Immunophenotyping of T lymphocytes by three-color flow cytometry in healthy newborns, children, and adults.
Reagents are now available which allow simultaneous assessment of three different fluorescence wave-lengths on most commercially available flow cytometers. Such three-color analyses provide more information than single- or dual-color analyses. The present study was undertaken in order to establish age-related differences in lymphocyte subpopulations by simultaneously measuring three surface antigens in newborns, children, and adults. A whole blood method was used to label cells with antibodies conjugated to FITC, PE, and perCP. We found that the percentage of lymphocytes expressing HLA-DR/CD28/CD8, HLA-DR/ CD38/CD8, CD95/CD45RO/CD8, CD95/CD45RO/CD4, CD95/CD4, and CD95/CD8 showed relative increases with age. The percentage of lymphocytes expressing CD28/CD8, CD38/CD8, and CD38/CD4 showed relative decreases with age, while the subset HLA-DR/CD38/ CD4 did not change. Three-color flow cytometry is a powerful tool to more precisely define lymphocyte subsets than the current two-color methods. We present values using a three-color panel in healthy newborns, children, and adults.
McCloskey
T W
TW
Division of Allergy/Immunology, North Shore University Hospital, New York University School of Medicine, Manhasset 11030, USA.
Cavaliere
T
T
Bakshi
S
S
Harper
R
R
Fagin
J
J
Kohn
N
N
Pahwa
S
S
D000293:Adolescent; D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D002648:Child; D002675:Child, Preschool; D005260:Female; D005434:Flow Cytometry; D015658:HIV Infections / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D007231:Infant, Newborn; D018655:Lymphocyte Count / Q000379:methods; D008297:Male; D008875:Middle Aged; D012307:Risk Factors; D016176:T-Lymphocyte Subsets / Q000145:classification* / Q000276:immunology
D015703:Antigens, CD