Vol.
1, /165-i
/70,
October
/995
Clinical
Liposome-encapsulated
Adjuvant
Muramyl
Immunotherapy
A Randomized
David
M.
Ilene
for Splenic
Multi-Institutional
Vail,2
D.
E. Gregory
Kurzman,
Richard
R. Dubielzig,
C. Helfand,
William
C. Kisseberth,
Cheryl
A
Joyce
Bruce
London,
R. Madewell,
Fidel,
Mona
Rosenberg
in
splenic
a large
Jr,
highly
Madison,
Specialists,
Houston,
Referral
Group,
Los
[D. M. V., E. G. M., I. D. K.,
Pathobiobogical
Sciences
[R. R. D.],
Wisconsin
53706;
Oakland
Texas
Angeles,
77034
[5. 5.]; and
California
90064
canine
Cancer
the
is
a naturally
MDP,
with
TNF-cx
to determine
the efficacy
This
study
was
HSA
and
without
gross
evidence
equivalent
(placebo
fobbowed
survival
necrosis
assessed
on
to
liposomes).
determine
a
small
subset
had significantly
(P
and
0.037)
overall
dogs receiving
placebo.
significantly
prolonged
overall
survival
ical stage
cantly
disease-free
or
subse-
survival
(P
=
II disease
greater
serum
of
prolonged
survival
and
receiving
disease-free
survival
compared
clinical
stage I disease
survival
(P = 0.026)
0.017) compared
tumor
Dogs
(P = 0.029)
Dogs with
disease-free
Dogs
dogs.
receiving
necrosis
not
IL-6,
among
other
in dogs
presently
had
and
2
To whom
Medical
requests
Sciences,
for reprints
2015
Linden
should
Drive
6/5/95.
Ltd., Basel,
be addressed,
West,
Madison,
a biposome
tissue
the
highly
as a model
OSA.
uptake
system
and
In humans,
monocyte
cytokines
of
in plasma
fur-
as well
tumonicidal
concentrations
of
(6, 9, 10).
therapeutic
metastatic
qualities
blinded
trials
L-MTP-PE
and overall
for
human
cancer,
evaluating
the
efficacy
expanded
our
We
have
metastatic
spontaneously
of
and uniformly
involving
was found
to
survival
times
(13, 14).
occurring
a national
of
trials
trial
L-MTP-PE
of
is
in
L-MTP-PE
occurring
HSA
often
splenic
HSA.
is a highly
metastatic
spontaneous
in the spleen
and occurring
more
L-
fatal
tumors
in
including
dog than in any other species
all cancers
in the dog. Large
cancer
commonly
(15-21).
HSA accounts
breed dogs, particularly
arising
in the
for 7% of
German
(P < 0.001)
The
abbreviations
TNF-a,
5/16/95;
accepted
by Ciba-Geigy
MTP-PE
to those
given alone or after chemotherapy
of the relevance
of spontaneously
underway
highly
and
in plasma
previously
used
liposome-encapsulated
revised
supported
within
endocytosis
with OSA,
intervals
with
most
of MTP-PE
In randomized
to other
with
half-life
100 dogs
metastasis-free
children
of MDP,
are similar
of a longer
(13-15).
in dogs
OSA
1).
that L-MTP-PE
can result in regression
in the Bl6 mouse melanoma
model (2,
another
significantly
when
Partly on the basis
(1-1
cell walls.
that
increases
increases
described
approximately
prolong
the
3
Received
4/7/95;
I This
study was
only
causes
and
and
derivative
in the circulation.
also
for OSA,
dogs,
with dogs with dinL-MTP-PE
had signififactor-a
We
cancer
times. The effects of L-MTP-PE
on serum
factor-a
and interleukin
6 activity
were
L-MTP-PE
=
12).
