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1990 76: 814-819
Tuftsin induces tissue factor-like activity in human mononuclear cells
and in monocytic cell lines
A Kornberg, R Catane, S Peller, S Kaufman and M Fridkin
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Tuftsin Induces Tissue Factor-Like Activity in Human Mononuclear Cells and in
Monocytic Cell Lines
By Abraham Kornberg, Raphael Catane, Shoshana Peller, Suzana Kaufman, and Mati Fridkin
Normal human monocytes and macrophages generate potent procoagulant activity (PCA) resembling tissue factor
(TF) in response to various stimuli. In this study we show
that tuftsin, a natural stimulator of many functions of
monocytesand macrophages, also stimulates a potent PCA
in mixed mononuclear cells and monocytes, and a mild PCA
in lymphocytes and cell lines of monocytic origin (U937and
THP). No activity was generated by several lymphoid cell
lines and HL-60 cells. The PCA resembled TF in that it
accelerated clotting through the extrinsic coagulation pathway and was inhibited by concanavalin-A and by monoclonal anti-TF antibodies. The induction of TF-like activity
by tuftsin was dose- and time-dependent. It was located in
the cell membrane and did not require T cells for expres-
sion. Generation of TF-like activity was prevented by
actinomycin D, while cytarabine had no effect on this
process, suggesting that expression of the activity depends on protein synthesis. Studies with various tuftsin
analogs suggest that tuftsin stimulates generation of TFlike activity, as well as other functions of monocytesvia the
same receptors. The results with the monocytic cell lines
show that tuftsin affects mainly mature cells. The induction
of TF-like activity in mononuclear cells by tuftsin constitutes an important link between mononuclear cells and the
immune and coagulation systems. It may play a major role
in the pathogenesis of thromboembolism and fibrin deposition in various inflammatory and immunologic disorders.
0 1990 by The American Society of Hematology.
T
In this study we show that tuftsin also stimulates the
generation of potent PCA resembling TF activity in monocytes, as well as mild PCA in lymphocytes and in cell lines of
monocyte-macrophage origin.
HE MONONUCLEAR cells of human peripheral blood
generate potent procoagulant activity (PCA) in response to a variety of stimuli, including lipopolysaccharide
(LPS),'.' l e c t i n ~ ,immunoglobulins
~
(Igs), immune complexes and aggregated I ~ s , ~the
. ' mixed lymphocyte reaction,6
complement,' tumor necrosis factor,' phorbol esters,' lymphokines, and others." The PCA resembles tissue factor
(TF). Both require factor VI1 for e ~ p r e s s i o n ~and
~ ~ ~are
'
inhibited by Con-A"112 and anti-TF a n t i b ~ d i e s . ~ .It
~ . 'is~
localized in the cell membra ne^'.''.'^ and generated mainly by
monocytes and macrophage^.'^.'^ Certain stimuli require
collaboration of T lymphocytes for optimal production of the
a ~ t i v i t y , ~ , 'while
~ ~ ' ' others are able to induce the activity
The expression of
without the assistance of these
TF-like activity may be of clinical significance in the pathogenesis of fibrin deposition in immunologic and inflammatory
disorders," as well as in disseminated intravascular coagulation and clot formation in gram-negative bacterial sepsis,2
cancer," and inflammatory bowel disease."
Tuftsin is a natural tetrapeptide that is liberated from the
parent carrier Ig by the action of two enzymes. One is in the
spleen, and the second is on the outer side of the plasma
membrane of phagocytic cells.2' It stimulates many functions
of monocytes, macrophages, and neutrophils, including
phagocytosis, pinocytosis, motility, chemotaxis, and immunologic, tumoricidal, and bactericidal activities.22
From the Department of Hematology, Assaf Harofeh Medical
Center, Zerifin; Department of Oncology and Radiation Therapy,
Hadassah University Hospital, Jerusalem; and Department of
Organic Chemistry. The Weizman Institute of Science, Rechovot,
Israel.
Submitted March 9, 1989;accepted April 6, 1990.
Address reprint request to Abraham Kornberg, MD, Department
of Hematology. University of Rochester Medical Center, 601
Elmwood Ave. Box 610, Rochester, NY 14642.
The publication costs of this article were defrayed in part by page
charge payment. This article must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C.section I734 solely to
indicate this fact.
o I990 by The American Society of Hematology.
