The issues of assessing reference ranges for the main subpopulations of blood lymphocytes in children at different ages

The age-related dynamics of main subsets of blood lymphocytes in children has been studied quite well, but the issue of the limits of reference values, especially at an early age, remains actual and important. The paper analyzes results of a direct study dataset of 624 blood samples of healthy children aged 1 week to 17 years and 11 months, residents of Moscow and the central Russian regions who underwent a clinical blood count during routine checkups. The main criterion for the complete inclusion of a child into the study was reliable information about the absence of acute and chronic health disorders. The absolute number of lymphocytes was determined as a complex of indices obtained of automated blood analysis, using 5 Diff technology of flow hemocytometry with fluorescent staining of nucleic acids. The percentage and absolute values of T lymphocytes, T helper cells, T cytotoxic cells, the ratio of helper/cytotoxic lymphocytes, as well as percentages and absolute numbers of B lymphocytes and natural killers were determined in residual blood samples by flow cytometry with 3-4 color staining. 14 age groups were formed with the number of cases from 40 to 64 in each group. The Kolmogorov–Smirnov method confirmed the normal distribution of the obtained numerical indicators, both in the general group and in all distinct age groups. The article provides detailed results of processing the data obtained using nonparametric (median, interquartile range, range from 10 to 90 centiles) and parametric (mean and standard deviation) statistics methods. Based on an estimate of the range of 10-90 centiles, the normal limits for the percentages and absolute values of the main lymphocyte subpopulations were calculated. We have been confirmed the well-known trend for a gradual decrease in the absolute number of lymphocytes and absolute values for all major subpopulations, as well as relatively higher indices of helper/cytotoxic ratio in early age groups. A higher percentage of B lymphocytes was detected in children in the age groups from 2 months to 2 years, which, together with a high total number of lymphocytes, determined fairly high absolute B cell counts in these age groups. The proposed normal ranges were validated on an independent group of 75 children aged 2 to 18 years. The individual indexes of the children in the validation group exceeded the reference values in 4 cases (5.3%, with acceptable level of discrepancies < 10%), which allows us to consider the norms validated. The most debatable findings concern a relatively high absolute level of В lymphocytes in young children which is, however, in good agreement with the pooled data of the world-wide meta-analysis data. The proposed normal ranges are designed in a form that is convenient for practical use.

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