stage,
were
and
MTP-PE
of metastases
Dogs
L-MTP-PE
bipo-
human
mechanisms
bipophibic
the
or
MDP3
dipabmitoylphosphati-
the monocyte/macrophage
its half-life
but
of
and
effects
free
as
destroy
of bacterial
enhances
It has been shown
of established
metastasis
of biposome-encapsulated
muramyl
tripeptide
phosphatidylethanobamine
(L-MTP-PE)
when used in combination
with splenectomy
and systemic
chemotherapy
for the treatment
of HSA in the dog. Thirtywere treated with splenectomy,
stratified
by clinical
and randomized
to receive doxorubicin/cyclophosphamide
chemotherapy
and either
L-MTP-PE
immunotherapy
dogs,
activity,
variety
Encapsulation
within
and
MTP
the addition
greatly
result
metastasis.
a
of
component
toxicity.
then prolongs
visceral
by
conjugate
by
such
to recognize
immunostimubant
Canine
splenic
hemangiosarcoma
(HSA)
is a spontaneous tumor
with high metastatic
potential.
Despite
surgical
excision,
most dogs die within
2 months
of diagnosis
as a
activated
It is a synthetic
has general
as
ability
occurring
natural
monocytes
cells
the
dylethanolamine.
of MTP-PE
overall
tumor
occurring,
may have potential
as
investigation
of anti-
immunomodubatons,
neoplastic
MTP-PE
ABSTRACT
of widespread
and
acquire
MTP,
Veterinary
Veterinary
[M. R.]
Macnophages
some-encapsulated
and lower
quently
spontaneously
chemoimmunotherapy.
(L-MTP-PE)
bipid
malignant,
HSA in the dog. Canine
HSA
animal
model
for additional
metastatic
Referral Services,
Bloomfield
Hills, Michigan 48302 [J. E. 0.];
School of Veterinary
Medicine,
University
of California-Davis,
Davis,
California
95616 [B. R. M., C. 0. R.]; Special Veterinary
Services,
Berkeley,
California
94704 [J. F.]; Greater
Houston
Veterinary
with
activities
compared
with plahas significant
antimetastatic
INTRODUCTION
Wisconsin-Madison,
two dogs
in the Dog:
and
of Veterinary
Medicine,
and the Comprehensive
Cancer
[D. M. V., E. G. M., I. D. K., S. C. H], University
of
designed
1165
Trial’
activity
Rodriguez,
Susaneck,
Departments
of Medical
Sciences
S. C. H., W. C. K, C. A. L.] and
School
Center
0.
Carlos
Steven
Hemangiosarcoma
and interbeukin
6 (P = 0.007)
cebo-treated
dogs. L-MTP-PE
E. Obradovich,
Research
Phosphatidylethanolamine
Clinical
MacEwen,
Stuart
Janean
Tripeptide
Cancer
coma;
Switzerland.
at Department
WI
53706.
4
of
tumor
HSA,
F. Shi,
production
saccharide
are:
muramyl
necrosis
factor
MDP,
muramyl
dipeptide;
L-MTP-PE,
tripeptide
phosphatidylethanolamine;
a; IL-6, interleukin
6; OSA,
osteosar-
hemangiosarcoma.
I. D. Kurzman,
and
E. G. MacEwen.
of interleukin
6 induced
in normal
dogs, submitted
in vitro and in vito
by muramyl
peptides
and lipopolyfor publication.
Downloaded from clincancerres.aacrjournals.org on June 18, 2017. © 1995 American Association for Cancer
Research.
1166
Adjuvant
Immunotherapy
shepherd
and retriever
excision
(i.e.,
diagnosis
The
because
addition
resulted
of
are predisposed.
to
This
study
was
the efficacy
splenectomy
and
other
doxorubicin
or without
times
for dogs
of
organs.
and cycbo-
vincristine),
with
splenic
HSA
therapy
that
Hospital
at the
collected
for assessment
and
4
in a randomized
when
manner
used
of
evabuated
on TNF-a
HSA
in the dog.
