0510:16p

I",..r"uliv"al Jmlrnal Mushrooms. Vol pp. 20"

The Antitumor Activity of Ganoderma

lucidum (Curt.:Fr.)P. Karst. (Ling Zhi)
(Aphyllophoromycetideae) Polysaccharides Is
Related to Tumor Necrosis Factor-a and Interferon-'Y
Dun-Hao Zhang and Zhi-Bin Un *
Department of Pharmacology, School of Basic Medical Sciences. Beijing Medical University, Beijing
100083, China

Iln all C(H-re"pUildellce should be addrcs!lCd

ABSTRACT: In the presentstudy. the antitumor activity I)f GL-B. a polysacchande isolated from Ganodenn"
/ucidum (Cun:Fr.)P. Karst. and its mechanism were stUdied in l'il'O and in vitro. The results were RSfollows:
( I) GL-B 50. 100. 200 I1g ml-1 inhibited the growth of implanted Sarcomu 180 in viJ..osignificantly and do.-e
dcpcndently. (2) GL-B diret:tly udded to the culture medium neither induced HL-6Q apoptoslSnor restr.sinedits
proliferation ill vitro. (3) The macrophageor T lymphocyte culture medium treated with GL-B (GL-B-M-CM
or GL-B- T-CM) 50, 100, and 2()()I1g ml-1 !iignificantly induced HL-60 apoptosis and inhibited it!i proliferdtion.
GL-B significantly increasedtumor necrosisfactor-a ( TNF-a) and interferon-)' (IFN-y) releasein dose-dependent
and lime-dependentinstance-'i.(4) As unu'eatedmacrophage!;and T Iymphl>cytesproduced little or no 'rNF-a and
IFN-y. and mlICrophngeculture medium with normal saline (N-M-CM) or T lymphocyte culture medium with nor-
mal saline (N- T-CM) did nllt inhibit HL-60 proliferation or induce its apoptosis, it seemedthat the antitumor ac-
tivity of GL-B was related to apoptosis induced by TNF-a-relea!ie from macrophages and IFN-y-releilse frOlI1
T lymphocytes.

KEY WORDS: Apoplosis. Ganodenn(llucidum. HL-60. interferon-yomacrophage.polysaccharides.proliferilii

Sarcoma 180. T lymphocyte. tumor necrosisfactor-a.

Ganoderma lucidum (Curt.: Fr.)P. Karst.
(Ling Zhi) has beenwidely usedas a medicine to
promote health and longevity in China for thou-
sandsof years.A seriesof experimentsin our lab-
oratory have demonstrated that G. lucidum has
various pharmacological effects, such as im-
munomodulatingones(Xia and Lin, .1989;Ma et
al.. 1991; Lei and Lin, 199.1.1992). Recently. G.
lucidum hasattracted great interestdue to the an-
titumor activity of its polysaccharide fraction.
Many investigators have demonstrated that the
polysaccharides isolated from Ganoderma /u-
cidum cou]d significantly inllibit the growth of'
implanted Sarcoma 180 and Lewis lung-carci-
noma in animal models in vitro (Sasaki, 1971;
Miyazaki and Nishiyama, ]981; Sone and Okuda.
]985; Maziyama et al., 1989). Although its anti-
tumor activity is beyond doubt, the mechanisms
remain unclear. Some scholars have suggested
that its antitumor activity may be mediated to

ABBREVIATIONS
Con A: concanavalinA: CY: cyclophosphamide:ELISA: enzyme-linked immunoabsorbentassay;
FCS: fetal calf serum; IFN-y. interferon-y: LPS: lipopolysaccharide; NBS: newborn bovine serum:
PHS: phosphate-bufferedsaline: PEC: peritoneal exudatecells: PI: propidinmiodide; TNF-a: tumor
necrosisfactor-a.

