International Journal of Psychophysiology 37 Ž2000.

219᎐232

Prolonged reduction of salivary immunoglobulin A

ž sIgA/ after a major academic exam

Renate Deinzer a,U , Christian Kleineidama , Renate Stiller-Winkler b,

Helga Idel b, Doris Bachg c
a
Institute for Medical Psychology, Uni¨ ersity of Dusseldorf,
¨ ¨
P.O. Box 101007, 40001 Dusseldorf, Germany
b
Institute for Hygiene, Uni¨ ersity of Dusseldorf,
¨ ¨
P.O. Box 101007, 40001 Dusseldorf, Germany
c
Diabetes Research Institute, Uni¨ ersity of Dusseldorf,
¨ ¨
P.O. Box 101007, 40001 Dusseldorf, Germany

Received 8 April 1999; received in revised form 30 September 1999; accepted 23 November 1999

Abstract

Objecti¨ e: In a previous study we observed a continuous reduction of salivary IgA concentration ŽwsIgAx. during a
period of academic stress. This reduction of sIgA concentration exceeded the stress period by at least 1 week. The
present study aimed to replicate and extend our previous finding. In particular, we wanted to examine the time of
recovery of wsIgAx alterations associated with academic stress. Method: Twenty-seven participants in a major medical
exam and 27 controls not participating in any exam during the study provided daily saliva samples Žimmediately after
awakening., from the 6th day prior to their last exam until the 14th day afterwards, for analysis of salivary IgA. Data
were averaged for the last weeks of exams and the first and second week after exams, respectively. Results: A
prolonged reduction of sIgA in exam students as compared to controls was observed. Fourteen days post-stress sIgA
concentrations of exam students were still significantly lower than control levels Ž Ps 0.004.. No recovery was
observable. At the same time exam students and controls did not differ in terms of self-reported stress and recovery.
Conclusions: Psychological and immunological stress effects may be dissociated, the latter considerably exceeding the
stress period. A closer look at the temporal dynamics of stress-induced immune alterations might increase our
understanding of psychoimmuno relationships. 䊚 2000 Elsevier Science B.V. All rights reserved.

Keywords: Immunoglobulin A; Stress; Academic exams; Recovery; Psychoimmunology; Long-term immunological effects; Alcohol
consumption

U
Corresponding author. Tel.: q49-211-81-13016; fax: q49-211-81-13015.
E-mail address: renate.deinzer@uni-duesseldorf.de ŽR. Deinzer..