to determine
and
with
chemotherapy
occurring
was
to eval-
in combination
IL-6
the
activities
were
h
after
Serum
AND
Subject
In
effect
line
was
7TD1
used
in this
cells
was
untreated,
stage
histologically
I (spleen
without
current
confirmed
not
evidence
in serum.
ruptured)
conditions,
trial
were
Randomization
entered.
in which
University
(I. D. K.,
all participating
treatment
count,
abdominal
from
ruptured)
were
Pretreatment
history,
physical
assay
4-6
After
All
surgery,
domized
dogs
to
(placebo
All
dogs
doxorubicin
m2) given
weeks
underwent
and
stratified
either
curve.
dog
complete
All
were
electrocardiogram,
treatment
of L-MTP-PE
mg/m2.
This
without
Liposome
MTP-PE
Ltd.
(Basel,
was
dose
was
freeze-dried
were
by addition
prepared
of 40
for
hydration,
the
eight
vial
liposome
and
Follow-Up
graphs
MTP-PE
dogs
administered
Evaluations.
before
examination,
were
contents
preparation
each
performed
(four
dogs
(4 mg/vial).
were
was
via
shaken
infused
the
1 mm
vigorously
for
through
a 10-jim
vein.
blood
counts
were
chemotherapy
treatment.
A complete
abdominal
ultrasound,
and thonacic
radioevery
receiving
2 months
L-MTP-PE
after
splenectomy.
and
four
For
receiving
were
died
Bresbow
and
Mantel-Cox
tests
difference
between
of clinical
stage
Bresbow,
Mantel-Cox,
significance
and
Cox
the
alive
causes
at the
Kaplan-
or lost to
compared
time
of
using
significance
the
to determine
In addition,
treatment
of
of progmes-
using
(23).
within
the effect
groups
using
the
hazard
tests
for
proportional
in serum
were
compared
groups
using
respectively.
identified
t
test
(between
of TNF-a
treatment
groups
repeated
t test
t
groups).
was
were
groups).
compared
using Pearson’s
significant.
24
The
between
2 test.
found,
groups)
dogs
the number
metastasis
(between
In
IL-6
between
and multiple
were
test (within
and
and
measures
differences
examination,
confirmed
metastasis
clinical
stage
was considered
one-way
activities
If overall
postmortem
independent
within
by use of the pained
histologically
organ
was evaluated
changes
time
treatment
of
curves
of
number
(22-24).
Overall
over
survival
and
development
that were
were
overall
as the num-
as a result
unrelated
groups
and
as the
generated
the
L-MTP-PE
defined
defined
for dogs
from
as the
on
dilutions.
the
euthanasia
accounts
containing
defined
to the
was
Treatment
independent
1
curves
which
on had
or
wells
survival
was
the
compared
proliferation
of surgery
time
to death
Survival
method,
complete
to allow
time
survival
(1.53
samples,
for sample
survival
of
2,3-bis[2-
was
was
between
(22).
were
to vials
the
surgery
ANOVA,
1-palmitoyl-2(1 g total bipid/
a peripheral
Complete
or
from
the
Disease-free
Disease-free
was
viability
test
of IL-6
adjusted
compared
plate
for an additional
the
maximal
analysis
2
(13).
from
ml saline
After
or without
performed
physical
were
dog
were
Overall
follow-up
of i.v.
on the first
in the
with
filter
double-
doses
studies
vial)
syringe
Meien
twice weekly
for 8
period.
The first
and
ratio
The
ran-
Preparation.
Lyophilized
liposomes
with
(CGP
19835A)
were supplied
by Ciba-Geigy
preparations
of days
metastasis.
(100 mgI
within
2
began
containing
dioleoyl-phosphatidybsenine
oleoyl-phosphatidylcholine
at a 3:7 molar
mm.
and
cycles
all subsequent
Liposomes
ben
equivalent
were
four
treatments
on previous
Switzerland).
groups.
disease.
stage
assignments
1 mg/m2;
based
placebo
sive
to receive
Liposome
gross
were
to 7TD1
The
samples
from
unit
50%
Analysis.
times
clinical
day of chemotherapy.
