|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 92 No. 3 (August 1), 1998:
pp. 834-841
Mutual Education Between Hematopoietic Cells and Bone Marrow Stromal
Cells Through Direct Cell-to-Cell Contact: Factors That Determine the
Growth of Bone Marrow Stroma-Dependent Leukemic (HB-1) Cells
By
Huijie Jiang,
Kenkichi Sugimoto,
Hitoshi Sawada,
Emi Takashita,
Maki Tohma,
Hiroyuki Gonda, and
Kazuhiro John Mori
From the Department of Molecular and Cellular Biology, Faculty of
Science, Niigata University, Niigata; and the Department of Internal
Medicine, Kokura Memorial Hospital, Kokurakitaku, Kitakyushu, Japan.
 |
ABSTRACT |
A stroma-dependent cell line (HB-1) was established from myelogenous
leukemic cells of CBA/N mouse. Characterization of the cells showed
that HB-1 proliferated on hematopoietic supportive stromal cells
(MS-10), but did not survive or proliferate on hematopoietic nonsupportive cells (MS-K). Direct contact between HB-1 and MS-10 appears to be necessary for HB-1 to proliferate on MS-10. We found that
interleukin-1 (IL-1) produced by MS-10 plays a major role in the
survival and proliferation of HB-1. IL-11 did not support the
proliferation of HB-1 cells by itself, but enhanced the proliferation of HB-1 cells in the presence of IL-1. The expression of IL-1 and
IL-11 was induced in MS-10 by the direct contact with HB-1 cells, and
the expression of IL-1 receptor type I (IL-1RI) and interleukin-11
receptor (IL-11R) was induced in HB-1 cells by the attachment of the
cells to MS-10. These findings show the existence of two-way
interactions between HB-1 and MS-10.
© 1998 by The American Society of Hematology.
| |
INTRODUCTION |
BIDIRECTIONAL INTERACTIONS between
hematopoietic cells and stromal cells in the marrow microenvironment
may be crucially important in hematopoietic regulation.1-4
The growth factor dependency and responses of acute myelogenous
leukemia (AML) progenitor cells mimic, in many ways, those
of normal hematopoietic precursor cells, including responses to factors
produced by bone marrow (BM) stromal cells and accessory
cells.3,4 Model systems of such interactions in vitro
have been very instructive in this regard. For example, the
interleukin-3 (IL-3)-dependent murine leukemia cell line DA-1 survives
and proliferates when cocultured with the hematopoietic-supportive stromal cell line MS-5 without addition of exogenous IL-3 or other growth factors.5 Coculture of DA-1 and MS-5 is associated
with expression of bcl-2 by the DA-1 cells and with expression of
granulocyte-macrophage colony-stimulating factor (GM-CSF) by the
MS-5 cells.6 There is considerable evidence that IL-1 also
plays a major role in hematopoietic regulation by modulating and
stimulating progenitor cell proliferation.7-9
In the present study, we examine the survival and proliferation of HB-1
murine AML cells cocultured with the MS-10 hematopoietic supportive
stromal cell line. We further examine the proliferative roles of IL-1
and other growth factors and the effects of coculture on expression of
growth-related genes.
| |
MATERIALS AND METHODS |
Mice.
Female mice of C57B16, CBA/H, CBA/N, and C3H/HeN were purchased from
Shizuoka Animal Farm (Hamamatsu City, Japan) and
maintained under specific pathogen-free conditions. Ten- to 12-week-old
mice were used for experiments.
Cells.
The MS-10 and MS-K stromal cell lines were established from BM of
C3H/HeN mice according to previously reported methods and were
maintained in -minimal essential medium (-MEM) supplemented with
10% (vol/vol) horse serum.5,10 MS-10 cells support the proliferation and differentiation of hematopoietic stem cells in vitro
as previously determined for the stromal line MS-5.10 The
HB-1 cell line was established from the spleen cells of CBA/N mice in
which myelogenous leukemia (M361) was induced by irradiation. HB-1
cells were maintained on MS-10 in -MEM supplemented with 10%
(vol/vol) fetal calf serum (FCS). For cell proliferation experiments, HB-1 cells suspended in -MEM supplemented with 10% (vol/vol) FCS
were overlaid and allowed to adhere onto MS-10 or MS-K cells. Suspended
HB-1 cells were counted daily. For further experiments, HB-1 cells
cultured on MS-10 cells were harvested and purified from MS-10 by
passage through a nylon-glass wool and G-10 column, yielding >98%
pure HB-1.11,12 Characterization studies showed that
HB-1 survived and proliferated on MS-10, but survival was not
supported by cytokines secreted by MS-10. HB-1 cells did not survive on
MS-K.
Growth stimulation of HB-1 cells by conditioned medium of MS-10
cells cocultured with HB-1 cells.
To clarify whether MS-10 produce and release soluble factor(s) that
support the proliferation of HB-1 cells, 15% of cocultured medium
(HB-1 cultured on MS-10 for 5 days) was added into the suspension
culture of HB-1 (1 × 105 cells/mL) in -MEM
supplemented with 10% (vol/vol) FCS. The number of HB-1 cells was
counted daily.