0167-8760r00r$ - see front matter 䊚 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 1 6 7 - 8 7 6 0 Ž 9 9 . 0 0 1 1 2 - 9
220 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232
1. Introduction
Secretory Immunoglobulin A is an antibody
found in high concentrations on all mucosal sur-
faces of the human body including the respiratory
tract. It is thought to serve as a first line of
defense against invading organisms and thus, to
play a key role in prevention of infectious disease
ŽTomasi, 1970, 1992; Ogra et al., 1971; Mestecky
et al., 1986; Jemmott and McClelland, 1989;
Mestecky and Russell, 1997; Lamm, 1998.. Stress
is thought to play a role in mucosal infections like
upper respiratory tract infection or periodontal
inflammation ŽMarcenes and Sheiham, 1992;
Cohen, 1995; Deinzer et al., 1998, 1999; Deinzer
et al., in press.. For these diseases it is known
that IgA is involved in the host’s defense against
their pathogens ŽTomasi, 1970, 1992; Ogra et al.,
1971; Ebersole et al., 1986; Mestecky et al., 1986;
Jemmott and McClelland, 1989; Ogawa et al.,
1991; Nieminen et al., 1993; Somer et al., 1993;
Mestecky and Russell, 1997; Lamm, 1998..
Secretory IgA, especially salivary IgA ŽsIgA.
became a focus of interest in psychoimmunologi-
cal research since it has been shown to be sensi-
tive to variations in subjective and objective stress
levels. Acute challenges like assessed seminar
presentations ŽEvans et al., 1994; Bristow et al.,
1997., competition stress in soccer coaches ŽKu-
gler et al., 1996., 100-min working sessions of air
traffic controllers ŽZeier et al., 1996. and a
laboratory computer game task ŽCarroll et al.,
1996., have all been shown to be associated with
sIgA increases. sIgA levels and secretion rates
were found to be related to desirability of daily
events ŽEvans et al., 1993; Stone et al., 1994., an
effect suggested to be mediated at least in part by
mood: negative mood tends to be associated with
lower sIgA ŽStone et al., 1987a,b, 1996.. De-
creases in sIgA have been observed at parturition
ŽAnnie and Groer, 1991. and during prolonged
periods of academic stress ŽJemmott et al., 1983;
Jemmott and Magloire, 1988; Deinzer and
¨
Schuller, 1998. although there is some inconsis-
tency in reports on academic stress effects ŽMc-
Clelland et al., 1985; Bosch et al., 1996; see Sec-
tion 4 of this paper..
By now, much is known about the immediate
effects of stress on salivary IgA. The significance
of stress-induced immune alterations, however,
would be better understood if we knew more
about their temporal dynamics. That is, when do
stress effects on sIgA peak and for how long do
they persist? Little is known about these ques-
tions, so far. In a recent paper we analyzed the
effects of a major academic exam on sIgA from
the 7th day prior to the exam until the 6th day
¨
afterwards ŽDeinzer and Schuller, 1998.. The most
striking result of this study was that sIgA concen-
tration not only decreased at exam days but
dropped further afterwards. Within the 6 days
post-stress period we assessed, sIgA concentra-
tion remained below baseline; no recovery was
observable. The present study aimed to replicate
and extend the results of our earlier study. Most
importantly, we now extended our post-stress as-
sessment period to 14 days, expecting sIgA con-
centration to recover at least in part within that
period.
Academic stress appears to be an ideal setting
for the assessment of temporal dynamics of
stress-induced immune alterations. The immedi-
ate immunological effects of academic stress are
well known ŽJemmott et al., 1983; Kiecolt-Glaser
et al., 1984; Glaser et al., 1985, 1990, 1991; Mc-
Clelland et al., 1985; Vassend and Halvorsen,
1987; Jemmott and Magloire, 1988; Mouton et al.,
¨
1989; Bosch et al., 1996; Deinzer and Schuller,
1998.. The stressor is of high ecological validity
and defined exam conditions guarantee some de-
gree of standardization. Subjects as well as ade-
quate control persons are easily available. Addi-
tionally, as the termination of the objective stres-
sor is clearly defined Ži.e. the exam or last exam
within a prolonged exam period., one can also
study immunological post-stress effects.
There have been suggestions that alterations
within the post-stress period may account for
development of disease ŽDeinzer, 1992. though it
is not clear today whether this should be due to
immunological alterations. However, the data of
our previous study strongly support the notion
that mucosal immune function is compromised
particularly during the post-stress period. The
 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232 221

aim of the present study was to add further data
relevant to this finding and thus, to improve our
understanding of immunological post-stress ef-
fects.
2. Methods
2.1. Subjects
Fifty-four medical students Ž27 exam students
and 27 controls. provided a full set of valid data
for this study. At the beginning of the study,
subjects were asked about their age, number of
semesters they studied medicine, exercise, regular
nicotine consumption, use of oral contraceptives
and whether they had a regular partner. These
data are provided in Table 1. Groups did not
differ on any of these variables.
Recruitment of subjects was done during their
seminars; in addition, fliers requesting their par-
ticipation were distributed throughout the cam-
pus. The students received monetary compensa-
tion ŽDM 50. for participation. Exclusion criteria
for participation were pregnancy, psychiatric dis-
orders, regular medication with known effects on
the immune system, and chronic diseases affect-
ing the immune system.
2.2. Academic stress ¨ s. control condition
This study took place during the 3-month
semester break after the winter-semester courses.
During that time, 27 of our subjects participated
in a major medical exam Žexam group. while the
remaining 27 Žcontrol group. did not participate
in any exam but rather enjoyed their vacation.
The medical exam under consideration consists of
a written part Ž2 days, 4 h each day. and an oral
part Ž2 h, two examiners. 1᎐3 weeks later. After
that students go on holidays for another 3᎐6
weeks. Exam results are given immediately after
the oral exam. Missing the exam impedes the
studies by at least 6 months. Additionally, only
two failures of the exam are allowed; afterwards
the student must discontinue his or her studies of
medicine. The failure rate of this exam varies
between 25 and 35%.
2.3. Sali¨ a samples and biochemical analyses
Subjects were instructed to give unstimulated
saliva samples by means of a Salivette 䊛 ŽSALI-
VETTE no. 51.1534, plain cotton roll; Sarstedt,
¨
Numbrecht, Germany. every morning immedi-
ately after awakening and before doing anything
else. This sampling time was chosen because of its