Liposomes
were given
weeks via a slow i.v. infusion
over a 5-8-mm
dose
and
a lipid
inducing
Results
survival
from
or
IL-6
One
in
IL-6
methosulfate
of
test
generated
IL-6.
added
50 jib 1 mg/mb
activity
pA of
wells
At that time,
incubated
containing
curve
100
human
assay.
phenazine
was
of 1 X i0
Then,
IN) were
was
hybrid-
to appropriate
CO2.
with
plate
IL-6
human
Statistical
and abdominal
(30 mg/m2)
plus iv. cycbophosphamide
together
once every
3 weeks,
beginning
of splenectomy.
standard
days
by
scheduled
the
metastasis.
Dogs
were
surgery
to be included.
L-MTP-PE
determine
standard
with
The
of the wells
of
splenectomy
for overt
tumor after
were
receive
liposome).
blind.
dogs
of the abdomen
to be free of gross
h. To
absonbance
amount
profile,
A total
for each
by adding
mixed
well.
mouse
of recombinant
curve
for
cell
for growth,
plate.
Indianapolis,
(XTT)
bioassay
The
on IL-6
added
concentrations
determined
salt
each
and thonacic
Treatment.
was
to each
jig/jib)
radiographs.
evaluation
required
cells
recombinant
examination,
were
for 44 h at 39#{176}C
in 5%
WI),
biochemistry
(9).
of a microtiten
a standard
liposome
serum
were
at 0, 1, 2,
fibrosancoma
previously.4
of sera
Mannheim,
to develop
Madison,
ultrasound
previously
to wells
A
mouse
is dependent
as described
Increasing
as to the
examination,
activities
Bioassays.
WEHI-164
which
dilutions
bide inner
was
by one investigator
of
IL-6
the
methoxy-4-nitro-5-sulfophenyb]-2H-tetrazolium-5-canboxani-
confacilities
Teaching
samples
biposome/chemotherapy
disease,
six veterinary
evaluation
IL-6
7TD1
This
blinded
and
line,
added
incubated
for this study.
of Wisconsin-Madison,
and
clinical
of complicating
eligible
dogs
veterinarians
a complete
blood
free
was performed
administered.
included
and
with
Medical
serum
combined
as described
cell
triplicate.
previously
HSA
or II (spleen
were
a multicentem
with
splenic
of metastasis,
disease
dogs
first
using
used,
serial
cells
Thirty-two
of TNF-a
TNF-a
oma
2-fold
METHODS
Populatioa.
of the Veterinary
of Wisconsin,
the
activity
(Boehminger
MATERIALS
patients
University
treatment.
TNF-a
designed
of dogs
placebo)
3,
has
16).
of spontaneously
a subset
surgical
2 months
doxonubicin/cycbophosphamide
for the treatment
of L-MTP-PE
lungs
typically
of L-MTP-PE
and
addition,
(15,
Despite
die within
the
(with
survival
days
dogs
metastasis
combinations
in median
HSA
most
of chemotherapy,
145-180
uate
breeds,
splenectomy),
phosphamide
only
for Canine
they
and the
undergoing
of organs
compared
frequency
treatment
with
using
the
of specific
groups
For all analyses,
and
P < 0.05
RESULTS
Subject
of the treatment
Demographics.
groups
Subject
are given
(Table
demographics
1). Nine
for both
dogs
Downloaded from clincancerres.aacrjournals.org on June 18, 2017. © 1995 American Association for Cancer
Research.
were
of
Clinical
Table 1
Subject
L-MTP-PE
Age (yr)
Weight
(kg)
Clinical
stage
I
10.2
27.2
(n
group
16)
=
(10.1);
4-14”
(25.7);
6.5-40”
Placebo
(n
Male-intact
S
2
Male-neutered
Female-neutered
3
8
8
6
II
Sex
0.8
16)
0.7
10 (9.6); 3-13”
31 (27.6);
10-50”
7
9
(mean);
0.6
0.5
0.4
I
0.3
0.2
range.