1521-9437/991$5.00
@ 1999 by Begell House. In.: 207

Medicirwl
Jan 16 05 10:19p
some extent by the activation of host immune
functions. However, such a hypothesis requires
substantialevidence. In thi~ article. we describe
our investigation of the antitumor activity of
GL-B, a mixture of partially purified polysaccha-
rides isolated from G. Lucidum.at a cellu]ar and
molecular level both in vn'o and in vitro.
MATERIALS AND METHODS
Animals
Inbred male and female 1-2-month-old (body
weight 18-25g) BALB/c mice were purchased
from the Departmentof Experimental Animals.
Beijing Medical University.
Drug
Ganoderma [u(.'idum polysaccharide (GL-B)
consists of seven fractions of polysaccharide iso-
lated from G, lucidllm. It is a yellowish and water-
soluble powder with molecular weights of 7,000
to 9,000 provided by the Departmentof Phyto-
chemistry, College of Phannacy, Beijing Medical
University (Li and He, 1991).
Cell Lines
HL-60 and Sarcoma 180were obtained from
Beijing Tumor I.nsritute.L929 cells were provided
by the Departmenlof Immunology, Beijing Med-
ical University. RPMI. 1640 powder was from
Gibco BRL. MTT, lipopolysaccharide(LPS), alld
concanavalinA (Con A) were from Sigma.
Preparation of T Lymphocytes and
Peritoneal Macrophages
Mice were killed and the spleens were chopped
with two ~Iides and filtered over a fine nylon mesh.
Cells were washed three times in Hank's balanced
salt solutjon containing 5% heat-inacrivated new-
born bovine serum (NBS). Cells were finally
suspended in RPMI-I640 supplemented with
100 IV ml-1 penicillin, 100~g.ml-1 streptomycin,
1 mmol.I-J sodium pyruvate, 2 mmol.I-1 L-gluta-
mine, 3.4 x 10-3 mmol.I-1 2-mercaptoethanol, and
10% NBS. Rat peritoneal exudate cells (PECs)
208
p.3
were harvested by peritoneal lavage using cold
Hank's solution containing 5% fetal calf serum
(FCS). PECs were washed twice and resuspended
in an RPMI 1640 medium containing 10% FC'S.
Peritoneal macrophages were further isolated
from the PECs by incubating the PECs in a 24-wel.l
plate at 37°C in a humidified atmo!.'Pherefor 2--4
h to allow for peritoneal macrophage adherence.
After this time, the nonadherent cells were re-
moved by washing three times with waTnl RPMJ
1640 medium. More than 95% of the adherent ce.ll
population was macrophages, as determined by
morphology and esterase staining.
Conditioned Media
To investigate the effect of GL-B on stimulat-
ing cytokine production by T lymphocytes and
macrophages, and the effect ofGL-B conditioned
media with T lymphocytes and macrophage", on
pruliferdtion and apoptosis of tumor cells, a pure
popu lation of macrophage.~or T Iymphocytes was
incubated separately in an RPMI 1640 medium
containing 10% NBS with or without various
concentrations of GL-B at 37°C tor 12-72. The
conditioned media, which were caJ]ed macro-
phage culture medium with GL-B (GL-B-
M-CM) and T lymphocyte culture medium with
GL-B (GL-B-T-CM), were then collected, fil-
tered, and stored at -70°C, respectively, for use.
Assay of Cytokines
Conditioned media from GL-B stimulated
macrophagesor T lymphocytes were assayedfor
activity of tumor necrosis factor (TNF-a) and in-
terferon-y(IFN-y). TNF-a was a.~sayedby bioas-
~y methodsusingL929 cells and IFN-y by solid-
phase enzyme-linked immunoabsorbent assay
(ELISA) a...described by the available kit.
Treatment of HL-60 Cells
HL-60 I.;ells,maintained in an RPMI 1640
mediumcontaining 10%FCS. werecultured at all
initiaJ concentration of Ixl05.mI-1 in the pres~
ence or absenceof 20% (voVvol) of GL-B stimu-
lating conditioned media or normal media. For
detecting the direct effect of GL-B on HL-60
Jan 16 05 10:23p
growth. the culture was treated with GL-B at dif-
ferent concentrations. Cultures were then incu-
bated at 37°C in a humidified atmospherefor 3
days.
Antitumor Experiment in
Tumor-Bearing Mice
Sarcoma 180cells were injected subderrnal.ly
into the axillary fossa of the right foreleg. The
mice were divided into severalgroups randomly.
Different doses of drugs were administered by
stomachtube.oncea day.The mice were killed 10
days later and the tumors were cut and weighed.
Analysis of Apoptosis
HL-60 cells were separated from the medium
by centrifugation and washed with phosphate-
buffered saline (PBS) after incubation. Hy-
podiploid DNA was analyzed using the method of
propidiumiodide (PI) labeling and flow cytome-
try. After washing, samples were resuspended and
placed in the dark at 4"C. PI fluorescence of indi-
vidual nuclei was analyzed using a FACScan flow
cytometer (Becton Dickinson).
Determination of Cells Proliferation
MTT (20 1.11.5 mg I-I every well) was added
to the cell culture 4 h before the end of incuba-
tion. After incubation,gave away supernatant,and
150 III of isopropanol was added and OD was
assessed using the Enzyme Labeling lru;trument
(Bio-Rad).
TABLE 1
Antitumor Effect of GL-B on Sarcoma 180 in BALB/c Mice (n = 10, x:l: SO)
Dose
Groups (mgokg--1)
23.61 :t 1.40
24.05:t 1.75
23.90 r 1.62
23.52:t 1.48
23.84:t 1.43
p.4
Statistical Analysis
Results were expressed as X ::!::SD and ana-
lyzed by I-test to compare the difference between
the groups with the Statistica software for Win-
dows@ (4.5. Statsoft Inc., 1993).
RESULTS
Antitumor Effect of GL-B on Locally
Implanted Sarcoma 180 In Vivo
Table I shows the antitumor effect of GL-B
on Sarcoma 180 in BALB/c mice in vivo. GL-B
50, 100,and 200.mg kg-l inhibited the growth of
Sarcoma 180 in a dose-dependentinstance.The
inhibitory ratesare 27.70%, 55.83%,and 66.70%,
respectively. The inhibitory rate of the GL-B
group treated with cyclopho!iphamide(CY) was
higher than that of the GL-B or the CY group.
Their inhibitory rateswere 80.98%,45.45%,and
75.95%, respectively(Table 2).
Effect of GL-B on Proliferation of HL-60
and Sarcoma 180 Cells In Vitro
On the basis of the above results, we further
added GL-B directly to the in vitro HL-60 and
Sarcoma 180 culture media. We unexpectedly
found that GL-B had no effect on proliferation of
either HL-60 or Sarcoma 180cells (Tables 3 and
4). Table 3 shows that GL-B 400 ~g'ml-J slightly
stimulated HL-60 cell proliferation.
Tumor weight Inhibitory rate
(mg) (%)
10.32 :!:2.60: 2.02:t 0.16
7.57:! 1.82" 1.46:t 0.61 27.70
6.33j :1.77"": 0.89 :t 0.45". 55.83
6.46 j 1.46".: 0.67 :t 0.47"" 66.70
5.84:1 1.68"" 0.44 j; 0.54"" 78.02
209
Jan 16 05 10:26p p.5