Table 1
Group differences on putatively intervening variables

Exam students Controls P Žtwo-tailed.

N 27 27
Age Žmean " S.D.. 24.6 Ž"3.7. 24.6 Ž"3.7. 0.98a
Semesters studied medicine 4.8 Ž"2.0. 4.6 Ž"1.7. 0.67a
Žmean " S.D..
Hours of exercise per week 2.8 Ž"2.7. 3.4 Ž"3.4. 0.46a
Žmean " S.D..
Sex Žfemalesrmales. 15r12 14r13 1.0b
Number of smokers 5 4 1.0b
Women using oral 8 11 0.25b
contraceptives
Subjects having a 19 14 0.26b
regular partner
a
Student’s t-test.
b
Fisher’s exact tests.
222 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232
good standardization with respect to preceding
activities and individual circadian rhythms. Previ-
ous studies of Hucklebridge et al. Ž1998. and our
group ŽStiller-Winkler et al., 1998. have shown
that sIgA concentration peaks at awakening,
shows a steep fall for the next 4 h and then
remains constant. The morning sIgA peak seems
to depend on the time of awakening rather than
the time of the day ŽHucklebridge et al., 1998..
Students were instructed to hold the cotton swab
of the Salivette 䊛 in the mouth for 5 min and not
to swallow during that time. Salivettes were
weighed before and after sampling and saliva
secretion Ždlrmin. was estimated from the dif-
ference between these weights. sIgA secretion
Žmgrmin. was calculated based upon concentra-
tion, saliva volume and sampling time Ži.e. 5 min..
Salivettes were stored at y20⬚C within 6 h of
sampling until analysis wstorage at room tempera-
ture for less than 48 h does not affect sIgA
measures ŽHennig, 1994.x. Salivary IgA concen-
tration was measured by a Behring nephelometer
analyzer ŽMarburg, Germany.. Samples were
thawed and spun for 10 min at 6000 rev.rmin.
For the sIgA determination specific human anti-
IgA serum ŽBehring, Marburg, Germany. was
used. The reliability of this nephelometric tech-
nique for repeated analyses is rtt s 0.98; unpub-
lished data from our laboratory show that these
results correlate with those of the radial im-
munodiffusion technique for determination of se-
cretory sIgA ŽBinding Site, Heidelberg, Germany.
of r s 0.85.
Salivary IgA analyses for one person were al-
ways run together and with the same reagents
and standard curve. The person who did the anal-
yses was blind to the condition of each subject
and to the study design.
2.4. Design
The study was subdivided into four study peri-
ods:
䢇 ‘baseline’ Ž3 days, 4 weeks prior to the written
exam.;
䢇 last week of exams Ž7 days, 6th day prior to
oral exam until day of oral exam.;
䢇 first week after exams Ž7 days, 1st᎐7th day
after oral exam.; and
䢇 second week after exams Ž7 days, 8th᎐14th
day after oral exam..
During these study periods subjects took saliva
samples every morning. Daily saliva samples were
thus taken during the baseline period and from
the 6th day prior to the oral exam until the 14th
day afterwards.
For each exam student there was a control
subject of the same sex who took saliva samples
at the same days as his or her exam partner did.
We took ‘baseline’ measures a few days after
the decision as to whether students were allowed
to participate in the examination or not. At that
time, most of the students were already preparing
for their exams. Thus, the ‘baseline’ period may
not be considered as real baseline in a sense that
exam students and controls were completely com-
parable in terms of stress. Rather, the ‘baseline’
period gives us some idea about sIgA at the
beginning of a prolonged examination period as
compared to the end of the stress period and the
time afterwards.
2.5. Self-reported stress and reco¨ ery
To validate the subjective significance of the
exam as stressor every subject filled out the recov-
ery-stress-questionnaire wRESTQ ŽErholungs-Be-
lastungs-Fragebogen.; Kellmann and Kallus, 1993;
Kallus, 1995a,bx at times indicated in Fig. 1. This
questionnaire assesses stress and recovery during
the last Ž3. days on 12 different scales, which can
take values between 0 and 6. The internal consis-
tencies of the subscales are all above 0.70; the rtt
after 24 h ranges from rtt s 0.79 Žsocial stress . to
rtt s 0.91 Žsocial relaxation.. The scales are:
䢇 general stress Žgeneral feeling of too much
stress to cope with.;
䢇 emotional stress Žfeelings of dissonance and
general irritation.;
䢇 social stress Žannoyance or anger at others.;
䢇 pressure Žunresolved conflicts and pressure.;
䢇 overfatigue Žbeing exhausted and tired.;
 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232 223