0.1
100
Table
2
Analysis
of diseas
e-free
clinical
L-MTP-PE
Placebo
surviva
1 and
Median
disease-free
survival
(days)
No. of
dogs
All
o verall
survival
Median
survival
(days)
P
value
P
277
143
0.029
15
17
212
122
0.017
355
148
0.017
8
232
0.69
425
0.46
7
166
L-MTP-PE
8
164
Placebo
9
96
treatment
Placebo
Clinical stage
and
of
the
and
the study
anabysis.
remainder
0.031
162
0.037
I
96
Data.
dogs
represented
Complete
died of HSA,
of complications
evaluation.
29
were
postmortem
of the dogs
dogs
and
still alive
#{176}_rl:.LLt.L______L4____l_Placebo
survival
survival
times
(P
that
survival
was 149 days, and
163 days. Table
2 sumoverall
0.029),
=
survival
times
of
with
clinical
stage
I, there
L-MTP-PE
significantly
stage
II
times
disease
(Figs.
was no significant
and placebo
groups
vivab
on overall
there
was
survival
a significant
longer
survival
times.
difference
disease-free
(P
0.017)
=
3 and
4).
survival
than
Within
However,
between
within
clinical
L-MTP-PE
(P
did dogs
clinical
difference
observed
between
with regard to disease-free
stage
and
300
400
500
700
600
I
800
treated
overall
with surgery
survival
duration
I
900
1000
for dogs with
chemo-
estimates
and doxorubicin/cycbophosphamide
therapy,
then randomized
to L-MTP-PE
or placebo.
L-MTP-PE
had significantly
longer
overall
survival;
marks,
censored observations.
cebo groups
and overall
with respect
survival
times
Patterns
Less
had
overall
200
Kaplan-Meier
2
HSA
stage
and
I
Survival Time (days post surgery)
Fig.
liver
I disease
100
0
the
sumII,
pba-
7).
(n
evaluations,
13 dogs),
=
Penitoneal
neum
(n
Dogs
P
receiving
0.029.
Tick
=
frequent
sites
and
one
bladder.
Overall,
had postmortem
(87%)
with
the number
of the
three
penitoneal
of metastasis
instance
often
(n
12),
panietab
included
each
of organs
stage
II disease
developing
common
and
of
(P
s.c.
=
0.08).
metastatic
sites
and
(n
tissue,
clinical
compared
metastasis
complete
involved
and
kidney
brain,
0.03 1)
=
the
(n
lung
visceral
omentum,
five dogs (55%)
with
evidence
of metastasis
clinical
most
both
organs,
(P
undergoing
most
surfaces
include
abdominal
survival
In dogs
metastasis
surfaces
involving
3),
to disease-free
(P
0.037).
of Metastasis.
postmortem
clinical
(Figs.
0.026)
1
for
A signifand pla=
0.037)
regardless
I
were
Of the 29 dogs
according
to clinical
stage and treatment
group.
difference
was observed
between
the L-MTP-PE
groups
with regard
to disease-free
survival
(P
overall
--
at the
I and 2). Approximately
37% of dogs receiving
L-MTP-PE
were disease-free
after 1 yean,compared
with only
15% of dogs receiving
placebo
biposomes.
Dogs with clinical
=
L-MTP-PE
(9 1%)
6 (21%)
died of other causes,
1
of chemotherapy,
and 1 died
manizes
stage
1000
11
examinations
that died.
was
the disease-free
In all,
and 3 (9%)
The overall
median
disease-free
the overall
median
survival
time
and
900
166
of the
Survival
interval,
for 24 (83%)
died, 20 (69%)
died as a result
without
necropsy
dogs
icant
cebo
:
.,
Treatment
time
800
0.9
breeds.
available
700
II
breed,
during
600
(days post surgery)
Survival
groups
Clinical
stage I
Clinical
stage II
Clinical
stage I
L-MTP-PE
died
500
400
stages
0.037
mixed
300
Fig.
