From www.bloodjournal.org by guest on July 31, 2017. For personal use only.
Childhood
Lymphoblastic
Establishment
of the
By
Leukemic
cells
leukemia
from
(ALL)
K562
as
had
normal
ratio:
cells
leukemic
cell
from
patient,
the
freshly
chemistry.
from
12
lines
cell
childhood
they
lines
cultured
cells
OKT-3
as
RECENT
focused
on
lymphocytes.’
Ia
.
OKT-6.
.
fresh
and
immunologic
cell
individuals
a natural
of killing
certain
sensitization.58
vious
Such
The
leukemic
activity,
those
mono-
childhood
marrow
acute
was
of NK
ALL
cell
of NK
cell
relapse
The
ALL.
They
WBC
larger
had,
present
studies
2 days
were
later.
no apparent
observation
pallor,
presented
g/dl,
The
cytes,
of
1% monocytes,
with
morphology
of the
positive
and
granular
FAB
classification.9
materials
esterase
red
cells
blood
determined
receptors
They
by
were
did
x-ray
the
the basis
of the hematologic
diagnosis
of T-ceII
solone
after
therapy
Blood,
with
Vol.
60,
rads)
both
and
C3
and
of ALL
(T-ALL)
,
she
irradiation.
6-mercaptopurine
No. 6 (December),
received
assay
receptors
(1gM,
was
were
L2
pallor
line
and
with
for autopsy
viral
FAB
of diagnosis
high
NK
blood.
was not granted.
and
99.5%
of the
Leukemic
the peripheral
she
hepatosplenomm
later,
and
When
at the time
of K562.
pyogenic
cytotoxic
follow-up
L2 morphology
observed
a
6 mo,
remission
1980.
was 420,000/cu
from
infections
activity
cells
for the
The
patient
She
had
during
the
of them.
and
IgG).
On
the clinical
cranial
and
cells
Leukemic
gradient
from
Hank’s
balanced
samples
diagnosis
of predni(L-Asp).
(2400
Soon
rads)
and
on maintenance
methotrexate
blood
when
were
blood.
the
markedly
were
blood
For
collected
the
stage
(>99.5%)
by
and
patient,
and
in the peripheral
washed
density
three
times
experiments,
I 2 children
as
of ALL
Ficoll-Hypaque
control
from
and
from
terminal
separated
solution.
relapse,
obtained
in the
increased
were
heparinized
salt
was
she was
cells
or
Department
cine,
(MTX)
of Pediatrics,
Matsumoto,
Supported
Culture
with
with
peripheral
ALL
at the time
control
leukemic
cell
preparations
Shinshu
University
School
of Medi-
Japan.
in part
and
Science,
by
and
grants
from
from
the
the
Ministry
Ministry
of
of
Education,
Public
Welfare,
Japan.
Submitted
a combination
METHODS
as
enlargement.
findings,
AND
for
cells
37#{176}C.lgG-Fc
IgD,
peripheral
above,
leukemic
of
Receptors
in 6.0%
described
blood
(PAS)
Peroxidase
of the
at
She was placed
1982
cell
acid-Schiff
L-asparaginase
(6MP)
blood
was made.
with
and
12.3
marrow
of them.
in 8.2%
immunologic
(VCR)
bone
no mediastinal
was induced
of
4% lympho-
negative.
found
immunoglobulins
a remission
(3000
The
cell
showed
vincristine
attaining
mediastinal
type
remission
(PRD),
August
be described
Permission
a partial
conventional
to
with
next
Preparation
Heparinized
hepatomegaly
lymphoblasts
in most
were
were
in 9.0%
of the chest
A complete
since
1980
the
temporarily
were
on December
During
time
she had
those
obtained
cyclophosphamide,
relapse
malaise,
a white
Periodic
rosette-forming
surface
and
The
found
activities
(SRBC)
observed
not bear
film
were
and
4% neutrophils,
lymphoblasts.
lymphoblasts.
general
a hemoglobin
mm,
mm with
95%
nonspecific
sheep
showed
37,000/cu
and 91%
with
(10 cm),
count
of 339,000/cu
hypercellular
An
blood
count
count
of
period.
Sample
2, 1979
splenomegaly
peripheral
a platelet
(WBC)
REPORT
on March
lymphadenopathy,
(8 cm).
each
3 mo,
MATERIALS
girl
and
lost
cell
of
to
phenotype
lineage.
CASE
A 9-yr-old
L-Asp.
count
than
episodes
fresh
identical
in February
and
cells
as will
a leukemia-derived
of the
VCR,
induced
9, 1980,
against
died
was
leukemic
and
but
lymphoblasts.
new
of several
for about
The
classification
of
she
peripheral
lymphoblasts.
was
twice,
in November
megaly.
with
a combination
no therapy
and
value).
virtually
to have marrow
vindesine,
Unfortunately,
normal
origin.
remission
occurred
are
a
enforcement
with
presented
cells
She was found
of PRD,
received
case of ALL that had leukemic
cells with NK activity
against
leukemia-derived
cell lines such as K562 and
Molt-3.
In addition,
we describe
the leukemic
cell lines
retaining
the activity
established
from
the patient,
which certainly
confirms
a new phenotype
of childhood
ALL
attained
drugs.
lines without
prewe report
a first
demonstrating
A partial
lymphocyte
characteristics
cells.
cyclic
against
of
respectively)
normal
functional
cultured
(63were
33.5%
SPI-802,
mitogen-induced
and
region
cytotoxicity
and
of normal
NK
combination
a child
cell
article,
and
and PRD.
12, 1979.
and
27.8%
method.
abnormal-
chromosomes
1 7.8%
and
received
6MP,
func-
have a lymphocyte
subpopulakiller (NK)
cell, which
is capable
tumor-derived
In this
and
to
cells
of
one of T-cell
with
Raji
phenotypic
were
attention
tions, of neoplastic
T cells from
lymphoblastic
leukemia
(ALL).
Normal
tion called
had
C3.
has been
functions
of neoplastic
Broder
et al.2 have dem-
For example,
against
marker
in SPI-801
and
cytoplasmic
chromosome
spontaneous
up
and
in hypotriploid
several
had
Ig
same
were
lysis:
value
surface
.
immunofluorescence
the
and
.
They
(38.2%
cells
human
.
11p
.
(percent
not
and
shared
chromosomes
-
6q
MoIt-3
T cell
.
Activity
Akabane
by the
lines
demonstrated.
cul-
and
Leu-7
much
YEARS,
suppressor
The
asialo-GM,
OKT-8.
as the
cyto-
leukemic
IgG-Fc,
and
lymphocyte
functions.
cells.
