KW, Lam CL, Yan C, Mak JC, Ooi GC, et al. Coriolus
versicolor polysaccharide peptide slows progressin of advanced
non-small cell lung cancer. Respir Med 2003;97:618–24.
[104] Ng TB. A review of research on the protein-bound polysaccha-
ride (polysaccharopeptide, PSP) from the mushroom Coriolus
versicolor (Basidiomycetes: Polyporaceae). Gen Pharmacol
1998;30:1–4.
[105] Ikekawa T. Cancer risk reduction by intake of mushrooms and
clinical studies on EEM. Int J Med Mushroom 2005;7:347.
[106] Min BS, Gao J, Nakamura N, Hattori M. Triterpenes from
the spores of Ganoderma lucidum and their cytotoxicity against
Meth-A and LLC tumor cells. Chem Pharm Bull 2000;48:
1026–33.
[107] Lotzova E, Tsubura E, Effector functions of cytokines in cancer,
in: advances in immunopharmacology 4, Hadden JW, et al.
1989; 245-250.
[108] Goldman R, Ferber E, Meller R, Zor U. A role for reactive
oxygene species in Zymosan and β-glucan induced protein
tyrosine phosphorylation and phospholipase A2 activation in
murine macrophage. Biochim Biophys Acta 1994;1222:265.
[109] Mizuno T. The extraction and development of antitumor-active
polysaccharides from medicinal mushrooms in Japan [Review].
Int J Med Mushroom 1999;1:9–30.
[110] Chang ST, Buswell JA. Ganoderma lucidum (Curt.:Fr.)
P. Karst. (Aphyllophoromycetideae)-A mushrooming medicinal
mushroom. Int J Med Mushroom 1999;1:139–48.
[111] Yap AT, Ng ML. Immunopotentiating properties of Lentinan
(1→3)-β-D-glucan extracted from culinary-medicinal mush-
room Lentinus edodes (Berk.) Singer (Agaricomycetideae). Int
J Med Mushroom 2003;5.
[112] Sarkar S, Koga J, Whitley RJ, Chatterjee S. Antiviral effect of the
extract of culture medium of Lentinus edodes mycelia on the
replication of herpes simplex virus 1. Antivir Res 1993;20:293–303.
[113] Matsuyama H, Mangindaan RE, Yano T. Protective effect of
Schizophyllan and Scleroglucan against Streptococcus sp.
infection in Yellow Tail (Seriola quinqueradiata). Aquaculture
1992;101:97–203.
[114] Oh KW, Lee CK, Kim YS, Eo SK, Han SS. Antiherpetic
activies of acidic protein bound polysaccharide isolated from
724 M.-F. Moradali et al. / International Immunopharmacology 7 (2007) 701–724
Ganoderma lucidum alone and in combinations with acyclovir
and vidarabine. J Ethnopharmacol 2000;72:221–7.
[115] Tsukagoshi S. Krestin (PSK). Cancer Treat Rev 1984;11:131–55.
[116] Ooi Vinsent EC. Pharmacological studies on certain mushroom
from China. Int J Med Mushroom 2001;3.
[117] El-Mekkawy S, et al. Anti-HIV-1 and anti-HIV-1-protease
substances from Ganoderma lucidum. Phytochemistry 1998;49:
1651–7.
[118] Chairul, et al. Applanoxidic acids A, B, C and D, biologically
active tetracyclic triterpenes from Ganoderma applanatum.
Phytochemistry 1991;30:4105–9.
[119] Toth JO, Luu B, Ourisson G. Ganoderic acid T and Z: cytotoxic
triterpenes from Ganoderma lucidum (Polyporaceae). Tetrahe-
dron Lett 1983;24:1081–4.
[120] Kohda H, Tokumoto W, Sakamoto K, et al. The biologically
active constituents of Ganoderma lucidum (Fr.) Karst, Hista-
mine release-inhibitory triterpenes. Chem Pharm Bull 1985;33:
1367–74.
[121] Morigiwa A, Kitabatake K, Fujimoto Y, Ikekawa N. Angioten-
sin converting enzyme-inhibitory triterpenes from Ganoderma
lucidum. Chem Pharm Bull 1986;34:3025–8.
[122] Lin CN, Tome WP. Novel cytotoxic principle of formosan
Ganoderma lucidum. J Nat Prod 1991;54:998–1002.
[123] Bok JW, Lermer L, Chilton J, Klingeman HG, Towers GH.
Antitumor sterols from the mycelia of Cordyceps sinensis.
Phytochemistry 1999;51:891–8.
[124] Keller AC, Maillard MP, Hostettmann K. Antimicrobial steroids
from the fungus Fomitopsis pinicola. Phytochemistry 1996;41:
1041–6.
[125] Menoli RCR, Mantovani MS, Ribeiro LR, Speit G, Jordão Q.
Antimutagenic effects of the mushroom Agaricus blazei Murrill
extracts on V79 cells. Mutat Res 2001;496:5–13.
[126] Moradali MF, Mostafavi H, Hedjaroude GA, Abbasi M, Ghods
S, Salahi A. Investigation of antibacterial potentiality of
methanol extraction from fungus Ganoderma applanatum.
World Conference on Magic Bullets Celebrating Paul Ehrlich
150th Birthday, Nürnberg, Germany; 2004.
[127] Chiu SW, Wang ZM, Leung TM, Moore D. Nutritional value of
Ganoderma extract and assessment of its genotoxicity and
antigenotoxicity using comet assay of mouse lymphocytes.
Food Chem Toxicol 2000;38:173–8.
[128] Shimizu A, Yano T, Saito Y, Inada Y. Isolation of an inhibitor of
platelet aggregation from a fungus, Ganoderma lucidum. Chem
Pharm Bull 1985;33:3012–5.
[129] Gottlieb D, Shaw PD. Antibiotics. Berlin: Springer-Verlag
Press; 1967.
[130] Fujii T, Ishida N, Maeda H, Mizutani I, Suzuki F, KS-2-A. US Patent
4163780, 1979.
[131] Ikekawa T, Ikeda Y, Yoshioka Y, Nakanishi K, Yokoyama E,
Yamazaki E. Antitumor polysaccharides of Flammulina
velutipes 2. The structure of EA-3 and further purification of
EA-5. J Pharmacobio-Dyn 1982;5:576–81.
[132] Zhuang C, Mizuno T, Ito H, Shimura K, Sumiya T. Chemical
modification and antitumor activity of polysaccharides from the
mycelium of liquid-cultured Grifola frondosa. Nippon Shoku-
hin Kogyo Gakkaishi 1994;41:733–40.
[133] Cho SM, Park JS, Kim KP, Cha DY, Kim HM, Yoo ID. Chemical
features and purification of immunostimulating polysaccharides
from the fruit bodies of Agaricus blazei. Korean J Mycol 1999;27:
170–4.
[134] Gutiérrez A, Prieto A, Martínez AT. Structural characterization
of extracellular polysaccharides produced by fungi from the
genus Pleurotus. Carbohydr Res 1996;281:143–54.