title: Immunophenotyping of blood lymphocytes in childhood. Reference values for lymphocyte subpopulations.
abstract: OBJECTIVE
  Immunophenotyping of blood lymphocytes is an important tool in the diagnosis of hematologic and immunologic disorders. Because of maturation and expansion of the immune system in the first years of life, the relative and the absolute size of lymphocyte subpopulations vary during childhood. Therefore we wished to obtain reference values for the relative and the absolute size of all relevant blood lymphocyte subpopulations in childhood.
  
  
  METHODS
  We used the lysed whole blood method for analysis of lymphocyte subpopulations in 429 blood samples from neonates (n = 20), healthy children (n = 358), and adults (n = 51). The following age groups were used: 1 week to 2 months (n = 13), 2 to 5 months (n = 46), 5 to 9 months (n = 105), 9 to 15 months (n = 70), 15 to 24 months (n = 33), 2 to 5 years (n = 33), 5 to 10 years (n = 35), and 10 to 16 years (n = 23).
  
  
  RESULTS
  Our results show that the absolute number of CD19+ B lymphocytes increases twofold immediately after birth, remains stable until 2 years of age, and subsequently gradually decreases 6.5-fold from 2 years to adult age. The CD3+ T lymphocytes increase 1.5-fold immediately after birth and decrease threefold from 2 years to adult age. The absolute size of the CD3+/CD4+ T-lymphocyte subpopulation follows the same pattern as the total CD3+ population, but the CD3+/CD8+ T lymphocytes remain stable from birth up to 2 years of age, followed by a gradual threefold decrease toward adult levels. In contrast to B and T lymphocytes, the absolute number of natural killer cells decreases almost threefold in the first 2 months of life and remains stable thereafter. Our study also showed that changes in the absolute size of lymphocyte subpopulations are not always consistent with changes in their relative size. This demonstrates that the relative counts of lymphocyte subsets do not reflect their actual size and are therefore of limited value.
  
  
  CONCLUSIONS
  On the basis of this study we strongly recommend that immunophenotyping of blood lymphocytes for the diagnosis of hematologic and immunologic disorders be based on the absolute rather than on the relative size of lymphocyte subpopulations. Our data can be used as age-matched reference values for blood lymphocyte immunophenotyping.
authors.affiliation: Department of Pediatrics, Sophia Children's Hospital/University Hospital Rotterdam, The Netherlands.
authors.forename: W M
authors.initials: WM
authors.lastname: Comans-Bitter
authors.forename: R
authors.initials: R
authors.lastname: de Groot
authors.forename: R
authors.initials: R
authors.lastname: van den Beemd
authors.forename: H J
authors.initials: HJ
authors.lastname: Neijens
authors.forename: W C
authors.initials: WC
authors.lastname: Hop
authors.forename: K
authors.initials: K
authors.lastname: Groeneveld
authors.forename: H
authors.initials: H
authors.lastname: Hooijkaas
authors.forename: J J
authors.initials: JJ
authors.lastname: van Dongen
mesh_terms: D000293:Adolescent; D000328:Adult; D000375:Aging / Q000097:blood; D016175:B-Lymphocyte Subsets; D002648:Child; D002675:Child, Preschool; D005260:Female; D005434:Flow Cytometry; D006402:Hematologic Diseases / Q000175:diagnosis; D006801:Humans; D007154:Immune System Diseases / Q000175:diagnosis; D016130:Immunophenotyping; D007223:Infant; D007231:Infant, Newborn; D018655:Lymphocyte Count; D008297:Male; D012016:Reference Values; D016176:T-Lymphocyte Subsets