The requirement of cell contact was investigated by culturing HB-1
cells (1 × 105/mL in -MEM supplemented with 10%
FCS) in the upper chamber of wells separated by Millicell-HA membranes
(0.45 µmol/L pore size; Millipore Products Division, Ashby Road,
Bedford, MA) from 2-day confluent MS-10 monolayers. The
number of HB-1 cells was counted every day.
Growth of HB-1 cells in the presence of interleukins.
Recombinant human IL-1 (rhIL-1, kindly provided by Dr T. Masuda,
Department of Medicine, Kyoto University, Kyoto, Japan) was added at a final concentration of 10 U/mL into the suspension culture of HB-1 cells. Recombinant murine IL-6 (rmIL-6, kindly provided
by Dr T. Hirano, Department of Medicine, Osaka University, Osaka,
Japan) was added at 10 U/mL. rhIL-11 (specific activity of
2 × 106 U/mg) was purchased from CosmoBio Co,
LTD (Tokyo, Japan) and used in culture at 20 U/mL of
rhIL-11. Conditioned medium of IL-3-producing STIL-3 leukemic T-cell
line (STIL-3 CM)13 was used to substitute rmIL-3. rmGM-CSF
(kindly provided by Kirin Brewery Co LTD, Tokyo, Japan)
was added at a concentration of 250 U/mL.
HB-1 cells, which were separated from coculture with MS-10 cells, were
transferred into plastic culture dishes and cultured in suspension in
-MEM supplemented with 10% (vol/vol) FCS. rhIL-1, rmIL-6, STIL-3
CM, rhIL-11, or rmGM-CSF was added in the cultured medium of HB-1
cells, respectively. The number of HB-1 cells was counted 6 days later.
To investigate the synergistic effect of IL-6, IL-3, IL-11, and GM-CSF
with IL-1 on the proliferation of HB-1 cells, HB-1 cells were
separated from MS-10 and cultured in suspension in -MEM supplemented
with 10% (vol/vol) FCS. Recombinant human IL-1 was added in the
culture medium of HB-1 cells, and rmIL-6, STIL-3 CM, rmIL-11, or
rmGM-CSF was added, respectively. The number of HB-1 cells was counted
6 days later.
Effect of anti-mouse IL-1 and anti-human IL-11
antibody on the growth of HB-1 cells.
To examine effects of IL-1 or IL-11 on the growth of HB-1 cells,
effect of anti-mouse IL-1 neutralizing antibody (purchased from
CosmoBio Co, LTD) or anti-human IL-11 neutralizing antibody (which
cross-reacts with murine IL-11, purchased from Funakoshi Co, LTD,
Tokyo, Japan) was diluted with serum-containing medium (from 1:20 to 1:500) for addition to the coculture of HB-1 cells and
MS-10. The number of HB-1 cells in suspension was counted every 2 days.
Extraction of total RNA and polymerase chain reaction (PCR).
Total RNA was extracted by the acid guanidinium
thiocyanate-phenol-chloroform method.14 Expression of
messenger RNA was determined by the reverse transcriptase-PCR (RT-PCR)
method as described previously.15 Briefly, 3 µg of total
RNA was reverse-transcribed using First-Strand cDNA Synthesis Kit
(Pharmacia, Uppsala, Sweden) and used for PCR under the
following conditions: final reaction volume of 20 µL with 5 pmol each
of two specific primers, at a final concentration of 200 µmol/L of
each of deoxyadenosine triphosphate (dATP), deoxythymidine triphosphate
(dTTP), deoxyguanosine triphosphate (dGTP), deoxycytidine
triphosphate (dCTP), 50 mmol/L KCl, 10 mmol/L Tris-HCl
(pH 8.3), 1.5 mmol/L MgCl2, 1 mg/mL gelatin, and 0.5 U Gene
Taq DNA polymerase (Nippon Gene, Tokyo, Japan).
PCR was performed using Program Temp Control System
(ASTEC, Fukuoka City, Japan) for 25 or 35 cycles under
the following conditions; 1 minute at 95°C, 2 minutes at 55°C,
and 3 minutes at 72°C. Amplified cDNA products were electrophoresed
on 2% (wt/vol) Nusieve 3:1 agarose gel (FMC Bio Products,
Rockland, ME) and stained with ethidium bromide.
A quantitative PCR analysis was performed to determine the mRNA levels
of  -actin, cytokines, and apoptosis-related factors.16 We estimated the amount of PCR products after defined cycles of PCR by
measuring 32P-labeled nucleotide incorporation. The amount
of amplified products of -actin increased approximately
logarithmically up to 30 to 35 cycles. To keep the reaction within
nonlimiting conditions and obtain detectable signal of the amplified
products of -actin, we chose 25 cycles of amplification. The mRNA
level of various cytokines was lower than that of -actin. Thus, 35 cycles of amplification was chosen.
