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Female hormones reduce neutrophil responsiveness in vitro.

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623
FEMALE HORMONES REDUCE NEUTROPHIL
RESPONSIVENESS IN VITRO
JILL P. BUYON, HELEN M. KORCHAK, LORENE E. RUTHERFORD,
M. GANGULY, and GERALD WEISSMANN
Neutrophils were preincubated with 17 betaestradiol and progesterone to determine the effects of
these hormones on chemotactic peptidestimulated superoxide anion (02-)
generation and degranulation. At
pharmacologic levels 17 beta-estradiol was more active
than progesterone with respect to inhibition of 02generation as well as degranulation. An increase of
preincubation time from 5 minutes to 25 minutes increased the percent inhibition. When 17 beta+stradiol
and progesterone were combined at levels which approximate those measured during gestation, there was
small but significant inhibition of 02-generation. Dexamethasone at equal molar concentration inhibited 02generation only after 25 minutes of preincubation and at
no time reached the level of inhibition attained by either
of the sex hormones alone. Both estradiol and progesterone at pharmacologic levels significantly inhibited betaglucuronidase and lysozyme release, whereas dexamethasone did not inhibit degranulation despite prolonged
preincubation. Neutrophils isolated from women during
various phases of the menstrual cycle and during the
-
From the Division of Rheumatology, Department of Medicine, New York University Medical Center, New York, New York.
Supported by grants from the National Institutes of Health
(AM1 1949, HL19721, AM07176, and A118676).
Jill P. Buyon, MD: Division of Rheumatology, Department
of Medicine; Helen M. Korchak, PhD: Division of Rheumatology,
Department of Medicine; Lorene E. Rutherford, BA: Division of
Rheumatology, Department of Medicine; M. Ganguly, PhD: Department of Obstetrics and Gynecology; Gerald Weissmann, MD:
Division of Rheumatology, Department of Medicine, New York
University Medical Center.
Address reprint requests to Jill Buyon, MD, c/o Gerald
Weissmann, MD, Department of Medicine, 550 First Avenue, New
York, NY 10016.
Submitted for publication July 13, 1983: accepted in revised
form January 27, 1984.
Arthritis and Rheumatism, Vol. 27, No. 6 (June 1984)
third trimester of pregnancy did not differ with respect
to chemotactic peptide-stimulated 02- generation.
These data suggest that inhibition of neutrophil responses requires the continuous presence of pharmacologic
levels of estradiol and progesterone.
More women than men suffer from rheumatoid
arthritis (RA) and systemic lupus erythematosus
(SLE). There is an increased incidence of both diseases in women of childbearing age. Generally, clinical
remissions of RA occur during pregnancy (1-4). The
effects of gestation on the clinical activity of SLE are
more variable. However, exacerbations often occur
during the postpartum period (43). The factors responsible for these observations are unknown. Roubinian et a1 (6) have demonstrated the beneficial and
adverse effects of sex hormones on the immunologic
abnormalities of murine lupus. Lahita et al (7) have
shown that estrogen metabolism is altered in SLE,
with increased production of active metabolites in
patients and first-degree relatives.
The neutrophil is an important effector cell in
both SLE and RA. In SLE the neutrophil has been
demonstrated to be impaired with respect to phagocytic and chemotactic activity, and these defects may
play a role in impaired host defenses (8-10). In RA
release by the neutrophil of proteolytic enzymes,
oxidation products of arachidonate and reactive oxygen species, may play a role in inflammatory synovitis.
Indeed, inhibitors of neutrophil responsiveness might
contribute both to the increased susceptibility to infection found in SLE ( I 1) and the decrease in inflammatory synovitis seen in pregnant RA patients. We
therefore investigated the effects of 17 beta-estradiol
(estradiol) and progesterone on two parameters of
624
BUYON ET AL
neutrophil function: superoxide anion (02-)generation and degranulation.