Two
cell
Their
73).
same
morphology.
the
blood
OKM1
N
onstrated
as
red
were
OKT-4
.
well
sheep
ities.
ALL
Activity:
the
Taro
determined
two
K562
in their
and
as
The
and
common
,
Killer
Retaining
Miyagawa,
established
the
markers.
of
0.3%).
±
essentially
cells
a 40:1
of control
were
Yukiaki
1gM
(147.4%
that
(0.1%
SPI-802)
were
leukemic
76.8%
than
ALL
at
Natural
Lines
cyte
against
assay
was
higher
and
activity
release
lysis
and
Kawai,
lymphoblastic
(NK)
With
Cell
Hiroshi
acute
killer
51Cr
(SPI-801
and
for
with
percent
immunologic
receptors
I
child
the
Leukemia
Komiyama,
natural
value)
harvested
tured
a
by
lymphocyte
leukemic
Atsushi
high
determined
effector:target
Leukemia
Address
of Pediatrics,
390.
February
reprint
1 1. 1982;
requests
Shinshu
accepted
to Dr.
University
Atsushi
School
July
1, 1982.
Komiyama.
Department
of Medicine,
Matsumoto,
Japan.
© 1982
by Grune
& Stratton,
Inc.
0006-4971/82/6006-0029$O1.OO/O
1429
From www.bloodjournal.org by guest on July 31, 2017. For personal use only.
1430
KOMIYAMA
(blasts
In
>98%)
were
addition,
healthy
by
obtained
in the same
heparinized
volunteers,
and
Ficoll-Hypaque
monocytes
peripheral
mononuclear
cells
centrifugation.
by
adherence
fashion
blood
obtained
(MNC)
were
MNC
to plastic
as described
was
were
surfaces
then
and
T-leukemia
above.
from
5
separated
depleted
used
of
line
respectively.
rabbits.
Terminal
Kit
of KOPT-KI
and
Anti-asialo
GM1
a normal
B-cell
line
antibody
was
prepared
deoxyribonucleotidyl
was determined
Assay
as normal
cell
1054,
by the
(Bethesda
transferase
Terminal
Transferase
Research
ET AL.
of SN
(TdT)
in
activity
Immunofluorescence
Lab.).
lymphocytes.
Chromosome
Establishment
of Cell
Leukemiccells
10%
fetal
calf
serum
25%
in RPMI
(FCS;
Flow
penicillin
1640 medium
Lab.,
(50
Rockville,
U/mI),
of
culture
(Falcon
flasks
pared
according
culture
medium
PHA-P
(Difco
l0
factor
cells/mI,
and
Lab.,
of Masucci
with
Lab.,
Detroit,
atmosphere
of 5% CO2 in air. The
and
cultures
Mich.)
The
established
establishment,
medium
were
without
stimulated
incubated
at
culture
I mo later
subcultures
FCS)
were
culture
MNC
from
cells.22
pre-
48-hr
from
Raji
tissue
cells.
of
from
1%
healthy
usually
every
cell
4 days
and
Effector
or presence
line
normal
g/ml
in culture
described
cytes
Morphology
and
For electron
in 0.05
the
cells
M cacodylate
buffer
dehydrated,
described.”’2
The
and
lead
Cytochemistry
examined
microscopy,
buffer,
tate
was
in
were
fixed
(pH
7.2)
and
embedded,
ultrathin
citrate
examined
2.5%
tetroxide
sectioned
were
stained
with
a Hitachi
as
in
uranyl
HS-8
ace-
were
nonspecific
and Other
formed
at 4#{176}C
and
Clark.’3
E-rosette
method’4
purified
rabbit
technique
cells
using
markers,
capping
OKT-3,
Co.,
N.J.),
and
IgG-F
were
(cALL),
Lab.),
(ab’)2
and
Dr. Shinpei
Nakazawa,
Tokyo.
The
anti-cALL
non-T,
non-B
and some
and anti-Ia
1gM-F
obtained.
anti-Ia
cell
antibody
of B-cell
were
ALL,
raised
AG,
were
in rabbits
with
supplied
Keio
by
University,
of non-T,
but none of T-ALL
as described
F.R.G.)
with
as described
75%
by Tsubota
tube.
also
the cell
mixtures
the labeling,
were
mixtures,
release)/(cpm
maximal
were
release
-
cell-mediated
done
on 51Cr-labeled
were
set up in triplicate.
incubated
equation
was
experimental
chicken
cpm
3 x
transferred
prepared
to
at a 40:1
spontaneous
blood
cells
and
some
at 37#{176}C
for 4, 8, 12,
used to express
release
cpm
cytotoxicity
red
as
in 2 ml
with 100
at 37#{176}C
for 60 mm,
After
medium
Cell
used
ratio,
were brought
to a I ml final
volume,
37#{176}Cfor 4 hr in a 5% CO2 incubator.
In
at
following
= l(cpm
Two
and
cell
ofihe
Cell
cell
lines
SPU-802,
suspension
Lines
were
established
respectively.
The
with a doubling
Contamination
with
detected
culture.
Cell
p.s, IgD-F
Marburg,
kindly
were
Hutchin-
cytotoxicity:
spontaneous
release))
(ADCC)
(CRBC).
x 100.
assay
All
was
assays
(SPI-801
and
by agar
mycoplasma
and
named
SPI-801
cells grew as a singletime of about 48 hr.
species
was
not
Becton
by immunization
KOPN-K,’8
reacted
al.’7
monocyte
(ab’)2
of Pediatrics,
of
et
Anti-common-ALL
antisera
line
EB
Monoclonal
(HNK-l;
anti-human
experimental
and 16 hr. The
percent
lysis
non-
Pharmaceuti-
anti-human
A (Behringwerke,
was raised
The
1 (Ortho
anti-Leu-7
Department
leukemia
0KM
excess.
medium.
Hansen,
Fred
Ill.)
Heights,
of 0. 1 ml of the culture
Establishment
antibodies.
Volger
monoclonal
commercially
anti-T-cell,
et al.’9
and
by
A.
Center,
SP!-802)
further
20
conventional
in antibody
monoclonal
of
CCRF-CEM,
John
Arlington
the culture
or
and several
were suspended
FCS, labeled
with
as
lympho-
PHA-MF
JM,
Wash.)
