Different nucleolar antigen expression in resting and proliferating human lymphocytes as studied by fluorescence microscopy and flow cytometry.
The aim of this study was to investigate the use of a nucleolar antigen to discriminate between proliferating and resting cells. Antinucleolar antibodies (Si87) were obtained from a scleroderma patient. The specificity of immunostaining was verified and morphological changes in nucleoli were monitored using a fluorescence microscope. Fluorescence of propidium iodide-stained DNA and nucleolar immunofluorescence were measured by flow cytometry. Following phytohaemagglutinin stimulation the number of nucleoli of normal human peripheral blood lymphocytes increased about 3-fold, accompanied by enlargement of nucleolar size. Simultaneously a mean increase in total immunofluorescence per cell by a factor of three was detected. The method developed and applied here allows a discrimination between resting and proliferating human lymphocytes on the basis of their nucleolar antigen content.
Dubben
H H
HH
University of Hamburg, Institute of Biophysics and Radiobiology, F.R.G.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000974:Antibodies, Antinuclear / Q000276:immunology; D000941:Antigens / Q000276:immunology*; D002455:Cell Division / Q000187:drug effects; D002466:Cell Nucleolus / Q000276:immunology*; D002478:Cells, Cultured; D003851:Deoxyribonucleases / Q000494:pharmacology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000502:physiology; D051379:Mice; D008856:Microscopy, Fluorescence; D010835:Phytohemagglutinins / Q000494:pharmacology; D012260:Ribonucleases / Q000494:pharmacology; D012595:Scleroderma, Systemic / Q000097:blood / Q000276:immunology
D000906:Antibodies; D000974:Antibodies, Antinuclear; D000941:Antigens; D010835:Phytohemagglutinins; D003851:Deoxyribonucleases; D012260:Ribonucleases