0006-497l/90/7604-O121%3.00/0
814
MATERIALS AND METHODS
Separation and identification of cells. Blood was collected in
heparin from normal volunteers. Mononuclear cells were isolated on
lymphoprep density gradient centrifugation (Mycomed, AS, Oslo,
Norway). The cell layer was centrifuged once at 140g and twice at
IOOgin phosphate-buffered saline (PBS) at room temperature for 10
minutes to remove the platelets. Monocytes (adherent cells) were
obtained by incubation of mixed mononuclear cells for 90 minutes at
37OC in a humidified atmosphere of 5% COz in air, either in large
Petri dishes (90 x 1 5 mm, Sterilin, Fettham, Middlesex, England,
10 mL per dish of 5.0 x lo6 cells/mL) or in small dishes (50 x 15
mm, 1 mL per dish of 2.0 x lo6 cells/mL) in RPMI 1640 supplemented with L-glutamine (Biological Industries, Beth Haemek,
Israel) and 10%heat-inactivated fetal calf serum (HI-FCS) (GIBCO,
Paisley, Scotland). Lymphocytes (nonadherent cells) were removed
from the dishes by aspiration with a Pasteur pipette. The dishes were
then washed twice with warm (37OC) media to remove the nonadherent cells. In part of the experiments the adherent cells were removed
by scraping the dishes with a rubber p~liceman,'~
washed twice in
PBS, and used in the incubation studies. In other parts of the
experiments the cells were incubated with various compounds in the
small Petri dishes, after which they were scraped and used in the
PCAassays.'
Monocytes were identified by positive staining with fluorideinhibited nonspecific esterases and phagocytic capability. The percentage of phagocytic cells was determined by quantitation of the
number of cells that ingested at least four latex particles, as
described by Edwards et
The mixed mononuclear cell population comprised 15% to 30% monocytes, the adherent cell population
85% to 95% monocytes, and the nonadherent cell population
consisted of 1% to 5% monocytes.
To remove T lymphocytes, a suspension of mononuclear cells was
incubated with sheep red blood cells (SRBCs) to allow the formation
of rosettes." The cell suspension was layered on lymphoprep and
centrifuged at 140g for 20 minutes. The T-depleted mononuclear cell
population was collected from the interface. The percentage of T
lymphocytes was determined by the formation of rosettes with
S R B C S . ' ~ .The
' ~ mixed mononuclear cells and the T-depleted cells
contained 72% f 5% and 3% + 2% T lymphocytes, respectively.
Blood, Vol 76, No4 (August 15). 1990: pp 814-819
-
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TUFTSIN, TISSUE FACTOR, AND MONOCYTES
- 1
815
The cell lines Raji, HD-Mar, REH, HL-60, U 937, and THP were
260
MONONUCLEAR
maintained by subculturing every 3 to 4 days in RPMI supplemented
0 240
with L-glutamine and 10%HI-FCS in 5% CO, in air.
Incubation of cells. After separation, cells were washed twice in
220
MONOCYTES
(ADHERENT)
PBS and incubated at 3.0 x lo6 cells/mL in the presence of the
following compounds, alone or in various combinations: tuftsin,
LYMPHOCYTES
tuftsin analogs, polymyxin-B (Teva Ltd, Tel Aviv, Israel), cytaraI180
( NON-ADHERENT )
bine (Upjohn, Kalamazoo, MI), actinomycin-D (Merck Sharp,
.E 160
Rahway, NJ), and LPS (from Escherichia coli 026:B6, Sigma, St
n
Louis, MO). Initially, incubations were performed in either RPMI
E 140
supplemented with 10% HI-FCS or PBS at 37OC in 5% CO, or at
+ 12D
room temperature. Because the results were very similar, later
2 100
studies were performed only in PBS at room temperature. The
a
lymphocytes were incubated in plastic tubes (Sterilin, Fettham) at a
73
80
final volume of 1 mL/tube. Initially, the monocytes were incubated
a,
E
60
either in tubes after scraping the Petri dishes or in Petri dishes and
then obtained by scraping.' Because the results of the two methods
TI 4 0
were similar, in later experiments the monocytes were first obtained
20
by scraping and then used in the incubation studies. In all experiments cells were used only if their viability, as determined by trypan
n
blue or eosin exclusion tests, exceeded 90%.