100
fashion
RESULTS
receptors
under
(MMC)
patients,
Dr.
in the
with
2 x 106 target
cells
10% heat-inactivated
ORBC
a Leitz-Dialux
and
and
Calif.),
Research
&
used
with
coated
detection
as described
OKT-8,
Sunnyvale,
antibodies
ALL
were
OKT-6,
Raritan,
(Bethesda
Koshiba
detected
For
with
monoclonal
was
by the
by an immunofluorescent
performed
plastic
Seattle,
PHA-MF,
normal
Molt-3,
Blood
10%
with
ALL
Sound
lines
same
treatment
by
Center,
Antibody-dependent
37#{176}C.
Recep-
detected
IgG-Fc
immunofluorescence
was
and
studied
1gM-coated
respectively.
detected
Kaplan
(ORBC)
rabbit
IgG.’5
several
OKT-4,
also
or indirect
antibodies
Dickinson,
(ab’)2
were
cells
and
direct
1gM
Monoclonal
blood
antibody
heat-aggregated
using
Cytoplasmic
and C3 were
complement,
conditions’6
microscope
red
of
was further
at 4#{176}C,
29#{176}C,
and
(IgG-Fc)
ox
serum
method
4 times
in the
supplied
cells
cell
prepared
As controls,
including
Puget
leukemic
with
Mo.)
and
as target
mitomycin-C
was
the other
target
MoIt-3,
used
harvested
of 24-hr
from
lines
Research
51Cr-labeled
the cultured
or
Louis,
(kindly
Lab.,
experiments,
2-aminoethyl
isothiouronium
SRBC
(E-rosette)
was percells
cell
1884
10 of the cells
stains
effector:target
incubated
to the
SRBC
of lgG
using
human
with
treated
by the cultured
anti-ORBC
cultured
surface
cytochemical
Markers
37#{176}C
according
formation
neuraminidase-treated
rosette
anti-T
Cell
formation
(AET)
tors for the Fc portion
of the
PAS
each
Spontaneous
rosette
bromide
hydrobromide
plus fresh
and
performed.
Immunologic
cal
esterase,
cells
cells. Briefly,
l640containing
and washed
routinely
using
leukemia
Cancer
son
St.
or presence
.sCi 51Cr (Amersham,
electron
microscope.
Peroxidase,
G-banding
as K562,
stimulation
preparation.
leukemic
Jurkat-F
effector
RPMI
previously
with
the
were
Con-A-MF
(Sigma,
Histocompatibility
glutaraldehyde
1% osmium
and
sections
and
with
and
smears.
freshly
(Con-A-MF),
PHA-MF
on
such
from
of 3-10-day
in the absence
cultured
on May-Giemsa-stained
the
and cells
lymphocytes.
of Con-A
lines
T lymphoblasts
included
patient
assayed
cell
normal
cells
the present
treated
PHA-MF.
Cell morphology
by
Assay
was
or lymphoma
PHA-induced
Con-A-MF,
Cell
performed
Cytotoxicity
10% concanavalin-A-MF
on November
After
were
cytotoxicity
Leukemia
absence
37#{176}C
in a humidified
in 2 of 20 flasks.
made
was
was initiated
Cell-Mediated
Cell-mediated
tissue
PHA-MF
et al.’#{176}
from
10%
M
at a cell
in 20 plastic
Calif.).
(RPMI
l0
(50 sg/ml),
(PHA-MF)
placed
to the method
volunteers.
9, 1980
were
Oxnard,
1640
Md.),
analyses
method.2’
containing
streptomycin
phytohemagglutinin-mitogenic
concentration
Analysis
Chromosome
weresuspended
2-mercaptoethanol,
and
Lines
in children.
a
by
non-B
The
et al.2#{176}
with
a
Morphology
and Cytochemistry
Morphological
characteristics
of both SPI-80l
and
SPI-802
cells
were similar
to those
of the original
leukemic
cells. As shown
in Fig. 1 , most cells had a
round
nucleus
with prominent
nucleoli
and relatively
voluminous
basophilic
cytoplasm.
The cytoplasm
contamed
several
to many vesicles and lacked azurophilic
granules.
Mitotic
figures
and bi- or trinucleated
cells
were
frequently
seen.
From www.bloodjournal.org by guest on July 31, 2017. For personal use only.
NK CELL
LEUKEMIA
AND
ITS CELL
LINES
1431
Table
1 . Cell Markers
on SPI-801
and SPI-802
Positivity
Percent
Normal
Peripheral
SPI-801
SPI-802
Lymphocytes
72-85
E-rosette
4#{176}c
56.2-66.5
60.0-68.4
(0.8-12.0)
(0.6-13.5)
29#{176}c
30.5-42.0
36.2-48.5
64-80
37#{176}c
30.0-40.4
34.0-44.5
2-5
(0.2-9.8)
(0.4-10.5)
ORBC
41.5-84.0
45.0-72.5
20-31
aggregatedlgG
92.0-98.0
94.5-98.5
24-36
19.2-31.6
18.6-34.5
IgG-Fc
C3
Fig. 1 .
May-Giemsa.
A
light
micrograph
of
SPI-802
cells
stained
with
OKT-3
0
OKT-4
OKT-6
OKT-8
OKM1
On electron
from
the two
microscopic
cell lines
diameter.
They
diffuse
chromatin
were polyribosomes
or few profiles
chondria
were
examination
were
generally
(Fig. 2), cells
15-18
j.z in
had a round
to oval nucleus
with
and 1-3 prominent
nucleoli.
There
throughout
the cytoplasm
and no
of rough
numerous
endoplasmic
reticulum.
in many cells. Golgi
tus was rather
small.
In some cells,
vesicles
and moderately
electron-dense
phous
material,
probably
lipid droplets,
strable.
Cytochemical
stains
for
AS-acetate
esterase
in the
Mitoappara-
electron-lucent
pools of amorwere demon-
peroxidase
and
fresh leukemic
naphtholcells were
negative.
PAS reaction
was positive
in about one-third
of the cells.
Both
SPI-801
and SPI-802
cells
were
totally
negative
for
peroxidase,
esterase,
and naphthol-AS-acetate
tion was granular-positive
in 42%
of SPI-802
cells.
Fig.
2.
An
electron
micrograph
of SPI-802
cells.
Note
a
round to oval nucleus
with several
nucleoli
and
abundant
polyribosomes
distributed
throughout
the
cytoplasm.
There
is no profile
of
well
developed
endoplasmic
reticulum.
