tion during a normal 24 h sleep-wake cycle was delayed by sleep deprivation, ... tered patterns in immune functions occurred indepen- dently of the cortisol ...
Effects
of sleep deprivation
on human
immune
functions H. MOLDOFSKY,’
R. DAVIDSON,
of the Toronto Hospital,
and Immunology,
Surgery,
J.
F. A. LUE,
Toronto Western Division
University
AND
R. GORCZYNSKI
Toronto, Ontario,
of Toronto, Toronto, Ontario,
Canada
M5T
Canada M5G
2S8 and Departments of Psychiatry, 1X5
METHODS ABSTRACT
Ten
The effect of 40 h of wakefulness on a variety of immunological parameters in the peripheral blood from 10 normal male subjects was studied. Sleep deprivation led to enhanced nocturnal plasma interleukin 1-like and interleukin 2-like activities. The rise in nocturnal response of lymphocytes to pokeweed mitogen stimulation during a normal 24 h sleep-wake cycle was delayed by sleep deprivation, but the response to the phytohemagglutinin mitogen was unaffected. With resumed nocturnal sleep, there was a prolonged decline in natural killer cell activity (measured as spontaneous cytolytic activity for human tumor cells) and return of an increased response to pokeweed mitogen. The altered patterns in immune functions occurred independently of the cortisol circadian rhythm, which remained unchanged. -MOLDOFSKY, H.; LUE, F. A.; DAVIDSON,J.
vation
on
R.; GoRczYNslu, human immune
1972-1977;
1989.
Key Words:
sleep
immunity
R. Effects functions.
of sleep depriFASEB J. 3:
lymphokines
natural killer
cells
RECENT STUDIES HAVE SHOWN a close connection between sleep and the immune system. A variety of peptides that have been shown to promote sleep in animals also have immunomodulatory effects. These agents include: factor S, muramyl peptides, interleukin 1 (IL 1), a-interferon, tumor necrosis factor (1), vasoactive intestine peptide (2-4), prostaglandin D2 (5), and delta sleep-inducing peptide (6). We have shown that aspects of cellular and humoral immune functions are altered with sleep in humans. Peripheral blood lymphocyte (PBL) proliferative responses to pokeweed mitogen (PWM), plasma IL 1-like, and IL 2-like activities were all greater during nocturnal sleep than during wakefulness. Circulating natural killer (NK) cell activity was
greater
determine
during
the
whether
daytime
these
than
immune
during
to sleep, we investigated the possibility vation would alter these patterns. 1972
sleep
measures that
(7).
normal
male
subjects
aged
19-2 7 years
To
are related sleep
healthy,
(mean age, 23 years) who maintained a regular sleepwake schedule were investigated for 5 consecutive days (Fig. 1). Subjects were physically and psychologically healthy as determined by physical examination, Cornell Medical Index, and sleep-wake questionnaires. They were requested to maintain a regular sleep-wake cycle during the week preceding the study and to abstain from the use of alcohol, nicotine, and caffeine during the study. The study was conducted in accordance with the Declaration of Helsinki. All subjects were required to complete the University of Toronto Human Review approved consent form. On the initial adaptation night subjects were screened to exclude those with sleep pathologies, including sleep apnea and nocturnal myoclonus. Upon awakening (day 2), serial venous blood samples (3 ml/sample) were taken every 2 h from 0800 to 2400 h and half-hourly from 2400 to 0730 h until the termination of the study (day 5). Blood was drawn using a manifold system connected to a heparinized drip (8). Blood samples were immediately taken to the laboratory for assessment of immune function. During this baseline period (day 2), subjects were trained and allowed 10 scheduled practices of a 20-mm battery of computerized tasks to assess performance and mood (9). Subjects slept at their usual bedtimes while being monitored polysomnographically. Beginning on day 3 (0930 h), subjects remained awake for 40 h while engaging in the computerized performance tasks hourly, participating in a variety of table games, and reading. Monitoring of subjects included electroencephalogram, electrooculogram, submental electromyogram, and electrocardiogram via portable recorder (Oxford, Mediog 9000) with constant laboratory observation. On day 4 (ca. 2300 h), subjects resumed their usual sleep habits but were allowed to extend their sleep period times. Polysomnographic recordings were scored manually according to standard criteria (10).