Primers for PCR.
The sequences for primer pairs are listed in
Tables 1 and
2. These sequences were synthesized by
Kurashiki Bouseki Co, LTD, Kurashiki City, Japan.
Lipopolysaccharide (LPS)-stimulated MS-5 cells were used as
positive control of IL-1 and IL-11 production; NFSA
cells17 were used as positive control of IL-1, IL-6,
IL-8, G-CSF, M-CSF, and GM-CSF gene expression; ConA-stimulated spleen cells of C3H/HeN were used as positive control of IL-2 and IL-4 gene
expression; WEHI-3 was used as positive control of IL-3 gene expression; and nonstimulated MS-5 cells were used as positive control
of IL-7 and stem cell factor (SCF) gene expression.
Thymine-adenine (TA) cloning and DNA sequencing.
Construction of T vectors, a rapid and general system for direct
cloning of unmodified PCR product, was performed with the method
described by Marchuk et al.18,19 DNA sequencing was performed with Bca BEST Dideoxy Sequencing Kit (TAKARA Co, LTD, Japan). Using this method, we checked all sequences of
the PCR products.
Northern blot.
Total cellular RNA was extracted by the acid guanidinium
thiocyanate-phenol-chloroform method.14 For Northern blots,
15 µg RNA were electrophoresed and transferred to a Hybond
N+ membrane as described in Paul et
al.21 Probes were labeled by 32P
using Bca BEST Labeling Kit. Hybridization was performed at 42°C, washed with 2× SSC (33.3 mmol/L NaCl, 33.3 mmol/L
C6H5O7Na3 2H2O, pH 7.0), 0.1% (wt/vol) sodium dodecyl sulfate (SDS), and the signal was detected by autoradiography with intensifying screens. The film
exposure was for 14 hours at  80°C.
Immunochemistry.
Anti-human IL-11 neutralizing antibody (which cross-reacts with murine
IL-11, produced in goats immunized with purified rhIL-11, purchased
from Funakoshi Co, LTD) was used for the detection of murine IL-11
molecules. Immunoprecipitations by anti-human IL-11 neutralizing
antibody were performed from the conditioned media of nonadherent BM
cells cultured on MS-10 for 6, 12, 24, 48, and 72 hours and media from
HB-1 cells cultured on MS-10, incubated overnight at 4°C, under
gentle rotation. Protein G-agarose was added to the system as an
immunoabsorbant and kept for 3 hours at 4°C. The precipitated
material was washed three times in 0.05% Tween-20 phosphate-buffered
saline (PBS) buffer and boiled for 3 minutes in the buffer containing
125 mmol/L Tris-HCl pH 6.8, 2% (wt/vol) SDS and 5% (vol/vol)
2-mercaptoethanol. The material was subjected to SDS-polyacrylamide gel
electrophoresis and Western blot transfer to polyvinylidene difluoride
membrane (Millipore, Tokyo, Japan) was performed. The filter was
blocked with 0.05% Tween 20-PBS buffer. Incubations were performed
with anti-human IL-11 antibody at a 1/400 dilution overnight in 0.05%
Tween 20-PBS buffer and detection with antigoat IgG-alkaline
phosphatase-conjugated antibody (Santa Cruz Biotechnology, Inc, Santa
Cruz, CA) was used for detection of the precipitate.
| |
RESULTS |
Growth of HB-1 cells on MS-10.
Rapid proliferation of HB-1 cells was supported by coculture on MS-10
cell monolayers, but not on MS-K monolayers
(Fig 1A). In an attempt to detect soluble
factors produced and released by MS-10 cells cocultured with HB-1
cells, which could support HB-1 proliferation, HB-1
cells were cultured with MS-10 cells or in culture wells separated from
MS-10 cells by Millicell membranes (Fig 1B). Neither condition
supported HB-1 survival, indicating that cell-to-cell contact was
required for HB-1 cell proliferation on MS-10.
View larger version (13K):
[in this window]
[in a new window]
| Fig 1.
Growth of HB-1 cells on hematopoietic supportive stromal
cells. (A) HB-1 cells were cultured on hematopoietic supportive stromal cells, MS-10 ( ) or hematopoietic nonsupportive stromal cells, MS-K
( ). The number of HB-1 cells in suspension was counted every day.
Data represent mean ± SE. (B) HB-1 cells were cultured on MS-10 cell
layer, but separated by Millicell-HA membrane () or cultured in the
presence of conditioned medium of the coculture (). HB-1 cells were
directly cultured on MS-10 cells as control (). The number of HB-1
cells was counted every day. Data represent mean ± SE.
|
|
Role of IL-1 and IL-11 in the proliferation of HB-1
cells.
MS-10 cells cocultured with HB-1 cells expressed IL-1 and IL-11,
as assessed by both RT-PCR and Northern blot analysis
(Figs 2A and
3A). IL-1 and IL-11 expression by MS-10
cells cultured alone was not detectable. HB-1 cells proliferated slowly
in suspension culture supplemented with rhIL-1
(Table 3). In contrast, rhIL-11, STIL-CM,
rmIL-6, and rmGM-CSF individually failed to support HB-1 proliferation.
rhIL-11 synergistically enhanced IL-1-stimulated proliferation.