"
Assay and characterization of procoagulant activity. After
Tuftsin (ua/ml)
incubation, cells were washed twice in PBS and resuspended in
veronal acetate buffered saline, pH 7.35.'2313*23
Procoagulant activity
Fig 1. The effect of various concentrations of tuftsin on
was determined using either intact cells or cells disrupted by
generation of procoagulant activity by mixed mononuclear cells,
sonification with ultrasonic vibration for 60 seconds in an ice bath
monocytes. and lymphocytes. PCA was assayed after 24 hours
(Sonifier B-12, Branson Sonic Power, Danburg, CT). To obtain the
incubation of 3 x l d / m L cells with different concentrations of
membrane-rich fraction, nuclei were sedimented at 400g and the
tuftsin. Modified-PT 31 veronal acetate buffer, RPMI 1640. or tuftsupernatant was centrifuged at 40,OOOg for 20 minutes at 2OoC
sin + RPMI 1640 in the absence of cells exceeded 150 seconds.
(Sorval, RC-5, DuPont, Newtown, CT)." Procoagulant activity was
The results are the mean f SD of 6 to 8 experiments.
measured by the modified prothrombin time (M-PT)3~6~8~'2~'4~'5~'9~20~23:
0.1 mL of pooled citrated plasma from at least 10 normal donors was
incubated with 0.1 mL cell suspension at 37OC for 1 minute, 0.1 mL
at 500 pg/mL tuftsin. The lymphocyte PCA was not affected
0.025 mol/L CaCl, was then added, and the clotting time recorded.
by the increasing concentrations of tuftsin. PCA was not
Each sample was run in duplicate. In some experiments PCA was
increased in either the untreated cells or in the controls with
assayed using plasma deficient in factors VII, VIII, IX (Pacific
tuftsin without cells during the same periods of incubation
Hemostasis, Ventura, CA), X (Diagnostica Stago, Ashieres-Sur(M-PT 142 to 156 seconds).
Seine, France), or XI (Merz-Dade, Dudingen, Switzerland).
PCA did not change during the first 6 hours of exposure to
The effect of monoclonal anti-TF antibodies (provided by Dr S.D.
a constant concentration of tuftsin (250 pg/mL) (Fig 2). The
Carson, University of Nebraska Medical Center, Omaha) on monoactivity increased significantly with M M C and monocytes to
nuclear PCA was studied by determination of M-PT of tuftsintreated mononuclear cells after incubation with monoclonal anti-TF
a maximal level a t 16 hours (P< .001), thereafter remaining
antibodies for 1 to 2 hours at 37°C.24
at the same plateau for up to 36 hours of incubation.
Reagents. Tuftsin was synthesized and provided by Abic Ltd
Lymphocytes generated significantly less PCA during the
(Ramat Gan,
The tuftsin analogs (Leu')-tuftsin, nitrosame incubation period (P < .001).
tuftsin, and (Des-Thr')-tuftsin were synthesized according to FridArtifacts due to LPS contamination of various reagents
kin et al.22*25
The compounds were dissolved in PBS, pH 7.4.
and solutions2 were excluded by incubating M M C and
Activated T F was purchased from Dade Diagnostica (Agnada,
tuftsin with polymyxin B. Although this substance binds to
Puerto Rico) and Concanavalin-A (Con-A) from Miles-Yeda (Rehothe
lipid A region of LPS and inhibits its ability to induce
vot, Israel). Statistical analysis was performed using the paired
monocyte PCA generati~n,~.'
elaboration of PCA was not
student's t-test.
f4L4;
U
0 0.05 015 5 50 100 250 500
RESULTS
Effect of tuftsin on procoagulant activity generation by
mixed mononuclear cells, lymphocytes, and monocytes.
Tuftsin in concentrations less than 5 pg/mL did not enhance
PCA generation, either in M M C or in purified cells (Fig 1).
Both monocytes and lymphocytes showed a significant increase in PCA expression after incubation with 5 NgfmL
tuftsin (P< .001; P = .007, respectively). However, the
effect on monocytes was significantly more prominent
(P< .001), increased progressively, and reached peak levels
inhibited by the addition of polymyxin B to the cultures of
M M C and tuftsin (M-PT without polymyxin B was ?O 12
seconds, and 62 * 8 seconds with polymyxin B).
Characterization and localization of procoagulant activity generated by mixed mononuclear cells. As tuftsin
proved to induce a similar increase in the levels of PCA in
M M C and monocytes, only M M C were used in further
experiments. Both PCA and commercial TF accelerated the
M-PT of r?wnal plasma and plasma deficient in factors VIII,
IX, and Xi, and had no effect on the M-PT of plasma
deficient in factors VI1 or X (Table 1).
