Hostname: page-component-78c5997874-8bhkd Total loading time: 0 Render date: 2024-11-06T09:55:42.680Z Has data issue: false hasContentIssue false
  • English
  • Français

The Use of CFSE-like Dyes for Measuring LymphocyteProliferation : Experimental Considerations and Biological Variables

Published online by Cambridge University Press:  17 October 2012

B.J.C. Quah ,
A.B. Lyons  and
C.R. Parish
B.J.C. Quah
Affiliation:
Department of Immunology, John Curtin School of Medical Research, Australian National University, Canberra, ACT, 2601, Australia
A.B. Lyons
Affiliation:
School of Medicine, University of Tasmania, Hobart, Tasmania, Australia
C.R. Parish*
Affiliation:
Department of Immunology, John Curtin School of Medical Research, Australian National University, Canberra, ACT, 2601, Australia
*
⋆ Corresponding author. E-mail:[email protected]
Get access

Abstract

The measurement of CFSE dilution by flow cytometry is a powerful experimental tool tomeasure lymphocyte proliferation. CFSE fluorescence precisely halves after each celldivision in a highly predictable manner and is thus highly amenable to mathematicalmodelling. However, there are several biological and experimental conditions that canaffect the quality of the proliferation data generated, which may be important to considerwhen modelling dye dilution data sets. Here we overview several of these variablesincluding the type of fluorescent dye used to monitor cell division, dye labellingmethodology, lymphocyte subset differences, in vitro versus in vivo experimental assays,cell autofluorescence, and dye transfer between cells.

Keywords

Cell proliferationCFSEFlow cytometry
Type
Research Article
Information
Mathematical Modelling of Natural Phenomena , Volume 7 , Issue 5: Immunology , 2012 , pp. 53 - 64
Copyright
© EDP Sciences, 2012