A small Golgi apparatus
and many mitochondria
are seen in
one of the cells ( x4800).
a-naphthyl
esterase.
ofSPl-801
butyrate
PAS reacand 50%
62-82
0
0
38-56
0
0
0-1
0
0
20-34
88.5-94.5
Leu-7
15-26
0
90.2-96.8
6-19
0
0
6-18
0
0
2-3
cALL
0
0
0
TceII
0
0
68-86
0
0
14-21
0
0
Human
monocyte
Surfacelg
Cytoplasmic
1gM
0-0.1
Ia
99.0-99.5
98.0-99.5
23-50
AsiaIo-GM
63.2-76.8
68.5-80.2
0
Data
those
obtained
obtained
Immunologic
using
using
neuraminidase-treated
AET-treated
and Other
Some
of the
with AET-treated
SRBC
are shown
and
SRBC are in parentheses.
Cell
Markers
fresh
leukemic
cells
formed
SRBC
at 4#{176}C
(positivity:6.2%)
rosettes
and
37#{176}C
(4.5%).
lgG-Fc
receptors
were demonstrated
in
10.2%
of the cells.
Cell markers
of the cultured
cells
were studied
several
times between
3 and I 8 mo after
the culture
initiation,
Table
1. Both SPI-80l
and
had
formation
receptors
and the results
are
and SPI-802
formed
for IgG-Fc
was dependent
and
C3.
on the SRBC
Their
shown
in
E-rosettes
E-rosette
treatment
and
From www.bloodjournal.org by guest on July 31, 2017. For personal use only.
1432
KOMIVAMA
temperature.
The
neuraminidase-treated
than
those
of the
SPI-801
with
AET-treated
at 29#{176}C
and
receptors
rosette
method
using
latter
method
IgG-Fc
method
and
and
the
Karyotypic
analyses
lines
monocyte
on SPI-801
The
essentially
1
U
I
(1J(
6
7
8
110
*
13
,.I,
I
They
OKT-
had
monoclonal
TdT
SPI-802
was
4
lysis
was
K562
and
against
Raji.
cytotoxicity
5
11
;1;
Lt
22
percent
value of normal
lympho± SD)
and higher
than
(IC
10
21
M
I)
#{149}
K562:
against
K562 but not against
Raji. The stimulation
of
effector
cells
with
PHA-MF
or Con-A-MF
had no
significant
augmentation
effects
on their
cytotoxicity
(p > 0.05),
although
in the separate
experiments
the
of these
16
against
147.4%
of the
± 6.5%,
mean
the absence
of any stimulation
against
Molt-3,
but they did not show the activity
Similarly
SPI-802
had
spontaneous
chromo-
I
activity
that of control
leukemic
cells from I 2 childhood
ALL
patients
(0.1%
± 0.3%).
The first cytotoxicity
assay of
the cultured
cells (exp. 2 in Table
2) was done on May
28, 1981.
SPI-801
cells
demonstrated
cytotoxicity
in
were
Their
NK
76.8%,
being
cytes
(52.1%
9
em.
abnormal
8p and
results
of the cytotoxicity
assay
are
2. The freshly
harvested
leukemic
cells
“
15
14
3. Un addi-
Representative
shown in Table
i;
ID
in Fig.
structurally
were probably
by the
more
3
2
as shown
several
which
Cytotoxicity
I)
.
‘I’
If
cells
had
two of
from 63 to
6q and
Cell-Mediated
antigens.
They
1gM. The two
chromosomes
the same.
demonstrated,
region,
ranging
of the cells analyzed,
l8p.
the culture
initiation,
respecwere analyzed
on SPI-801
and
on SPI-802.
were
and
1 lp
were
in hypotriploid
majority
detected
receptors.
OKT-6,
cALL
and T-cell
Ig or cytoplasmic
3 and 5 mo after
Fifty metaphases
30 metaphases
IgG-Fc
OKT-4,
vast
affinity
were
asialo-GM,
on the cell surfaces.
in the cultured
cells.
Analysis
two cell
low
somes were
67. In the
tion,
many
chromosomes,
in
were
immunofluorescence
anti-human
Chromosome
done
lively.
percentages
aggregated
IgG. When
studied
between
I 6 and I 8 mo of culture,
antibodies,
and lacked
did not have surface
cell lines had
not detectable
their
receptors
than 92% of the cells possessed
were not reactive
with OKT-3,
8, anti-Leu-7,
The
with
higher
4#{176}Cwere
56.2%-66.5%
in SPl-802,
and they
37#{176}C,indicating
for SRBC.
the
of cells rosetted
were apparently
SRBC.
former
cells
at
and 60.0%-68.4%
lower
by
percentages
SR BC
ET AL.
‘ii
#{149}#{149}l#{149}
17
,t’1t4
12
#{149}
18
II
x
V
Fig. 3.
A representative
karyotype
of
the cultured
cell lines. This karyotype
is of a
cell from SPI-801
demonstrated
by the Gbanding
method.
Note 6q and 1 1 p . There
are several
marker
chromosomes
(M).
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NK CELL
LEUKEMIA
AND
ITS CELL
LINES
1433
Table
2.
Results
of Cytotoxicity
Assay
Cytotoxicity
Target
K562
Effector
Experiment
Exp.
Effector
Cells
1
Were
Fresh
Cells
Stimulated
Cells
MoIt-3
Raji
Percent
Percent
cpm
With
Lysis
Percent
Lysis
cpm
cpm
Lysis
None
1,699
±
12
76.8
ND
ND
None
1,268
±
113
52.1
ND
ND
leukemic
cells
Normal
lymphocytes
Exp. 2t
SPI-801
SPI-802
None
596
±
13
8.5
1,630
±
120
15.3
390
±
10
0
PHA-MF
464
±
17
4.2
1,706
±
100
16.9
383
±
11
0
Con-A-MF
652
±
14
10.3
1,309
±
170
371
±
9
0
None
826
±
103
16.0
367
±
15
0
PHA-MF
834
±
127
16.3
339
±
17
0
406
±
16
0.5
433
±
10
1.2
Con-A-MF
Normal
574
±
ND
7.8
22
47.8
184
±
286
390
±
16
0
860
±
50
0
386
±
25
0
0
892
±
36
0
401
±
8
0
0
910
±
68
0
392
±
30
None
1,807
MoIt-3
None
340
±
30
0
JM
None
338
±
41
0
None
322
±
18
None
345
±
16
±
8.6
2,815
40.1
lymphocytes
CCRF-CEM
Jurkat-F
#{149}EXp1 was
1884
done on November
Effector
and target
cells were
mixed
at 40:
1 and
incubated
maximal
release:
tExp.