depri-
‘To whom correspondence should be addressed: Department Psychiatry, Toronto Western Division of the Toronto Hospital, Bathurst St., Toronto, Ontario, Canada M5T 2S8. 0892-6638/89/0003-1972/$01
of 399
.50. © FASEB
Adaptation/xcresfliflg Day
Blood
sa.pling
2
Day
3
Day
4
begins
extended
sampling
sleep
ends
5 Blood
clock time
q 2 hr
(B sa.p.)
#{149}.-
q 1/2 hr
(16)
2400
0800
#{149}
0600
1. Protocol summary. Subjects were observed for 5 consecutive days with blood drawn every 2 h from 0800 to 2400 h and halfhourly from 2400 to 0730 h from morning of day 2 until day 5. Subjects from
slept their usual time on days 1 and 2, but remained awake arising on day 3 until normal time of sleep on day 4, then resumed sleep until desired awakening. Subjects remained awake, with hourly mood and performance testing and continuous monitoring.
KILLER
and
IL
2-like
activity,
as
described
previously
(7).
Briefly, PBL were obtained by ficoll/Hypaque separation of whole blood. Cells were resuspended in aminimal essential medium (a-MEM) supplemented with 1 x 10-6 M 2-mercaptoethanol, 10% heat-inactivated fetal calf serum, and 10% heat-inactivated commercial human AB serum before use. This serum mixture has proved to be optimal in our hands for mitogen proliferation and human NK assays. All assays were performed in triplicate on PBL within 60 mm of sampling. Plasma samples were stored at 4#{176}C throughout the sampling period and were assayed together, again in triplicate, some 24-48 h after termination of the test. For all assays shown, SEM was less than 10-15%. LYMPHOCYTE
PROLIFERATION
supernatant
These assays are described perimental groups containing
elsewhere
(11).
Briefly,
ex-
x iO ficoll/Hypaque separated PBL were incubated in triplicate for 72 h in 250 el a-MEM with and without addition of phytohemagglutinin (PHA) (Weilcome Labs, Research Triangle Park, N.C.), final concentration, 1% or PWM (Gibco Labs, Chagrin Falls, Ohio), final concentration, 1%. Thereafter, individual wells were pulsed with 1 tCi/well [3H]thymidine and cells were harvested for analysis in a well-type /3-scintillation counter 16 h later. Background counts per minute (cpm) varied from 500 to 1000 in stimulated cultures over the course of the experiment. Data were expressed as arithmetic mean cpm over background. Within a 24-h period, cpm for PHA and PWM stimulation have been found to vary for any one individual by 30-60%; from individual to individual, the z cpm varied from 15,000-40,000 (PHA) or 6000-15,000 (PWM) in different experiments. However, when repeated samples (within 5 mm)
SLEEP DEPRIVATION
AND
IMMUNE
1
FUNCTION
analyzed,
the assays
15%.
CELL
of
INTERLEUKIN
ACTIVITY
individual
1-LIKE
IL 1-like activity
wells.
From
the
curve
ACTIVITY
in the plasma
(7).
Briefly,
was assayed
as described
1 x 106 mouse thymocytes were plates for 72 h in the presence
incubated in microtiter of concanavalin A (ConA) (Sigma, St. Louis, Mo.), 5 eg/ml. Control cultures received thymocytes and ConA only or, in addition, a standard source of IL 1 (obtained as
the
supernatant
confluent line,
16-18
of
lipopolysaccharide-stimulated
cultures
of the mouse macrophage tumor Thereafter, samples were pulsed for 1 tCi 3H-tritiated thymidine (3HTdR)
P388D1).
h with
(New England harvested, and tillation counter.