When added to cocultures of HB-1 on MS-10, anti-mouse IL-1
neutralizing antibody blocked proliferation of HB-1; anti-human IL-11
neutralizing antibody slightly inhibited HB-1 proliferation (Fig 4).
View larger version (34K):
[in this window]
[in a new window]
| Fig 2.
PCR analysis of the expression of cytokines in MS-10
cells and their receptors in HB-1 cells by the coculture of MS-10 and HB-1 cells. (A) Expression of IL-1 and IL-11 in MS-10 cells was analyzed by RT-PCR after coculture with HB-1 cells. Each lane represents MS-10 cells alone (lane 3), MS-10 cells cocultured with HB-1
cells (lane 4), and HB-1 cells cocultured with MS-10 cells (lane 5),
respectively. LPS-stimulated MS-5 cells were used as positive control
(lane 2). Marker was 100 bp ladders (lane 1). (B) Expression of IL-1
receptor type I and IL-11 receptor in HB-1 cells was analyzed by RT-PCR
after coculture with MS-10 cells or after IL-1 stimulation. Each
lane represents HB-1 cells separated from MS-10 cells for 5 days (lane
3), HB-1 cells cocultured with MS-10 cells (lane 4). HB-1 cells
separated from MS-10 cells for 5 days were reseeded on MS-10 cells
again and harvested 5 days later (lane 5) or cultured in the presence
of IL-1 and harvested 5 days later (10 U/mL) (lane 6). C3H/HeN
spleen cells were used as positive control (lane 2). Marker was 100 bp
ladders (lane 1).
|
|
View larger version (35K):
[in this window]
[in a new window]
| Fig 3.
Northern blot analysis of the expression of ILs in MS-10
cells or their receptors in HB-1 cells by the coculture of MS-10 and
HB-1 cells. (A) Result of hybridization with the
32P-labeled probes for IL-1 and IL-11. A total of 15 µg of total RNA of each sample was loaded in each lane. Each lane
represents HB-1 cells separated from MS-10 cells for 5 days (lane 3);
HB-1 cells cocultured with MS-10 cells (lane 4); HB-1 cells separated from MS-10 cells for 5 days then reseeded on MS-10 cells again and
harvested 5 days later (lane 5); or cultured in the presence of 10 U/mL
IL-1 and harvested 5 days later ( lane 6 ); MS-10 cells cocultured
with HB-1 cells (lane 7); and MS-10 cells alone (lane 8).
LPS-stimulated MS-5 cells were used as positive control (lane 2). MS-5
cells were used as negative control (lane 1). The level of -actin is
shown as control. (B) Result of hybridization with the
32P-labeled probes for IL-1RI and IL-11R. A total of 15 µg of total RNA from each sample was loaded in each lane. Each lane
represents HB-1 cells separated from MS-10 cells for 5 days (lane 3);
HB-1 cells cocultured with MS-10 cells (lane 4); HB-1 cells separated from MS-10 cells for 5 days then reseeded on MS-10 cells again and
harvested 5 days later (lane 5); cultured in the presence of 10 U/mL
IL-1 and harvested 5 days later (lane 6); MS-10 cells cocultured
with HB-1 cells (lane 7); MS-10 cells alone (lane 8). C3H/HeN spleen
cells and NIH3T3 cells were used as control. The level of -actin is
shown as control.
|
|
View larger version (18K):
[in this window]
[in a new window]
| Fig 4.
Effect of neutralizing antibodies on the growth of HB-1
cells. HB-1 cells were cultured on MS-10 with or without anti-IL-1 neutralizing antibody (A) or anti-IL-11 neutralizing antibody (B). The
number of HB-1 cells was counted every 2 days.
|
|
Expression of growth factor genes by MS-10 cocultured with
nonadherent BM cells.
Coculture with nonadherent BM cells for 48 hours resulted in increased
expression of genes for SCF, G-CSF, M-CSF, and GM-CSF by MS-10 cells,
assessed by quantitative PCR16
(Fig 5). Basal expression of IL-6 and IL-7
was unchanged. Expression of genes for IL-1, IL-1, IL-2, IL-3,
IL-4, and IL-8 by MS-10 cells cultured alone or cocultured with BM
cells was not detected. A transient expression of the IL-11 gene was
observed between 6 and 24 hours of coculture, but was undetectable
after 48 hours (Fig 6). Immunoreactive IL-11 was found in the coculture
supernatant at 24 hours (Fig
7). No such transient expression of IL-1
was observed.
View larger version (38K):
[in this window]
[in a new window]
| Fig 5.