Incubation of Con-A with tuftsin-treated mononuclear
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816
v
200
220
180
--
1
-
.-
Y
MIXED MONONUCLEAR
CELLS
140
"
6
I
.
'
Sonicated-tuftsin
Culture media
Whole
Supernatant
Pellet
Pellet
-
Modified PT (4
69 f
152
60*
146 f
164*
10
* 14
6
12
15
Mononuclear cells were incubated with 250 pg/mL tuftsin for 24
hours. The cells were disrupted and the supernatant and pellet were
obtained after centrifugation at 40,OOOg as described in Materials and
Methods. Results are mean i SD of 3 to 5 experiments.
'The procoagulant activity was assayed after 24 hours of incubation of
the pellet of the disrupted cells with 250 pg/mL tuftsin.
100
0
Cell Fraction
Intact
Sonicated
J--+-----r
0
Table 2. Generation of Procoagulant Activity by Various
Fractions of Tuftsin-Treated Mononuclear Cells
Cell
nonocvrES
(AMRENT 1
LYMPHOCYTES
( NOW-ADHERENT)
.Ic
-
KORNBERG ET AL
"
" "
12 18 24 30
Incubation Time (hrs)
""-
36
Fig 2. Time course of generation of procoagulant activity by
tuftsin-treated mixed mononuclear cells, monocytes, and lymphocytes. Cells, 3 x l d / m L , were cultured with 250pg/mL tuftsin fw
various periods of time. The results are the mean i SD of 6 t o 8
experiments.
cells for 1 hour resulted in dose-related inhibition of PCA
(Table l ) , suggesting that Con-A preferentially binds to
monosyl or glycosyl residues.l',l2 The PCA was reduced
significantly by incubation of tuftsin-treated MMC for 1 to 2
hours with 10 and 40 pg/mL monoclonal anti-TF antibodies
(Table 1).
Experiments with various cell fractions (Table 2) demonstrate that the TF-like activity of the membrane-rich pellet
was the same as that of the intact cells. Low activity was
found in the supernatant mainly containing cytosol and in the
culture media or PBS. No activity was generated on incubation of disrupted MMC with tuftsin.
Effect of T lymphocytes and DNA and protein synthesis
inhibitors on the generation of TF-like activity by mixed
mononuclear cells. Depletion of T lymphocytes from
72% * 5% to 3% * 2% did not prevent the generation of
TF-like activity by MMC exposed to tuftsin for 24 hours
(Table 3). Incubation of MMC with tuftsin in the presence of
10 pgjmL actinomycin-D reduced the generation of TF-like
activity by 93%,while 5 pg/mL cytarabine did not inhibit its
expression.
Effect of tuftsin analogs on the generation of procoagu[ant activity by mixed mononuclear cells. The effect of
various tuftsin analogs on the generation of the TF-like
activity by MMC is compared with that of tuftsin in Table 4.
(Leu')-tuftsin, which stimulates phagocytosis to the same
extent as t u f t ~ i n , induced
~ ~ . ~ ~slightly less activity at concentrations of 5 pg/mL and 250 pg/mL. Nitro-tuftsin, a tuftsin
analog that does not affect phagocyt~sis,~~
failed to induce
generation of TF-like activity. (Des-Thrl)-tuftsin, which has
been shown to exert an inhibitory effect on tuftsin-stimulated
phagocyt~sis,~~
also inhibits the generation of TF-like activity by tuftsin.
Table 1. Characterization of Tuftsin-Induced Mononuclear Procoagulant Activity
Inhibitor
Modified PT
Substrate
Plasma
(pg/mL)
(5)
Thromboplastin (dilution 1: 16)
Noma1
Normal
VI1 deficient
X deficient
Vlll deficient
IX deficient
XI deficient
Normal
-
Mononuclear cells
Mononuclear cells
Mononuclear cells
Mononuclear cells
Mononuclear cells
Mononuclear cells
Mononuclear cells
Mononuclear cells
Normal
-
Con-A
16
64
128
Monoclonal anti-TF antibodies
2
10
40
53t
69 f
143 k
162 t
60f
68t
64 i
4
10
14
16
12
8
10
102 f 8
116 f 10
132 i 10
55* 4
1182 8
136 i 6
Generation of mononuclear procoagulant activity was assayed after incubation of 3 x 106/mLcells with 250 pg/mL tuftsin for 24 hours. The effect of
Con-A and monoclonal anti-TF antibodies was determined after 1 to 2 hours of incubation with the tuftsin-treated cells at 37°C.The results are the
mean k SD of 3 to 5 experiments.
