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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 By now, much is known about the immediate


effects of stress on salivary IgA. The significance
of stress-induced immune alterations, however,
Secretory Immunoglobulin A is an antibody would be better understood if we knew more
found in high concentrations on all mucosal sur- about their temporal dynamics. That is, when do
faces of the human body including the respiratory stress effects on sIgA peak and for how long do
tract. It is thought to serve as a first line of they persist? Little is known about these ques-
defense against invading organisms and thus, to tions, so far. In a recent paper we analyzed the
play a key role in prevention of infectious disease effects of a major academic exam on sIgA from
ŽTomasi, 1970, 1992; Ogra et al., 1971; Mestecky
the 7th day prior to the exam until the 6th day
et al., 1986; Jemmott and McClelland, 1989;
¨
afterwards ŽDeinzer and Schuller, 1998.. The most
Mestecky and Russell, 1997; Lamm, 1998.. Stress
striking result of this study was that sIgA concen-
is thought to play a role in mucosal infections like
tration not only decreased at exam days but
upper respiratory tract infection or periodontal
dropped further afterwards. Within the 6 days
inflammation ŽMarcenes and Sheiham, 1992;
post-stress period we assessed, sIgA concentra-
Cohen, 1995; Deinzer et al., 1998, 1999; Deinzer
tion remained below baseline; no recovery was
et al., in press.. For these diseases it is known
observable. The present study aimed to replicate
that IgA is involved in the host’s defense against
and extend the results of our earlier study. Most
their pathogens ŽTomasi, 1970, 1992; Ogra et al.,
importantly, we now extended our post-stress as-
1971; Ebersole et al., 1986; Mestecky et al., 1986;
sessment period to 14 days, expecting sIgA con-
Jemmott and McClelland, 1989; Ogawa et al.,
1991; Nieminen et al., 1993; Somer et al., 1993; centration to recover at least in part within that
Mestecky and Russell, 1997; Lamm, 1998.. period.
Secretory IgA, especially salivary IgA ŽsIgA. Academic stress appears to be an ideal setting
became a focus of interest in psychoimmunologi- for the assessment of temporal dynamics of
cal research since it has been shown to be sensi- stress-induced immune alterations. The immedi-
tive to variations in subjective and objective stress ate immunological effects of academic stress are
levels. Acute challenges like assessed seminar well known ŽJemmott et al., 1983; Kiecolt-Glaser
presentations ŽEvans et al., 1994; Bristow et al., et al., 1984; Glaser et al., 1985, 1990, 1991; Mc-
1997., competition stress in soccer coaches ŽKu- Clelland et al., 1985; Vassend and Halvorsen,
gler et al., 1996., 100-min working sessions of air 1987; Jemmott and Magloire, 1988; Mouton et al.,
traffic controllers ŽZeier et al., 1996. and a ¨
1989; Bosch et al., 1996; Deinzer and Schuller,
laboratory computer game task ŽCarroll et al., 1998.. The stressor is of high ecological validity
1996., have all been shown to be associated with and defined exam conditions guarantee some de-
sIgA increases. sIgA levels and secretion rates gree of standardization. Subjects as well as ade-
were found to be related to desirability of daily quate control persons are easily available. Addi-
events ŽEvans et al., 1993; Stone et al., 1994., an tionally, as the termination of the objective stres-
effect suggested to be mediated at least in part by sor is clearly defined Ži.e. the exam or last exam
mood: negative mood tends to be associated with within a prolonged exam period., one can also
lower sIgA ŽStone et al., 1987a,b, 1996.. De- study immunological post-stress effects.
creases in sIgA have been observed at parturition There have been suggestions that alterations
ŽAnnie and Groer, 1991. and during prolonged within the post-stress period may account for
periods of academic stress ŽJemmott et al., 1983; development of disease ŽDeinzer, 1992. though it
Jemmott and Magloire, 1988; Deinzer and is not clear today whether this should be due to
¨
Schuller, 1998. although there is some inconsis- immunological alterations. However, the data of
tency in reports on academic stress effects ŽMc- our previous study strongly support the notion