Fig. 1. Means and S.E.M.s of the recovery-stress-questionnaire. The questionnaire assesses stress and recovery for the last 3 days.
Group by time of measurement interactions were significant on an alpha-level of at least 2% for all but the social stress scales.
Results of t-tests for independent measures: U PF 0.05; UU P F 0.01; and UUU P F 0.001
224 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232
䢇 lack of energy Žbeing unable to concentrate;
postponing important tasks.;
䢇 somatic complaints Žheadaches, general so-
matic discomfort.;
䢇 success Žhaving success, making important de-
cisions.;
䢇 social relaxation Žhaving laughed, spent nice
hours with friends.;
䢇 somatic relaxation Žfeeling of somatic relax-
ation and efficiency.;
䢇 general well-being Žbeing in a good temper.;
and
䢇 sleep Žrelaxing and calm sleep..
According to our earlier results ŽDeinzer and
¨
Schuller, 1998. we expected students to show
increased levels of subjective stress and reduced
levels of recovery during the exam period and to
recover to control levels within a few days after
the exam.
2.6. Medication and alcohol consumption
To be able to control for influences due to
medication or alcohol consumption we instructed
students to report every use of medication and
the amount of alcoholic beverages they con-
sumed. Students received a little diary where they
noted every pharmaceutic drug they used includ-
ing the product’s name and the dosage they had
applied as well as the type and volume of al-
coholic beverages they had consumed. Students
were asked to make diary entries every evening
before they went to sleep. Data on alcohol con-
sumption were processed in units of one glass Ž0.2
Table 2
Group differences on mean daily alcohol consumption
Baseline
Exam students 0.33" 0.490
Controls 0.86" 0.88
U
t Žd.f.. t Ž49. s 2.7
P Žtwo-tailed. 0.009
U
Different Ns are due to inaccurate recordings of some subjects on single days; daily alcohol consumption is expressed in units
of one glass Ž0.2 l. of wine.
l. of wine Ži.e. approx. 16 g alcohol. according to
the guidelines of Feuerlein et al. Ž1991..
2.7. Statistical analyses
To avoid singular matrices in repeated mea-
sures analysis of variance and to improve clarity
of presentation we averaged ᎏ and thus aggre-
gated into one measure for each period ᎏ the
daily sIgA measures of the four study periods.
Averaging across study periods also allows for
compensation of low-volume saliva samples at
single days which are not analyzable. Indeed,
some subjects provided insufficient saliva samples
at some days. However, all cases provided at least
three sufficient samples per 7-day study period
Žwhich indeed was the criterion to be included
into analyses ., and only six cases per group pro-
vided in one period less than six sufficient sam-
ples.
Differences across periods and group by peri-
ods interactions were examined by repeated mea-
sures ANOVAs and Greenhouse᎐Geisser correc-
tions for sphericity were applied to significance
statements. Below, original degrees of freedom
and epsilon values are presented. A significant
group by study period interaction was expected if
sIgA recovered during the study. For exploratory
reasons, several t-tests were run on sIgA data.
For these comparisons we also computed effect
sizes Ž d .. According to Cohen Ž1992. effects sizes
of ds 0.2, 0.5 and 0.8 can be considered as small,
medium and large, respectively.
Outlying data Ži.e. data exceeding their respec-
tive group means at a given study period by more
Last week First week Second week
of exams after exams after exams
0.24" 0.30 1.22" 1.63 0.68" 0.22
0.97" 0.91 0.88" 0.66 0.86" 0.76
t Ž51. s 3.9 t Ž47. s 1.0 t Ž47. s 0.7
- 0.001 0.328 0.49
 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232
than 2 S.D.. were excluded prior to analyses. The
exclusion of outlying data is necessary within small
sample sizes to avoid statistical artifacts. A single
outlying value can considerably affect the magni-
tude and even the direction of group differences.
Four subjects per group were to be excluded from
analyses due to outlying data. Inclusion of these
subjects did not change the overall picture of
results.
Kolmogorov᎐Smirnov᎐Goodness of Fit Tests
were computed to test for normality; for these
tests an alpha-level of 20% was considered to be
significant. Square root transformations were per-
formed for variables which were not normally
distributed. This was necessary for baseline mea-
sures of sIgA concentration and secretion, only.
To make sure that group differences in sIgA
were not due to differences in medication all
analyses were also run restricted to those subjects
who never took any medication. The results of
these analyses were, however, virtually the same
as the results of the complete group. Thus, stress
effects were not due to differences in medication
of the two study groups.
Since groups differed in terms of daily alcohol
consumption prior to exams Žbut not afterwards;
Table 2. we examined the relationship between
mean daily alcohol consumption and sIgA levels.
However, Spearman rank correlations revealed
no significant relations at any time ŽTable 3..
3. Results
The exam was a potent stressor in terms of
psychological self-report as can be seen from Fig.
Table 3
Spearman rank correlations between mean daily alcohol consumption and sIgA
Baseline
␳ y0.1545
U
Valid N 48
P 0.29
Žtwo-tailed.
U
Different Ns are due to inaccurate recordings of alcohol consumption and outlying sIgA data.
225
1 which shows the results of the recovery-stress-
questionnaire. Exam students reported more
stress and less recovery in the last week of exams
while control group values remained constant.
Within 3 days post-examination exam students
recovered to control values in all but the success
scales. At that time they even reported less lack
of energy, less somatic complaints and more so-
cial relaxation than controls.
Fig. 2 shows sIgA concentrations and secretion
rates throughout study periods and Table 4 gives
the corresponding statistics for group compar-
isons at each study period. Since study groups did
not differ with respect to baseline values, these
were included as covariates for repeated mea-
sures ANOVAs analyzing recovery of sIgA from
prior to exams until 2 weeks after exams. A
significant group by study period interaction was
expected if there was any recovery of sIgA within
2 weeks post-stress. However, this was neither
found for sIgA concentration Ž F2,43 s 0.84, ␧ s
0.83, Ps 0.43. nor for secretion rates Ž F2,43 s
0.25, ␧ s 0.88, Ps 0.76. although the main effect
for groups was significant for concentration Ž F1,43
s 4.96, Ps 0.03. but not for secretion data Ž F1,43
s 1.94, Ps 0.17.. Table 4 shows a general trend
of effect sizes for secretion data to be smaller
than those for concentration data; but even for
secretion data medium effect sizes for group com-
parisons were observed in the weeks after exams.
At the last day of the study, 14 days after exams,
sIgA concentrations Žmean " S.D.: exam stu-
dents: 23.52" 30.78 mgrdl; controls: 64.52"
68.31 mgrdl; t Ž39. s 2.50; Ps 0.005; ds 0.71.
and secretion rates Žexam students: 0.08" 0.11
mgrmin; controls: 0.19" 0.23 mgrmin; t Ž39. s
Last week First week Second week
of exams after exams after exams
0.2191 y0.0172 0.0652
48 46 45
0.14 0.91 0.67
226 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232