TABLE 2
Antitumor Effect of GL-B on Sarcoma 180 in BALB/c Mice (n = 10,.i:t. SD)

Dose Body weight (g) Tumor weight Inhibitory rate

Groups (mgokg-1) Origin difference (mg) (%)

NS -23.14:t 1.62 9.58 :t 2.11 1.87:t 0.78

GL-B 100 22.79:t 1.41 7.22 :t 1.73 1.02 t 0.51.M 45.45
CY 20 22.87 ;t 1.55 6.65 :t 1.42 0.45:t O.48*'d 75.95
Gl-B +CY 100+20 23.35:!: 1.34 6.70:1: 1.36 0.36:!: 0.17** 80.98
.p<O.05vs. N$; ..p< 0.01 va. NS
Ap < 0.05 va. OB + CY; Mp < 0.01 vs. OL-B + CY

TABLE 3 Effect of GL-B on Inducing HL-60 Cells

Effect of Gl-B on Proliferation of Hl-60 Cells Apoptosls In Vitro
In Vitro (n = 8, x :I: SD)

Concentration
Table 5 indicatesGL-B 50. .100.200 ~g mil
Groups (j1g'ml-1) 00570 nm cannot induce HL-60 cell apoptosis;the propor-
tions of apoptic cells were 0.87 :t 0.24%. 0.65 :t
APMI 1640 -0.346 :t 0.041 0.31 %, 0.63 :t 0.45%, respectively.The positive
GL-B 50 0.384:t 0.137 control drug Etopeside Vp-16 significantly in-
100 0.402 j: 0.44
ducedHL-60 cell apoptosis;theapoplic cells wa1'
200 0.418:t 0.063
400 0.465:t 0.139' 67.70 :t 4.04%.
Vp-16
- 80 0.241 j: 0.067"'
'p< 0.05 Ys. RPMI 1640; "p< 0.01 VS. RPMI1640