MATERIALS AND METHODS
Preparation of cell suspensions. Heparinized blood
was obtained fram normal female volunteers. Purified preparations of neutrophils were isolated using Hypaque/Ficoll
gradient (12) followed by dextran sedimentation and hypotonic lysis of erythrocytes (13). This procedure allowed
studies of cell suspensions containing 98 ? 2% neutrophils
with few contaminating erythrocytes or platelets. The cells
were suspended in a buffered salt solution consisting of Na’
(150mM), K ’ (5 mM), Ca’k (1.3 mM), Mg2‘ (1.2 mM), C1(155 mM), and HEPES (10 mM), pH 7.45. Release of
cytoplasmic lactate dehydrogenase (LDH) was used as an
indicator of cell death (14). Rupture of cells by detergent
(triton X-100) led to release of total LDH, beta-glucuronidase, and lysozyme (14).
Superoxide anion generation. Superoxide anion generation was monitored by determination of superoxide dismutase-inhibitable reduction of cytochrome C in the presence of cytochalasin B. Duplicate reaction mixtures
containing 1.25-1.5 x lo6 neutrophils, 75 nmoles horse heart
ferricytochrome C (type III), and the various steroids at
designated concentrations in a final volume of 1 ml were
preincubated for 0-25 minutes at 37°C. Cytochalasin B ( 5 pg/
ml) was added and cells were incubated at 37°C for an
additional 5 minutes before exposure to stimuli (e.g.,
FMLP). Five minutes after stimulation, cells were spun
down at 4°C at 1,000g in a Sorvall RC-3 centrifuge and the
supernatants were collected. Absorption at 551.2 nm was
determined in a Beckman model 25 spectrophotometer and
the nmoles of superoxide anion generated per lo6 cells were
calculated as previously described (15). Results of steroid
incubations are expressed as the percent of the FMLPstimulated response (assumed to be 100%).
Lysasomal enzyme release (degranulation). For the
determination of enzyme release, neutrophils (3.5-5 x lo6)
were preincubated with the various steroids, after which
cytochalasin B (5 pg/ml) was added and the mixture incubated for an additional 5 minutes. Appropriate stimulus was
added and after a 5-minute incubation at 3 7 T , the cell
suspensions were centrifuged at 1,000g in a Sorvall RC-3
centrifuge for 5 minutes at 4°C. Aliquots of the supernatants
were taken for standard determinations of beta-glucuronidase (16) and lysozyme (17). Total enzyme activity and
residual cell-associated activity were measured in selected
reaction mixtures after cells were lysed by the addition of
0.2% triton X-100. Appropriate control experiments were
performed to determine whether there was preferential loss
of enzyme activity in resting or treated cells or whether test
reagents interfered with enzyme assays. Results are expressed as the percent of maximally stimulated enzyme
release from control cell suspensions. All experiments were
done in duplicate.
Xanthine oxidase-hypoxanthhe generation of superoxide anion. Duplicate tubes containing a I-ml solution of
xanthine oxidase ( I : 100 dilution), hypoxanthine ( 2 mg/100
ml) (100 nmoles), and ferrieytochrome C (75 nmoles) were
prepared with one of the following: dexamethasone 4 pg/ml,
estradiol 3 pg/ml, or progesterone 3 pg/ml. Tubes were
incubated at 37°C for 5 minutes. Superoxide generation was
monitored as above and expressed as nmoles cytochrome C
reduced.
FMLP binding. Specific binding of FMLP to the
plasma membrane of the neutrophil was determined using
the silicone oil centrifugation assay as described by Mackin
et al (18). Neutrophils (5-10 x lo6) were preincubated with
the sex hormones in the presence of cytochalasin B (5 pg/ml)
at designated concentrations in a final volume of 100 pI for
0-25 minutes at 37°C. Nonspecific binding was determined
in the presence of excess unlabeled FMLP (10-4M). ’HFMLP (10 7M)was then added and the cells were incubated
at 37°C for an additional 5 minutes. An aliquot was then
layered over 150 pl versilube F50 silicone fluid in a 400-pI
plastic microcentrifuge tube, and spun down for 30 seconds
in a Beckman Model B microcentrifuge. Tube tips were
excised and pellets were resuspended in 10 ml of scintillation
fluid.