(MoIt-3),
2 was done on May 28, 1 98 1 . Effector and target cells were mixed at 40: 1 and incubated
and 389 (Raji); cpm maximal release: 3416 (K562),
5675 (MoIt-3),
and 4019 (Raji).
Cpm:
ND,
2
9, 1 980.
ND
expressed
1 04.
Fresh
leukemic
as mean
±
cells
were
obtained
from
the
present
4 hr;
cpm
spontaneous
0
release:
359;
cpm
(K562),
900
patient.
for 4 hr; cpm spontaneous
release:
333
SD.
not done.
14 mo
SPI-802
treatments
of normal
lymphocytes
with the PHA-MF
and Con-A-MF
increased
the values
of their
percent
lysis
K562
against
from
52.0%
±
4.7%
to 85.8%
±
6.0%
and 73.6%
± 2.3%,
respectively.
Leukemia
cell
lines of Molt-3,
JM, CCRF-CEM,
and Jurkat-F
1884
did not have cytotoxic
activity
against
K562,
Molt-3,
and Raji.
The next
whether
activity
activity
SPU-802
cyte
for
experiment
was
performed
to
examine
mixtures
from
5 volunteers,
although
we failed
lymphocyte
The
to
the culture
SPI-802
value)
cultured
initiation,
was
4.6%
and
cells
percent
lysis
by
(9.2%
of normal
(10.4%),
respectively.
5.2%
did
not
show
significant
levels
of
ADCC
against
antibody-coated
CRBC
in a single
assay:
percent
lysis by SPI-801
was 0.8% ± 0.4% and
that by SPI-802
was I .0% ± 0.2% as compared
with
the
the cultured
cells
had the same
cytotoxic
as cytotoxic
T cells and whether
they had the
against
a target
of normal
cells. SPI-80l
and
were stimulated
with MMC-treated
lympho-
after
and
value
(mean
±
of
normal
lymphocytes
of
3 1 .2%
5.4%
±
SD).
DISCUSSION
The
most
freshly
striking
finding
in our
leukemic
harvested
cells
a leukemia-derived
cell
patient
had
line
is that
NK
of K562.
the
activity
In the
study their HLA-DR,
and then the cytotoxicity
of the
cells was assayed
on PHA-induced
normal
T lymphoblasts
derived
from the counterpart
of the stimulator
other
lymphocytes.
They
apparent
cytotoxicity
percent
lysis were
stage of the illness
or its chemotherapy.
Fortunately,
we were
able
to establish
the leukemia
cell lines
retaining
the activity
and to confirm
the results
of
lymphocyte
value).
did
not
in the
less than
To further
demonstrate,
however,
experiment:
values
of
2.6%
(4.6%
of normal
investigate
characteris-
tics of the cytotoxic
activity
of SPI-801
and SPU-802,
a
kinetic
study was performed
with Molt-3
as a target
in
which percent
lysis was evaluated
at 4 time points
over
a period
of
16 hr.
As
shown
in
Fig.
4, the
highest
percent
lysis was observed
at 1 2 hr: percent
lysis by
SPI-801
and SPU-802
was 33.4%
(83.5%
of normal
lymphocyte
value) and 29.2% (73.0%),
respectively.
In
the
latest
4-hr
assay
for cytotoxicity
on a K562
target
against
any
1 2 ALL
such
NK
patients,
the
activity,
and
leukemic
this
studies
on the fresh cells.
Cellular
and
functional
was
cells did not have
not related
to the
similarities
between
the
fresh leukemic
cells and cultured
cells indicate
that the
cell lines
were
derived
from
the leukemic
cells.
A
decrease
of the cytotoxic
activity
of the cultured
cells
as compared
with that of the fresh leukemic
cells may
be explained
activity
during
lines.23’24
the
culture
by
Abnormal
usual decrease
as demonstrated
karyotypes
in
of the cytolytic
in T killer cell
our
cultured
cell
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KOMIYAMA
1434
ability
to kill certain
cultured
cytotoxic
T cells, are not under
tibility
complex
restriction.39
140
20
derived
stronger
lines,
from
such as K562 and Molt-3,
than cell lines
normal
lymphocytes.7
Of tumor-derived
cell
derived
cell
and C3. The cultured
cells
were
and lacked surface
Ig and cytoplasmic
excluding
stage of
the possibility
maturation.
associated
rum
10
antigens
and monoclonal
8
12
16
and SPI-802
against
normal
lymphocytes
tested
for cytolysis
1 6-hr period.
Values
on the
although
The
finding
SPI-802,
indicative
fresh
It
we failed
to perform
chromosome
cells, support
their leukemic
that
the
two
shared the same
of an outgrowth
neoplastic
derived
two
clone
of independent
a 6q
anomaly
leukemic
origin,
with lymphoid
ALL.25
lines,
SPI-801
and
is
of a
from a single cell rather
than
normal
NK cells. In addition,
in our cell lines certainly
favors
their
as it is an anomaly
closely
associated
malignancies
and is not infrequent
in
spontaneous
The
blasts
in the
supports
their
cell
ananature.
chromosome
abnormalities
of the leukemic
cells
absence
growth
of
non-B
T-cell
growth
nature.
Another
leukemic
lympho-
factor
also
proof of the
of
leukemic
origin is the presence
ofasialo-GM1
in a high
density
on the cultured
cells, because the glycosphingolipid
has been
demonstrated
to be a cell surface
marker
of ALL lymphoblasts.26
NK cells are considered
to be non-T,
non-B lymphocytes.6’27’29
Most NK cells have low affinity
receptors
for SRBC30’3’
and avid receptors
for IgG-Fc,30’32’33
and
some of them have receptors
for C3.34 Recent
studies
with
monoclonal
antibodies
have demonstrated
that
NK
gens29’35’36
cells
such as OKT-3,
the presence
positive
for
cells
do not express
T-cell-associated
OKT-4,
and
anti-
OKT-6
despite
of E-receptors,36’37
and most of them are
OKMI29’3537
or Leu-7
(HNK-1).38
Some
express
an
Ia
antigen.29’37
NK
cells
as defined
by anti-T
antibodies
such
heteroantise-
as OKT-3,
They
did
monocyte
have
OKT-
not react
antibodies,
with
but
fresh
and
against
against
Molt-3
as well but not against
Raji and normal
lymphoblasts.
Such phenotypic
characteristics
and targetcell-dependent
spontaneous
cytotoxicity
of the freshly
harvested
and cultured
cells described
here are, therefore, identical
to those of NK cells.
is of
receptors
lines,
lyses
cALL1gM,
they were B cells at any
did not express
T-cell-
possessed
OKMI
and Ia antigens.