Effects of GL-B-M-CM and GL-B-T-CM on

TABLE 4 Proliferation of HL-60 Cells In Vitro
Effect of GL-B on Proliferation of Sarcoma
180 Cells In Vitro (n = 8, x f: SD) Tables 6 and 8 show that every concentration
of GL-B-M-CM and GL-B- T-CM significantly
Concentration
Groups (J1Q"mt-1) CD 570 nm inhibited proliferation of HL-60 cells in "ilro.
Compared with RPMI 1640and N-M-CM or N-
RPMI1640 - 0.369 j: 0.108 T-CM groups,p < 0.05 orO.OI.
GL.B 50 0.337 j: 0.045
100 0.405 :t 0.0520.0570.093
200 0.411 :t
400 0.442 :t
y~"--~ 0.244 :t 0.037 Effect of GL-B-M-CMand GL-B-T-CM on
'p< 0.01 VS. RPMt 1640. Inducing Apoptosis of HL-60 Cells In Vitro

TABLE 5 Table 8 shows that50. 100. and 200 Jlg.ml-1

Effect of GL-B on Apoptosis of HL-60 Cells of GL-B-M-CM significantly induced HL-60
After 72 h of In Vitro Incubation~n = 3, x:t: SD) cells apoptosis;the proportions of apt>ptoticcells
were 18.81 :i: 0.93%. 20.98 :i: 1.57%. 23.00 :i:
Concentration Apoptotic cells 0.56%. respectively.comparedwith the N-M-CM
Groups (~g.ml-1) (%) group. 7.44:1:1.07%. P<O.Ol.
RPMI1640 1.94:t 0.41
Table 9 shows that 50, ]00. and 200 Jlg.ml-J
GL-B 0.87 t 0.24 of GL-B- T-CM significantly induced HL-60 cell
0.65 :t 0.31 apoptosis;the proportions of apoptotic cells were
0.63 :t 0.45 19.39:t 1.13%..2i,94:t0.84%, 22.85:t 1.49%,re-
67.70:t 4.04'. spectively, compared with the N- T-CM group,
7.75 :t 1.14%,p<0.OI.

0!10
 lO:29p p.6

TABLE 6 TABLE 7
Effect of GL-B-M-CM on Proliferation of Effect of GL-B- T-CM on Proliferation of
HL-60 Cells In Vitro (n = 8, x:t SO) HL~60 Cells In Vitro (n = 8, x:I: SO)

Concentration Concentration
Groups mgoml-1 OD 570nm Groups mgoml-1 OD 570 nm
RPMI1640 RPMI1640 0.604 j; 0.130
N-M-CM N- T-CM - 0.577:t 0.141
GL-B-M-CM GL-B- T-CM 50 0.476:t 0.123',1.
100 0.439:t 0.162'M
200 0.266 i 0.1 02" ~
Vp-16 80 0.208:t 0.116" ~

'p< 0.05 YS RPMI1640; '.p<0.01 YS. RPMI1640:

4p < 0.05 Y5. N-S-CM; Mp < 0.01 YS. N-S-CM

TABLE 8 TABLE 9
Effect of GL-B-M-CM on Apoptosis of HL.60 Effect of GL-B-T-CM on Apoptosis of HL-60
Cells In Vitro (n = 3, x:t SD) Cells In Vitro (n = 3, x::I: SO)
Concentration Apoptotlc cells Concentration Apoptotic cells
Groups mg-ml-1 (%) Groups mg'ml-1 (%)
RPMI1640 -1.29:!: 0.46 RPMI1640
N-M-CM -7.44:t 1.07- N.T-CM
GL-B-M-CM 50 18.81 :t 0.93"-d GL-B-T-CM
100 20.98 :t 1.57"" M
200 23.00 :t 0.56-- M
yp-16- ~ 71.45:1:2.43"",1A
"p<O.05 YS. RPMI164Q;""p<0.O1 Y5. RPMI1640;
Ap < 0.05 Y5. N-M-CM; 6Ap < 0.01 Ys. N-M-CM.

FIGURE 1. Effect of GL-B on apoptosis of HL-60 cells FIGURE 2. Effect of Gl-B-M-CM on apoptosis of
incubated for 72 h in vitro. Hl-60 cells incubatedfor 72 h in vitro-
(A) Control: (8) GL-B 50 ~g.ml-1: (C) GL-B 100 (A) Control; (8) Gl-B 50 ~g-ml-l; (C) Gl-B 100
~g.mr 1; (D) GL-B 200 ~g.mt1; (E) Vp-16 80 ~g.ml-l. ~g-ml-1; (0) Gl-B 200 1J9-ml-1;(E) Vp-16 80 J.19-m.l.

