A whole-blood assay for qualitative and semiquantitative measurements of CD69 surface expression on CD4 and CD8 T lymphocytes using flow cytometry.
A whole-blood flow cytometry-based assay was utilized to assess CD4 and CD8 T-lymphocyte activation in response to phytohemagglutinin (PHA) stimulation. T-lymphocyte activation was assessed by qualitative (percent CD69) and semiquantitative (anti-CD69 antibody binding capacity) measurements of CD69 surface expression. Whole-blood samples from 21 healthy and 21 human immunodeficiency virus (HIV)-infected (<500 absolute CD4 counts per mm3) individuals were stimulated with 20 microg of PHA per ml for 18 to 24 h. The proportions of activated CD4 and CD8 T lymphocytes expressing CD69 (percent CD69) and the levels of CD69 expression on each T-lymphocyte subset (anti-CD69 antibody binding capacity) were measured. By using this assay system, T-lymphocyte activation was impaired in both CD4 and CD8 T-lymphocyte subsets of HIV-infected individuals. The proportions of CD69-positive CD4 and CD8 T lymphocytes were 43 and 27% lower, respectively, in samples from HIV-infected individuals compared to samples from healthy individuals. Similarly, the levels of CD69 expression on each activated CD4 and CD8 T-lymphocyte subset were 48 and 51% lower, respectively. These results suggest that both qualitative and semiquantitative measurements of CD69 surface expression by flow cytometry can be used to assess T-lymphocyte activation.
Lim
L C
LC
McLendon Clinical Laboratories, Clinical Immunology Laboratory, University of North Carolina Hospitals, Chapel Hill 27514, USA.
Fiordalisi
M N
MN
Mantell
J L
JL
Schmitz
J L
JL
Folds
J D
JD
D015703:Antigens, CD / Q000097:blood*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000097:blood*; D017252:CD3 Complex / Q000097:blood; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018956:CD5 Antigens / Q000097:blood; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods*; D015658:HIV Infections / Q000276:immunology; D006801:Humans; D037181:Lectins, C-Type; D008213:Lymphocyte Activation; D010835:Phytohemagglutinins / Q000494:pharmacology; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D018956:CD5 Antigens; D037181:Lectins, C-Type; D010835:Phytohemagglutinins; D000099192:CD69 Antigens