Induction of cytokines in MS-10 cells by the coculture
with nonadherent BM cells. Nonadherent BM cells were cocultured with MS-10, and the expression of cytokines was analyzed by quantitative RT-PCR, MS-10 alone (lane 2); MS-10 cocultured with nonadherent BM
cells for 48 hours (lane 3); nonadherent BM cells (lane 4); nonadherent
BM cells cocultured with MS-10 for 48 hours (lane 5). Lane 1 is a
positive control.
|
|
View larger version (29K):
[in this window]
[in a new window]
| Fig 6.
Induction of IL-11 production in MS-10 cells by the
coculture with nonadherent BM cells. MS-10 cells were cocultured with nonadherent BM cells for 6, 12, 24, and 48 hours. Expression of the
genes for IL-1 and IL-11 in MS-10 cells was analyzed by quantitative RT-PCR, MS-10 alone (lane 2); MS-10 cocultured with nonadherent BM
cells for 6 hours (lane 3); 12 hours (lane 4); 24 hours (lane 5); or 48 hours (lane 6); nonadherent BM cells (lane 7). Lane 1 is a positive
control.
|
|
View larger version (121K):
[in this window]
[in a new window]
| Fig 7.
Detection of IL-11 molecules in conditioned media of the
coculture of nonaderent BM cells and MS-10. MS-10 cells were cocultured with nonadherent BM cells for 6, 12, 24, 48, and 72 hours. IL-11 molecules were analyzed by immunochemistry. Negative controls (lane 1, conditioned medium of MS-10 alone and lane 8, 10% FCS in -medium).
Positive control (lane 2, conditioned medium of HB-1 cultured on
MS-10). Conditioned medium of MS-10 cocultured with nonadherent BM
cells for 6 hours (lane 3); 12 hours (lane 4); 24 hours (lane 5); 48 hours (lane 6); and 72 hours (lane 7). M lane 1 is molecular-weight
marker.
|
|
Expression of IL-1 and IL-11 receptor genes by HB-1 cells.
The proliferative effects of IL-1 and IL-11 on HB-1 cells imply that
the factors' respective receptors are expressed constitutively or
induced by contact with MS-10. Accordingly, IL-1RI gene was not
expressed in HB-1 cells cultured alone, but was expressed when HB-1
cells were cultured on MS-10 (Figs 2B and 3B). Basal expression of
IL-11R on HB-1 cells was increased by coculture on MS-10, and addition
of IL-1 to the suspension culture of HB-1 cells induced expression
of IL-1RI and increased IL-11R expression.
| |
DISCUSSION |
Intercellular interactions between hematopoietic cells and BM stromal
cells play a crucial role in hematopoiesis.1,2 The
existence of two-way interactions between DA-1 cells and hematopoietic supportive stromal cells was demonstrated by Takashita et
al.6 For example, we previously showed that contact between
DA-1 factor-dependent leukemia cells and the supportive stromal cell
line, MS-5, induced the expression of GM-CSF by the stromal cells and
of bcl-2 by the leukemia cells.6 In preliminary studies to
characterize HB-1 cells, direct contact between HB-1 and MS-10 was
found to be necessary for the survival and proliferation of HB-1 cells. Separation of HB-1 from MS-10 cells resulted in apotosis of HB-1 cells,
although no difference was found in Fas ligand (FasL), tumor necrosis
factor (TNF) receptor p55, TNF receptor p75, interleukin-1 converting enzyme (ICE) or bcl-2 between HB-1 cells cultured alone and
cultured on MS-10 (data not shown).
The aim of the present study was to elucidate factors that determine
the dependency of HB-1 cells on MS-10. We found that rhIL-1 was able
to substitute partially for stroma contact in supporting the survival
and proliferation of HB-1. Suspension culture of HB-1 cells in the
presence of IL-1 resulted in the survival and slow growth of the
cells. Addition of IL-11 to HB-1 cells in suspension culture resulted
in a significant enhancement of the growth of HB-1 cells. Expression of
IL-1RI and IL-11R was induced on HB-1 cells and expression of IL-1
and IL-11 was induced in MS-10 by coculture of MS-10 and HB-1 in direct
contact. Expression of IL-1RI and IL-11R was also induced in HB-1 cells
by IL-1. These findings indicate strongly that two-way interactions
exist between HB-1 and MS-10, as previously shown for DA-1 and
MS-5.6
Contact between HB-1 and MS-10 was necessary for the induction of
IL-1 and IL-11 in MS-10 cocultured with HB-1 cells. Expression of
IL-1 was detected in MS-10, but not in HB-1 cells. The medium from
coculture of MS-10 cells and HB-1 cells did not support the survival of
HB-1 cells. When HB-1 cells were cultured on MS-K or in transwells
above MS-10, neither of the stromal cells released IL-1 or IL-11
into the culture medium (data not shown). Thus, our data suggest that
coculture of HB-1 and MS-10 induces expression of membrane-bound IL-1
on MS-10 cells.
Induction of IL-1 production by MS-10 cells may represent a first
step in the mutual education of hematopoietic cells and stromal cells.
