The phytohemagglutinin response of human peripheral blood lymphocytes as a function of donor age: A re-examination using BrdU-Hoechst flow cytometry

D. Schindler, M. Kubbies, R. E. Priest, H. Hoehn, P. S. Rabinovitch

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Mononuclear cells were isolated from peripheral blood by a standard Ficoll-Hypaque technique from 127 healthy donors, ranging in age from newborns to 86 years of age. As a measure of their in vitro growth response, the fraction of noncycling cells was determined at 48 and 72 h after phytohemagglutinin (PHA) exposure by means of BrdU-Hoechst flow cytometry. This technique provides an optimal assay system for the non-cycling cell fraction, since all cycling cells will have incorporated BrdU thereby quenching the fluorescence of the Hoechst 33258 fluorochrome. Lymphocytes from prepubertal donors showed significantly decreased non-cycling cell fractions, as did lymphocytes from an additional group of 14 adults with hypogonadism due to the 45, XO condition (Turner-Syndrome). Much to our surprise, we found no definitive correlation between donor age and the non-cycling fraction of cells from the adult lymphocyte donors. Nor did we find any age-related increase in the variance of the non-cycling cell fraction. These observations suggest that the previously reported age-related decline in the PHA response of human PBL may reflect an increasing delay, rather than an overall diminution, of the PHA response as a function of donor age.

Original languageEnglish
Pages (from-to)253-263
Number of pages11
JournalMechanisms of Ageing and Development
Volume44
Issue number3
DOIs
StatePublished - Sep 1988
Externally publishedYes

Keywords

  • Aging
  • BrdU-Hoechst
  • Flow cytometry
  • Lymphocytes

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