The cells were counted for 10 minutes in a Beckman
Model LS 7000. Specific binding was calculated as the
difference between total binding with ’H-FMLP ( w ~ M )and
nonspecific binding with ’H-FMLP (lO-’M) in the presence
of excess unlabeled FMLP (10-4M). Results of steroid
incubations are erpressed as the percent of specific binding
with FMLP alone (assumed to be 100%). All experiments
were done in duplicate.
Materials. Cytochalasin B was purchased from Aldrich Chemical Co. (Milwaukee, WI). Progesterone, 17
beta-cstradiol, dexamethasone, cytochrome C (type III),
FMLP, xanthine oxidase (grade I11 from buttermilk), Micrococcus lysodeikricus, and phenolphthaleine glucuronic acid
were obtained from Sigma Chemical Co. (St. Louis, MO).
’H-FMLP was purchased from New England Nuclear (Boston, MA). Superoxide dismutase was obtained from Miles
Laboratories (Elkhart, IN). Phorbol myristate acetate was
from Chemalog (South Plainfield, NJ). Lactate dehydrogenase “Statzyme” kit was obtained from Worthington Biochemical Corp. (Freehold, NJ). “Pantex immuno-direct estradiol 1251” kit was from Bio-Analysis. Inc. (Santa Monica,
CA). “Coat-A-Count Progesterone” kit was obtained from
Diagnostic Products Corp. (Los Angeles, CA).
Dexamethasone, progesterone, and estradiol were
initially dissolved in 100% ethanol. Serial dilutions were
made in HEPES buffer. Final ethanol concentration did not
exceed 0. I%. Measurement of serum estradiol and progesterone was performed by radioimmunoassay according to
the method of Abraham (19).
Statistical analysis. Statistical analysis was carried
out using Student’s t-test.
RESULTS
Effects of estradiol and progesterone on superoxide anion generation. Initial studies were carried out to
determine whether estradiol altered the generation of
0 2 - by cytochalasin B-treated neutrophils. FMLP at
10-7M was used as the stimulus. Cells were preincubated with the hormone for 5 minutes. A dose-response curve was generated which revealed that estradiol at 3 pg/ml significantly inhibited 03-production
625
FEMALE SEX HORMONES AND NEUTROPHILS
by 38% ( P < 0.001). At 1.5 pdml 02-generation was
inhibited by 32% (P < 0.01). There was no effect on
02-generation at levels ranging from 0.3 pg/ml to
0.0003 pg/ml (Figure 1). When progesterone at 3 pg/ml
was preincubated for 5 minutes with the cells, 02generation was inhibited by 17% ( P < 0.001). No
effects were observed at lower concentrations. When
estradiol and progesterone were used in combination
(0.3 pglml each, values 1 log order lower than in the
previous experiments), there was 15% inhibition of
02-generation (P < 0.001) (Table 1).
Effects of increased preincubation with estradiol
and progesterone on superoxide anion generation. Nelson et a1 (20) have suggested that when glucocorticoids
are incubated with neutrophils, a longer time of preincubation enhances inhibition of neutrophil responsiveness. The increase of preincubation did not result in
cell death (20). We have investigated different times of
preincubation and found a positive correlation between length of preincubation and percent inhibition of
Table 1. Effects of 17 beta-estradiol and progesterone on superoxide anion generation by 10 -7MFMLP-stimulated neutrophils
after 5 minutes meincubation
Hormone (n)
17 beta-estradiol 3 &ml (16)
17 beta-estradiol 1.5 &ml (3)
Progesterone 3 pghl (14)
17 beta-estradiolO.3 &ml
L progesterone 0.3 &ml
(6)
Inhibi!ion*
38 2 3
32 2 3
17 2 2
15 2 2
Pt
co.001
<0.01
<0.001
<O.OOl
* Expressed as % inhibition of control which was equal to an
average of 31 nrnoles cytochrome C reduced/106 cells. Values
represent mean 2 SE.