Clelland et al., 1985; Bosch et al., 1996; see Sec- that mucosal immune function is compromised
tion 4 of this paper.. 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 2.2. Academic stress ¨ s. control condition
relevant to this finding and thus, to improve our
understanding of immunological post-stress ef-
fects. 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
2. Methods remaining 27 Žcontrol group. did not participate
in any exam but rather enjoyed their vacation.
The medical exam under consideration consists of
2.1. Subjects 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
Fifty-four medical students Ž27 exam students
weeks. Exam results are given immediately after
and 27 controls. provided a full set of valid data
the oral exam. Missing the exam impedes the
for this study. At the beginning of the study,
studies by at least 6 months. Additionally, only
subjects were asked about their age, number of
two failures of the exam are allowed; afterwards
semesters they studied medicine, exercise, regular
the student must discontinue his or her studies of
nicotine consumption, use of oral contraceptives
medicine. The failure rate of this exam varies
and whether they had a regular partner. These
between 25 and 35%.
data are provided in Table 1. Groups did not
differ on any of these variables.
Recruitment of subjects was done during their 2.3. Sali¨ a samples and biochemical analyses
seminars; in addition, fliers requesting their par-
ticipation were distributed throughout the cam-
pus. The students received monetary compensa- Subjects were instructed to give unstimulated
tion ŽDM 50. for participation. Exclusion criteria saliva samples by means of a Salivette 䊛 ŽSALI-
for participation were pregnancy, psychiatric dis- VETTE no. 51.1534, plain cotton roll; Sarstedt,
orders, regular medication with known effects on ¨
Numbrecht, Germany. every morning immedi-
the immune system, and chronic diseases affect- ately after awakening and before doing anything
ing the immune system. 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 oral exam until day of oral exam.;
activities and individual circadian rhythms. Previ- 䢇 first week after exams Ž7 days, 1st᎐7th day
ous studies of Hucklebridge et al. Ž1998. and our after oral exam.; and
group ŽStiller-Winkler et al., 1998. have shown 䢇 second week after exams Ž7 days, 8th᎐14th
that sIgA concentration peaks at awakening, day after oral exam..
shows a steep fall for the next 4 h and then
remains constant. The morning sIgA peak seems During these study periods subjects took saliva
to depend on the time of awakening rather than samples every morning. Daily saliva samples were
the time of the day ŽHucklebridge et al., 1998.. thus taken during the baseline period and from
Students were instructed to hold the cotton swab the 6th day prior to the oral exam until the 14th
of the Salivette 䊛 in the mouth for 5 min and not day afterwards.
to swallow during that time. Salivettes were For each exam student there was a control
weighed before and after sampling and saliva subject of the same sex who took saliva samples
secretion Ždlrmin. was estimated from the dif- at the same days as his or her exam partner did.
ference between these weights. sIgA secretion We took ‘baseline’ measures a few days after
Žmgrmin. was calculated based upon concentra- the decision as to whether students were allowed
tion, saliva volume and sampling time Ži.e. 5 min.. to participate in the examination or not. At that
Salivettes were stored at y20⬚C within 6 h of time, most of the students were already preparing
sampling until analysis wstorage at room tempera- for their exams. Thus, the ‘baseline’ period may
ture for less than 48 h does not affect sIgA not be considered as real baseline in a sense that
measures ŽHennig, 1994.x. Salivary IgA concen- exam students and controls were completely com-
tration was measured by a Behring nephelometer parable in terms of stress. Rather, the ‘baseline’
analyzer ŽMarburg, Germany.. Samples were period gives us some idea about sIgA at the
thawed and spun for 10 min at 6000 rev.rmin. beginning of a prolonged examination period as
For the sIgA determination specific human anti- compared to the end of the stress period and the
