Simultaneous cytometric analysis for the expression of cytoplasmic and surface antigens in activated T cells.
A method of two-colour immunofluorescence staining has been developed to allow the simultaneous analysis of both surface and cytoplasmic antigens. This involves the use of direct fluorochrome antibody conjugates for cell-surface antigen staining, followed by cell permeabilization and the staining of cytoplasmic antigens with biotinylated antibodies and streptavidin-fluorochrome conjugates. Fluorochrome-antibody conjugates bound to cell-surface epitopes were found not to be affected by the subsequent permeabilisation and cytoplasmic staining. This method was used to examine the surface phenotype of T cells expressing a cytoplasmic antigen, STA. STA is a unique determinant detected in activated human T cells by the monoclonal antibody K-1-21, which also recognizes a cross-reactive conformation-dependent epitope on human free kappa light chains. Cytometric analysis showed that STA is found in both Leu 2a+ cytotoxic/suppressor T cells and Leu 3a+ helper/inducer T cells but is not induced in the Leu 15+ population which contains suppressor T cells. STA was also shown to be an activation antigen in murine T cells.
Hayden
G E
GE
Clinical Immunology Research Centre, University of Sydney, N.S.W., Australia.
Walker
K Z
KZ
Miller
J F
JF
Wotherspoon
J S
JS
Raison
R L
RL
D000818:Animals; D000941:Antigens / Q000032:analysis*; D000954:Antigens, Surface / Q000032:analysis* / Q000276:immunology*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000276:immunology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D008213:Lymphocyte Activation; D008722:Methods; D051379:Mice; D010641:Phenotype; D013601:T-Lymphocytes / Q000276:immunology / Q000648:ultrastructure*; D013950:Thymus Gland / Q000166:cytology; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7
D000941:Antigens; D000954:Antigens, Surface; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7