t P values represent significance of variation from control.
generation. Estradiol inhibited 02-generation by
38% after 5 minutes and 46% after 25 minutes ( P <
0.001). Progesterone inhibited 02-generation by 17%
after 5 minutes, compared with 44% after 25 minutes
of preincubation ( P < 0.007) (Figure 2). When estradi02-
Z 40
.c
2
W
3
c
W
W
.‘0 30
c
4
a,
’0
.x
2
x 20
=I
v)
(6)
*
T
20
Lc
0
c
.-0
-2 l o
1
-
10.
r
c
c
H
s
17-P Estradiol
( 3pg/rnl)
[17- p Est r ad iol Ip,,
,,
Figure 1. Dose-response curve of the effect of 17 beta-estradiol on
superoxide anion generation by FMLP
M)-stimulated neutrophils with 5 minutes preincubation. FMLP-stimulated response is
used as the control response (0% inhibition) which was equal to an
average of 31 nmoles cytochrome C reduced/106 cells. Vertical bars
indicate *SE from the mean.
Progesterone
( 3pg /rnl)
Dexornethosone
(4pg/rnl)
Figure 2. Effect of 17 beta-estradiol, progesterone. and dexamcthasone on superoxide anion generation by FMLP ( lO~’M)-stimulated
neutrophils after 5 minutes and 25 minutes preincubation. FMLPstimulated response is used as the control response (0% inhibition).
At S minutes preincubation, this was equal to an average of 31
nrnoles cytochrome C reduced; at 25 minutes preincubation. it was
equal to an average of I5 nmoles cytochrome C reduced. * = P <
0.001 versus control; ** = P < 0.007 versus control; NS = not
significant. Vertical bars indicate zSE from the mean; n values in
parentheses.
626
01 and Progesterone were used in combination at 0.3
pdml each, the inhibition of 02-remained at 15%. At
30 minutes LDH release was <5%, indicating cell
viability. The suppressive effect of estradiol on O2
generation was rapid, whereas that of progesterone
was more time-dependent.
Dexamethasone (4 pg/ml), a known in vivo
antiinflammatory agent, did not inhibit 02-generation
after 5 minutes of preincubation. After 25 minutes of
preincubation, dexamethasone inhibited 02-generation by 32% ( P < 0.001). Inhibition of neutrophil
function by dexamethasone was never as great as that
exerted by the sex hormones, in numerous side-byside experiments. Control experiments were done with
the cells preincubated in 0.1% ethanol alone. There
was no significant inhibition of 02-generation with
preincubation for 5-25 minutes.
To assure that these hormones were not acting
as scavengers of 02-,we used a cell-free system to
generate O2 by the enzymatic conversion of hypoxanthine to xanthine. The addition of estradiol, progesterone, or dexamethasone at the highest concentrations used in our initial experiments did not suppress
02-generation. As expected, superoxide dismutase
inhibited 02-generation.
Effects of estradiol and progesterone on release of
beta-glucuronidase. We next investigated the effect of
these steroids on neutrophil degranulation using betaglucuronidase release as a marker of the azurophil
granule contents. There was no significant inhibition
by estradiol at 3 pg/ml, progesterone at 3 pg/ml, or
dexamethasone at 4 pg/ml when they were preincubated with the cells for 5 minutes. When preincubation
was extended to 25 minutes, estradiol at 3 pg/ml
significantly inhibited beta-glucuronidase release by
30% ( P < 0.01), whereas progesterone inhibited betaglucuronidase by 15% ( P < 0.05). Dexamethasone had
no effect on enzyme release (Figure 3). When progesterone and estradiol were combined at 0.3 pg/ml each,
there was no inhibition of enzyme release with preincubation up to 25 minutes. These findings contrast
with inhibition of 02- generation attained with the
same combined concentration of the sex hormones.
