A simultaneous three-color T cell subsets analysis with single laser flow cytometers using T cell gating protocol. Comparison with conventional two-color immunophenotyping method.
We describe a method for simultaneous analysis of CD3, CD4, and CD8 positive cells from whole blood utilizing single laser flow cytometers. All three T cell values are attained from a single test tube. CD4 and CD8 positive cells are identified only if they are CD3 positive. Thus the values obtained by this method for T helper/inducer and T cytotoxic/suppressor cells can be reported directly as a percentage of T lymphocytes. Analysis for CD4 and CD8 positive cells is accomplished, by first gating on CD3 positive T lymphocytes, hence the approach is referred to as a T gating method. As the third dye, conjugated to anti-CD3 monoclonal antibodies (MAbs), we utilized peridinin chlorophyll protein (PerCP), a new red fluorochrome. The proposed method may prove to be practical for monitoring disease progression in AIDS, where longitudinal T helper/inducer and T cytotoxic/suppressor cell enumeration must be performed unambiguously by a simple, reproducible, and fast method.
Mandy
F F
FF
Division of Infectious Immunology, Bureau of Laboratories and Research Services, Health and Welfare Canada, Ottawa, Ontario.
Bergeron
M
M
Recktenwald
D
D
Izaguirre
C A
CA
D017252:CD3 Complex / Q000032:analysis; D015704:CD4 Antigens / Q000032:analysis; D016516:CD4-CD8 Ratio; D016827:CD8 Antigens / Q000032:analysis; D002338:Carotenoids; D004396:Coloring Agents; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D015800:Protozoan Proteins; D016176:T-Lymphocyte Subsets / Q000276:immunology*
D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D004396:Coloring Agents; D015800:Protozoan Proteins; C073227:peridinin chlorophyll-a protein, Dinophyceae; D002338:Carotenoids