Skip to main content
British Journal of Cancer logoLink to British Journal of Cancer
. 1981 Jun;43(6):793–803. doi: 10.1038/bjc.1981.117

Fluorescence polarization with FDA in leukaemic cells: a clear difference between myelogenous and lymphocytic origins.

H Tsuda, H Maeda, S Kishimoto
PMCID: PMC2010726  PMID: 6941807

Abstract

Intracellular fluorescence polarization (IFP) values of normal human lymphocytes and leukaemic cells from newly diagnosed patients were determined from fluorescence polarization using fluorescein diacetate (FDA). Thirty healthy donors and 40 patients with various types of leukaemia (20 myelogenous and 20 lymphocytic) were included in the present studies. The result was that myeloid cells had about twice the polarization value of lymphocytic cells. The use of FDA for the determination of IFP appears to be useful for differential diagnosis, at least between acute myelogenous and lymphocytic leukaemias. These 2 types of leukaemia also showed a pronounced difference in fluorescence intensity when treated with FDA, perhaps owing to a difference in uptake velocity. The previously described membrane microviscosity using 1,6-diphenyl-1,3,5-hexatriene (DPH), however, did not show such a difference between these 2 leukaemias. The fluorescein-binding protein(s) was also investigated in order to clarify its effect on IFP, but there seemed little evidence for the existence of any such dyebinding protein(s). The advantages of the present method, using FDA, reside in its simplicity, rapidity and considerable sensitivity, requiring a small sample of blood usually less than 5 ml.

