Download Prolonged Disease-free Period in Patients with Advanced Epithelial

Vol.
1, 501-507,
May
1995
Clinical
Prolonged
Disease-free
Epithelial
Ovarian
Period
Cancer
Tumor-infiltrating
Kazuyuki
Fujita,
Koichi
and
Adoptive
Shoji
two
Kodama,
Takeshi
Tanaka2
Department
of Obstetrics
Niigata
and
Gynecology,
was
(5)
ited
that
more
patients
Thirteen
did not show
any
with
detectable
epithelial
lesion
ovarian
after
cancer,
who
hardly
cisplatin-containing
chemotherapy
following
primary
operation,
were treated
with adoptive
transfer
of tumor-infiltrating
lymphocytes
(TIL group). Eleven patients
with almost
equivalent
conditions of disease, who were treated with only chemotherapy
following
primary operation,
served as a control group. The
median time of follow-up
was 36 (range, 23-44)
months in
the TIL group and 33 (range, 14-48)
months in the control
group. The estimated
3-year overall survival rate of diseasefree patients in the TIL group and in the control group was
100%
and 67.5%, respectively.
A significant
difference
was
noticed between
the overall survival
rate of the ilL group
and the control
group (P < 0.01). Furthermore,
the estimated 3-year disease-free
survival
rate of the patients in the
TIL
group and in the control
group
was 82.1 % and 54.5%,
respectively.
The disease-free
survival
rate of patients in the
TIL group and in the control group was significantly
different (P < 0.05). These results
suggest
that the adoptive
transfer
of TILs after all chemotherapy
has been finished
might
be one promising
method to achieve complete
cure of
advanced
epithelial
ovarian cancer.
to treat
can
cancers
achieve
lymphoma,
agents
as a supplement
cures
and
of certain
prolong
( 1-4).
However,
even
vanced
condition
cannot
lives
now,
have
been
used
to surgery
diseases,
be completely
20 years
and radiotherapy
such
of patients
epithelial
for over
as leukemia
with
ovarian
ovarian
improve
cancer
overcome
with
by the
primary
PATIENTS
7/19/94;
This
was
work
accepted
supported
2/3/95.
in part by a grant-in-aid
from
the Ministry
Health and Welfare for the Comprehensive
10-Year Strategy
Control.
2 To whom
requests for reprints should be addressed.
use lasts
the
infusion
This
3-5
with
alone
exhib-
result
to metastatic
the duration
only
adoptive
5 of 7 patients
of tumor.
rate of patients,
suggests
melanoma
of response
months,
we treated
ilLs
can
13 patients
cancer
with the adoptive
transfer
cisplatin-containing
chemotherapy
AND
of advanced
Gynecology
They
stage
Stage
were
to
with
of cryoprefollowing
a
METHODS
of either
Center,
or
April
1989
documented
and March
1992,
epithelial
ovarian
43 pacancer
(International
Federation
of Obstetrics
II, III, or IV) were enrolled
in this
treated
cology
in the
Department
Niigata
Niigata
of Obstetrics
University
City
Hospital,
Hospital.
and
Niigata
Eligibility
and
study.
criteria
GyneCancer
were
as
follows:
age not less than 18 years; Eastern
Cooperative
Oncology Group
performance
status
of 3 or lower (able to perform
minimal
self-care);
life expectancy
more
than 2 months;
no
chemotherapy
or radiotherapy
for 4 weeks
prior to entry into
protocol;
adequate
bone marrow
function
(WBC
count
>4000/
mm3;
platelet
count
> 100,000/mm3),
hepatic
function
(total
bilirubin
mine
concentration
>2.0
concentration
gave
Good
were
excluded
chemotherapy.
tumor
<
informed
Clinical
mass was
following
from
ination,
after
mg/dl),
1.5 mg/dl);
consent
cryopreservation
considered
renal
function
(creat-
infection.
to the Japanese
Guidelines.
All
Gov-
In 19 patients,
re-
clinically
detected
after completion
of
a primary
operation.
These
patients
other
and were
24 patients
detected
by CF scan,
and/or MRI, 1 1 patients
were
and
and no active
according
Practice
this study
In the
was
obtained
I
TIL
by
et a!.
melanoma
with
(6),
cancer
is similar
However,
since
Patients.
Between
tients with histologically
cells
Received
with
decrease
tumors
operation.
and
con-
treated
in single
epithelial
ovarian
served TILs after
and
ad-
cancer
a 50%
solid
Rosenberg
of metastatic
trials
ovarian
to ilLs.
from
rIL-2.
treated
clinical
the survival
sidual
tumor
chemotherapy
cancer
with
In our
by ilLs
ernment
Chemotherapeutic
generated
in
(1-4).
improve
the survival
rate of patients
in this manner.