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Références

Weston, S.A., Parish, C.R.. New fluorescent dyes for lymphocyte migration studies. Analysis by flow cytometry and fluorescence microscopy. J Immunol Methods, 133 (1990), no. 1, 87–97. CrossRefGoogle Scholar
Lyons, A.B., Parish, C.R.. Determination of lymphocyte division by flow cytometry. J Immunol Methods, 171 (1994), no. 1, 131–7. CrossRefGoogle ScholarPubMed
Lyons, A.B.. Divided we stand : tracking cell proliferation with carboxyfluorescein diacetate succinimidyl ester. Immunol Cell Biol, 77 (1999), no. 6 , 509–15. CrossRefGoogle ScholarPubMed
Nordon, R.E., Ginsberg, S.S., Eaves, C.J.. High-resolution cell division tracking demonstrates the FLt3-ligand-dependence of human marrow CD34+CD38- cell production in vitro. Br J Haematol, 98 (1997), no. 3, 528–39. CrossRefGoogle ScholarPubMed
Parish, C.R., Glidden, M.H., Quah, B.J., Warren, H.S.. Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation. Curr Protoc Immunol, 84 (2009), 4.9.1–4.9.13. Google Scholar
Lastóvicka, J., Budinsky, V., Spisek, R., Bartunkova, J.. Assessment of lymphocyte proliferation : CFSE kills dividing cells and modulates expression of activation markers. Cell Immunol, 256 (2009), no. 1-2, 79–85. CrossRefGoogle Scholar
Quah, B.J., Parish, C.R.. New and improved methods for measuring lymphocyte proliferation in vitro and in vivo using CFSE-like fluorescent dyes. J Immunol Methods, 379 (2012), no. 1-2, 1–14. CrossRefGoogle Scholar
Quah, B.J., Warren, H.S., Parish, C.R.. Monitoring lymphocyte proliferation in vitro and in vivo with the intracellular fluorescent dye carboxyfluorescein diacetate succinimidyl ester. Nat Protoc, 2 (2007), no. 9, 2049–56. CrossRefGoogle Scholar
B.J. Quah, C.R. Parish. The use of carboxyfluorescein diacetate succinimidyl ester (CFSE) to monitor lymphocyte proliferation. Journal of visualized experiments : JoVE, 44 (2010).
Morrison, R.N., Lyons, A.B., Nowak, B.F., Hayball, J.D.. Snapper (Pagrus auratus) leucocyte proliferation is synergistically enhanced by simultaneous stimulation with LPS and PHA. Fish Shellfish Immunol, 16 (2004), no. 3, 307–19. CrossRefGoogle ScholarPubMed
Holyoake, T., Jiang, X., Eaves, C., Eaves, A.. Isolation of a highly quiescent subpopulation of primitive leukemic cells in chronic myeloid leukemia. Blood, 94 (1999), no. 6, 2056–64. Google ScholarPubMed
Ueckert, J.E., Nebe von-Caron, G., Bos, A.P., ter Steeg, P.F.. Flow cytometric analysis of Lactobacillus plantarum to monitor lag times, cell division and injury. Lett Appl Microbiol, 25 (1997), no. 4, 295–9. CrossRefGoogle Scholar
Hodgkin, P.D., Lee, J.H., Lyons, A.B.. B cell differentiation and isotype switching is related to division cycle number. J Exp Med, 184 (1996), no. 1, 277–81. CrossRefGoogle Scholar
Gett, A.V., Hodgkin, P.D.. Cell division regulates the T cell cytokine repertoire, revealing a mechanism underlying immune class regulation. Proc Natl Acad Sci U S A, 95 (1998), no. 16, 9488–93. CrossRefGoogle Scholar
Hasbold, J., Lyons, A.B., Kehry, M.R., Hodgkin, P.D.. Cell division number regulates IgG1 and IgE switching of B cells following stimulation by CD40 ligand and IL-4. Eur J Immunol, 28 (1998), no. 3, 1040–51. 3.0.CO;2-9>CrossRefGoogle ScholarPubMed
Hasbold, J., Hodgkin, P.D.. Flow cytometric cell division tracking using nuclei. Cytometry, 40 (2000), no. 3, 230–7. 3.0.CO;2-E>CrossRefGoogle ScholarPubMed
Blake, S., Hughes, T.P., Mayrhofer, G., Lyons, A.B.. The Src/ABL kinase inhibitor dasatinib (BMS-354825) inhibits function of normal human T-lymphocytes in vitro. Clin Immunol, 127 (2008), no. 3, 330–9. CrossRefGoogle ScholarPubMed
Blake, S.J., Lyons, A.B., Hughes, T.P.. Nilotinib inhibits the Src-family kinase LCK and T-cell function in vitro. J Cell Mol Med, 13 (2009), no. 3, 599–601. CrossRefGoogle ScholarPubMed
Fraser, C.K., Blake, S.J., Diener, K.R., Lyons, A.B., Brown, M.P., Hughes, T.P., Hayball, J.D.. Dasatinib inhibits recombinant viral antigen-specific murine CD4+ and CD8+ T-cell responses and NK-cell cytolytic activity in vitro and in vivo. Exp Hematol, 37 (2009), no. 2, 256–65. CrossRefGoogle ScholarPubMed
Nordon, R.E., Nakamura, M., Ramirez, C., Odell, R.. Analysis of growth kinetics by division tracking. Immunol Cell Biol, 77 (1999), no. 6, 523–9. CrossRefGoogle Scholar
