# Evaluating Pediatric Reference Ranges for Extended Immunophenotyping from a Finnish Cohort against Published References

## TL;DR

This study establishes pediatric reference ranges for immune cell subsets and finds differences in immune responses between children and adults.

## Contribution

The study provides new pediatric reference values for immunophenotyping and identifies age-related differences in lymphocyte function.

## Key findings

- Children have lower lymphocyte mitogen responses compared to adults, likely due to higher naïve lymphocyte counts.
- IEI patients show distinct immune cell patterns, including increased transitional B cells and γδ T cells.
- Reference ranges from this study can improve pediatric immunological diagnostics.

## Abstract

Flow cytometric immunophenotyping of lymphocytes and dendritic cells, and functional lymphocyte mitogen response tests are used in the diagnostics of inborn errors of immunity (IEI), especially in pediatrics. These routinely used tests lack sufficient age-matched reference values in children. We established reference values for lymphocyte and dendritic cell subsets for four age groups from 68 healthy children under 12 years of age. These values were then compared to prior publicly available articles and 46 clinical samples from children with confirmed IEI diagnosis. Mitogen response results were also compared between 27 children and 177 adults. In the literature review, we found considerable variability in lymphocyte subset definitions and statistical approaches. Most IEI patients had increased transitional and naïve B, and decreased memory B cells. CHH patients had increased γδ T and DNTs. Lymphocyte stimulation via FASCIA method provides weaker stimulation results in children than in adults, which seems to result from a larger proportional count of naïve lymphocytes in children. The established reference values can be used in diagnostics of pediatric immunological conditions in laboratories that use similar analytic methods. Lower lymphocyte mitogen response results in children need to be taken into consideration when interpreting the results of lymphocyte functional tests.

The online version contains supplementary material available at 10.1007/s10875-025-01959-y.