The
cultured
cells
had spontaneous
cytotoxicity
K562.
The cultured
cells had the cytotoxicity
(hr)
Fig. 4.
Kinetics
of NK activity
of SPI-801
MoIt-3.
SPI-801
(#{149}-#{149}). SPI-802
(O-O).
( D-G
). and Jurkat-F
1 884 ( 0
0 ) were
against
MoIt-3
targets
at 4 time points over a
represent
means
±
SD.
that
They
and OKT-8.
or anti-human
4, OKT-6,
anti-Leu-7
Time
NK
(leukemia)
times
UgG-Fc,
negative
human
cell linestt
and, unlike
any major histocompaNK
activity
is many
lines, Raji is an exception,
as it is virtually
resistant
to
NK activity.7’40
The freshly
harvested
leukemic
cells as
well as the cultured
cells had receptors
for SRBC,
30
that
tumor
against
ET AL.
interest
that
our
cultured
for neuraminidase-SRBC
for AET-SRBC.
neuraminidase-SRBC3’
cells
but
As NK cells
and the AET
tors
had
modest
avid
recep-
have affinity
for
E-rosette
forma-
tion is rather
specific
for T cells,’3
this may be further
evidence
to support
that the cultured
cells are of NK
cell origin.
One of our unexpected
results
was the
absence of a Leu-7 antigen
on our cell lines. It is well
known
that lymphocyte
surface
antigens
change
with
maturation.4’
It
is likely,
therefore,
that
the
expression
on NK cells38 is restricted
to mature
the circulation,
and our patient’s
leukemic
cells
earlier
stages
Alternatively,
of maturation
not to express the antigen.
since NK cells are heterogeneous
in the
surface
markers,28’29’3537’42
leukemic
Leu-7
cells
were
derived
from
population
of N K cells.
-
Interleukin
2 (IL-2)
NK cell activity.43
Our
augmented
normal
NK
of our cell lines, however,
MF treatment.
Further
mine whether
the absence
response
to IL-2
during
the leukemic
ture
Leu-7
cells in
were at
there
is a possibility
an
0KM
that
1
the
and
been
shown
to augment
MF containing
IL-2,
indeed,
cell activity.
The NK activity
was not augmented
by the
studies
are needed
to deteror marked
decrease
of their
has
resulted
from
transformation
the
cell abberation
or long-term
cul-
in vitro.
Evidence
has been presented
on the morphological
association
of human
NK cells with
large granular
lymphocytes
(LGL).30’44
LGL
have relatively
abun-
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NK CELL
LEUKEMIA
AND
ITS CELL
LINES
1435
dant cytoplasm
with many mitochondria,
plasmic
reticulum,
and electron
dense
patient’s
leukemic
cells had abundant
many mitochondria
mic reticulum
as
granules.
leukemic
the
The
cells
during
in
rabbit
antiserum
are
leukemic
structures
our recent
was
of endoplasazurophilic
or
LGL
the
(data
not
lymphocyte
as ADCC
however,
NK
cells
cells
of
the proliferation
of a malignant
clone
derived
a single
cell of lymphocyte
lineage.
Pandolfi
et
al.4 reported
an interesting
patient
phocytic
leukemia
whose
leukemic
but not NK
activity.
The
patient
the
the view
with chronic
lymcells had ADCC
is, therefore,
in
leukemic
that
by the
leukemic
a chest
made.
NK
and
x-ray
film
enlargement
patients
failed
in our
with
such
cells,
the
diagno-
It was an unexpected
to demonstrate
patient,
clinical
because
features
the patient
have thymic
enlargement.45’46
cells are thymus-independent
cells,42’47
absence
of thymic
enlargement
despite
kemic
infiltration
in our patient
appears
UgG-Fc
receptor-bearing
lymphocytes.
With
respect
to the relationship
between
effector
cells for NK and
ADCC
activity,
of interest
is the demonstration
of
ALL
with such functions,
since
the leukemia
results
from
from
of
of lymphocytes.
was initially
that
T-ALL
In recent
Neville33
has clearly
demonstrated
and
K cells
are distinct
subsets
subsets
formation
finding
IgG-Fc
with
(K cells).32
supports
certainly
mediastinal
population
effector
functions
of a lymphocyte
population
activity
as demonstrated
in
As our patient’s
leukemic
cells had a function
of N K
activity,
it is of interest
to determine
clinical
characteristics of the disease.
Our patient
had massive
leukemic
infiltration,
high blast cell counts,
and a poor prognosis. On the basis
of the hematologic
findings
and
E-rosette
possibility.
in the
expansion
or ADCC
are distinct
sis of T-ALL
think it is surprising
that our cell lines had
but not ADCC,
since NK cells have been
in the same
receptors
with
ours
patients
K cells
between
LGL
and
results
that anti-SPI-802
reactive
with
these
transformation
of the former
in favor
One may
NK activity
that
the
several
cells and
shown)
years,
contrast
cells. The clonal
with either
NK
cells comprise
several
lymphocyte
populations
in the morphology.
The morphological
resem-
blance
leukemic
found
and no or few profiles
do LGL,
but lacked
endoOur
with
absence
of cytoplasmic
granules
in the
with NK activity
suggests
that they lost
granules
that NK
different
sparse
granules.
cytoplasm
most
as those
of
Since
NK
however,
the
massive
leuconceivable.
NK cells play important
roles in host defense
virus infections48’49
as well as tumor
development
against
and
metastasist47’5#{176} in vivo.
sodes of infections
were
In our patient,
apparent
epinot found during
the observa-
tion
however,
period.
It is unclear,
of infections
leukemic
cells.
will
disclose
due
was
Further
the
whether
to the NK
studies
on such
clinical
the absence
activity
patients
characteristics
of the
as ours
of ALL
of NK
cell origin.
REFERENCES
I.
Fu
SM,
Induction
of in vitro
human
2.
Chiorazzi
leukemic
Broder
Muul
N,
differentiation
TA:
evidence
for
T,
adult
T-cell
B-cell
differentiation.
4.
Pandolfi
leukemia
Kiessling
mouse.
mia
Immunol
6.
5:1 12-1
Herberman
reactivity
allogeneic
tumors.
7. Jondal
selective
effector
cells.
K,
C,
DAG,
Gralnick
acute
leukemias.
9.
of two
T
N EngI
J
Uchino
ADCC
Effect
Smith
ML,
not
10.
killer
for mouse
according
JR.
with
chronic
activity.