Effects of estradiol and progesterone on release of
lysozyme. Lysozyme release was used as a marker of
the specific and azurophil granule contents. Estradiol
at 3 pg/ml inhibited lysozyme release by 24% after 5
minutes of preincubation ( P < 0.04). Progesterone at a
similar concentration also inhibited lysozyme release
by 24% after 5 minutes of preincubation ( P < 0.02).
After 25 minutes of preincubation, estradiol did not
BUYON ET AL
(4)
*
T
17-P Estradiol
(3pg / m l )
7
**
p vs Control c0.05
Progesterone
( 3pg / m 1 )
,
Dexomethasone
(4pg/ml)
Figure 3. Effect of 17 beta-estradiol, progesterone, and dexamethasone on beta-glucuronidase release by FMLP (10~7M)-stimulated
neutrophils after 25 minutes preincubation. FMLP-stimulated response is used as the control response (0% inhibition) which was
equal to an average of 20% release of enIyrne. Vertical bars indicate
+_SEfrom the mean; n values in parentheses.
significantly inhibit lysozyme release; marked variability was noted among donors. However, progesterone
consistently inhibited lysozyme release, reaching 40%
after 25 minutes preincubation ( P < 0.003). Dexamethasone did not significantly inhibit lysozyme release
after 5 minutes preincubation nor after 25 minutes
preincubation.
Effects of estradiol and progesterone on binding
of FMLP. These experiments were performed to determine the influence of sex hormones on binding of
FMLP to the plasma membrane of the neutrophil.
Despite the large variation in specific binding of individual donors, in the presence of estradiol at 3 pg/ml
there was a trend toward increased specific binding of
FMLP after both 5 minutes and 25 minutes of preincubation. There was no significant difference in the
specific binding of FMLP in the presence of progesterone at 3 pg/ml after 5 or 25 minutes af preincubation.
Likewise, when estradiol and progesterone were combined at 0.3 pdml each, there was no significant
difference in specific binding. The combination of
progesterone and estradiol was only studied after 5
minutes of preincubation, in keeping with previous
experiments, in which there was no augmentation of
627
FEMALE SEX HORMONES AND NEUTROPHILS
inhibition of 02- generation with longer times of
preincubation (Table 2).
Relationship between hormone levels and superoxide anion generation. To determine whether different levels of circulating hormones in vivo would affect
02-generation by neutrophils, stimulated neutrophils
were examined. These were obtained from women
during various phases in their menstrual cycle and
women in their last trimester of pregnancy.
Nine premenopausal women were studied once
per week for 4 consecutive weeks. Cells were isolated
as in previous experiments with no addition of serum.
Levels of estradiol and progesterone were determined
at the time of each experiment. When serum estradiol
levels were plotted with respect to FMLP-stimulated
02- generation, no correlation was found between
ability of neutrophils to generate O2 and circulating
levels of estradiol (r = 0.335) (Figure 4). There was
also no significant correlation found when progesterone levels were plotted with respect to FMLP-stimulated 02-generation. Similar results were obtained
when the tumor-promoting agent, phorbol myristate
acetate, was used to stimulate O2 production.
Neutrophils were next isolated from 9 women
in their third trimester of pregnancy. There was no
correlation between the ability of their neutrophils to
generate 0 2 - and circulating hormone levels. The
levels measured for estradiol ranged from 14-30 ng/ml
(mean 22 k 2 SE). The levels measured for progesterone ranged from 63-400 ng/ml (mean 230 2 40 SE).
The estradiol levels were I log order lower than those
used in initial experiments in which the combination of
estrddiol and progesterone inhibited neutrophil 02generation. The highest progesterone levels were
equal to the concentrations used in those experiments.
However, despite the high circulating levels of hormones measured in these volunteers during the third
trimester of pregnancy, the quantity of 02-generated
rn
0
O
r :0.335
A
A
0
VO
V JT
UT
1
~
Table 2.
