Flow cytometric DNA-quantification of three-color immunophenotyped cells for subpopulation specific determination of aneuploidy and proliferation

Citation
U. Oelschlaegel et al., Flow cytometric DNA-quantification of three-color immunophenotyped cells for subpopulation specific determination of aneuploidy and proliferation, J IMMUNOL M, 253(1-2), 2001, pp. 145-152
Citations number
12
Categorie Soggetti
Immunology
Journal title
JOURNAL OF IMMUNOLOGICAL METHODS
ISSN journal
00221759 → ACNP
Volume
253
Issue
1-2
Year of publication
2001
Pages
145 - 152
Database
ISI
SICI code
0022-1759(20010701)253:1-2<145:FCDOTI>2.0.ZU;2-T
Abstract
A method is described for three-color immunophenotyping and simultaneous DN A-quantification using a flow cytometer equipped with a 488-nm argon laser and a mercury lamp (UV). The approach includes reproducible immunophenotypi ng comparing antigen expression before and after cell manipulation for DNA- measurement. The coefficients of variation after DNA-staining (CV = 3.13 fo r T-cells in peripheral blood and CV = 3.38 for T-cells in bone marrow) wer e adequate for exact DNA-analysis. For aneuploidy detection, a true interna l standard was established measuring, for example, the DNA-content of T-cel ls in B-cell disease simultaneously with the DNA-content of the malignant c ells. Using this method, aneuploidies could be unequivocally detected in 17 out of 24 patients with multiple myeloma. Furthermore, intratumor heteroge neities in DNA-content and antigen expression could be recognized, allowing an exact separation of tumor cells and normal hematopoiesis. The study als o demonstrated the importance of exact immunophenotypic characterization of lymphocyte subpopulations and the determination of their specific prolifer ation, for example after proliferation induction in cell cultures. Future s tudies should address the applicability of this rather simple multiparamete r approach for simultaneous immunophenotyping and DNA-measurement especiall y in the detection of minimal amounts of aneuploid cells after chemotherapy . (C) 2001 Elsevier Science B.V. All rights reserved.