Background: As the frequency of allergic disease and especially the food allergy in children is still increasing - we undertook studies to determine the specific sensitivity of lymphocytes T in order to improve the diagnostics and monitoring of treatment. The usefulness of evaluation CD69 and PCNA expression of T cells from children with cow's milk allergy (CMA) in these procedures was examined.
Aim: The study on usefulness of lymphocytes CD69 and PCNA detection for diagnosis of CMA in children and for monitoring of the treatment.
Patients and methods: The subjects in this study included 26 children from 1.5 to 15 years old with CMA who were under clinical care of our Immunological Department. Patients were examined (medical history of the child, physical examination, specific food provocation) and laboratory investigations were made before therapy and after 6 months of elimination diet. Allergen-specific IgE antibody was determined. Children's T cell cultures were stimulated by beta-lactoglobulin, alpha-casein, whole cow's milk, phytohaemoglutinin PHA and without stimulation. Expression of T cells CD69 and PCNA from patients with CMA and 10 healthy children were analysed by use of flow cytometry. 10 children with CMA after 6 months of elimination diet and next 2 weeks with cow's milk provocation were examined also for CD69 and PCNA expression in lymphocytes from peripheral blood.
Results: After 6 months of elimination diet it seems that the expression of CD69 receptor has tendency to fall, these differences, however, are not statistically significant. But after next 2 weeks with cow's milk diet the expression of CD69 was increased with statistically significance (p=0.04). T cells PCNA expression of children with CMA was lower than in healthy children in PHA stimulated cultures. Our study indicates that PCNA expression of T cells from patients with CMA increased even more after 6 months of elimination diet in all cultures but there was statistically significant differences only after stimulation with cow's milk and PHA (p=0.05 and p=0.04 - respectively). Subsequently through 2 weeks children were provoked with cow's milk and after that PCNA expression in vivo lowered (no statistically significant differences).
Conclusions: CD69 expression of T lymphocytes was significantly higher after specific stimulation of children with cow's milk allergy (p=0.04). The evaluation of CD69 expression of children with CMA may be also useful in monitoring of treatment. Estimation of PCNA expression suggests the efficiency of T cells connected with DNA synthesis related to the stage of allergy disease and demands further investigations.