Full text

PDF
793

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andreeff M., Darzynkiewicz Z., Sharpless T. K., Clarkson B. D., Melamed M. R. Discrimination of human leukemia subtypes by flow cytometric analysis of cellular DNA and RNA. Blood. 1980 Feb;55(2):282–293. [PubMed] [Google Scholar]
  2. Bennett J. M., Catovsky D., Daniel M. T., Flandrin G., Galton D. A., Gralnick H. R., Sultan C. Proposals for the classification of the acute leukaemias. French-American-British (FAB) co-operative group. Br J Haematol. 1976 Aug;33(4):451–458. doi: 10.1111/j.1365-2141.1976.tb03563.x. [DOI] [PubMed] [Google Scholar]
  3. Blecher T. E., Bisby R. H. Mononuclear cell-membrane "fluidity": a study in some haematological malignancies. Br J Cancer. 1977 Dec;36(6):763–769. doi: 10.1038/bjc.1977.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bodey G. P., Rodriguez V. Approaches to the treatment of acute leukemia and lymphoma in adults. Semin Hematol. 1978 Jul;15(3):221–261. [PubMed] [Google Scholar]
  5. Brodrick J. W., Glaser C. B., Largman C., Geokas M. C., Graceffo M., Fassett M., Maeda H. Interaction of chymotrypsinogens with alpha 1-protease inhibitor. Biochemistry. 1980 Oct 14;19(21):4865–4870. doi: 10.1021/bi00562a025. [DOI] [PubMed] [Google Scholar]
  6. Cercek L., Cercek B. Application of the phenomenon of changes in the structuredness of cytoplasmic matrix (SCM) in the diagnosis of malignant disorders: a review. Eur J Cancer. 1977 Sep;13(9):903–915. doi: 10.1016/0014-2964(77)90166-9. [DOI] [PubMed] [Google Scholar]
  7. Clarkson B. D., Dowling M. D., Gee T. S., Cunningham I. B., Burchenal J. H. Treatment of acute leukemia in adults. Cancer. 1975 Aug;36(2):775–795. doi: 10.1002/1097-0142(197508)36:2+<775::aid-cncr2820360824>3.0.co;2-v. [DOI] [PubMed] [Google Scholar]
  8. Classification of acute leukemia. Ann Intern Med. 1977 Dec;87(6):740–753. doi: 10.7326/0003-4819-87-6-740. [DOI] [PubMed] [Google Scholar]
  9. Collard J. G., De Wildt A. Localization of the lipid probe 1,6-diphenyl-1,3,5 hexatriene (DPH) in intact cells by fluorescence microscopy. Exp Cell Res. 1978 Oct 15;116(2):447–450. doi: 10.1016/0014-4827(78)90467-6. [DOI] [PubMed] [Google Scholar]
  10. Durr F. E., Monroe J. H., Schmitter R., Traul K. A., Hirshaut Y. Studies on the infectivity and cytopathology of Epstein-Barr virus in human lymphoblastoid cells. Int J Cancer. 1970 Nov 15;6(3):436–449. doi: 10.1002/ijc.2910060315. [DOI] [PubMed] [Google Scholar]
  11. Epstein M., Norman A., Pinkel D., Udkoff R. Flow system fluorescence polarization measurements on fluorescein diacetate-stained EL4 cells. J Histochem Cytochem. 1977 Jul;25(7):821–826. doi: 10.1177/25.7.70456. [DOI] [PubMed] [Google Scholar]
  12. Forman E. N., Padre-Mendoza T., Smith P. S., Barker B. E., Farnes P. Ph1-positive childhood leukemias: spectrum of lymphoid-myeloid expressions. Blood. 1977 Apr;49(4):549–558. [PubMed] [Google Scholar]
  13. Gordon D. S., Hutton J. J., Smalley R. V., Meyer L. M., Volger W. R. Terminal deoxynucleotidyl transferase (TdT), cytochemistry, and membrane receptors in adult acute leukemia. Blood. 1978 Dec;52(6):1079–1088. [PubMed] [Google Scholar]
  14. Greaves M., Janossy G. Patterns of gene expression and the cellular origins of human leukaemias. Biochim Biophys Acta. 1978 Oct 27;516(2):193–230. doi: 10.1016/0304-419x(78)90008-2. [DOI] [PubMed] [Google Scholar]
  15. HUMMEL J. P., DREYER W. J. Measurement of protein-binding phenomena by gel filtration. Biochim Biophys Acta. 1962 Oct 8;63:530–532. doi: 10.1016/0006-3002(62)90124-5. [DOI] [PubMed] [Google Scholar]
  16. Hinuma Y., Konn M., Yamaguchi J., Wudarski D. J., Blakeslee J. R., Jr, Grace J. T., Jr Immunofluorescence and herpes-type virus particles in the P3HR-1 Burkitt lymphoma cell line. J Virol. 1967 Oct;1(5):1045–1051. doi: 10.1128/jvi.1.5.1045-1051.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Inbar M., Shinitzky M., Sachs L. Microviscosity in the surface membrane lipid layer of intact normal lymphocytes and leukemic cells. FEBS Lett. 1974 Jan 15;38(3):268–270. doi: 10.1016/0014-5793(74)80069-4. [DOI] [PubMed] [Google Scholar]
  18. Janossy G., Hoffbrand A. V., Greaves M. F., Ganeshaguru K., Pain C., Bradstock K. F., Prentice H. G., Kay H. E., Lister T. A. Terminal transferase enzyme assay and immunological membrane markers in the diagnosis of leukaemia: a multiparameter analysis of 300 cases. Br J Haematol. 1980 Feb;44(2):221–234. doi: 10.1111/j.1365-2141.1980.tb01204.x. [DOI] [PubMed] [Google Scholar]
  19. Klein E., Klein G., Nadkarni J. S., Nadkarni J. J., Wigzell H., Clifford P. Surface IgM-kappa specificity on a Burkitt lymphoma cell in vivo and in derived culture lines. Cancer Res. 1968 Jul;28(7):1300–1310. [PubMed] [Google Scholar]
  20. Lozzio C. B., Lozzio B. B. Cytotoxicity of a factor isolated from human spleen. J Natl Cancer Inst. 1973 Feb;50(2):535–538. doi: 10.1093/jnci/50.2.535. [DOI] [PubMed] [Google Scholar]
  21. Maeda H. Assay of proteolytic enzymes by the fluorescence polarization technique. Anal Biochem. 1979 Jan 1;92(1):222–227. doi: 10.1016/0003-2697(79)90649-3. [DOI] [PubMed] [Google Scholar]
  22. Maeda H., Ishida N., Kawauchi H., Tsujimura K. Reaction of fluorescein-isothiocyanate with proteins and amino acids. I. Covalent and non-covalent binding of fluorescein-isothiocyanate and fluorescein to proteins. J Biochem. 1969 May;65(5):777–783. doi: 10.1093/oxfordjournals.jbchem.a129077. [DOI] [PubMed] [Google Scholar]
  23. McCaffrey R., Harrison T. A., Parkman R., Baltimore D. Terminal deoxynucleotidyl transferase activity in human leukemic cells and in normal human thymocytes. N Engl J Med. 1975 Apr 10;292(15):775–780. doi: 10.1056/NEJM197504102921504. [DOI] [PubMed] [Google Scholar]
  24. Minowada J., Onuma T., Moore G. E. Rosette-forming human lymphoid cell lines. I. Establishment and evidence for origin of thymus-derived lymphocytes. J Natl Cancer Inst. 1972 Sep;49(3):891–895. [PubMed] [Google Scholar]
  25. Petitou M., Tuy F., Rosenfeld C., Mishal Z., Paintrand M., Jasnin C., Mathe G., Inbar M. Decreased microviscosity of membrane lipids in leukemic cells: two possible mechanisms. Proc Natl Acad Sci U S A. 1978 May;75(5):2306–2310. doi: 10.1073/pnas.75.5.2306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosenthal S., Canellos G. P., DeVita V. T., Jr, Gralnick H. R. Characteristics of blast crisis in chronic granulocytic leukemia. Blood. 1977 May;49(5):705–714. [PubMed] [Google Scholar]
  27. Shinitzky M., Inbar M. Difference in microviscosity induced by different cholesterol levels in the surface membrane lipid layer of normal lymphocytes and malignant lymphoma cells. J Mol Biol. 1974 Jan 5;85(4):603–615. doi: 10.1016/0022-2836(74)90318-0. [DOI] [PubMed] [Google Scholar]
  28. Shinitzky M., Inbar M. Microviscosity parameters and protein mobility in biological membranes. Biochim Biophys Acta. 1976 Apr 16;433(1):133–149. doi: 10.1016/0005-2736(76)90183-8. [DOI] [PubMed] [Google Scholar]
  29. Thomas J. A., Buchsbaum R. N., Zimniak A., Racker E. Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ. Biochemistry. 1979 May 29;18(11):2210–2218. doi: 10.1021/bi00578a012. [DOI] [PubMed] [Google Scholar]
  30. Udkoff R., Norman A. Polarization of fluorescein fluorescence in single cells. J Histochem Cytochem. 1979 Jan;27(1):49–55. doi: 10.1177/27.1.438505. [DOI] [PubMed] [Google Scholar]
  31. Yamane I., Tomioka F. The concomitant effect of unsaturated fatty acid supplemented to medium on cellular growth and membrane fluidity of cultured cells. Cell Biol Int Rep. 1979 Sep;3(6):515–523. doi: 10.1016/0309-1651(79)90087-0. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES