Background. The use of easily accessible biomarkers for assessing young patients’ health is weighty. This cohort study is aimed at measuring stress/immune biomarkers in the saliva of healthy school-age children and comparing subgroups according to age, sex, and stress perception. Material and Methods. 503 children under 12 years old (
8.7
±
1.3
) were included with anthropometric evaluation (height, waist, hip circumference, body weight, and body mass index (BMI)). Levels of opiorphin (OPI), free cortisol, alpha-amylase (sAA), and secreted immunoglobulin (sIgA) were determined by quantitative assays (ELISA) in unstimulated saliva. Unpaired
t
-test, Welch test, and Mann–Whitney
U
test were applied for appropriate group comparisons, and the correlation between variables was analyzed with Spearman’s rank coefficient. Results were considered significant at
p
<
0.05
. Results. sIgA and sAA exhibited significant differences depending on age and sex: IgA (ng/mL):
86
±
68.6
vs.
104.9
±
72.1
for (6-7 y.o.) and (8-11 y.o.), respectively, and
108.1
±
80.1
vs.
94.6
±
62.2
for male and females, respectively; sAA (U/mL):
78.9
±
54.4
vs.
100.5
±
81.2
for (6-7 y.o.) and (8-11 y.o.). No difference related to age or sex between groups was observed for cortisol and OPI. However, OPI levels were higher and correlated to prior stress exposure in children (
0.31
±
0.4
vs.
0.26
±
0.5
ng
/
mL
,
p
=
0.031
). sAA was negatively correlated to low mood self-declaration in children in the last two weeks (
r
=
−
0.10
,
p
=
0.045
). Conclusions. sIgA and sAA can be used as sex- and age-related biomarkers in children 6-12 y.o., which is not the case for free cortisol and opiorphin. However, OPI reflected previous exposure to stress, suggesting its use for evaluating stress-related changes in children