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817
TUFTSIN, TISSUE FACTOR, AND MONOCYTES
Table 6. Generation of Procoagulant Activity by Tuftsin-Treated
Cell Lines
Table 3. Effect of D N A and Protein Synthesis Inhibitors and
Depletion of T Lymphocytes on Generation of Procoagulant
Activity by Tuftsin-Treated Mononuclear Cells
~~
Modhd PT (s)
~
Calls
Cytarabine
(pglmL1
Mononuclear
-
Mononuclear
Mononuclear
T-cell-depleted mononuclear
-
5
Actinomycin D
(pg/mL)
10
-
Modified PT
(5)
69 +
64+
134 f
702
Cell Line
5 pg/mL Tuftsin
250 pglmL Tuftsin
120 + 10
172 i 18
>200
>200
128i 9
125 f 8
170 + 15
188 + 22
2180
83 + 10
86+ 6
~~
10
8
12
6
Generation of mononuclear procoagulant activity was assayed after
incubation of 3 x 106/mL cell in the presence of 250 pg/mL tuftsin with
and without the inhibitors and T lymphocytes for 24 hours. The results
are mean + SD of 4 to 6 experiments.
Eflect of tuftsin on generation of procoagulant activity by
cell lines. Tuftsin in various concentrations had no effect
on the generation of TF-like activity by HL-60 cells and by
the lymphoid cell lines Raji (B type), HD-Mar (T type), and
REH (pre-B) (Table 5). It induced a modest but significant
increase in the expression of the activity in the cell line of
monocyte-macrophage origin U937 and T H P a t concentrations of 5 pg/mL and 250 pg/mL (P = .001).
Mononuclear cells
Raji ( 6 )
Hd-Mar (TI
REH (pre-8)
u 937
THP
69+ 8
Generation of procoagulant activity by cell lines was assayed after
incubation of 3 x 106/mL cells with tuftsin for 24 hours. The cell lines
were defined according to Drexler et ai.'' The results are mean + SD of 3
to 4 experiments.
different effects of actinomycin D and cytarabine on PCA
generation.
Tuftsin-induced TF-like activity exhibits several unique
properties. First, the main inducers of the activity in monocytes such as LPS, lectins, and immune complexes all require
collaboration of T lymphocytes for optimal e f f e ~ t , ~ - ' ~ - ' ~
whereas the action of tuftsin was not prevented by the
removal of T cells. However, it has been claimed that there
DISCUSSION
are two pathways for the generation of TF-like activity, one
Stimulated monocytes generate PCA after exposure to a
of them being relatively T-cell independent.I2." Second, the
wide range of inducers in vitro,'.'' while various monocytic
activity appears relatively late in tuftsin-treated M M C and
functions are activated by tuftsin.21*22Therefore, it is not
monocytes compared with its expression by other i n d ~ c e r s . ~ , ~ , ~
surprising that the tetrapeptide also stimulates PCA generaFor instance, MMC expressed significant increase in the
tion in monocytes. Moreover, LPS, the most potent stimulus
activity 4 hours after stimulation with 10 pg/mL LPS
for the generation of monocyte TF-like activity' as well as
(M-PT 122 * 8 and 82 * 10 seconds, respectively) that
other inducers belonging to the immune system, such as
increased after 8 hours (M-PT 64 * 8 seconds), and reached
complement, lymphokines, and immune complexes, augpeak levels after 16 hours (M-PT 44 f 6 seconds). Despite
ments different functions of monocyte^.^^^^^ It seems that
these differences, the results with the monoclonal anti-TF
there is a general link between induction of monocyte TF-like
antibodies prove conclusively that tuftsin-induced PCA resemactivity and activation of a number of functions of monobles TF. Finally, most of the studies attribute the bulk of
cytes.
