Two-color flow cytometric analysis of preterm and term newborn lymphocytes

Abstract

Immune system maturation proceeds postnatally in humans. Therefore, newborns, especially those of a lower gestational age, are not fully immunocompetent and are more likely to acquire perinatal infections. In order to investigate the neonatal immune system status, the major lymphocyte subpopulations were studied in newborns of different gestational age, comparing term newborns and adults. The cord blood from 66 newborns and the peripheral blood from 23 adults were analyzed using fluorochrome labelled monoclonal antibodies and two-color flow cytometry. The newborns were divided into three groups according to their gestational age. Ten prematures were under 32 weeks of gestation, 35 were of 32-37 weeks and there were 21 term newborns. The percentage of cytotoxic T lymphocytes (CD4 CD8+) was lower in term newborns as compared to the adult controls (17.8 versus 30.3%), and so were the percentages of activated T lymphocytes (CD3+Ia+; 0.3 versus 3.7%), cytotoxic non-MHC restricted T lymphocytes (CD3+CD16+CD56+; 0.2 versus 1.8%) and NK cells (CD3-CD16+CD56+; 4.8 versus 15.5%). On the contrary, the proportions of unlabelled cells were increased in term cord blood. The expression of CD45R0 marker on neonatal lymphocytes was very low (1%). In comparison to the higher-gestation newborns, the lower gestation prematures had reduced percentages of T lymphocytes (CD3+; 43 versus 65%), mostly helper T lymphocytes (CD4+CD8-; 35 versus 50%), and increased percentages of unlabelled cells. The percentages of NK cells (CD3+CD16+CD56+) and B lymphocytes (CD3-CD19+; CD3-Ia+) did not differ among the tested newborn groups. There were no significant differences in major lymphocyte subpopulations between the group of highest-gestation prematures and the group of term newborns that differed significantly when compared to adults. The lowest-gestation newborns showed the most immature lymphocyte phenotype with the highest percentages of unlabelled cells.