;lll
 16 05 10:31p p.?

level during 24-48 h, and subsidedafter 72 h. The

TNF-a level in the supernatant of 12.5-400
Jlg.ml-1 GL-B cultured with macrophagesrosedur-
ing 24 h as the dose increased(Fig. 5).

Effect of GL-B on IFN-y Production

of T Lymphocytes

Figure 6 shows that the IFN-y level in the su-

pernatantof 200 ~g'ml-1 GL-B cultured with T
lymphocytesrose in 24 h and subsidedafter 48 h.
The IFN-y level in the supernatantof 12.5-2()()
~g.mI-1 GL-B cultured with T lymphocytes rose
during 24 h as the dose was increased to 400
~g'mi-i. At that level it started to subside(Fig. 7).

FIGURE 3. Effect of GL-B- T-CM on apoptosis of

HL-60 cells incubated for 72 h in vitro.
(A) Control; (B) GL-B 50 J.1g.ml-l; (C) GL-B 100
~g.ml-1; (0) GL-B 200 J.1g.ml-1;(E) Vp-16 80 J.1g.ml-'.
Effect of GL-B on Macrophage
TNF-a Production
f'lgure 4 shows that the TNF-a level in the su-
pernalant of 200 ~g'mJ-1 GL-B cultured with
macrophages rose in 24 h, remained at the highest
DISCUSSION
In the presentstudy,we investigatedthe antitu-
mor activity of GL-B, a purified polysaccharide
isolated from Ganodenna lucidum. The results
showed thal GL-B inhibited the growth of im-
planted Sarcoma 180 significantly and in a dose-
dependent, combination immunotherapy, GL-B
with a chemotherapysuch as cyclophosphamide
achieved the best effect. On the ba is of the pre-
ceding in vivo results, we added GL-B directly to

u zu 40 60 ISU
Time (hour)

t'IGURE: 4. E:ftect of 200 j.lg'ml-1 GL-B on TNF-a induction in the super-

natant of murine peritoneal macrophages cultured with lPS after 3-72 h of
incubation (n = 4, x:t SD).

~l~

Jan
JAn 16 05 10:34p p.B

FIGURE 5. Effect of different doses of GL-B on TNF-a induction in the super-

natant of murine peritoneal macrophages cultured with LP5 after 24 h of incuba-
tion (n = 4. x:!: SO).

the in vitro cultures of Sarcoma] 80 and HL-6()
cells. Unexpected results showed that directly
adding GL-B to the in vitro tumor cell culturescan-
not inhibit their proliferation. evenat the very high
concentrationof 400 Jlg.ml-l. GL-B also hasno ef-
fect on inducing tumor cell apoptosismeasuredby
using a FACScan flow cytometer. It :\eems (hat
GL-B alone hasno direct effect on proliferation of
tumor cells.
In a further study,GL-B was first addedto the
culture medium of macrophagesand T lymph()-
cytes, then the conditioned GL-B-M-CM and GL-

-
80

FIGURE 6. Effect of 200 Jig.ml-1 GL-B in IFN.'YInOuCtlonin the supernatant or

murine spleen cells cultured with ConA after 3-72 h of incubation.

;ll~
 lO:37p
B- T-CM media were added to the in vitro H.l.-60
culture medium as a drug. The results showed that
both GL-B-M-CM and GL-B-T-CM significantly
inhibited proliferation of HL-60; the FACScan
flow cytometer also found that both of them sig-
nificantly induced HL-6() cell apoptosis.
To reveal further the mechanism of the antitu-
mor effect of GL-B-M-CM and GL-B- T-CM. the
leve.JofTNF-a in GL-B-M-CM and .lFN-yin GL-
B- T -CM were ac;sayedby bioassay and ELISA. re-
spective.Jy.TNF-a and IFN-y were known to play
important roles in suppressing tumor cell growth
and inducing apoptosis of many different kinds of
tumor cells (Thomson, 1991; Malorni and
Rainaldi, 1996). Many experiments showed that
TNF-a and lFN-y cooperated with each other in
inducing tumor cell apoptosis (Volm and Mattern,
1995; Rawadi et aJ.. 1996: Sveinbjinsson and
Rushteldt; 1997). Our experimental results
showed that the levels ofTNF-a in GL-B-M-CM
and IFN-y in GL-B-T-CM rose significantly in a
do5e-dependent mode. Moreover. our l"e!;ults
showed that there was a positive correlation be-
tween the level of TNF-a in GL-B-M-CM and
lFN-y in GL-B- T -CM and the antitumor effect of
GL-B-M-CM and GL-B-T-CM.
In the present study. we investigated the anti-
tumor activity and mechanism of GL-B. The re-
sults were a.'i follows: first, GL-B did inhibit the
214
p.9
growth of implanted Sarcoma 180 ill vi~'o signifi-
cantly and dose-dependently. Second. GL-B di-
rectly added to a culture medium neither induced
HL-60 apoptosis nor restrained its proliferation in
~'itro.Third. the macrophage or T lymphocyte cul-
ture medium treated with GL-B significantly in-
duced HL-60 apoptosis and inhibited its prolifer-
ation. GL-B significantly increased TNF-(X and
IFN-y release dose- and time-dependently. TNF-
(Xand IFN-y were known to play imp<)rtant roles
in suppressing tumor cell growth and inducing
apoptosis of rumor ceJIs. Finally. as untreated
macrophages aJ1dT lymphocytes produced little
or no TNF-(X and IPN-y. and N-M-CM or N- T-CM
did not inhibit HL-60 proliferation or induce its
apoptosis, it seemed that the antitumor activity of
GL-B was related to promoting TNF-(X release
from macrophages and IFN -y release from T lym-
phocytes.
REFERENCES
Lei L. ~., and Lin Z. 8. 1991. Effects of Gan()dennap<)ly-
saccharideson the activity of DNA polymerasea in spleen
cells stimulated by alloanligens in mice in vitro. J Beijill,,?
Med uni\'. 23,329-333.
1.e.1
L. S., and LiD Z. B. 1992. Effects of GalllJdemlo poly-
saccharideson T cell subpopulationsand pn)(]uction of in-
0510:40p p.IO

rerleukjn 2 in mixed lymphocyte resp<Jnse. Acta Pharmaceut Sasaki T. 1971. Antitumor polysaccharidesfr.)m some pI.,ly-
Sin;ca. 27, 331-335. porace. Gullodermu (PeN.) Pal. and Phellimls l.,l/l/t~US
Li R. Z., and He Y. Q. 1991. Antiaging principle of GantJ- (Berk. ex CUrl.) Aoshtma. Chern Phurm 81111. 19,821.
den/la Itlcidum and active component. J Beijing Med Un;v. Sone Y.. and Okuda R. J985. Structuresand anlilumoraCliv-
23,473-475. ity of polysaccharides isolated from fruitiJlg b{~y und Ihe
Ma Li, LiD Z. B.. Li R. Z. et al. 1991. Effects of Ganodenmt g«)wing cullure or mycelium of Ganodenlla lilcit/ilm. Agric
polysaccharides on IIJ-2 production by mQUsesplenocytes BioI Chem. 49, 2()41-2653.
in I'itro. J Beijing Med Uni!." 23.412-416. Sveinbjinsson B., and Rushfeldt C. 1997. Cytutoxi..,effet:I.1f
Malomi W.. and Rainaldi G. 1996. Tumor necrosis factor u cYlokine!; on murine colOIl c:u'cinomucells involve!; TNF-
is a powerful apoptotic inducer in Iymphl-)idleukemic cells mediated apoploslS. Bicchem Bi('phy.~Re.~Commllll. 233,
expressingtheP 170glucoprotein. Int J Cancel;67.238-247. 27(}-275.
Maruyama M.. etat. 1989. AntitUmor activity of Sarcodonas- Thomson A. 1992. The Cytokille Hankbook. A.:ademic Pt-es~.
perotIt.' (Berk.) 5.lto and Ganodennctluc;dum (Fr.)Karst. J pp.241-256.
Phafwa...obiod)'n. 12. 118. Volm M., and Mattern J. 1995. Isolation of Oap_C{. a novel
Miyazaki T.. and Nishjima M. 1981. Studieson fugal poly- mediator of interferon-gammu induced cell death. J Bioi
saccharides 17. Structural examination of a water-soluble. Cnem. 270, 27932-27936.
antitumor polysaccharide of Ganudenna luL';dum. Chefn Xia D., and Lin Z. B. 1989. Effects of GallO/lelma polysac-
Pharm Bu/l, 29.3611-3616. charides on immune function in mice. J Beijlllg Med {!ni.\
Rawadi {;.. et al. 1996. Effects of MYt.'op/a,~Inaformentans
on 21,533-5.C{6.
the myelomonocytic lineage. J lInlmtnol. 156,670-678.

215