Sequential flow cytometric analysis of cell-cycle related changes in LFA-1 (CD18/CD11a) expression by trisomy 21 (Down's syndrome) lymphoblastoid cells.
Lymphocyte function associated antigen 1 (LFA-1) is a heterodimeric leucocyte adhesion molecule comprising non-covalently associated 95 kD, CD18 and 180 kD, CD11a subunits. Lymphoblastoid cell-lines (LCL) derived from persons with Down's syndrome (Trisomy 21) exhibit increased expression of LFA-1 compared with normal LCL. Although this is probably due to a gene-dosage related increase in the synthesis of CD18, cell-cycle differences between Trisomy 21 (T21) and normal LCL could also influence LFA-1 expression. We have therefore analysed expression of CD18 on G1 and G2M cells using sequential flow cytometry. T21 and normal LCL were co-stained with the DNA-binding vital dye HO342 (Hoechst 33342), and with a CD18 monoclonal antibody. The LCL were first sorted on the basis of HO342 staining into G1 and G2M populations, and these fractions then analysed for CD18 expression. Irrespective of the stage of the cell-cycle, expression of CD18 was increased on T21 compared with normal LCL. Although more CD18 was detected on both T21 and normal G2M compared with G1 cells, the relative density of CD18 in G2M was less than in G1 because G2M cells were larger.
McMurray
B P
BP
Department of Medical Genetics, St Mary's Hospital, Manchester, U.K.
Taylor
G M
GM
Williams
A
A
D'Souza
S
S
Harris
R
R
D015703:Antigens, CD / Q000032:analysis*; D000943:Antigens, Differentiation; D001562:Benzimidazoles; D002453:Cell Cycle; D002460:Cell Line; D002469:Cell Separation; D002891:Chromosomes, Human, Pair 21; D004247:DNA / Q000032:analysis; D004314:Down Syndrome / Q000235:genetics / Q000276:immunology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D015870:Gene Expression; D006801:Humans; D016169:Lymphocyte Function-Associated Antigen-1; D016028:Receptors, Leukocyte-Adhesion; D013601:T-Lymphocytes / Q000276:immunology* / Q000648:ultrastructure; D014314:Trisomy
D015703:Antigens, CD; D000943:Antigens, Differentiation; D001562:Benzimidazoles; D005456:Fluorescent Dyes; D016169:Lymphocyte Function-Associated Antigen-1; D016028:Receptors, Leukocyte-Adhesion; D004247:DNA; C017807:bisbenzimide ethoxide trihydrochloride