Although IL-1RI was not dectectable on HB-1 cells cultured alone, it is
plausible that HB-1 expresses low levels of IL-1RI, which is
undetectable by the present PCR technique and that IL-1 stimulated an
enhanced expression of IL-1 receptors on HB-1 (analogous to the effect
of IL-2 on the expression of its receptors on
lymphocytes20). We observed that coculture of MS-10 with BM
cells also resulted in the transient production of IL-11 (at both mRNA
and protein levels), which is consistent with reports that human IL-11
supports the growth of hematopoietic cells in long-term BM culture and
acts synergistically with other growth factors to stimulate multiple
lineages of hematopoietic cells in vitro.21-25 Yanai et
al26 reported that the survival, self-renewal, and
differentiation of hematopoietic stem cells was
controllable separately. In a similar stepwise control
mechanism, IL-1 may support the survival and slow growth of HB-1
cells, which is then accelerated by additional signals from IL-11.
In previous studies, the nonsupportive MS-K cell line was characterized
by comparison to the hematopoietic supportive line MS-5 (analogous to
MS-10).4 The major differences between these two cell lines
are cell-to-cell contact through which extracellular matrix-bound SCF
appears to be transferred from MS-5 to hematopoietic cells.5 Although MS-K did adhere a small number of HB-1
cells, MS-K did not support the proliferation of HB-1, suggesting a
difference in the expression of integrin(s) and/or cytokine(s)
between MS-10 and MS-K. We have tested antibodies to some of the known
adhesion molecules such as very late activation antigen (VLA)-4,
vascular cell adhesion molecule (VCAM)-1, lymphocyte
function-associated antigen (LFA)-1, intercellular adhesion molecule
(ICAM)-1, CD44, Mac-1, and VLA-5 to elucidate if any of
the antibodies block hematopoietic cell adherence, but have found none
so far. We are now trying to obtain monoclonal antibodies against
MS-10, hoping to find the candidate molecule(s). Using the model of
MS-10 and MS-K, specific novel integrin(s) the adherence of
hematopoietic cells to stromal cells is expected to be
found.
| |
FOOTNOTES |
Submitted January 21, 1997;
accepted February 23, 1998.
Supported in part by grants from Kirin Brewery Co, LTD, and from Sankyo
Co, LTD, Tokyo, Japan.
Address reprint requests to Kazuhiro John Mori, MD,
Department of Molecular and Cellular Biology, Faculty of Science,
Niigata University, Niigata 950-21, Japan.
The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" is accordance with 18 U.S.C. section 1734 solely to indicate this fact.
| |
ACKNOWLEDGMENT |
We are grateful to Dr T. Masuda (Department of Medicine, Kyoto
University) and Dr T. Hirano (Department of Medicine, Osaka University)
for providing us with rhIL-1 and rmIL-6.
| |
REFERENCES |
1.
Roberts RA,
Spooncer E,
Parkinson EK,
Lord BI,
Allen TD,
Dexter TM:
Metabolically inactive 3T3 cells can substitute for marrow stromal cells to promote the proliferation and development of multipotent haemopoietic stem cells.
J Cell Physiol
132:203,
1987[Medline]
[Order article via Infotrieve]
2.
Yamazaki K,
Roberts RA,
Spooncer E,
Dexter TM,
Allen TD:
Cellular interactions between 3T3 cells and interleukin-3-dependent multipotent haemopoietic cells: A model system for stromal-cell-mediated haemopoiesis.
J Cell Physiol
139:301,
1989[Medline]
[Order article via Infotrieve]
3.
Wolf NS:
The hematopoietic microenvironment.
Clin Haematol
8:469,
1979[Medline]
[Order article via Infotrieve]
4.
Roberts R,
Gallagher J,
Spooncer E,
Allen TD,
Bloomfield F,
Dexter TM:
Heparan sulphate bound growth factors: A mechanism for stromal cell mediated haemopoiesis.
Nature
332:367,
1988
5.
Suzuki J,
Fujita J,
Taniguchi S,
Sugimoto K,
Mori KJ:
Characterzation of murine hemopoietic-supportive (MS-1 and MS-5) and non-supportive (MS-K) cell lines.
Leukemia
6:452,
1992[Medline]
[Order article via Infotrieve]
6.
Takashita E,
Sugimoto K,
Adachi Y,
Mori KJ:
Induction of bcl-2 gene expression by intercellular information from hemopoietic supportive stromal cells to DA-1 cells.
J Cell Physiol
161:367,
1994[Medline]
[Order article via Infotrieve]
7.
Rodriquez-Cimadevilla J,
Beauchemin V,
Villemeuve L,
Letendre F,
Shaw A,
Hoang T:
Coordinate secretion of interleukin-1 and granulocyte-macrophage colony-stimulating factor by the blast cells of acute myeloblastic leukemia: Role of interleukin-1 as an endogenous inducer.
Blood
76:1481,
1990[Abstract/Free Full Text]
8.
Estrov Z,
Kurzrock R,
Estey E,
Wetzler M,
Ferrajoli A,
Harris D,
Blake M,
Gutterman JU,
Talpaz M:
Inhibition of acute myelogenous leukemia blast proliferation by interleukin-1 (IL-1) receptor antagonist and soluble IL-1 receptors.