In an attempt
to eliminate
minimal
residual
tumor
and
patients
INTRODUCTION
cells
patients
is detected
3 years
suspensions
regression
than
obtained
ABSTRACT
about
objective
ovarian
that epithelial
in sensitivity
of the disease
within
in 1 1 of 20
of ilLs.
epithelial
Niigata
recurrence
are lymphoid
observed
transfer
951 [T. T.], Japan
and
single-cell
reported
[A. T.];
and
of
of the patients
culturing
of Obstetrics
and Gynecology,
Niigata University
School
1, Asahimachi-dori
[K. F., H. I., Ko. T., S. K., Ke. T.];
of Obstetrics
and Gynecology,
Niigata City Hospital
Center,
treatment,
thirds
ilLs3
Takahashi,
Department
of Medicine,
Department
Cancer
501
Advanced
Transfer
ventional
Ikarashi,
Hirokazu
Tokunaga,
Kenichi
after
with
Research
Lymphocytes1
Takakuwa,
Akiteru
in Patients
Cancer
treated
with
in whom
ultrasonogram,
whose
TILs
or consent
as a control
no
residual
internal
examwere < 1 X i0
to receiving
group,
second-line
ilLs
was
not
and the remaining
of
for Cancer
The abbreviations
used are: ilL, tumor-infiltrating
lymphocyte;
rIL-2,
recombinant
interleukin
2; C-FDA, carboxyfluorescein
diacetate;
CT,
computed
tomography;
MRI, magnetic resonance
imaging.
3
Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer
Research.
502
Adoptive
Immunotherapy
Using
Cryopreserved
TIL
.13 patients
performed
until
4
Primar
or Incompleted)
Viable
cells
were
at the
density
containing
5%
24 patients
residual
the
tumor
DMSO,
4
1 1 patients
control
13 patients
group
NoTILs
TIL
1 x 10
>
ery rate
as
group
the
cells
number
Clinical
protocol
for ovarian
of
patients
cancer
3-h
in stages
II, III, and
IV.
three
another
before
cells
exceeded
received
(TIL
group;
Treatment
sues
with
treatment
Fig.
1).
Design.
resected
reached
TIL
100
in liquid
tion,
TILs were
at a primary
approximately
after
operation.
5
X
units/mI
rIL-2,
nitrogen
for several
the
obtained
When
10
after
TILs
were
from
the
2-3
months.
of chemo-
completion
weeks
frozen
of
and
After
cancer
number
tis-
C-FDA
mented
with
patients
for
20
ml
25%
mm
15-30
HBSS
opera-
5-fluorouracil-cyclophosphamide-Adriamycin-cisplatin
or
cyclophosphamide-Adniamycin-cisplatin
was administered
all patients.
5-Fluorouracil-cyclophosphamide-Adniamycin-cisplatin
350
was
mg/m2
on day
administered
according
to the following
cyclophosphamide
on day 1, 40 mg/m2
1, 50 mg/m2
orouracil
cisplatin
as a continuous
on day
infusion
phamide-Adriamycin-cisplatin
administration.
to five
serum,
were
All
performed
and
did not show
of chemotherapy
for
this
was
not
any
study.
After
visited
the
rence,
and
internal
serum,
and
hospital
examination
detectable
lesion
the primary
recovery
to the
regularly
examination,
toxicity
by
patients.
for
ultrasonogram
after
due
infusions
for TIL
After
early
tumor
or
CT
last
of
pa-
scan
in
were
For
a recov-
and cultured
S weeks
after
before
of
thawing),
administra-
administered
i.v.
were
and
washed
0.2%
to
FITC-labeled
in phenol
sodium
azide
mAb,
incubated
anti-HLA-DR
(Becton
at
Dickinson
In vitro
cytotoxicity
of TILs
fresh, frozen
targets
of autologous
and 1(562 cells.
Cryopreserved
digestion
were thawed
10% human
AB serum
was prepared
incubation
with
labeled
with
Sigma
C-FDA
three
(50
Co.,
times
with
and cultured
in
for 24 h. Single-
from monolayers
of viable
trypsin
at 37#{176}C
and washed
Chemical
incubated
was
or allogetumor
cells
with
various
ig/ml
diluted
St. Louis,
HBSS.
with
MO)
RPM!
for 60 mm
The cells
numbers
tumor
in the
(5
X
of effector
Japan),
and
an automated
Compact
MT).
as
C-FDA
in the surviving
fluorescence
microscope
Results
follows:
C-FDA
count - test
spontaneous
C-FDA
C-FDA
licates
were
three-step
were
<10%.