1.92; Ps 0.033; ds 0.59. of exam students were
still considerably lower than the respective con-
trol levels.
Table 5 shows the correlation between aggre-
gated sIgA measures for exam students and con-
trols. sIgA was fairly stable within control stu-
dents while in exam students lower correlations
were found between last week of exams and
post-exam weeks. This is what one would expect if
an increasing part of intra-individual variance is
determined by situational Že.g. stress . influences
Žsee the discussion on state-trait influences on
intra-individual variation in Deinzer et al., 1995..
To assess the relation between subjective stress
and reduction of sIgA concentration the three
stress scales best differentiating between exam
students and controls Žgeneral stress, pressure
and overfatigue. were aggregated into one mea-
sure describing subjective stress during the last
weeks of exams. This measure equals the mean of
the three scales at days y3 and 0. We compared
exam students in the upper and lower tertile of
this measure. Decrease of sIgA concentration
from baseline to the last week of exams was
considerably larger within highly stressed exam
students Župper tertile: 38.96" 61.2 mgrdl; lower
tertile: 20.25" 27.6 mgrdl.. This difference, how-
ever, missed the intended level of significance by
far w t Ž16. s 0.84, Ps 0.20x which presumably is
due to the low sample size available for this
comparison; the effect size d of this comparison
equaled ds 0.39. A simultaneous comparison of
the three tertiles via ANOVA revealed a similar
result: Cohen’s effect size f ŽCohen, 1992. was
medium to large Ž f s 0.33.; the level of signifi-
cance, however, was even worse Ž F2.20 s 1.05, Ps
0.37. which is due to the fact that ANOVAs
examine non-directed hypotheses, only. Secretion
rates again revealed a considerably smaller effect
size Župper terzil: 0.058" 0.17 mgrmin; lower
terzil: 0.048" 0.08 mgrmin; t Ž15. s 0.20, Ps
0.41; ds 0.10; ANOVA: F2,19 s 0.92, Ps 0.41;
f s 0.31..
Pearson and Spearman correlations revealed
no significant relationship between the time of
the oral exam and sIgA concentrations or secre-
tion rates for any group at any time. Žy0.27F r F
0.21; y0.25F ␳ F 0.30..