Blood
79:1938,
1992[Abstract/Free Full Text]
9.
Estrov Z,
Black RA,
Sleath PR,
Harris D,
Van Q,
laPushin R,
Estey EH,
Talpaz M:
Effect of interleukin-1 converting enzyme inhibitor on acute myelogenous leukemia progenitor proliferation.
Blood
86:4594,
1995[Abstract/Free Full Text]
10.
Itoh K,
Tezuka H,
Sakoda H,
Konno K,
Nagata K,
Uchiyama U,
Uchino H,
Mori KJ:
Reproducible establishment of hemopoietic supportive stromal cell lines from murine bone marrow.
Exp Hematol
17:145,
1989[Medline]
[Order article via Infotrieve]
11.
Julius MH,
Simpson E,
Herzenberg A:
A rapid method for the isolation of functional thymus-derived murine lymphocytes.
Eur J Immunol
3:645,
1973[Medline]
[Order article via Infotrieve]
12.
Collins LS,
Dorshkind K:
A stromal cell line from myeloid long-term bone marrow cultures can support myelopoiesis and B lymphopoiesis.
J Immunol
138:1082,
1987[Abstract/Free Full Text]
13.
Sawada H,
Sugimoto K,
Aramaki K,
Mori KJ:
Hemopoietic features of splenectomized mice bearing IL-3 producing T cell leukemia.
Leuk Res
13:1131,
1989[Medline]
[Order article via Infotrieve]
14.
Chomczynski P,
Sacchi N:
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol chloroform extraction.
Anal Biochem
162:156,
1987[Medline]
[Order article via Infotrieve]
15.
Kawasaki ES:
Amplification of RNA
, in Innis MA,
Gelfand DH,
Sininsky JJ,
White TJ
(eds):
PCR Protocols, a Guide to Methods and Amplifications.
San Diego, CA, Academic
, 1990
, p 21
16.
Yokoi H,
Natsuyama S,
Iwai M,
Nada Y,
Mori T,
Mori KJ,
Fujita K,
Nakayama H,
Fujita J:
Non-radioisotopic quantitative RT-PCR to detect changes in mRNA levels during early mouse embryo development.
Biochem Biophys Res Commun
195:769,
1993[Medline]
[Order article via Infotrieve]
17.
Murray D,
Basic I,
Milas L,
Meyn RE:
DNA cross-linking following exposure to cis-platinum in primary and serially passaged cultured cells derived from two murine fibrosarcomas.
Cancer Chemother Pharmacol
20:133,
1987[Medline]
[Order article via Infotrieve]
18.
Marchuk D,
Drumm M,
Saulino A,
Collins FS:
Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products.
Nucleic Acids Res
19:5,
1991[Abstract/Free Full Text]
19. Treco DA: Current Protocols in Molecular Biology. New York, NY,
Wiley, 1987
20.
Janssen RA,
Heijn AA,
de The TH,
Leij L:
Poor induction of interleukin-2 receptor expression on CD8bright+ cells in whole blood cell cultures with CD3 mAb. Implications for immunotherapy with CD3 mAb.
Cancer Immunol Immunother
38:53,
1994[Medline]
[Order article via Infotrieve]
21.
Paul SR,
Bennett F,
Calvetti JA,
Kelleher K,
Wood CR,
O'Hara RM,
Leary AC Jr,
Sibley B,
Clark SC,
Williams DA,
Yang Y-C:
Molecular cloning of a cDNA encoding interleukin 11, a stromal cell-derived lymphopoietic and hematopoietic cytokine.
Proc Natl Acad Sci USA
87:7521,
1990
22.
Morris JC,
Neben S,
Bennett F:
Molecular cloning and characterizaton of murine interleukin-11.
Exp Hematol
24:1369,
1996[Medline]
[Order article via Infotrieve]
23.
Hilton DJ,
Hilton AA,
Raicevic A,
Rakar S,
Harrison-Smith M,
Gough NM,
Begley CG,
Metcalf D,
Nicola NA,
Willson TA:
Cloning of murine IL-11 receptor alpha-chain; requirement for gp130 for high affinity binding and signal transduction.
EMBO J
13:4765,
1994[Medline]
[Order article via Infotrieve]
24.
Miyajima A,
Miyatake S,
Schreurs J,
De Vries J,
Arai N,
Yokota T:
Coordinate regulation of immune and inflammatory responses by T cell-derived lymphokines.
FASEB J
2:2462,
1988[Abstract]
25.
Tsuji T,
Sugimoto K,
Yanai T,
Takashita E,
Mori KJ:
Induction of the gene expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) in the bone marrow cells and marrow-fractionated cell populations by interleukin-3 (IL-3): IL-3-mediated positive feed-back mechanism of granulopoiesis.
Growth Factors
11:71,
1994[Medline]
[Order article via Infotrieve]
26.