I
Kaplan-Meier
disease-free
survival
duration
estimates
for dogs
with HSA treated
with surgery
and doxorubicin/cycbophosphamide
chemotherapy,
then randomized
to L-MTP-PE
or placebo.
Dogs receiving
L-MTP-PE
had significantly
longer
disease-free
survival;
P = 0.037.
Tick marks,
censored observations.
value
188
127
different
200
Disease-Free
16
16
All
1167
0.9
8
8
aMedian
Research
demographics
Treatment
Parameter
Cancer
pemito-
diaphragm.
=
3), heart
skin,
and
stage I disease
with 13 dogs
A comparison
and
the frequency
by
treatment
Downloaded from clincancerres.aacrjournals.org on June 18, 2017. © 1995 American Association for Cancer
Research.
of
and
1168 Adjuvant
Immunotherapy
for Canine
HSA
0.9’
0.8’
Dl’
0.7’
.
0.6
J
g
0.5’
0.4#{149}
a
I
#{176}
0.3’
0.2
:
0.1’
:
0’
I
I
I
I
I
I
I
100
200
300
400
500
600
700
Disease-Free
Survival
observations.
clinical
stage
groupings
clinical
stage
I disease
stage
=
II disease
dogs
that
frequently
receiving
time
when
Dogs
receiving
activity
the placebo
No
and
no
IL-6
Activities.
change
TNF-a
receiving
ity,
L-MTP-PE
PE-treated
at
0.007)
and
had
than
3 (P
higher
compared
were
serum
with
observed
appeared
to
be
administration.
serum
were
IL-6
those
0.013)
=
dogs
in
over
similar
returning
Dogs
activities
were
group
groups
was
reported
weights,
under
dogs).
HSA
because
were
elsewhere
sex
similar.
chemotherapy
and
equal
two
by clinical
representation
was
stage
L-MTP-PE
has
less
frequent
with
dogs
did
liposomes
had
disease-free
and
after
splenectomy
(15,
16).
In contrast,
dogs
for
cheme-
in
stage
had
means.
stage
I dogs
this
explain
this
however,
this
makes
opportunity
stage
the
for
are
OSA
(13,
and, again,
is combined
14)
The
use
of
to a
they show
a
with che-
for the antimetastatic
activby the patterns
of metastasis
dogs.
had
overall
survival
times
when
compared
stage
I dogs
receiving
Although
disease-free
receiving
and
overall
placebo,
was
not
Dogs
with
significant.
stage
direct
stage
The
cause(s)
likely
L-MTP-PE
have potential
advantages.
enhance
on suppress
the
I disease
were
Certain
effects
less
to therapy;
stage
II disease
spleen
with
extension.
to enjoy
concomitantly
II
of the metastatic
does not appear
to
because
penitoneab
are more
times
their
subsequent
of the tumor-bearing
pro-
survival
unlike
metastasis
sense,
significantly
Dogs
better
with
is
subsewith
outcomes
(i.e., longer
disease-free
and overall
survival
times
frequent
metastasis);
therefore,
banger patient
numbers
required
to establish
a difference
in this group.
may
may
im-
previously
L-MTP-PE-tmeated
intuitive
I disease
for
with
I disease
pemitoneal
by the rupture
findings
reported
is unresolved.
A comparison
stage I and stage II disease
difference.
days
stage
difference
to develop
277
for dogs
These
metastatic
II disease.
longer
nate for placeboof
reported
the benefits
with
the
mate in the
Dogs
with clinical
stage
II disease
developed
fewer
sites of metastasis
dogs.
In particular,
hepatic
metasta-
and
for this difference
pattern
between
clinical
HSA.
Dogs
combination
time
with
clinical
with
L-MTP-PE
defined
antitumor
a 15%
any
extend
in dogs
disease-free
likely
between
to only
is the longest
by
they
after
of disease-
compared
survival
survival
motherapy.