H: “Natural”
specificity
Ortaldo
cells
Moloney
leuke-
to genotype.
Eur
Nunn
ME,
of mouse
Holden
HT,
lymphoid
cells
Lavrin
against
ofeffector
DH:
Natural
syngeneic
cells.
Leclerc
that
C, Targan
target
JC,
Komiyama
S: Human
cells.
Nature
McVay-Boudreau
natural
killer
spontaneous
killer
272:62-64,
14.
L, Shen
cells in nonimmune
cells
1978
FW,
Kaplan
cell
to the
of the
H,
maturation
of the
generation
level
neutrophil
A, Saitoh
of
NK
of similar
blastogenesis
in
1980
Nakahata
T,
Miyagawa
Y,
in a neutrophil
impaired
ME,
H, Kawai
function.
H, Tsukada
of bactericidal
leukemia.
of human
Ferrarini
defect
J Pediatr
Am
Clark
ability
J Hematol
C:
An
8:197-203,
improved
T lymphocytes.
M, Akabane
of leukemic
T:
in
1980
rosetting
J Immunol
cells
assay
Meth
5:131-135,
G:
Lymphocyte
for
Dickler
with
16.
Tonda
in human
GP,
Risso
lymphoid
A,
Viale
leukemias.
Eur
J Immunol
1975
I 5.
cyte
M,
receptors
globulin
Cantor
spleen
and
and
Galton
1976
Morosawa
assessment
acute
membrane
5:89-93,
M, Spina
G,
I974
and
Int J Cancer
Flandrin
for the classification
I 24:2458-2463,
abnormality
149:1260-
1979
detection
17, 1975
A,
M-T,
Med
S: Disappearance
correlated
J Immunol
T: Abnormal
childhood
13.
E, Argov
cytotoxicity
Quantitative
J
Klein
J Exp
33:451-458,
of lymphocytes
morphologic
12.
in the
Daniel
C: Proposals
explanation
Komiyama
94:19-25,
Blood
MG,
in vivo.
D,
Sultan
Br J Haematol
cultures.
Akabane
growth
Catovsky
HR.
Masucci
activated
tumor
JM,
after
I I.
1978
T-cell
NK
of
1975
M,
Bennett
effect
normal
10:24-34,
a suppressor
but
distribution
H:
mitogen-induced
RB,
of
II. Characterization
evidence
prevent
1264, 1979
a suppressor-cell
Immunopathol
Slease
with
and
for tumor-derived
8. Kasai
H: Direct
populations
of
1978
Goldman
of an interaction
E, Wigzell
cells
RB,
cytotoxic
16:230-239,
DM,
R, Klein
Specificity
M,
of
on pokeweed
with
I. Cytotoxic
cells.
SR:
148:1570-1578,
Takatsuki
Characterization
1980
Harris
synthesis
J, Durm
suppressor
Immunol
F, Strong
56:653-660,
5.
K,
cells
lymphocytic
JP,
nonspecific
Sagawa
Clin
RB:
Med
requirement
of human
leukemia
Herberman
J Exp
Characterization
the
cells in the development
Med 298:66-72,
1978
3. Uchiyama
Halper
D, Whang-Peng
Waldmann
leukemia;
HG,
and immunoglobulin
B lymphocytes.
5, Poplack
L,
Kunkel
HB,
Ross GD,
populations,
Kunkel
B lymphocytes.
Winchester
in Ross
HG:
Interaction
J Exp
RJ:
NR,
Med
Methods
Friedman
of
aggregated
136:191-196,
for enumerating
H (eds):
Manual
1972
lymphoof Clinical
-y-
From www.bloodjournal.org by guest on July 31, 2017. For personal use only.
KOMIYAMA
1436
Immunology
(ed 2). Washington
biology,
I 980,
17.
Volger
Cooper
MD:
LB.
Crist
Cell
leukemia.
N Engl
H, Kubonishi
Comparative
19.
34. Jondal
Micro-
H,
ER,
Lawton
Research
Sinks
1977.
New
cells
(eds):
York,
guish
LF:
cell
leukemia
DS
Janossy
Al,
J,
Pressman
leukemia
cell
20.
I ); antibody
leukemia
Tsubota
Ri,
human
specific
antigen.
T,
JM,
527
of human
Advances
in
antibodies.
37.
antiserum
carries
Cancer
the
non-B
Inst
lymphatic
21.
leukemias
2:971-972,
Svedmyr
lines
tious
EA,
acute
Dennert
for strong
Proc
lymphoblastic
28.
29.
Ortaldo
I,
by
flow
30.
large
RB:
West
icity
Med
Cannon
line;
Immunol
T,
Kasai
M,
302:674-677,
Wigzell
1980
H: “Natural”
with
specificity
of
the
killer
HD,
SO,
antigens
of human
Henney
for
Eur
natural
Timonen
T,
on
purified
highly
antibodies.
JR.
Herberman
and
NK
N,
153:569-582,
GB,
Characteristics
to natural
of
HD,
Bonnard
characterization
of
GD,
Herberman
lymphocytes
against
effector
cells.
L,
murine
Jondal
natural
M,
M:
J
Target-effector
killer
system.
natural
cell
CS,
Klein
E, Klein
to natural
killer
Specific-
killer-target
hybrid.
J Exp
G: Increased
cells
by superinfection
Schlossman
SF:
suppressor
after
Med
sensitiv-
induction
or sodium
303:370-373,
L,
of
butyrate.
J
BR:
J Exp
Med
Kuribayashi
killercell
On
I:
DE,
Nature
Tumors.
the
immune
in human
RB
New
heterogeneity
Gillis
and
(ed):
York,
of
1981
5: Interleukin
291:335-338,
Morphology
in Herberman
the
I 54:750-762,
K, Kern
activity.
E, Timonen
of
subsets
1980
Bloom
cells.
Regulation
I-lymphocyte
2
1981
surface
properties
Natural
of
Cell-Mediated
Academic,
factors
1980,
pp
173-
lymphoblastic
Lampkin
BC: Surface
leukemia.
markers
and
LW,
L, Sen
L, Aur
JV: Initial
prognostic
factors
cyte
rosette
formation
in
cells
Borella
Blood
Habu
Okumura
109
50:671-682,
5,
K,
Fukui
J Med
of NK
Quinnan
children
with
In
acute
K,
vivo
SL,
Mauer
lymphoblastic
and
enhancement
127:34-38,
Manischewitz
antibody-dependent
Kasai
effects
J Immunol
GV,
George
and lymphoblast-erythro-
Shimamura
N:
activity
mice.