IgA serum ŽBehring, Marburg, Germany. was time afterwards.
used. The reliability of this nephelometric tech-
nique for repeated analyses is rtt s 0.98; unpub- 2.5. Self-reported stress and reco¨ ery
lished data from our laboratory show that these
results correlate with those of the radial im- To validate the subjective significance of the
munodiffusion technique for determination of se- exam as stressor every subject filled out the recov-
cretory sIgA ŽBinding Site, Heidelberg, Germany. ery-stress-questionnaire wRESTQ ŽErholungs-Be-
of r s 0.85. lastungs-Fragebogen.; Kellmann and Kallus, 1993;
Salivary IgA analyses for one person were al- Kallus, 1995a,bx at times indicated in Fig. 1. This
ways run together and with the same reagents questionnaire assesses stress and recovery during
and standard curve. The person who did the anal- the last Ž3. days on 12 different scales, which can
yses was blind to the condition of each subject take values between 0 and 6. The internal consis-
and to the study design. 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:
2.4. Design
䢇 general stress Žgeneral feeling of too much
The study was subdivided into four study peri- stress to cope with.;
ods: 䢇 emotional stress Žfeelings of dissonance and
general irritation.;
䢇 ‘baseline’ Ž3 days, 4 weeks prior to the written 䢇 social stress Žannoyance or anger at others.;
exam.; 䢇 pressure Žunresolved conflicts and pressure.;
䢇 last week of exams Ž7 days, 6th day prior to 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; l. of wine Ži.e. approx. 16 g alcohol. according to
postponing important tasks.; the guidelines of Feuerlein et al. Ž1991..
䢇 somatic complaints Žheadaches, general so-
matic discomfort.;
2.7. Statistical analyses
䢇 success Žhaving success, making important de-
cisions.;
䢇 social relaxation Žhaving laughed, spent nice To avoid singular matrices in repeated mea-
hours with friends.; sures analysis of variance and to improve clarity
䢇 somatic relaxation Žfeeling of somatic relax- of presentation we averaged ᎏ and thus aggre-
ation and efficiency.; gated into one measure for each period ᎏ the
䢇 general well-being Žbeing in a good temper.; daily sIgA measures of the four study periods.
and Averaging across study periods also allows for
䢇 sleep Žrelaxing and calm sleep.. compensation of low-volume saliva samples at
single days which are not analyzable. Indeed,
According to our earlier results ŽDeinzer and some subjects provided insufficient saliva samples
¨
Schuller, 1998. we expected students to show at some days. However, all cases provided at least
increased levels of subjective stress and reduced three sufficient samples per 7-day study period
levels of recovery during the exam period and to Žwhich indeed was the criterion to be included
recover to control levels within a few days after into analyses ., and only six cases per group pro-
the exam. vided in one period less than six sufficient sam-
ples.
Differences across periods and group by peri-
2.6. Medication and alcohol consumption
ods interactions were examined by repeated mea-
sures ANOVAs and Greenhouse᎐Geisser correc-
To be able to control for influences due to tions for sphericity were applied to significance
medication or alcohol consumption we instructed statements. Below, original degrees of freedom
students to report every use of medication and and epsilon values are presented. A significant
the amount of alcoholic beverages they con- group by study period interaction was expected if
sumed. Students received a little diary where they sIgA recovered during the study. For exploratory
noted every pharmaceutic drug they used includ- reasons, several t-tests were run on sIgA data.
ing the product’s name and the dosage they had For these comparisons we also computed effect
applied as well as the type and volume of al- sizes Ž d .. According to Cohen Ž1992. effects sizes
coholic beverages they had consumed. Students of ds 0.2, 0.5 and 0.8 can be considered as small,
were asked to make diary entries every evening medium and large, respectively.
before they went to sleep. Data on alcohol con- Outlying data Ži.e. data exceeding their respec-
sumption were processed in units of one glass Ž0.2 tive group means at a given study period by more

Table 2
Group differences on mean daily alcohol consumption

Baseline Last week First week Second week


of exams after exams after exams

Exam students 0.33" 0.490 0.24" 0.30 1.22" 1.63 0.68" 0.22
Controls 0.86" 0.88 0.97" 0.91 0.88" 0.66 0.86" 0.76
U
t Žd.f.. t Ž49. s 2.7 t Ž51. s 3.9 t Ž47. s 1.0 t Ž47. s 0.7
P Žtwo-tailed. 0.009 - 0.001 0.328 0.49
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.
R. Deinzer et al. r International Journal of Psychophysiology 37 (2000) 219᎐232 225

than 2 S.D.. were excluded prior to analyses. The 1 which shows the results of the recovery-stress-
exclusion of outlying data is necessary within small questionnaire. Exam students reported more
sample sizes to avoid statistical artifacts. A single stress and less recovery in the last week of exams
outlying value can considerably affect the magni- while control group values remained constant.
tude and even the direction of group differences. Within 3 days post-examination exam students
Four subjects per group were to be excluded from recovered to control values in all but the success
analyses due to outlying data. Inclusion of these scales. At that time they even reported less lack
subjects did not change the overall picture of of energy, less somatic complaints and more so-
results. cial relaxation than controls.
Kolmogorov᎐Smirnov᎐Goodness of Fit Tests Fig. 2 shows sIgA concentrations and secretion
were computed to test for normality; for these rates throughout study periods and Table 4 gives
tests an alpha-level of 20% was considered to be the corresponding statistics for group compar-
significant. Square root transformations were per- isons at each study period. Since study groups did
formed for variables which were not normally not differ with respect to baseline values, these
distributed. This was necessary for baseline mea- were included as covariates for repeated mea-
sures of sIgA concentration and secretion, only.
sures ANOVAs analyzing recovery of sIgA from
To make sure that group differences in sIgA
prior to exams until 2 weeks after exams. A
were not due to differences in medication all
significant group by study period interaction was
analyses were also run restricted to those subjects
expected if there was any recovery of sIgA within
who never took any medication. The results of
2 weeks post-stress. However, this was neither
these analyses were, however, virtually the same
found for sIgA concentration Ž F2,43 s 0.84, ␧ s
as the results of the complete group. Thus, stress
effects were not due to differences in medication 0.83, Ps 0.43. nor for secretion rates Ž F2,43 s
of the two study groups. 0.25, ␧ s 0.88, Ps 0.76. although the main effect
Since groups differed in terms of daily alcohol for groups was significant for concentration Ž F1,43
consumption prior to exams Žbut not afterwards; s 4.96, Ps 0.03. but not for secretion data Ž F1,43
Table 2. we examined the relationship between s 1.94, Ps 0.17.. Table 4 shows a general trend
mean daily alcohol consumption and sIgA levels. of effect sizes for secretion data to be smaller
However, Spearman rank correlations revealed than those for concentration data; but even for
no significant relations at any time ŽTable 3.. 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-
3. Results
dents: 23.52" 30.78 mgrdl; controls: 64.52"
68.31 mgrdl; t Ž39. s 2.50; Ps 0.005; ds 0.71.
The exam was a potent stressor in terms of and secretion rates Žexam students: 0.08" 0.11
psychological self-report as can be seen from Fig. mgrmin; controls: 0.19" 0.23 mgrmin; t Ž39. s

Table 3
Spearman rank correlations between mean daily alcohol consumption and sIgA

Baseline Last week First week Second week


of exams after exams after exams

␳ y0.1545 0.2191 y0.0172 0.0652


U
Valid N 48 48 46 45
P 0.29 0.14 0.91 0.67
Žtwo-tailed.
U
Different Ns are due to inaccurate recordings of alcohol consumption and outlying sIgA data.
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 considerably larger within highly stressed exam
still considerably lower than the respective con- students Župper tertile: 38.96" 61.2 mgrdl; lower
trol levels. tertile: 20.25" 27.6 mgrdl.. This difference, how-
Table 5 shows the correlation between aggre- ever, missed the intended level of significance by
gated sIgA measures for exam students and con- far w t Ž16. s 0.84, Ps 0.20x which presumably is
trols. sIgA was fairly stable within control stu- due to the low sample size available for this
dents while in exam students lower correlations comparison; the effect size d of this comparison
were found between last week of exams and equaled ds 0.39. A simultaneous comparison of
post-exam weeks. This is what one would expect if the three tertiles via ANOVA revealed a similar
an increasing part of intra-individual variance is result: Cohen’s effect size f ŽCohen, 1992. was
determined by situational Že.g. stress . influences medium to large Ž f s 0.33.; the level of signifi-
Žsee the discussion on state-trait influences on cance, however, was even worse Ž F2.20 s 1.05, Ps
intra-individual variation in Deinzer et al., 1995.. 0.37. which is due to the fact that ANOVAs
To assess the relation between subjective stress examine non-directed hypotheses, only. Secretion
and reduction of sIgA concentration the three rates again revealed a considerably smaller effect
stress scales best differentiating between exam size Župper terzil: 0.058" 0.17 mgrmin; lower
students and controls Žgeneral stress, pressure terzil: 0.048" 0.08 mgrmin; t Ž15. s 0.20, Ps
and overfatigue. were aggregated into one mea- 0.41; ds 0.10; ANOVA: F2,19 s 0.92, Ps 0.41;
sure describing subjective stress during the last f s 0.31..
weeks of exams. This measure equals the mean of Pearson and Spearman correlations revealed
the three scales at days y3 and 0. We compared no significant relationship between the time of
exam students in the upper and lower tertile of the oral exam and sIgA concentrations or secre-
this measure. Decrease of sIgA concentration tion rates for any group at any time. Žy0.27F r F
from baseline to the last week of exams was 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᎐232 227

4. Discussion similar to what we observed in our previous study.


We suggest this effect to be due to some impreci-
The present study replicates and extends re- sions in assessment of saliva secretion adding
sults of a previous study from our laboratory error variance to the data. The reliability of saliva
¨
ŽDeinzer and Schuller, 1998.. During a prolonged secretion measures very much depends on proce-
period of academic stress, sIgA concentrations dural control: subjects are not allowed to swallow
after awakening were reduced as compared to during assessment; they should not move the cot-
baseline and to control values. Furthermore, no ton swab or even chew on it during assessment in
recovery of sIgA concentration could be observed order to avoid stimulation of saliva flow; and they
within 14 days post-stress even though students should strictly keep the given sampling duration.
recovered very soon in terms of self-reported In a field study like ours, however, it is not
stress and recovery. Recovery in sIgA concentra- possible to perfectly control saliva sampling pro-
tion showed a tentative relation to subjective cedures. This, however, can adversely affect the
stress responses at the time of the exams: those reliability of sIgA secretion data. A comparison of
subjects showing the largest stress responses the re-test correlations given in Table 5 supports
showed the lowest recovery of sIgA concentra- this notion: within control students re-test corre-
tion. lations of secretion data tend to be smaller than
Secretion data mirrored concentration data al- those of concentration data. Irrespective of these
though effect sizes appeared to be smaller. This is considerations we run another analysis to make

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.
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 alterations in the compliance of exam students
after exams were not merely due to differences in with respect to sampling time. There is a rapid
saliva volume: an ANCOVA with the covariates, fall in sIgA immediately after awakening ŽHuckle-
IgA concentration at baseline, saliva volume at bridge et al., 1998; Stiller-Winkler et al., 1998..
baseline and saliva volume in the second week Thus, if there was a systematic delay of early
after exams, revealed a highly significant group morning saliva sampling this might explain our
effect on sIgA concentration in the second week results. Exam students and controls, however, re-
after exams Ž F41,1 s 7.02; Ps 0.011. ported perfect compliance with respect to sam-
The group differences we observed were not pling time. Furthermore, in an recent, yet unpub-
due to differences in medication. In our earlier lished, study from our lab saliva samples were
¨
studies ŽDeinzer and Schuller, 1998. we had also taken in the afternoon under experimenters’ con-
found that daily sleep has no effect on sIgA trol. In that study we got very similar results
levels. Several other alternative explanations can including a prolonged reduction of sIgA in exam
also be excluded: our study groups did not differ students but not in controls.
with respect to age, sex, number of semesters Taken together, it seems to be justified to as-
studied, exercise, smoking, partnership, and use sume that it was academic stress which induced
of oral contraceptives ŽTable 1.. Furthermore, the prolonged reduction of sIgA secretion we
daily alcohol consumption does not seem to have observed. The pathway, however, by which stress
any influence on sIgA concentration ŽTable 3.. affects sIgA for such a long period of time re-
One might argue that effects may be due to 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 the objective stressor; they persist even if the
interesting direction. These authors have shown, stressor is removed. Second, stress-induced alter-
that sIgA after awakening is related to cortisol ations of sIgA are not confined to the perception
secretion. They furthermore cite data indicating of stress; they persist even if subjects recover to
that sIgA secretion is influenced beside others by control levels in terms of self-reported stress and
glucocorticoids ŽSabbadini and Berczi, 1995.. Un- recovery. This has implications both for basic and
fortunately, no cortisol data are available from clinical research. Future research on stress effects
our subjects. There are, however, numerous data on sIgA should consider that effect sizes might be
indicating that prolonged stressors result in alter- largest not during stress but afterwards. The time
ations of the hypothalamus pituitary adrenocorti- of recovery of stress-induced alterations pre-
cal ŽHPA. axis. Future studies should, therefore, sumably varies from stressor to stressor. It is not
analyse whether there is a relationship between clear yet on which factors it depends ᎏ duration
long-term effects of stress on HPA functioning of the stressor or its intensity or both. This would
and sIgA alterations. be an interesting question for future studies and
The study of Hucklebridge et al. Ž1998. also might help to improve our understanding of
brings up another question: are the post-stress stress᎐immune relationships.
effects we observed restricted to awakening sam- There seems to be some disagreement whether
ples of sIgA or is the whole diurnal sIgA cycle academic stress increases or decreases sIgA. Sev-
affected? Preliminary data from our lab indicate eral studies have now been published reporting a
effect sizes to be larger in the morning than in decrease ŽJemmott et al., 1983; Jemmott and Ma-
the afternoon. This matter, however, warrants ¨
gloire, 1988; Deinzer and Schuller, 1998. while
further exploration. others report increases ŽMcClelland et al., 1985;
At least two things can be learned from our Bosch et al., 1996.. However, closer inspection of
observation, so far. First, stress-induced alter- Bosch’s and McClelland’s data reveals, that they
ations of sIgA are not confined to the presence of 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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