Target
of
Conversion
using
effector
The
units
count
-
in medium
to determine
respectively.
triplicate
of
(maximum
=
incubated
cells.
to lytic
cells was
photometer
C-FDA
cells
at
was
to percentage
cytotoxicity
X-100
were used
C-FDA
counts,
determined
titrations
converted
count)/(maximum
count).
alone or with 2% Triton
spontaneous
and maximum
with
were
percentage
10)
cells
37#{176}C
for 3 h. After incubation,
C-FDA
in the supernatant
masked
with
5 mM calf hemoglobin
(Wako
Pharmaceuticals,
percentages
recur-
marker
then
cytotoxicity
of thawed
cultivation
discharge,
detection
for
consid-
to the
and
( Reitz
completion
were
were
FCS
Cytotoxicity.
medium;
Ltd., Tokyo,
counted
with
chemo-
described
operation
from
check
and
eligibility
the patients
were given
rIL-2 was used only
administered
tients
the
and
at 37#{176}C
and washed
were
three
for tumor
marker
in
CT scan, and/or MRI
of operation
matched
following
chemotherapy
cycle,
TILs simultaneously.
and
the effect
who
for
After
albumin
1%
obtained
by enzymatic
RPM! 1640 containing
1640
5-fluorouracil
and
Japan).
Vitro
medium
5-flu-
cyclophos-
chemotherapy
examination,
check
by ultrasonogram,
patients
mg/m2
The
HBSS,
and 2 weeks
an appropriate
assessed
by using
neic tumor
cells
cell suspension
cells by brief
schedule:
Adniamycin
350
1-5.
excluded
underwent
to evaluate
Only
above
ered
regimen
patients
cycles.
Internal
and examination
therapy.
1, and
on days
to
at 37#{176}C
(with
TILs (1 X 10)
anti-CD56,
Tokyo,
In
cryopreserved
the primary
Ltd.),
months.
at 4#{176}C
for 45 mm, washed
twice,
and resuspended
in 0.5 ml
medium
for fluorescence-activating
cell sorting
analysis.
The
mAbs
used
were
anti-CD3,
anti-CD4,
anti-CD8,
anti-CD16,
Japan,
cultivation
Product
several
iO#{176}.
Just
the
7.5%
with
simultaneously.
containing
with
anti-CD25,
of cells
cells;
10k’
the phenotype
of TILs was determined
by flow
and in vitro cytotoxicity
of TILs was assessed
by a
assay.
The
TILs
in 100 ml sterile
saline
supple-
4#{176}C,
stained
patients
X
serum
with
1 X
in
of cultivation,
5
AB
to plain
cells/ml
i05
2 weeks.
freezing
Flow Cytometry.
13
X
weeks
for
The
(7).
removed
(Plancer
times
cells.
elsewhere
2
rapidly
isolated
i07
of TILs,
red-free
therapy
of
human
thawed
for
(3 weeks
X
were
2-3
nitrogen
culture
atmosphere
described
freezer
liquid
washed
above
2-6
approximately
were
70-95%),
cytometry
from
they
cultivation
or
No consent
1
in
(7).
of initially
was
After
in 30%
from
complete
number
the cells
reached
isolated
elsewhere
in anti-CD3-coated
with
was
rIL-2.
frozen
performed
in a humidified
sample
a programmed
described
cells/ml,
used
were
placed
at a density
TILs
was
described
IL-2,
activation,
cryopreserved
tion
Fig.
of
administration,
excluded
iO
tumor
with
then
using
were
and
(Costar)
medium
were
counted
at 37#{176}C.The
of CD3
number
TILs
(-)
TILs
methods
2 X
each
therapy
of TILs.
to the
medium
plates
complete
(+)
CO2
from
1 week
6-well
tumor
of
additional
detected.
exogenous
culture
After
19 patients
then
from
cells
complete
residual
according
free
viable
No
was
Infusion
tissue
medium
4
and
cancer
plates
Chemotherapy
4 weeks.
lesion
Culture
Operation
(Completed
every
a recurrent
the
All
measurements
SDs
was
of
the
performed
Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer
Research.
repas
Clinical
Table
I
Phenotypic
flow
cytometry
analysis
and
characteristics
of killing
of fresh
cancer
tissues
The phenotype
of TILs was determined
by fluorescence-activated
cell
Dickinson
fluorescence-activated
cell sorter.
Fluonescein
isothiocyanate-labeled
appropriate
dilution.
Leu-4 recognizes
mature
T cells (CD3);
Leu-3 recognizes
class Il-restricted
helper/inducer
T cells (CD4);
HLA-DR
recognizes
B cells,
cells and neutrophils
(CD16);
anti-IL-2
receptor
recognizes
low affinity
interleukin
associated
with natural
killer activity
(CDS6).