Effect of 17 beta-estradiol and progesterone on binding of
lO-’M FMLP to the plasma membrane
Preincubation time*
Hormone (n)
5 minutes
25 minutes
17 beta-estradiol 3 pg/ml (4)
Progesterone 3 pg/ml (4)
17 beta-estradiol 0.3 pg/ml
+ progesterone 0.3 pg/ml (4)
158 -t 21
102 2 29
132 ? 21
165 t 19
237 ? 74“
-
ND
* Expressed as % of specific binding of ’H-FMLP (10- ’ M ) . Values
represent mean t SE. ND = Not done.
t Not significant ( P < 0.081), Student’s I-tailed t-test.
0
1
200 300 400 500
Estrogen Levels (pg /ml)
100
Figure 4. Relationship between the serum 17 beta-estradiol levels
of normal females during the menstrual cycle and superoxide anion
generation by their neutrophils (polymorphonuclear leukocytes.
PMN) upon stimulation by FMLP (lO-’M).
by neutrophils isolated from this environment was
equal to the quantity of 02-generated by neutrophils
(31 nmoles reduced cytochrome C) isolated from women during the menstrual cycle, when hormone levels
are 10-100 times lower.
DISCUSSION
Sex hormones influence both inflammation and
susceptibility to infection, although the exact role of
these hormones has not been elucidated. With respect
to inflammation, Mueller and Kappas (21) treated 11
RA patients with estrogens and noted diminution in
swelling, warmth, and discomfort in involved joints.
They suggested that estrogens ameliorated the disease
by suppression of lymphocyte metabolism.
In conflict with those findings is a recent report
628
by Allen et al (22) in which implanted subcutaneous
capsules of beta estradiol in rats were found to enhance experimental arthritis induced by extracts of
cell walls. The authors postulated that estrogens depress the clearance and sequestration of streptococcal
cell wall fragments by the reticuloendothelial system,
thereby increasing the number of fragments that disseminate to synovial tissue which increases the severity of reactive arthritis.
In regard to infection, Toivanen (23) demonstrated that under carefully controlled conditions, estrogens potentiated staphylococcal infection in mice,
while progesterone had no significant effect. Prom the
above it is clear that the mechanisms by which sex
hormones modify inflammation and infection remain
poorly understood.
Despite the central role of neutrophils in the
inflammatory response, few studies have been done to
determine the effects of sex hormones on these cells.
Dillard and Bodel (24) reported that estradiol reduced
release of pyrogen (interleukin-1?) from mixed leukocytes after phagocytosis of dead staphylococci, whereas progesterone augmented release of endogenous
pyrogen. These initial investigations were later expanded by Bodel et al (25), who found that the
presence of either estradiol or progesterone led to
decreased oxygen consumption of neutrophils following phagocytosis of staphylococci. They also reported
that preincubation of the cells with estradiol resulted
in diminished degranulation, whereas progesterone
appeared to enhance degranulation as measured by
release of acid phosphatase. We have confirmed the
inhibitory effects of these hormones on the respiratory
burst which, in our experiments, was stimulated by a
synthetic chemopeptide. However, both estradiol and
progesterone decreased release of granule contents
from the neutrophil as measured by release of betaglucuronidase and lysozyme.
These observations suggest that the sex hormones interfere with neutrophil activation. The precise mode of action is unknown. In response to a
stimulus, such as FMLP, the neutrophil transforms
molecular oxygen to the highly reactive free radical
superoxide anion, presumably by means of a complex
generating system the terminal moiety of which is
available to the external plasma membrane (26). Skubitz et a1 (27) have demonstrated that progesterone at 3
pg/ml does not inhibit the binding of FMLP to the
membrane of leukemic promyelocytes. Our data are in
agreement with these findings: sex hormones did not
interfere with the binding of FMLP to the plasma
BUYON ET AL
membrane of mature granulocytes. Indeed, estradiol
appeared to increase specific binding of FMLP.