Nuclear, Boston, Mass.), radioactivity was counted All groups were set up
Data
were
equals minus of IL
cpm (culture cpm (culture 1-like activity
first
cultures with preparation.
expressed
as
cpm,
6.7 Ci/mM, in a /3-scinin triplicate.
where
with ConA and putative with ConA only). Thereafter, were evaluated from the t
various
INTERLEUKIN
ASSAYS
and
immunologic
(effector:target vs. specific 5tCr release), the activity of each sample was expressed as lytic units/106 PBL (1 lytic unit defined as that required to give 20% lysis of I x iO target in 4 h).
previously
Blood samples were assayed for lymphocyte responses to mitogen stimulation, NK activity, IL 1-like activity,
of the
The NK activity assay has been described (12). In brief, 1 x iO 51Cr-K562 target cells were incubated in triplicate in 300 tl medium with individual PBL samples at ratios of 100:1, 60:1, and 30:1. After 4 h, specific 51Cr release was measured by sampling an aliquot (50 tl) of the
Figure
processed
in any
did not exceed
NATURAL
sleep
were
variation
described
wak#{149}tulness
Resu.ed
Blood
one individual
maximum Baseline
Day
Day
from
1
amounts
of a standard
cpm IL 1) units
cpm in IL I
2 ASSAY
IL 2 was assayed by using an IL 2-dependent human NK cell line (RMG-D4). Serial dilutions of plasma were added to 5 x 10 cells in 200 tl medium in flatbottomed microculture plates. At 48 h, cells were pulsed with 1 tCi [3H]thymidine for 16 h. Data were again expressed as cpm over cultures receiving no IL 2 (background range in these experiments: 300-700 cpm). Experiment-to-experiment variation with commercial IL 2 has given peak stimulation ranging from 4000 to 11,000 cpm. However, the dose of commercial material needed to give peak response remained constant (15 tl). PLASMA
CORTISOL
Cortisol levels were assayed by using a radioimmunoassay Products tectable
bilities
Corp., level was
were
in triplicate (RIA)
Los Angeles, Calif.). 0.2 tg/dl. Intraand
4.1 and
8.8%,
for all samples kit (Diagnostic Minimum interassay
devaria-
respectively.
1973
Technical difficulties Each subject provided and all assays were response assays were
TABLE
resulted in some loss of data. more than 76 blood samples, carried out in triplicate. Mitogen obtained
from
all 10 subjects;
Da y2
four
subjects were missing three or more points (i.e., 13, 5, 3, 3). NK assays were obtained for eight subjects, with four subjects missing three or more samples (i.e., 20, 21, 4, 3). IL 1 assay was completed for six subjects, with
three
subjects
5, 3).
IL
missing
2 assay
one subject
three
was
missing
or more
completed
samples
for
21 samples.
five
(i.e.,
subjects,
Samples
14, with
for the various
assays were lost for a variety of technical reasons (e.g., sporadic contamination of cultures, inactivation of IL
standard,
and failure
of proliferation
of cell lines).
values for the various assays were random. The of freedom (df) were appropriately adjusted statistical analyses.
1
Lost
degrees for all
To permit comparison of assays among subjects, the values obtained over the 76-h study for each parameter were standardized by using z statistical transformations. To determine the effects of day and time, data for each parameter were averaged for 12 time blocks (Mhours) (four 4-h blocks between 0700 and 2259 h: i.e., 0700-1059 h, 1100-1459 h, 1500-1859 h, 1900-2259 h, and
eight
1-h blocks
day. Two-way
between
2300
and
(3 days and 12 Mhours)
0659
h) for each
repeated-measures
analysis of variance (ANOVA) was carried parameter. This analysis was also repeated ing the time base for latencies to the various
out for each after adjustsleep stages
(1, 2, slow wave sleep [SWS] and rapid eye [REM]). To accommodate the more limited
1. Comparison of sleep physiology features on days 2 and 4 in means (and standard deviations)4
expressed
movement IL 1 and
Sleep Sleep
period latency,
time, mm
REM
latency,
Stage 1, mm Stage 2, mm Stages 3 and REM, mm
mm
462.7 25.0 109.0 28.2 252.6 60.0 92.6
mm
4, mm
Da y4
(22.7) (16.0) (65.6) (8.9) (33.5) (24.0) (24.2)
(95.2) (3.2) (23.8) (10.9) (78.5) (30.3) (40.6)
628.6k
6.3’ 76.7 22.8 341.3’ 103.1’ 140.9’
Subjects’ baseline sleep (day 2) values are typical for age group. There were no significant differences between adaptation/screening (day 1) and baseline (day 2). Resumed sleep (day 4) after a night of sleep loss resulted in significantly longer sleep period time, stages 2, 3, 4, REM, and shortened sleep latency. bDiffers from baseline value, P < 0.0001, paired I test, d.f = 9. ‘Differs from baseline value, P < 0.01, paired test, d.f = 9.
that of baseline day 2 was less than on days 3 and 4, and sleep deprivation day 3 was less than resumed sleep day 4 (SNK, P < 0.05). Response to PWM stimulation was generally higher during nighttime than daytime (F = 47.3, d.f = 11,99, P < 0.0001). During sleep
deprivation, the nocturnal rise in activity did not occur until 0400-0559 h (F 6.4, d.f 22,188, P < 0.0001) as compared with increased activity from 2400 to 0559 h =
during during
baseline resumed
=
sleep on day sleep on
2 and from 2400 to 0559 h day 4 (SNK, P < 0.05)
(Fig. 2). The mitogen response was greater during all sleep stages than during daytime wakefulness (F 9.8, d.f 6,47, P < 0.0001), with maximum response occurring during SWS and minimum response during stage 1, REM, and wakefulness after sleep onset. =
=
IL 2 data sets, the repeated-measures ANOVA was performed on only nine nighttime blocks (i.e., 1900-0659 h). To further examine the effects of sleep stages, another two-way (2 days and 7 stages) repeated-measures ANOVA was carried out using average values from each stage (daytime wake, nighttime wake, 1, 2, 3, 4,
and REM). This latter analysis grouped 0700 and 2300 h as daytime wake. When significant, comparison
the Student-Newman-Keuls was employed.
data between F values were (SNK)
multiple
higher
stage (F
physiology
Table
1 shows
2.1,
d.f
and
day
of the sleep
was reduced
sleep).
of day 2
with there was a prolongation of sleep period time, stage 2, SWS, and REM. There were no significant differences in sleep physiology parameters between the adaptation and baseline nights.
day 2, sleep latency
4 (resumed
physiology Compared
on day 4 and
blood
lymphocyte
mitogen
stimulation
Pokeweed
mitogen
Vol. 3
June 1989
=
with 4.3,
mean
d.f
=
=
cell
h) after
onset
on days
to
response increas2,18, P < 0.05);
of
2 and
4
activity
progressively
decreased
over the 3 days of
observed; however at nighttime on days 2 and 4 there was greater NK cell activity during the 4 h before onset of sleep than during the remaining time asleep (F 4.2, df 8,56, P < 0.0001). There was a persis=
tent reduction in NK cell activity from 2300 to on day 4 during resumed sleep, compared with mum rise in NK cell activity from 2400 to 0059 sleep onset on baseline day 2 (F = 1.7, df = P < 0.05), followed by the nocturnal decline in
response
A day effect was observed, ing over the 3 days (F
1974
responsiveness
0600-0800
at any other time 11,99, P < 0.05).
the study (F = 7.4, df = 2,14, P < 0.01), i.e., mean NK activity on baseline day 2 was higher than on days 3 and 4, and higher on day 3 than on resumed sleep day 4 (SNK, P < 0.05). No particular daytime pattern was
=
Peripheral
no differences over the 3 days However, PHA response was
than
killer
NK activity aspects
showed stages.
6 h (approximately
1 sleep
Natural
Sleep
mitogen response
PHA stimulation or during sleep
=
RESULTS
(baseline)
Phytoliemagglutinin
(Fig.
3). However,
sleep
stages.
The FASEB Journal
there
were no differences
0559
22,136, activity
among
MOLDOFSKY
h
a maxih with
the
ET AL.
Dsy
2,
Bas.lins
s1s.p
Interleukin
1-like activity
1.0
Despite potential difficulties in measuring plasma IL 1 activity because of suspected inhibitors associated with the biologic assay used in this study (13), peaks of plasma IL 1-like activity occurred in five of the six sUbjects at various times during baseline sleep. In contrast to the previous study (7) where IL I activity was observed with SWS after sleep onset, these grouped data did not show any specific diurnal or sleep stage-related pattern. Mean nighttime plasma IL 1-like activity was higher during sleep deprivation and resumed sleep compared with baseline (F 4.6, df 2,10, P < 0.05).
z s C 0
r e -0.
=
=
-1.
-1.
Interleukin
2-like
activity
-1.
Once
-1.
more,
1 h during -1.
3, !xtended
IL
part
2-like
activity
of sleep,
was
as in the
greater initial
for study
(7). Mean nighttime plasma IL 2-like activity was highest from 0100 to 0159 h on all three nights (F 2.4, df 8,32, P < 0.05). Furthermore, the plasma IL 2like activity was higher during 0100-0159 h of day 3 sleep deprivation in comparison with that during 2400-0059 h of both day 2 baseline sleep and day 4 resumed sleep (F 2.2, c/f 16,58, P < 0.01).
-2.
Day
plasma the early
=
wakefulness
=
z
=
=
$
Plasma
C 0
cortisol
The pattern of cortisol production showed a typical daytime peak (0700-1100 h) and nocturnal trough (2300-0300 h). This circadian rhythmicity for all subjects was not altered by either sleep deprivation or by
r
e
the Day
4, Rssuaed
recovery
sleep
for
any
individual.
sleep
1.#{149}
DISCUSSION
0.
z
Despite
0..
with and
$ C 0
r e -I.’ -1.
7-li.
11-15
15-19
19-23
1357
time (Mhour) Figure 2. Summary of PBL response
to PWM. Change in cpm for each sample was z transformed within subjects by setting mean to 0 and standard deviation (SD) to 1. Transformed values were then grouped into 12 time blocks (Mhours) and two-way (3 days and 12 Mhours) repeated-measures ANOVA was performed. Mean cpm over background for all 10 subjects was 10737 for day 2, 11911 for day 3, and 13210 for day 4. One SD was 5553, 4723, and 3624 for days 2, 3, and 4, respectively. Response was higher on days 3 and 4 than on day 2 (F = 4.3, d.f = 2,18, P < 0.05), higher during nighttime than daytime (F = 47.3, d.f - 11,99, P < 0.0001); nocturnal increase was delayed until 0400 h during the sleep deprivation night (F = 6.4, d.f = 22,188, P < 0.0001). Bars indicate maximum and minimum SEM. PWM response vs. time (n 10). SLEEP DEPRIVATION
AND
IMMUNE
FUNCTION
great
regard illness,
scientific
advances
in the
past
30 years
to sleep physiology and its role in health the function of sleep remains unresolved.
The theory that sleep is a restorative process for tissue anabolic activities to counter catabolic functions has been controversial, with evidence presented both in favor of (14) and in opposition to the theory (15). This study largely confirms our initial observations that aspects of the cellular and humoral immune functions are related to sleep (7). If sleep serves as a restorative process, then the observed increased proliferative response of peripheral blood lymphocytes during baseline sleep may be an indicator of this process. This study shows a potential for increased responses to PWM, a B cell stimulator, during sleep. However, it also confirms our previous findings that responses to PHA, a T cell stimulator in vitro, do not show increased activity with onset of sleep (7). In fact, no
special
diurnal
pattern
was
observed
except
that
PHA hours
mitogen response increased inexplicably for a few in the early part of the morning, approximately 6 h after the onset of sleep. As in the first report, NK cell activity shows a particular pattern of increased activity before sleep and a precipitous drop after onset of sleep (7). This decline in peripheral blood NK cell 1975
Day 2, Baseline
sleep
sleep might relate to reduced cellular which would not be consistent with the theory. On the other hand, in support of the
1.I
function
1.6
production, restorative
with
1.4
theory,
i.2
NK cells into tissues
during
sleep
of pathogenic 1.0
z 0.4
fl
S 0.2
r e
o.c
TI
-
-0.2 -0.4
Day 3, Sleep
deprivation
1.0
0.$
40 h
PWM
W
Whereas
during
a reduction
-0. 2
and lysis
the
24-h
mean
PWM
sleep
deprivation
in the number
re-
or whether
the
is accompanied
of B cells or their
resumption
of sleep
by
functional
would
sociated with restoration of B cells and/or tional capabilities. Similarly, the significance
be as-
their funcof the per-
sistent decline in NK cell activity with resumption of sleep after extended wakefulness remains to be established.
On
the
other
hand,
the
lack
of a specific
sleep
4
deprivation suggests
-0.0
4,
Resuced
Sleep
effect that
T cell
on stimulation activity
with
or certain
PHA subsets
mitogen of T cell
function may not be as readily affected by 40 h of wakefulness. If, as suggested by animal studies (16-19), IL 1 has sleep-inducing properties, then sleep deprivation might be expected to alter IL 1 levels. OUr previous analysis
z S
r
of the
surveillance
cells.
of wakefulness.
activity
C 0
be redistribution
sponse showed an overall increase in activity from baseline day 2 to day 4, 24-h mean NK cell activity was decreased over the same time interval. Furthermore, a specific nocturnal change occurred during sleep deprivation. The expected rise in response to stimulation by PWM commencing with sleep on the baseline night was delayed until 0400-0559 h on day 3. With resumed sleep, the nocturnal rise in response to stimulation with PWM was reestablished. It is as yet unknown whether the kinetic delay in response to stimulation with
z
-0.
may
for immune
Sleep deprivation is a technique that has been commonly used to assess the function of sleep (15). This study shows that aspects of the immune cellular functions, i.e., PWM and NK cell activity, are altered with
0.0
C 0
there
0.
(7) led us to anticipate
an increase
in IL 2 activity
after
increased IL 1 activity. The observed increase in plasma IL 1-like and IL 2-like activities during the night of sleep deprivation might reflect changes in these interleukins within the central nervous system. IL 1 has been shown to be a product of astroglial cells (20). The normal brain produces IL 1/3 mRNA, and receptors
0.
e
are widely
distributed
throughout
the brain
(21).
A net-
work of IL 1 immunoreactive fibers has been observed in the human hypothalamus (22). Furthermore, IL 1-like biologic activity and IL 1/3 RIA measurements are augmented 7-11
11-15
15-19
19-23
1357
time (Mhour) Figure 3. Summary
of NK
activity.
Percent
lysis
for each
in
the
cerebrospinal
fluid
of
cats
while
asleep as compared with wakefulness (23). A frequent confounding influence to the interpretation of the physiological effects of sleep deprivation sample
the possibility
that
the emotional
stress
is
of the disruptive
experience may contribute to the observed changes. was z transformed within subjects by setting mean to 0 and stanThis idea was raised by Home (24) in his review of the dard deviation (SD) to 1. Transformed values were then grouped studies of assessments of aspects of the iminto 12 time blocks (Mhours) and two-way (3 days and 12 Mho#{252}rs) two earlier repeated-measures ANOVA was carried Out. Mean percent lysisfor all eight subjects was 27.9% for day 2, 26.3% for day 3, and 20.9% for day 4. One 5D was 11.2%, 12.9%, and 11.6% for days 2, 3, and 4, respectively. NK activity was higher on day 2 than on days 3 and 4 (F = 7.4, d.f 2, P < 0.01), and was lower during resumed sleep on day 4 from 2400 to 0059 h than during baseline sleep on day 2 (F 1.7, a’.f = 22,136, P < 0.05). Bars indicate maximum and minimum SEM. NK cell activity vs. time (n = 8).
1976
Vol. 3
June 1989
mune
system
study involved phagocytosis
after
human
sleep
deprivation.
The
first
72 h of sleep deprivation. The reduced by polymorphonuclear granulocytes and
increased interferon production by lymphocytes were accompanied by raised adrenocortical and adrenomedullary activities (25). The second study involving 64 h of wakefulness with subjects in constant light and
The FASEBJournal
MOLDOFSKY ET AL.
in visual and world showed
isolation from the surrounding response to PHA but no changes in endocrine function (26). These studies are also flawed by the reliance on single morning blood samples taken before and after sleep deprivation and a
reduced
several days later. The diurnal fluctuations observed in our previous (7) and current study suggest that earlier studies using single sample techniques are likely to be difficult to interpret. Although the experimental procedures
may
be stressful
(e.g.,
repeated
blood
sampling),
of stress,
subjects.
showed
a normal
Furthermore,
circadian
no day-to-day
pattern
change
2
during
nocturnal
sleep deprivation
in immune
functions
that have been
ety of stressful circumstances (27, 28) may expected disordered sleep that accompanies tionally distressing situations or the diurnal
of the immune
13.
secretion at the changes
reported
12.
occurred
were unrelated
to any changes in the pattern of cortisol night. On the other hand, it is possible that
10.
for all
in this circadian pattern throughout the study. Finally, the changes in PWM, PHA, NK cell activity, IL 1, and IL
9.
11.
there was no clinical or physiological evidence that the subjects who participated in this study were distressed. The patterns of plasma cortisol, a physiological indicator
chronobiological
auditive
in a varirelate to the these emofluctuation
14. 15. 16.
functions. 17.
We
thank Dr. A. W. MacLean
Angus
and R. Heslegrave
testing
protocol,
research
for statistical
for providing assistants
advice,
computerized
M. Gregoris,
Drs. R. G.
performance
research
was
Environmental cal Research
supported
by
Medicine Council
the
contract of Canada
Defence
and
Civil
no. 02E.97711-5-8483 grant
no.
MA
for asThis
Instmtute and
19. Shoham,
Krueger,
of
Medi-
7733.
REFERENCES
20.
21.
1. Krueger, J. M., Toth, L. A., Cady, A. B., Johannsen, L., and Obal, F. (1988) Immunomodulation and sleep. Sleep Peptides: Basic and Clinical Approaches (Inoue, S., Schneider-Helmert, D., eds) pp. 95-129, Japan Sci. Soc. Press/Springer Verlag, Berlin 2. Drucker-Colin, R., Bernal-Pedraza, J., Fernandez-Cancino, F., and Oksenberg, A. (1984) Is vasoactive intestinal polypeptide (VIP) a sleep factor? Peptides 5, 837-840 3. Obal, E, Sary, G., Alfoldi, P., Rubicsek, G., and Obal, E (1986) Vasoactive intestinal polypeptide promotes sleep without effects on brain temperature in rats at night. Neurosci. Leti. 64, 236-240 4. Riou, F., Cespuglio, R., and Jouvet, M. (1982) Endogenous peptides and sleep in the rat. III. The hypnogenic properties of vasoactive intestinal polypeptide (VIP). Ne’uropeptides 2, 265-277 5. Ueno, R., Honda, K., Inoue, S., and Hayaishi, 0. (1983) Prostaglandin D2 a cerebral sleep-inducing substance in rats. Proc. Nail. Acad. Sci. USA 80, 1735-1737 6. Yehuda, S., Shredny, B., and Kalechman, Y. (1987) Effects of DSIP, 5-HTP and serotonin on the lymphokine system in a preliminary study. mt. j Neurosci. 33, 185-197 7. Moldofsky, H., Lue, F. A., Eisen, J., Keystone, E., and Gorczynski, R. M. (1986) The relationship of interleukin-1 and immune functions to sleep in humans. Psychosom. Med. 48, 309-318 8. Weitzman, E. D., Czeisler, C. A., Zimmerman, J. C., Ronda, J. M., and Knauer, R. S. (1982) Blood sample collection in
SLEEP DEPRIVATION
AND
IMMUNE
FUNCTION
analytical
18. Susic, V., and Totic, S. (1987) Short interleukin-l on sleep and temperature
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Receivedfor publication January 20, 1989. Accepted for publication March 21, 1989.
1977