Table 4
Salivary immunoglobulin A during study periods
U
Baseline Week prior First week Second week
to exams after exams after exams

Concentration (mgr dl)

Exam students 5.56" 3.3 35.66" 31.7 29.01" 27.0 25.04" 20.5
Žmean " S.D..
Controls 6.07" 3.2 50.49 " 41.6 39.30" 26.0 46.47" 30.1
Žmean " S.D..
t Žd.f.. t Ž44. s 0.53 t Ž44. s 1.36 t Ž44. s 1.32 t Ž44. s 2.82
UU
P 0.299 0.0915 0.098 0.004
d 0.16 0.40 0.38 0.82

Secretion
Exam students 0.29" 0.17 0.093" 0.088 0.070" 0.068 0.078" 0.083
Žmean " S.D..
Controls 0.30" 0.19 0.111" 0.114 0.101" 0.072 0.111" 0.082
Žmean " S.D..
Žmgrdl.
t Žd.f.. t Ž44. s 0.29 t Ž44. s 0.62 t Ž44. s 0.149 t Ž44. s 1.37
UU
P 0.387 0.270 0.072 0.089
d 0.09 0.18 0.44 0.39
U
Square root transformed data.
UU
One-tailed significance.
 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232
4. Discussion
The present study replicates and extends re-
sults of a previous study from our laboratory
¨
ŽDeinzer and Schuller, 1998.. During a prolonged
period of academic stress, sIgA concentrations
after awakening were reduced as compared to
baseline and to control values. Furthermore, no
recovery of sIgA concentration could be observed
within 14 days post-stress even though students
recovered very soon in terms of self-reported
stress and recovery. Recovery in sIgA concentra-
tion showed a tentative relation to subjective
stress responses at the time of the exams: those
subjects showing the largest stress responses
showed the lowest recovery of sIgA concentra-
tion.
Secretion data mirrored concentration data al-
though effect sizes appeared to be smaller. This is
Fig. 2. Means and S.E.M.s of sIgA concentration Ža. and secretion rates Žb.. Results of t-tests for independent measures:
q
P F 0.10; U PF 0.05; and UU PF 0.01.
227
similar to what we observed in our previous study.
We suggest this effect to be due to some impreci-
sions in assessment of saliva secretion adding
error variance to the data. The reliability of saliva
secretion measures very much depends on proce-
dural control: subjects are not allowed to swallow
during assessment; they should not move the cot-
ton swab or even chew on it during assessment in
order to avoid stimulation of saliva flow; and they
should strictly keep the given sampling duration.
In a field study like ours, however, it is not
possible to perfectly control saliva sampling pro-
cedures. This, however, can adversely affect the
reliability of sIgA secretion data. A comparison of
the re-test correlations given in Table 5 supports
this notion: within control students re-test corre-
lations of secretion data tend to be smaller than
those of concentration data. Irrespective of these
considerations we run another analysis to make
228 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232
Fig. 2. Ž Continued.
sure that group differences in the second week
after exams were not merely due to differences in
saliva volume: an ANCOVA with the covariates,
IgA concentration at baseline, saliva volume at
baseline and saliva volume in the second week
after exams, revealed a highly significant group
effect on sIgA concentration in the second week
after exams Ž F41,1 s 7.02; Ps 0.011.
The group differences we observed were not
due to differences in medication. In our earlier
¨
studies ŽDeinzer and Schuller, 1998. we had also
found that daily sleep has no effect on sIgA
levels. Several other alternative explanations can
also be excluded: our study groups did not differ
with respect to age, sex, number of semesters
studied, exercise, smoking, partnership, and use
of oral contraceptives ŽTable 1.. Furthermore,
daily alcohol consumption does not seem to have
any influence on sIgA concentration ŽTable 3..
One might argue that effects may be due to
alterations in the compliance of exam students
with respect to sampling time. There is a rapid
fall in sIgA immediately after awakening ŽHuckle-
bridge et al., 1998; Stiller-Winkler et al., 1998..
Thus, if there was a systematic delay of early
morning saliva sampling this might explain our
results. Exam students and controls, however, re-
ported perfect compliance with respect to sam-
pling time. Furthermore, in an recent, yet unpub-
lished, study from our lab saliva samples were
taken in the afternoon under experimenters’ con-
trol. In that study we got very similar results
including a prolonged reduction of sIgA in exam
students but not in controls.
Taken together, it seems to be justified to as-
sume that it was academic stress which induced
the prolonged reduction of sIgA secretion we
observed. The pathway, however, by which stress
affects sIgA for such a long period of time re-
mains subject to future studies. The data of Huck-
 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232 229

Table 5
Spearman rank Žbelow diagonals. and Pearson correlations Žabove diagonals. between sIgA measures at different times

Baseline Last week First week Second week

of exams after exams after exams

Concentration
(mgr dl)
Baseline Exam 0.80 0.11 0.40
Controls 0.64 0.57 0.54
Last week of exams Exam 0.83 0.17 0.39
Controls 0.73 0.71 0.70
First week after exams Exam 0.35 0.38 0.64
Controls 0.59 0.80 0.66
Second week after exams Exam 0.45 0.51 0.80
Controls 0.55 0.73 0.90

Secretion (mgr min)

Baseline Exam 0.78 0.31 0.37
Controls 0.66 0.57 0.47
Last week of exams Exam 0.86 0.61 0.56
Controls 0.64 0.63 0.58
First week after exams Exam 0.39 0.51 0.80
Controls 0.57 0.80 0.54
Second week after exams Exam 0.38 0.51 0.81
Controls 0.43 0.71 0.62