Our data would suggest that both estradiol and
progesterone have direct cellular effects on the respiratory burst since these hormones do not act as
scavengers of 02-generation, in the manner of ceruloplasmin. With respect to degranulation, evidence from
in vitro systems suggests that estradiol enhances the
stability of artificial membranes such as those of
liposomes (28). Thus, the actions of estradiol and
progesterone in vivo may be contingent upon more
primary alterations in the biosynthesis of various
activating proteins and respiratory coenzymes as well
as alterations of membrane phospholipids.
The in vitro studies reported here demonstrate
that estradiol and progesterone significantly inhibit
neutrophil responses. The effect of estradiol on 02inhibition was rapid, whereas that of progesterone
required prolonged preincubation. With respect to
degranulation, inhibition of beta-glucuronidase release
was time-dependent with both steroids, whereas the
inhibition of lysozyme release did not appear to be
time-dependent . Clearly our doses exceed measured
serum levels.
During the menstrual cycle, estradiol peaks
immediately before ovulation at 0.5 ng/ml, and progesterone peaks immediately after ovulation at 28 ng/ml
(29). During pregnancy, estradiol levels continue to
rise and reach 30 ng/ml before parturition, whereas
progesterone reaches its maximal concentration of 400
ng/ml at 36 weeks (30). Only by combining the hormones could significant, albeit small, inhibition of 02be attained with estradiol at a level 1 log order higher
than that of the third trimester and with progesterone
at a level equal to that of the third trimester. Dexamethasone at 4 pg/ml did not inhibit 02-generation to
the extent achieved by either of the sex hormones at
equal concentration.
Our studies on neutrophils from women during
various phases of the menstrual cycle and gestation
demonstrate that the hormonal environment in vivo
does not alter the in vitro responses of neutrophils to
various external stimuli. These data suggest that neither hormone causes permanent changes in the neutrophi1 and that continuous contact of the cells with
estradiol and/or progesterone may be necessary for
inhibition of cellular responses. These observations
corroborate the findings of Bodel et a1 (25) that cells
from women during their third.trimester of pregnancy
exhibit a reduced respiratory burst after phagocytosis
when compared with cells from the same women
FEMALE SEX HORMONES AND NEUTROPHILS
postpartum. Bodel’s experiments were done with the
cells suspended in autologous serum, which was absent in our studies.
Estradiol w a s more active than progesterone
with respect t o inhibition of neutrophil activation.
Estradiol inhibited superoxide anion generation at
concentrations of 1.5 pdml. When estradiol and progesterone were combined at 0.3 pg/ml each, levels
which begin t o approximate those measured during
gestation, 02-was significantly decreased. Degranulation was inhibited by both hormones only at higher
concentrations, i.e., 3 pg/ml.
In the neutrophil, metabolites of the respiratory
burst and granule constituents contribute t o microbial
killing as well as tissue damage (15). Were the sex
hormones t o inhibit neutrophil function in vivo t o
the extent w e have found in vitro, one might expect
inflammation t o be ameliorated during pregnancy (during which these hormones approach the inhibitory
concentrations used in our experiments). Similarly,
were abnormalities in the handling or metabolism of
these hormones to be documented in conditions such
as SLE, one might expect increased susceptibility to
infection.
Since normal estrogen metabolism is deviated
in SLE to estrogenic species which may exceed the
biologic activity of their parent compounds (7), it will
be important t o study the 16-hydroxylated estrogen
metabolites in SLE with respect t o their inhibition of,
neutrophil function. In the absence of such studies,
our data d o not yet provide a n adequate explanation
for the effects of pregnancy or estrogen metabolism on
host defenses and inflammation.
ACKNOWLEDGMENTS
We would like to thank Ms Suzette Tenhet and Ms
Phyllis Kronhaus for assistance ir? manuscript preparation,
and Dr. Paul Aisen for helpful advice.
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