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Clinical and Diagnostic Laboratory Immunology, May 1998, p. 392-398, Vol. 5, No. 3
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Whole-Blood Assay for Qualitative and Semiquantitative Measurements of CD69 Surface Expression on CD4 and CD8 T Lymphocytes Using Flow Cytometry

Lony C. L. Lim, Michelle N. Fiordalisi, Janet L. Mantell, John L. Schmitz,* and James D. Folds

McLendon Clinical Laboratories, Clinical Immunology Laboratory, University of North Carolina Hospitals, Chapel Hill, North Carolina 27514

Received 11 July 1997/Returned for modification 20 October 1997/Accepted 9 February 1998

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 µg 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.

* Corresponding author. Mailing address: McLendon Clinical Laboratories, Clinical Immunology Laboratory, University of North Carolina Hospital, CB#7600, 1035 East Wing, 101 Manning Dr., Chapel Hill, NC 27514. Phone: (919) 966-8453. Fax: (919) 966-0486. E-mail: jschmitz.dhl1{at}mail.unch.unc.edu.

Clinical and Diagnostic Laboratory Immunology, May 1998, p. 392-398, Vol. 5, No. 3
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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