De Boer, R.J., Ganusov, V.V., Milutinovic, D., Hodgkin, P.D., Perelson, A.S.. Estimating lymphocyte division and death rates from CFSE data. Bull Math Biol, 68 (2006), no. 5, 1011–31. CrossRefGoogle ScholarPubMed
Hawkins, E.D., Hommel, M., Turner, M.L., Battye, F.L., Markham, J.F., Hodgkin, P.D.. Measuring lymphocyte proliferation, survival and differentiation using CFSE time-series data. Nat Protoc, 2 (2007), no. 9, 2057–67. CrossRefGoogle ScholarPubMed
Nordon, R.E., Ko, K.H., Odell, R., Schroeder, T.. Multi-type branching models to describe cell differentiation programs. J Theor Biol, 277 (2011), no. 1, 7–18. CrossRefGoogle ScholarPubMed
Lee, H.Y., Hawkins, E., Zand, M.S., Mosmann, T., Wu, H., Hodgkin, P.D., Perelson, A.S.. Interpreting CFSE obtained division histories of B cells in vitro with Smith-Martin and cyton type models. Bull Math Biol, 71 (2009), no. 7, 1649–70. CrossRefGoogle ScholarPubMed
S.J. Blake, T.P. Hughes, A.B. Lyons. Drug-interaction studies evaluating T-cell proliferation reveal distinct activity of dasatinib and imatinib in combination with cyclosporine A. Exp Hematol, (2012).
Banks, H.T., Sutton, K.L., Thompson, W.C., Bocharov, G., Doumic, M., Schenkel, T., Argilaguet, J., Giest, S., Peligero, C., Meyerhans, A.. A new model for the estimation of cell proliferation dynamics using CFSE data. J Immunol Methods, 373 (2011), no. 1-2, 143-60. CrossRefGoogle ScholarPubMed
Banks, H.T., Sutton, K.L., Thompson, W.C., Bocharov, G., Roose, D., Schenkel, T., Meyerhans, A.. Estimation of cell proliferation dynamics using CFSE data. Bull Math Biol, 73 (2011), no. 1, 116–50. CrossRefGoogle ScholarPubMed
Parish, C.R.. Fluorescent dyes for lymphocyte migration and proliferation studies. Immunol Cell Biol, 77 (1999), no. 6, 499–508. CrossRefGoogle Scholar
Zeng, R., Spolski, R., Casas, E., Zhu, W., Levy, D.E., Leonard, W.J.. The molecular basis of IL-21-mediated proliferation. Blood, 109 (2007), no. 10, 4135–42. CrossRefGoogle ScholarPubMed
Wallace, P.K., Tario, J.D. Jr., Fisher, J.L., Wallace, S.S., Ernstoff, M.S., Muirhead, K.A.. Tracking antigen-driven responses by flow cytometry : monitoring proliferation by dye dilution. Cytometry, Part A, 73 (2008), no. 11, 1019–34. CrossRefGoogle ScholarPubMed
Singh, Y., Dyson, J., Garden, O.A.. Use of SNARF-1 to measure murine T cell proliferation in vitro and its application in a novel regulatory T cell suppression assay. Immunol Letters, 140 (2011), no. 1-2, 21–9. CrossRefGoogle Scholar
Fulcher, D.A., Lyons, A.B., Korn, S.L., Cook, M.C., Koleda, C., Parish, C., Fazekas de St Groth, B., Basten, A.. The fate of self-reactive B cells depends primarily on the degree of antigen receptor engagement and availability of T cell help. J Exp Med, 183 (1996), no. 5, 2313–28. CrossRefGoogle ScholarPubMed
Lyons, A.B.. Pertussis toxin pretreatment alters the in vivo cell division behaviour and survival of B lymphocytes after intravenous transfer. Immunol Cell Biol, 75 (1997), no. 1, 7–12. CrossRefGoogle Scholar
Lyons, A.B.. Analysing cell division in vivo and in vitro using flow cytometric measurement of CFSE dye dilution. J Immunol Methods, 243 (2000), no. 1-2, 147–54. CrossRefGoogle ScholarPubMed
Lyons., A.B., Hasbold, J., Hodgkin, P.D.. Flow cytometric analysis of cell division history using dilution of carboxyfluorescein diacetate succinimidyl ester, a stably integrated fluorescent probe. Method Cell Biol, 63 (2001), 375–98. CrossRefGoogle Scholar
A.B. Lyons, K.V. Doherty. Flow cytometric analysis of cell division by dye dilution. Current protocols in cytometry, (2004), Chapter 9 : Unit 9 11.
Lyons, A.B., Watkins, M., Simpson, C.C., Konrad Muller, H.. Modulation of lymphocyte migration to the murine spleen after marginal zone macrophage phagocytosis of blood-borne particulate material. Immunol Invest, 35 (2006), no. 1, 75–92. CrossRefGoogle Scholar
Quah, B.J., Barlow, V.P., McPhun, V., Matthaei, K.I., Hulett, M.D., Parish, C.R.. Bystander B cells rapidly acquire antigen receptors from activated B cells by membrane transfer. Proc Natl Acad Sci U S A, 105 (2008), no. 11, 4259–64. CrossRefGoogle ScholarPubMed
Chaudhri, G., Quah, B.J., Wang, Y., Tan, A.H., Zhou, J., Karupiah, G., Parish, C.R.. T cell receptor sharing by cytotoxic T lymphocytes facilitates efficient virus control. Proc Natl Acad Sci U S A, 106 (2009), no. 35, 14984–14989. CrossRefGoogle ScholarPubMed