lebridge et al. Ž1998. might, however, point to an
interesting direction. These authors have shown,
that sIgA after awakening is related to cortisol
secretion. They furthermore cite data indicating
that sIgA secretion is influenced beside others by
glucocorticoids ŽSabbadini and Berczi, 1995.. Un-
fortunately, no cortisol data are available from
our subjects. There are, however, numerous data
indicating that prolonged stressors result in alter-
ations of the hypothalamus pituitary adrenocorti-
cal ŽHPA. axis. Future studies should, therefore,
analyse whether there is a relationship between
long-term effects of stress on HPA functioning
and sIgA alterations.
The study of Hucklebridge et al. Ž1998. also
brings up another question: are the post-stress
effects we observed restricted to awakening sam-
ples of sIgA or is the whole diurnal sIgA cycle
affected? Preliminary data from our lab indicate
effect sizes to be larger in the morning than in
the afternoon. This matter, however, warrants
further exploration.
At least two things can be learned from our
observation, so far. First, stress-induced alter-
ations of sIgA are not confined to the presence of
the objective stressor; they persist even if the
stressor is removed. Second, stress-induced alter-
ations of sIgA are not confined to the perception
of stress; they persist even if subjects recover to
control levels in terms of self-reported stress and
recovery. This has implications both for basic and
clinical research. Future research on stress effects
on sIgA should consider that effect sizes might be
largest not during stress but afterwards. The time
of recovery of stress-induced alterations pre-
sumably varies from stressor to stressor. It is not
clear yet on which factors it depends ᎏ duration
of the stressor or its intensity or both. This would
be an interesting question for future studies and
might help to improve our understanding of
stress᎐immune relationships.
There seems to be some disagreement whether
academic stress increases or decreases sIgA. Sev-
eral studies have now been published reporting a
decrease ŽJemmott et al., 1983; Jemmott and Ma-
¨
gloire, 1988; Deinzer and Schuller, 1998. while
others report increases ŽMcClelland et al., 1985;
Bosch et al., 1996.. However, closer inspection of
Bosch’s and McClelland’s data reveals, that they
found the same phenomena that we report in the
230 R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232

present study: several days after the exams sIgA Appendix A. Means and S.D. of daily sIgA
concentrations are below exam values. Indeed, measures (mgrr dl)
Bosch and McClelland took their ‘baseline’ sam-
ples several days ŽMcClelland et al., 1985. and 2
weeks ŽBosch et al., 1996. after exams. Not having Exam students Controls
included control groups into their studies these Mean S.D. Mean S.D.
authors believed that the post-stress decrease of
Baseline
sIgA they observed was indicative for a stress- Day 1 49.28 70.44 50.59 62.25
induced sIgA increase. The majority of studies Day 2 36.67 27.18 35.74 40.87
thus supports the notion that sIgA reaches a Day 3 32.35 48.23 54.34 54.24
minimum some time after termination of pro-
longed periods of stress, like major academic ex- Last week of exams
Day 4 45.93 77.85 39.29 62.13
ams. This does, however, not preclude that exami- Day 5 49.43 86.07 45.42 51.11
nation itself Žas acute stressor. induces a phasic Day 6 34.89 48.70 39.12 51.01
sIgA increase ŽEvans et al., 1994; Bristow et al., Day 7 26.62 30.03 77.15 94.58
1997. although tonic sIgA levels are below con- Day 8 30.24 41.51 54.26 64.72
trol values. To examine this hypothesis we are Day 9 27.58 34.35 49.51 62.03
Day 10 37.87 60.89 56.65 68.62
currently assessing both phasic and tonic sIgA
alterations in subjects under academic stress. First week after exams
In summary, this study is one of the first to Day 11 16.12 21.61 36.62 42.02
demonstrate prolonged immunological stress ef- Day 12 21.84 28.64 42.99 45.82
fects far exceeding the stress period. This has Day 13 23.26 32.62 52.09 61.59
far-reaching implications for future research. We Day 14 25.65 39.38 31.60 39.37
Day 15 40.40 52.72 29.34 34.81
do not know the mechanisms inducing long-last- Day 16 44.49 67.12 43.50 55.86
ing immunological stress effects. We suggest, Day 17 30.17 43.62 45.09 50.23
however, that the post-stress effects we observed
are not restricted to sIgA. In a recent study we Second week after exams
observed prolonged stress and post-stress effects Day 18 21.00 16.58 51.68 57.60
Day 19 28.00 31.24 40.24 39.58
on interleukin 1␤ concentrations in the gingival Day 20 25.43 26.47 53.00 57.70
crevice ŽDeinzer et al., 1999.. Future studies Day 21 22.24 32.96 20.67 32.50
should investigate which other parameters are Day 22 21.16 38.59 39.64 47.75
affected. Furthermore, the clinical relevance of Day 23 31.77 33.29 55.10 54.88
these immune alterations should be studied in Day 24 23.52 30.78 64.52 68.31
more detail. This might be done by assessing
immune reactions after controlled antigen chal-
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