Yanai T,
Sugimoto K,
Takashita E,
Aihara Y,
Tsurumaki Y,
Tsuji T,
Mori KJ:
Separate control of the survival, the self-renewal and the differentiation of hemopoietic stem cells.
Cell Struct Funct
20:117,
1995[Medline]
[Order article via Infotrieve]
27.
Zsebo KM,
Williams DA,
Geissler EN,
Broudy VC,
Martin FH,
Atkins HL,
Hsu RY,
Birkett NC,
Okino KH,
Murdock DC,
Jacobsen FW,
Langley KE,
Smith KA,
Takeishi T,
Cattanach BM,
Galli SJ,
Suggs SV:
Stem cell factor is encoded at the S1 locus of the mouse and is the ligand for the c-kit tyrosine kinase receptor.
Cell
63:213,
1990[Medline]
[Order article via Infotrieve]
28.
Tsuchiya M,
Asano S,
Kaziro Y,
Nagata S:
Isolation and characterization of the cDNA for murine granulocyte colony-stimulating factor.
Proc Natl Acad Sci USA
83:7633,
1986[Abstract/Free Full Text]
29.
Harrington MA,
Edenberg HJ,
Saxman SM,
Pedigo LM,
Daub R,
Broxmeyer HE:
Cloning and characterization of the murine promoter for the colony-stimulating factor-1 encoding gene.
Gene
102:165,
1991[Medline]
[Order article via Infotrieve]
30.
Miyatake S,
Otsuka T,
Yokota T,
Lee F,
Arai K:
Structure of the chromosomal gene for granulocyte-macrophage colony-stimulating factor: Comparison of the mouse and human genes.
EMBO J
4:2561,
1985[Medline]
[Order article via Infotrieve]
31.
Tokunaga K,
Taniguchi H,
Yoda K,
Shimizu M,
Sakiyamo S:
Nucleotide sequence of a full-length cDNA for mouse cytoskeletal beta-actin mRNA.
Nucleic Acids Res
14:2829,
1986[Free Full Text]
32.
Lomedico PT,
Gubler U,
Hellmann CP,
Dukovich M,
Giri JG,
Pan YCE,
Collier K,
Semionow R,
Chua AO,
Mizel SB:
Cloning and expression of murine interleukin-1 cDNA in Escherichia coli.
Nature
312:458,
1984[Medline]
[Order article via Infotrieve]
33.
Gray PW,
Glaister D,
Chen E,
Goeddel DV,
Pennica D:
Two interleukin 1 genes in the mouse: Cloning and expression of the cDNA for murine interleukin 1-beta.
J Immunol
137:3644,
1986[Abstract]
34.
Yokota T,
Arai N,
Lee F,
Rennick D,
Mosmann T,
Arai KI:
Use of a cDNA expression vector for isolation of mouse interleukin 2 cDNA clones: Expression of T-cell growth-factor activity after transfection of monkey cells.
Proc Natl Acad Sci USA
82:68,
1985[Abstract/Free Full Text]
35.
Fung MC,
Hapel AJ,
Ymer S,
Cohen DR,
Johnson RM,
Campbell HD,
Young IG:
Molecular cloning of cDNA for murine interleukin-3.
Nature
307:233,
1984[Medline]
[Order article via Infotrieve]
36.
Sideras P,
Bergstedt-Lindquist S,
Severnson E,
Noma Y,
Naito T,
Azuma C,
Tanabe T,
Kinashi T,
Matsude F,
Yaoita Y,
Honjo T:
IgG1 induction factor: A single molecular entry with multiple biological functions.
Adv Exp Med Biol
213:227,
1987[Medline]
[Order article via Infotrieve]
37.
Van Snick J,
Cayphas S,
Szikora JP,
Renauld JC,
Roost EV,
Boon T,
Simpson RJ:
cDNA cloning of murine interleukin-HP1: Homology with human interleukin 6.
Eur J Immunol
18:193,
1988[Medline]
[Order article via Infotrieve]
38.
Lupton SD,
Gimpel S,
Jerzy R,
Brunton LL,
Hjerrild KA,
Cosman D,
Goodwin RG:
Characterization of the human and murine IL-7 genes.
J Immunol
144:3592,
1990[Abstract]
39.
Tekamp-Olson P,
Gallegos C,
Bauer D,
McClain J,
Sherry B,
Fabre M,
Deventer SV,
Cerami A:
Cloning and characterization of cDNAs for murine macrophage inflammatory protein 2 and its human homologues.
J Exp Med
172:911,
1990[Abstract/Free Full Text]
40.
Sims JE,
March CJ,
Cosman D,
Widmer MB,
MacDonald HR,
McMahan CJ,
Grubin CE,
Wignall JM,
Jackson JL,
Call SM,
Friend D,
Alpert AR,
Gillis S,
Urdal DL,
Dower SK:
cDNA expression cloning of the IL-1 receptor, a membrane of the immunoglobulin superfamily.
Science
241:585,
1984
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
|
Abstract
Introduction
Abstract