RJA,
1977
H,
Iamaoki
in nude
and
N EngI
1976
Dow
murine
J Immunol
KY,
in acute
Simone
48.
a
I, Wong
AM,
growth
of human
NK
1980
killer
Tsukimoto
Reduction
killer
1981
Kay
reactivity
RB:
(HNK-l).
of the solubilized
Reid
cells,
prognostic
leukemia.
J Immunol
relationship
with
1981
of human
cycle
J Med
Against
46.
RB:
human
a comparison
I 27:448-452,
antigen
lines
and
NK
45.
J
killer
Herberman
effectors;
nature
antibody-depen-
185
mouse
cell.
for I cell
and
its loss in a somatic
EL,
Saksela
human
(NK)
antibody
and
and
natural
Immunity
killer
1978
monoclonal
JE:
of
infection.
J
Nagai
Y,
GM,.
of transplanted
tumor
1981
The
role
cell-mediated
cytomegalovirus
M,
anti-asialo
of
natural
killer
cytotoxicity
Exp
Med
during
I 50:1549-1554,
I 979
Bonnard
GD,
Fc receptor
Herberman
lymphocytes
antibody-dependent
Neville
43.
44.
in
W: Evidence
killer
J Immunol
reactivity
viral
Minato
natural
Oku-
detected
monoclonal
1981
151:614-627,
augments
PC: Phenotypes
with
a monoclonal
B, Patarroyo
murine
and
Kung
A differentiation
by
lymphoid
N EngI
42.
1977
I 18:2058-2066,
distinct
Cancer
marker
J Med
H,
lymphocytes
WH,
of human
33.
T cells.
pp 579-580
CM:
response-Inducer
47.
cytotoxic
Kay
and
Human
Hirano
40:58-68,
with
J Exp
cell
32.
T,
Heterogeneity
Sharrow
T, Ortaldo
118:355-361,
parison
for a
distin-
lymphocytes.
1981
Ahrlund-Richter
41. Reinherz
specific
B cells
Blazar
Med
which
1979
ity of human
1981
Natural
myeloid
complex
beings.
1980,
cells
Immunol
granular
K cells.
31.
structure
294:245-248,
HS:
cytometry
Timonen
human
xenogeneic
Exp
1975
1977
characteristics
of surface
127:2401-2409,
with
I
15:596-605,
I
detected
(ADCC)
in the human
ity and
WE,
Newman
(CTL).
Balch
JC,
Biddison
natural
identified
Roder
cytotoxic
populations
JA,
within
the Epstein-Barr
infec-
1975
JR.
Hansen
150:471-481,
for B
with
of alloreactive
in
N FigI
Cytotoxic
Koren
Cell
Determination
cells
Inst
specific
of T cells
as a cell-surface
E, Pross
Il.
5:1 17-121,
populations.
7:487-492,
T, Oda
GM,
cells;
MA,
Failure
to cooperate
H: Clones
leukemia.
mouse.
Kall
RS:
Chromosomes
R, Klein
leukemia
Immunol
interaction
72:1622-1626,
Elsevier/North-Holland,
T: Asialo
Kiessling
the
The
K, Ohashi
acute
in
39.
of
1978
Nakahara
Moloney
T,
chromo-
are present
Sci USA
Allen
J Immunol
York,
K, Tada
cells
M,
cells
B and
as a functional
antibodies
and
127:2575-2580,
127:1024-1029,
Han
Cancer
human
J Cancer
Monoclonal
KA,
cytotoxicity
Abo
of cells
for human
virus
Acad
Hengartner
AA:
New
mura
27.
J Nail
effector
antigens
Eur
CG,
Sandberg
Leukemia.
26.
Nail
G, Dc Rose
272:617-618,
25.
LF,
lines
Int
cells
cell
lymphocytes
K cells
40.
M: Cytotoxic
histocompatibility
Fathman
Nature
type.
technique
by Epstein-Barr
response.
24.
Sinks
on
cell
1980
killer
LD,
I
38.
PC:
NK
J lmmunol
cellular
and
61:987-991,
markers
markers
against
Clouse
natural
cytolytic
1971
transformed
humoral
disease
banding
Jondal
mononucleosis.
23.
5,
of surface
with
M: A rapid
Lancet
22.
Kung
human
JM,
Fast
dent
Ph’
1977
Seabright
somes.
J, Nakazawa
D: Correlation
Surface
subpopulations.
288:394-396,
from
Rabbit
that
to a non-T,
J Nail
Minowada
Pressman
59:845-850,
and
Zarling
between
Immunol
Higby
cell
lymphocyte
36. Zarling
I978
cell
for
Nature
lines;
Elsevier/North-
D:
line
H:
Cytotoxicity
and heterogeneity
non-B
(NALM-
lymphoblastic
M, Pross
VI.
I975
35.
MF,
J, Freeman
J, Yohn
marker
1978
S. Greaves
of fresh
AR,
of childhood
leukemia-lymphoma
Minowada
a non-T,
chromosome
Pearl
pp 363-366
Koshiba
against
for
298:872-878,
T, Higby
human
P. Hilgeis
Leukemia
1978,
J Med
expression
in Bentvelzen
Holland,
Society
phenotype
T, Nakazawa
of
marker
DE,
a new
I, Han
markers
of the
patients,
Bockman
J, Tsubota
surface
analysis
WM,
leukemia;
Minowada
G, Koshiba
American
lymphocytes.
Pre-B-cell
lymphoblastic
18.
DC,
pp 213-228
ET AL.
cellular
RB:
active
A functional
cytotoxicity.
J
49.
corn-
HB:
Deficient
Immunol
vitro
and
tions.
1977
ME:
Human
subpopulations
125:2604-2609,
killer
of
Fc
1980
cells
and
receptor-bearing
natural
lsaacs
cytotox-
in natural
killer
lymphocytes.
cells;
50.
J
NK
D, Clarke
in vivo
Lancet
2:950-952,
284:622-624,
JE,
in tumour
1980
Tyrrell
DAJ,
of leucocyte
in children
lalmadge
cells
JR.
production
with
Webster
interferon
recurrent
ADB,
Valman
(interferon-a)
respiratory
tract
in
infec-
1981
Meyers
growth
KM.
and
Prieur
metastasis
DJ,
Starkey
in beige
JR:
mice.
Role
Nature
of
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1982 60: 1429-1436
Childhood lymphoblastic leukemia with natural killer activity:
establishment of the leukemia cell lines retaining the activity
A Komiyama, H Kawai, Y Miyagawa and T Akabane
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