Additional
evidence
ity of L-MTP-PE
is represented
quent
that
spontaneously
occurring
and cyclophosphamide
placebo
1).
treatment
overall
survival
times similar
to those reported
previously
dogs with splenic
HSA
receiving
similar
combination
motherapy
median
treated
L-MTP-PE
with
were
in the dog (15-2
of the
Stratification
nearby
groups.
demonstrates
in dogs
with
doxomubicin
for HSA
distributions
study
chemotherapy
prolongation
tumor
with a different
histogenesis,
favorable
outcome
when
L-MTP-PE
longed
=
with
significant
dogs
portant,
than
under
combination
compared
The
splenic
Dogs
administration.
for the population
in ensuring
the two treatment
This study
activity
receiving
and
study
successful
in
group
treated
sis
signifi-
at 2 (P
800
Time (days post surgery)
free
and overall
survival
times
when
placebo
group
(37%
1 year disease-free
toward
in the L-MTP-
that
of the placebo
h after
demographics
to those
ages,
.
700
observed
in this study.
treated
with L-MTP-PE
than did placebo-treated
time
DISCUSSION
The
.
600
experienced
counterparts,
Subject
.
500
L-MTP-PE
splenectomy
for
however,
peaked
at 2 h
administration.
significantly
activity
3 h and
by 4 h after
group
greater
was
lipid equivalent
(Fig. 6). In contrast,
showed
increased
serum
IL-6 activ-
placebo
values
cantly
IL-6
L-MTP-PE
peaked
baseline
had
receiving
(P < 0.001).
receiving
which
with
activity
L-MTP-PE,
administration
in serum
receiving
,
400
4
Kaplan-Meier
overall
survival
duration
estimates
for dogs with
HSA stratified
by clinical
stage.
Dogs with clinical
stage I disease
had
significantly
longer
overall
survival;
P = 0.017. Tick marks,
censored
observations.
L-MTP-PE
0.01)
=
In dogs
in serum
Dogs
5).
2 h after
group
.
300
L-MTP-PE-tneated
(Fig.
changes
in dogs
dogs
(P
compared
showed
increased
serum
TNF-a
activity,
which
and returned
to near baseline
values by 4 h after
TNF-a
,
200
Fig.
ceiving
with
dogs
organs
0.03)
=
Dogs
with
metastasis
(P
and received
II disease
in fewer
(P
3.
were
penitoneal
stage
metastasis
TNF-a
over
Table
than
develop
clinical
in the liven
biposomes,
noted
in
likely
placebo.
Serum
placebo
less
had
developed
L-MTP-PE
and less
presented
to subsequently
Dogs
0.04).
are
were
,
100
(days post surgery)
3
censored
0
Survival
Kaplan-Meier
disease-free
survival
duration
estimates
for dogs
with HSA stratified
by clinical
stage.
Dogs with clinical
stage I disease
had significantly
longer
disease-free
survival;
P = 0.026. Tick marks,
Fig.
-
and less
would be
doxorubicin
chemotherapy
agents
of immunothemapeutic
Downloaded from clincancerres.aacrjournals.org on June 18, 2017. © 1995 American Association for Cancer
Research.
Clinical
Table 3
No.
Grouping
dogs
clinical
All
All
Me tastatic
No. of
organs
of
dogs
involved”
24
1.8 (0-5)
site s in dogs with p ostmortem
P
value
Peritoneal
metastasis
13 (54%)
0.36
10
1.4 (0-5)
Placebo
All treatment
groups
Clinical stage I
14
2.0 (0-4)
Clinical
stage II
Clinical
stage I
L-MTP-PE
Placebo
Clinical
stage II
L-MTP-PE
15
2.1 (0-4)
4
5
6
9
Placebo
a Mean
(range).