One lytic unit is defined
as the
% of Positive
Patient
CD3
TIL cultures
CD4
CD8
with CD3CD4
CD25
No. of
cells injected
(X lOu)
CD56
dominant
epithelial
Killing
activity
(LUS(J107
Autologous
cells)
K562
Allogeneic
group)
10.7
99.1
2.3
38.1
ND”
1.0
52.1
ND
53.6
43.2
99.4
2.6
44.3
ND
2.8
83.3
17.8
64.5”
15.2’
7
99.3
80.3
16.7
99.2
1.2
92.8
9.0
3.5
16.2
10.5
10.7”
20.6
11
98.3
80.5
14.7
98.0
8.0
14.3
32.5”56.2’
with CD3CD8
0.8
cells in excess
of 50%
1
99.1
36.0
68.3
92.9
2
5
6
99.9
99.1
98.3
90.8
98.3
95.2
34.9
8.5
21.9
9.2
10.3
28.7
68.2
91.7
76.8
80.5
87.2
66.3
81.8
90.6
99.0
99.9
98.9
98.0
12
99.2
20.4
82.1
13
99.9
37.6
60.8
not done.
tumor
tumor
tumor
tumor
ND,
Fresh
C Fresh
d Fresh
e Fresh
1Fresh
Fresh
I’ Fresh
Fresh
J Tumor
tumor
tumor
tumor
tumor
cells
g
I
described
cells
cells
cells
cells
from
from
from
from
patient
patient
patient
patient
cells
cells
cells
cells
from
from
from
from
from
patient
patient
patient
patient
patient
I 1.
previously
number
(8).
of effector
cells
65.4
(CD8
1.6
dominant
9.4
1.5
45.2
group)
ND
4.4
52.3
ND
ND
2.9
2.4
1.8
3.5
2.0
2.0
66.7
100.2
90.9
95.2
121.0
104.9
16.5
28.1
22.3
28.3
18.4
26.4
54.l 22.7
43.5” 66.7’
58.1’ 8.2k
45.0’ 217h
48.6’ 19.4’
72.6” 52.9’
0.9
1.1
4.0
10.6
3.5
7.0
66.5
92.0
50.5
90.8
90.3
33.9
ND
ND
3.5
21.2
4.1
9.1
99.9
1.0
65.2
20.0
1.0
98.9
21.3
58.0’
84.8
0.1
26.7
11.4
2.6
69.3
18.6
736h542j
7.
10.
4.
8.
6.
9.
One
lytic
unit
50%
was
lysis
defined
as
of 5 X i03
the
variety
of
allogeneic
target
target
cells,
tumor
of the same
killer-sensitive
Survival
of TILs
when the patient
between
survival
Kaplan-Meier
data
until
is still
curves
were
obtained
the death
alive.
were
of patients
or
curves
using
and
the
Survival
calculated
higher
two
against
autologous
in the CD8
in the
RESULTS
CD4
cytotoxicity
r!L-2
and
from
advanced
stage
Infusion
freshly
ovarian
markers,
rate,
and
cytotoxic
observed
with
preparations
70-95%
by the dye exclusion
After
recovery
was
against
in cell
two
test
from
against
activity
of
(10.5-28.3
compared
surface
group
were
the
group),
cells
lymphocytes
expression
cells
were
were
mainly
culture
in excess
TIL
period
activated
HLA-DR
50%
cytotoxic
cells
13
(CD8’
ilLs
(CD4
were
(Table
In 9 of
4 preparations,
of
preparations
for
T lymphocytes,
antigen.
CD3CD8
and in the remaining
13 propagated
the
were
of the
Phenotypes
with
cinoma
dominant
tested
activity
preparations
lytic
units/107
The
serous
a
nomas,
to those
group
mucinous
The
years.
cystadenocarcinomas,
and
two
endometnioid
patient
as
K562
cell
(Table
cytotoxic
cells
generally
ages
In the
13 patients
two
low
fresh
in the
in the control
serous
of the patients
group
two
endo-
adenocar-
ranged
(Table
undifferentiated
adenocarcinomas
TIL
group
cystadeno-
undifferentiated
control
varied
it was
and allogeneic
2), seven
as
The
cytotoxic
The
cystadenocarcinomas,
one
7,
well
types.
high
types.
K562
of 1 1 patients
and
included.
55)
of
preparations
cells
autologous
of
prepara-
tumor
however,
both
at least
killing
of eight
showed
to
was
from
cell
characteristics
similar
with
Preferential
against
cells);
adenocarcinomas,
were
13
tumor
tested
that against
to 67 (median,
at least
against
all
three
1 1 and
allogeneic
4). In the TIL
carcinomas,
metrioid
patients
regard
compared
preparation
and
natural
With
and one of three
cell
fresh
the
activity
autologous
tumor
some
with
cells.
Patients.
activity.
allogeneic
from
activity
tumor
the
All
13 patients
TILs
of the
by
during
to be viable
cryopreservation.
change
and these
CD3CD4
of
tumors
major
as judged
dominant
propagated
In all 13 preparations,
no
phocytes,
preparations,
We
against
line.
in seven
In the
tumor,
and
cytotoxic
types.
group
group.
low
cells
cell
observed
dominant
was
in the 13 patients
are presented
in Table
1. All
TIL cultures
consisted
of mainly
CD3
T lym-
growth
cultured
TILs
rIL-2 expanded
resected
cancer.
of TILs were confirmed
when
recovered
from
cryopreservation,
of TILs.
was
dominant
the
tumor
tumor
cells
type,
cell
study,
autologous
ovarian
autologous
leukemia
in this
tumor
fresh
histological
cultured
killing
allogeneic
tions
including
K562
preferential
from
method.
Culture
65.3’
2.
cells.
last contact
differences
ND
5.
mediating
Statistical
Analysis.
the day of administration
503
ovarian
88.1
b
once
from
99.9
9
10
group).
propagated
99.9
8
with
TILs
3
4
TIL cultures
a
CD16
of 50% (CD4
of cultured
Research
sorting
analysis
performed
with a 488-nm
argon
laser on a Becton
mAbs were purchased
from Becton
Dickinson
Japan and used at the
class I-restricted
cytotoxic/suppressor
T cells (CD8);
Leu-2 recognizes
macrophages,
and activated
T cells; Leu-1 ic recognizes
natural
killer
2 receptor(CD2S);
and Leu-19
recognizes
the major subset of cells
number
of effector
cells mediating
50% lysis of 5 X tO3 target cells.
cells
HLA-DR
cells in excess
cells
Cancer
from
3),
seven
adenocarci-
were
included.
Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer
Research.
28
504
Adoptive
Immunotherapy
Using
T able 2
ilL
Cryopreserved
Characteni
sties
of the
patients
treated
with
adoptive
transf
en of cultu red
ilLs
after
chemotherapy
Prognosis
Age
No. of
chemotherapy
courses
(yr)/
performance
Patient
status
Clinical
diagnosis
1
58/0
Ovarian
Histopathology
cancer
stage
Previous
treatment
Serous
Regimen
Outcome
Outcome
disease
Observed
period
(mo)
of
Surgery
(N’)
FCAP”
5
NED
NED
44
cystoadenocarcinoma
Surgery
(B1C)
CAP
5
NED
NED
39
cystoadenocarcinoma
lie
48/0
Ovarian
cancer
Mucinous
3
56/0
stage
Ovarian
stage
IV
cancer
lIe
Endometrioid
adenocarcmoma
Surgery
(Bi)
FCAP
3
NED
NED
36
5
49/0
Ovarian
cancer
Endometrioid
adenocarcinoma
Surgery
(Bi)
FCAP
5
NED
REC
39
6
28/0
stage lie
Ovarian
cancer
stage lib
Mucinous
cystoadenocarcinoma
Surgery
(A)
FCAP
3
NED
NED
36
7
51/0
Ovarian
cancer
Undifferentiated
Surgery
(Bi)
FCAP
5
NED
NED
37
stage
Ovarian
stage
Ovarian
stage
Ovarian
IlIc
cancer
ha
cancer
IV
cancer
2
8
55/0
10
48/0
12
31/1
adenocarcinoma
Serous
cystoadenocarcinoma
Surgery
(A)
FCAP
3
NED
NED
31
Serous
cystoadenocarcinoma
Surgery
(Bi)
FCAP
5
NED
REC
33
Serous
cystoadenocarcinoma
Surgery
(Bi)
CAP
5
NED
NED
32
Serous
cystoadenocarcinoma
Surgery(A)
CAP
5
NED
NED
25
stage IlIc
14
67/0
15
64/0
16
55/1
19
Ovarian
stage
Ovarian
stage
Ovarian
stage
Ovarian
57/1
cancer
lIe
cancer
Ilic
cancer
Mucinous
cystoadenocarcinoma
Surgery
(B2”)
CAP
5
NED
NED
38
Serous
cystoadenocarcinoma
Surgery
(Bi)
CAP
5
NED
NED
23
Serous
cystoadenocarcinoma
Surgery
(B2)
CAP
5
NED
NED
24
lib
cancer
stage Ilic
REC,
a
No
I,
FCAP,
macroscopic
tumor
residuum.
5-fluorouracil-cyclophosphamide-Adniamycin-cisplatin;
recurrence
of disease.
Largest diameter
of tumor
residuum
was less than 2 cm.
Largest diameter
of tumor
residuum
was larger than 2 cm.
NED,
no evidence
of disease;
d
The
ages
years.
of the
Primary
patients
tumor
ranged
from
reduction
42
surgery
to 68
(median,
revealed
that
49)
residual
( Fig.
3). The difference
patients
in the
tumor mass remained
in 9 of 13 patients
in the TIL group and in
6 of 11 patients
in the control
group
(“Residuum,”
Table
4).
However,
by completion
of cisplatin-containing
chemotherapy,
group
was
were
detected
group
after
macroscopic
completion
these
by
residual
15 patients
examinations
CT
scan,
and/or
infused
into
plications
i3
such
tumor
mass
and
they
such
as internal
MRI.
When
patients
were
as nausea,
entirely
diagnosed
disappeared
to be free
examination,
the
of
had
the
of disease
autologous
group,
vomiting,
in
in the
ultrasonography,
cultured
TIL
no
hepatitis,
sion, or respiratory
distress
due
ability
and loss of intravascular
Survival
Analysis.
patients.
erage,
The
33.6)
(average,
TILs
were
remarkable
com-
oliguria,
to increased
fluid were
34.6)
overall
group
and
tively
(Fig.
of patients
group
3-year
Survival
observation
months
3-year
and
cyclophosphamide-Adniamycin-cisplatin;
hypoten-
capillary
observed
permein any
in
months
survival
2). The
the
rate
and
between
group
and
that
23
and
the overall
of patients
14
The
patients
100%
for
to 44
from
group.
of disease-free
was
available
from
group
control
group
difference
in the TIL
were
ranged
ilL
in the
in the control
data
period
all
control
group
was
82.1%
and
54.5%,
the disease-free
and
that
significant
in the vaginal
22 and
TIL
and
residual
‘ Table
disease-free
4).
was
ence
between
76.2%
TIL
group
these
rate
in the
and
33.3%,
that
control
the
respectively
patients
group
had
operation
(‘ ‘ Re-
estimated
group
survival
of the
significant
TIL
Before
nine
the primary
patients,
the disease-free
and
statistically
For
the ilL
administration.
in the
after
lesions
from
chemotherapy,
patients
tumor
survival
group
six
ilL
control
Recurrent
of two patients
from
rate of
in the
(P < 0.05).
stump
32 weeks
group
survival
of patients
and
(Fig.
3-year
in the
control
4). The
differ-
rate of the patients
patients
in the
control
in the
group
was
(P < 0.05).
(av-
to 48
in the
TIL
respec-
survival
rate
in the control
was statistically
significant
(P < 0.01).
The estimated
disease-free
survival
rate of the patients
in the TIL group
in the
group
of cisplatin-containing
siduum,’
DISCUSSION
Although
estimated
67.5%,
between
TIL
statistically
macroscopic
patient.
24
CAP,
C
respectively
imen
chemotherapy
is reported
epithelial
ovarian
an advanced
isolated
with
tumors
sion
(7).
the median
about
solid
tumors
by culturing
The
shown
to express
administration
with
a variety
cisplatin-containing
2 years
treatment
rate
for patients
(4).
specific
of TILs
of human
reg-
in the
survival
is only
and
in patients
cancer,
the
effective
stage
from
rIL-2
with
to be relatively
ilLs
have
of
at
been
single-cell
suspensions
lysis
of autologous
mediates
tumors
tumor
(9);
regres-
however,
Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer
Research.
in
Clinical
Tab le 3
Characteristi
cs of the patients
treated
withou
t
adoptive
tran sfer of cul tured ilLs
Cancer
Research
505
afte r chemotherapy
Prognosis
Age (yr)/
No. of
performance
Previous
diagnosis
Histopathology
Observed
chemotherapy
Regimen
courses
Outcome
of
period
Patient
status
Clinical
1
58/0
Ovarian cancer
stage lie
Serous
cystoadenocarcinoma
Surgery
(A”)
FCAP”
5
NED
NED
2
46/0
Ovarian
Serous
cystoadenocarcinoma
Surgery
(B2’)
CAP
5
NED
REC,
Serous
eystoadenoeareinoma
Surgery
(A)
CAP
3
NED
NED
33
cancer
treatment
Outcome
disease
(mo)
48
DOD
30
stage Ilie
3
43/0
4
49/0
5
49/1
6
42/0
7
42/0
Ovarian
cancer
stage
Ovarian
stage
Ovarian
stage
Ovanna
stage
lIe
cancer
lie
cancer
lib
cancer
11th
Endometrioid
adenocarcinoma
Surgery
(A)
FCAP
5
NED
NED
28
Endometnioid
adenocareinoma
Surgery
(A)
FCAP
5
NED
NED
47
Serous
eystoadenocareinoma
Surgery
(BV’)
FCAP
5
NED
NED
42
cancer
Serous
eystoadenocarcinoma
Surgery
(Bi)
FCAP
5
NED
DOD
39
Serous
eystoadenocareinoma
Surgery
(Bi)
FCAP
5
NED
REC,
DOD
24
Undifferentiated
adenocareinoma
Serous cystoadenocareinoma
Surgery
(B2)
FCAP
5
NED
REC,
DOD
29
Surgery
(B2)
FCAP
5
NED
REC
47
Undifferentiated
adenocareinoma
Surgery
(A)
CAP
5
NED
NED
14
Ovarian
stage
8
68/0
9
68/0
10
47/0
11
50/0
REC,
a
No
b
FCAP,
macroscopic
IV
Ovarian
cancer
stage
Ovarian
stage
Ovarian
stage
Ovarian
stage
Ilie
cancer
Ilie
cancer
IIIe
cancer
lie
tumor
residuum.