TF-like activity induced by various stimuli to monocytes and
The nature of PCA and the process of its induction by
macrophage^.'^^'^ In that respect it was found that purified
tuftsin resembles, in several aspects, the PCA generated in
lymphocytes bind less than 5% tritiated tuftsin, as compared
response to other stimuli: (1) Properties of TF, as defined by
with polymophonuclears and monocytes." In light of these
its dependence on factors VI1 and X for e x p r e s s i ~ n ~ ~ ~findings,
~ ~ ~ ' ~the
~ ~origin
~ ; of the activity in the preparations of
binding to C O ~ - A " , 'and
~ ~ inhibition
~~;
by monoclonal anti-TF
tuftsin-treated purified lymphocytes is unclear. The possibila n t i b o d i e ~ . ~(2)
~ ~Localization
~ ' ~ . ~ ~ to the cell membranes as
ity of contamination with minute amounts of monocytes in
demonstrated by the potent PCA of intact cells and of the
our studies cannot be ruled 0 ~ t . INevertheless,
~
it was
sediment containing the membrane-rich fraction of the
demonstrated by cytologic assay that lymphocyte-like cells
disrupted cell^.^.'^*'^^'^ (3) Requirement of R N A and protein
can express TF-like activity after stimulation with immune
synthesis for full expression of PCA, as indicated by the
c~mplexes,~
and that pure lymphocytes can bind '251-tuftsinyltyrosine," indicating the existence of a subpopulation of
Table 4. Effect of Tuftsin Analogs on Mononuclear Procoagulant
lymphocytes that possess tuftsin's receptor site, and, thereActivity Generation
fore, can be ~timu1ated.I~
Modified PT ( 5 )
The dose-response relationship between the concentrations
Analog
5 pglmL
250 pg/mL
of tuftsin and the generation of TF-like activity is different
from its effect on other functions of monocytes. A significant
Tuftsin
Thr-Lys-Pro-Arg
118 + 8
58 i 5
increase in the activity was noted with 5 pg/mL, which
(Leu')-tuftsin
Leulys-Pro-Arg
125 + 1 1
72 2 8
reached peak levels in the presence of 500 pg/mL. In
Nitro-tuftsin
Lev-Lys-ProrArg-Nitro
154 + 14 162 + 12
Tuftsin +
contrast, the in vitro effects of tuftsin on phagocytosis,
(Dis-Thr'l-tuftsin (Des-Thr'I-Lys-Pro-Arg
147 2 9 124 -f: 10
chemotaxis, migration, and cytotoxicity of monocytes were
shown to be evident already at concentrations as low as
Generation of mononuclear procoagulant activity was assayed after
to
pg/mL, becoming maximal at 1 to 10 pg/mL and
incubation of 3 x 106/mL cells with tuftsin analogs for 24 hours. The
results are mean + SD of 3 to 4 experiments.
decreasing at higher concentration^.^^^^^ Tuftsin concentra'v9.
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818
KORNBERG ET AL
tions in the blood range between 0.2 and 0.5 pg/mL,” which
poses the question regarding the clinical significance of our
studies. However, it is plausible that during inflammatory or
immunologic disorders the concentration of tuftsin in the
blood, and especially in tissue, is increased and reaches levels
that induced in vitro the generation of TF-like activity.
Moreover, tuftsin was degraded during incubation to free
amino acids as well as peptidic fragments as shown by amino
acid analysis and by high pressure liquid chromatography
(HPLC), respectively. Degradation was usually maximal
after 16 hours and amounted to 30% to 95% of the peptide. It
is also known that in consequence to its association with
specific receptor sites on phagocytic cells, tuftsin is internalized by the cells and degraded by a battery of various
peptidases.22The results of the experiments with the various
tuftsin analogs suggest that tuftsin induces TF-like activity
in monocytes, as well as affecting other functions of the cells
via the same receptor sites. Because tuftsin is part of the CH,
domain of the Fc-fragment,” and since immune complexes
and aggregated Igs induce the activity in monocyte^,^.^ it is
not unlikely that the tuftsin binding sites on monocytes are
related in some way to the Fc-receptor.
Our results with the monocyte-macrophage cell lines are in
agreement with other studies on the response of such cell
lines, including PU 5, WEHI 265, TG-PEG, and 5774, to
various stimuli.” The lack of response of the tuftsin-treated
B, T and pre-B type cell lines may support the conclusion that
the monocyte-macrophage is the principal cell responsible
for the generation of TF-like activity.
Tuftsin, a natural tetrapeptide of the immune system,
plays a significant role in the defense of the body and in
inflammatory and immunologic p h e n ~ m e n a . ~TF-like
’ , ~ ~ activity induced by tuftsin in monocytes is an additional
example of the link between the immune and coagulation
systems, which may have a significant importance in the
pathogenesis of fibrin deposition in inflammatory and immunologic disorders, and in activation of coagulation during
septicemia and malignancy.
ACKNOWLEDGMENT
We thank Drs H. Ben-Bassat, E. Fibach, and A. Treves for
providing the cell lines, and Dr S.D. Carson for providing the
monoclonal anti-tissue factor antibodies.
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