1.2 (0-5)
9
4 (40%)
9 (64%)
Lung
metastasis
P
value
7 (29%)
0.22
3 (33%)
10 (67%)
2.0 (0-5)
0.6 (0-2)
0.29
2 (50%)
1.0 (0-3)
2.7 (1-4)
0.01
5 (50%)
0.24
1.00
2 (20%)
5 (36%)
0.40
0.04
4 (44%)
3 (20%)
0.20
0.07
2 (50%)
0.76
7 (50%)
0. 1 1
3 (33%)
10 (67%)
2 (50%)
0.34
1 (20%)
0 (0%)
2 (33%)
2(40%)
3 (50%)
0.03
8 (89%)
0.26
0 (0%)
7 (78%)
0.11
3 (33%)
1500
I
*
:
Placebo
D
L-MTP.PE
75-
I
‘‘
U....
<
P
value
12 (50%)
100-
.
0
50-
E
4)
1000
E
Placebo
0
L-MTP-PE
T
500#{149}
C1)
25-
I
I
1
F:T[.F:
r!.
I
I
0
1
2
4
3
Time Post Injection
(hours)
such
that
as L-MTP-PE.
doxomubicin
monocytes
induced
rise in circulating
toxic
potential
dren
with
hanced
monocyte-mediated
and
activity
TNF-a
activities
In
dogs
of both
and
IL-6
25).
the
Furthermore,
in studies
with
(26,
effect
cytokines
activities
en-
27).
in serum.
been
there
Similar
observed
IL-6 activity
cantly
0.013)
dogs
receiving
induction
with
respect
immunological
in
identical
doses
to tumorcidal
activity
these findings
suggest
that L-MTP-PE
immunomodubation,
and that dogs with
results
of this
antimetastatic
clini-
with surgery
mally
splenic
and
randomized
to L-
of L-MTP-PE4
Induction
of circulating
TNF-a
and IL-6
also has
strated
in human
patients
with various
cancers
L-MTP-PE
(8, 28-30).
Although
the significance
The
significant
increases
then
Dogs receiving
L-MTP-PE
had signifigreater serum IL-6 activity at 2 h (P = 0.007) and 3 h (P =
after administration
than did dogs receiving
placebo.
of similar
in-
i
(hours)
of dogs with HSA treated
chemotherapy,
4
(I)
after
was
in
Serum
6
doxorubicin/cycbophosphamide
MTP-PE
or placebo treatment.
serum
for a 4-h period
L-MTP-PE,
have
a
of chil-
on
Fig.
cabby healthy
cyto-
doxomubicin
of L-MTP-PE
was determined
receiving
laboratory
of canine
After chemotherapy,
with enhanced
administered
cytotoxicity
of dogs,
IL-6
administration.
creased
serum
(10,
L-MTP-PE
In a subset
TNF-a
work by our
the activation
by L-MTP-PE
(10).
monocyte
populations
occurs
OSA,
Previous
enhance
can
3
Time Post Injection
Fig. 5 Serum TNF-a activity
of dogs with HSA treated
with surgery
and doxorubicin/cycbophosphamide
chemotherapy,
then randomized
to
L-MTP-PE
or placebo treatment.
Dogs receiving
L-MTP-PE
had significantly
(I) greater
serum TNF-a activity at 2 h after administration
than did dogs receiving
placebo;
P < 0.001.
agents,
revealed
1169
stages
L-MTP-PE
,
Research
examinations
P
value
Liver
metastasis
Cancer
results
splenic
been
(9).
demon-
treated
with
of cytokine
is not entirely
clean,
in some form of
HSA are capable
responses.
study demonstrate
that L-MTP-PE
activity
in highly
malignant,
has
mini-
chemotherapy
responsive,
spontaneously
occurring
HSA in the dog. Canine
HSA may have relevant
poten-
Downloaded from clincancerres.aacrjournals.org on June 18, 2017. © 1995 American Association for Cancer
Research.
1170 Adjuvant
Immunotherapy
for Canine
HSA
tiab as a model
for additional
investigation
therapy
and the role immunomodubation
of
may
antimetastatic
play in such
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D M Vail, E G MacEwen, I D Kurzman, et al.
Clin Cancer Res 1995;1:1165-1170.
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