5-fluorouraeil-cyelophosphamide-Adriamycin-eisplatin;
Largest
d
diameter
Table 4
to tumor
Characteristics
residuum
was
o f patients
ilL
No.
of patients
macroscopic
tumor
CMacroscopic
clinical
critical
trials,
point
chemotherapy
with regard
injection
agglutinin,
(n=13)
7
4
2
5
5
4
5
9
6
7
3
2
1
7
0
2
2
P<O.Ol
‘___i
control
6o
group
(n=ll)
1
3
Y_I
faa
Srt
4
Pr.o1
2
Overall survival rate of patients with ovarian cancer stage II, III,
and IV who were diagnosed
as having no evidence
of disease after
completion
of chemotherapy.
Fig.
of ilLs
is limited
(6). The
in combination
with
especially
and the
drugs.
reported
killer
group
group
11
51.1 ± 9.5
of responses
transfer
(10)
that
the
of cellular
by delayed-type
natural
TIL
residuum.
immunoactivation
as demonstrated
cyclophosphamide-Adriamyein-eisplatin;
cancer
Control
is to establish
adequate
conditions,
to timing
for infusion
of lymphocytes
we
CAP,
operation.
the duration
of anticancer
induces
ov arian
residuum.
of adoptive
Recently,
TILs
tumor
of disease;
2 em.
operation
Histopathology
Serous
Mucinous
Endometrioid
Undifferentiated
a Diagnosed
at the primary
No
with
no evidence
than 2 cm.
than
group
Completed”
Residuumc
b
less
13
51.4 ± 10.8
Age (yr)
Clinical stage”
II
III
IV
Status of primary
our
NED,
recurrence
of disease; DOD, dead of disease.
CLargest diameter of tumor residuum was larger
cytolytic
adoptive
transfer
immunity
in patients,
hypersensitivity
activity
of
to phytohemagainst
K562
cells,
and
blood.
percentages
These
of cells
findings
bearing
suggest
mor regression
was mediated
immunity.
Cisplatin-containing
tered
after
noactivation
Additionally,
adoptive
transfer
of cellular
drug-resistant
the CD8
antigen
in peripheral
the possibility
that
in part by the
chemotherapy,
activated
cellular
if it is adminis-
of ilLs,
immunity
tumor
interferes
observed
with
tu-
the immu-
in patients
induced
by ilLs.
cells
possess
increased
often
Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer
Research.
506
Adoptive
Immunotherapy
Using
Cryopreserved
TIL
so
L,
80
C
C
3
60
60
L,
C
control
group
(n=l
1)
40
S
I
control
20
0
3
Years
Fig.
3
Disease-free
survival
completion
Start
with
as having
and sensitivity
examined
investigate
the
setting,
cytotoxicity
TILs
ovarian
cancer
no evidence
stage
of disease
II,
after
when
major
the
change
markers;
slightly
against
autologous
TILs
period
was
but still
tumor
of patients
the completion
after
As
shown
with
and
valuable
cytotoxie
was
recognized
in patients
with
tumors
in the
was
detected
lose
transfer
1, ilLs
demonstrated
surface
out
patients
operation,
after
re-
CD3CD8
(c)
cells
of CD56
Ten
or mixtures
or CD16
of 1 1 preparations,
had
some
well
as autologous
cytotoxic
cells
in addition
A significant
of both,
TILS
were
with
activity
tumor
the exception
against
cells,
present
allogeneic
and several
ilLs
a small
number
to anticancer
cells
substantial
num-
in some
of preparation
tumor
could
7,
cells
as
kill K562
part of the clinical
with measurable
treated
gous
TILs
new
study,
no
significant
injected,
and
nomodulation
in patients
and that of
sitivity
to phytohemagglutin
patients
in the control
group.
One of our interests
was to
evaluate
whether
adoptive
transfer
of ilLs prolongs
the diseasefree survival
time of patients.
In this study the difference
be-
transfer
of ilLs
tween
rate of disease-free
the disease-free
patients
survival
and that of patients
in the control
icant. It has been well documented
of a primary
operation
epithelial
ovarian
cancer
rate
of patients
group
in the TIL
group
group was statistically
signifthat the degree of completion
influences
prognosis
of advanced
stages,
tients receive
several
courses
of chemotherapy
clinical
study of patients
with stage III epithelial
5-year survival
rate in cases with no macroscopic
of patients
with
although
the pa(14-16).
ovarian
residual
In our
cancer,
tumor
protocol
degree
tumor cells. Therefore,
by their characteristics.
in the TIL
survival
ease,
in primary
TILs
which
ilLs
mass,
rate
can
elim-
are refractory
were administered
and tumor
regres-
direct
tumor
few reports
our
rence. One of two
ilL group showed
and
was
trials,
tumor
with
allogeneic
tumor cells.
noted
between
the overall
to autologous
difference
of
carried
survival
that
cells
patients
was
tumor
lysis mediated
have dealt with
combined
differences
among
patients
in the TIL group despite
different
transferred
ilLs,
such as the ratio of CD4
:CD84
of cells
cases.
five
at the time
by
the
analysis
on ilL-treated
patients.
This article
is the
of the survival
rate of patients
with epithelial
ovarian
chemotherapy.
In this
or
in patients
drugs.
prognostic
first report
cancer
and
disease-free
tumor
after
of recurrence
the analysis
indicating
of residual
and
tumor
residual
group,
sion was assumed
to reflect
infused
ilLs
(4, 5). However,
killing
in the CD8
in the CD4
CD3CD4
while
also
mate
in the ilL
41%,
group
when
residual
of disease
no sign
no residual
increased
preference
since (a) histocompatible
cells were not targeted
in our assay,
we could not clarify
the cell type responsible
for the cytotoxic-
bers
observed
cm,
study,
macroscopic
a significantly
was
<2
Furthermore,
with
had macroscopic
in the TIL
had
In a large
to the patients
considered
the cytotoxicity
of the mixed cell activities,
of mainly
who
of cryopreserved
dominant
group.
However,
we
against
tumor cells to be a measure
consisted
on
who
In this
patients
group
activity
of eight cultures
three
cultures
TILs
in four
in the control
tumors
22%.
4
tPrsSocol
and had no evidence
with
cm,
operation.
of autologous
tumor cells in seven
dominant
group
and in one of
(b) Cultured
>2
primary
of chemotherapy.
in Table
84%,
fro. Slat
of patients
operation
was
(7). These
findings,
along with
of TILs (6), led us to initiate
the
adoptive
survival
primary
to
3 months.
in cell
Disease-free
after
We
might
exceeds
4
tumor
of chemotherapy.
ilLs
timing
and
rate
cells
covery
from cryopreservation
the results
of the clinical
trial
TILs
suitable
grow
in growth
decreased
treatment
not
of cultivation
noted
fresh
with
could
of
Fig.
completion
(1 1-13).
of cryopreservation
can be used
since
3
Y_1
to immunotherapy
influence
whether
clinical
ity.
(n=6)
(,O_
4
Protocol
of chemotherapy.
immunogenieity
also
from
of patients
III, and IV who were diagnosed
No
group
‘20
while
of killing
activity
with
ilLs
and
were
observed
conditions
of
cells, number
against
autolo-
we could not predict
the effect of
We also estimated
whether
immuinduced
by ilLs
influences
the recur-
patients
with recurrent
disease
that was in the
very little change
in delayed-type
hypersenand
compared
in the other
with
patient,
natural
patients
killer
with
activity
nonrecurrent
immunoactivation
was
disclearly
induced
relation
(data not shown).
An explanation
between
the clinical
outcome
transfused
a variety
ilLs
might be that infused
ilLs
are still a mixture
of
of cells and that many unsolved
issues remain
regard-
ing
4
the function
K. Tanaka,
of CD4
unpublished
and
CD8
for
after
and
cells
the
and
the lack of corphenotype
of
of class
data.
Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer
Research.
I and
Clinical
class
II MHC-restricted
quite
lation
small to evaluate
and disease-free
investigate
the parameter
This
TILs
study
after
method
cells.
suggests
all chemotherapy
to obtain
Since
the number
the correlation
survival
rate,
favorable
of patients
was
between
immunomodufurther
study is needed
to
to predict
effectiveness
of ilLs.
transfer
of cryopreserved
has
been
finished
is one
feasible
outcomes.
1934-1939,
manuscript.
9. Rosenberg,
Clin.
10.
Oneol.,
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