An optimized multi-parameter flow cytometry protocol for human T regulatory cell analysis on fresh and viably frozen cells, correlation with epigenetic analysis, and comparison of cord and adult blood

Journal of Immunological Methods

Volumn 387, Issue 1-2, 2013, Pages 81-88

  Nettenstrom, L ^a   Alderson, K ^a   Raschke, E E ^b   Evans, M D ^a   Sondel, P M ^a   Olek, S ^b   Seroogy, C M ^a  

^a UNIVERSITY OF WISCONSIN SCHOOL OF MEDICINE AND PUBLIC HEALTH   (United States)

^b EPIONTIS GMBH   (Germany)

Author keywords

Cord blood; Demethylation; Epigenetics; Flow cytometry; Foxp3; T regulatory cell

Indexed keywords

ADULT; ARTICLE; CELL VIABILITY; CONTROLLED STUDY; CORRELATIONAL STUDY; DEMETHYLATION; EPIGENETICS; FLOW CYTOMETRY; GENETIC ASSOCIATION; HUMAN; IMMUNE RESPONSE; METHODOLOGY; MULTIPARAMETER FLOW CYTOMETRY; NORMAL HUMAN; PERIPHERAL BLOOD MONONUCLEAR CELL; PRIORITY JOURNAL; PROCESS OPTIMIZATION; REGULATORY T LYMPHOCYTE; UMBILICAL CORD BLOOD;

ADULT; CELL SURVIVAL; CELLS, CULTURED; CRYOPRESERVATION; DNA METHYLATION; EPIGENESIS, GENETIC; FETAL BLOOD; FLOW CYTOMETRY; FORKHEAD TRANSCRIPTION FACTORS; HUMANS; INFANT, NEWBORN; LEUKOCYTES, MONONUCLEAR; LINEAR MODELS; MIDDLE AGED; POLYMERASE CHAIN REACTION; SEQUENCE ANALYSIS, DNA; T-LYMPHOCYTES, REGULATORY; YOUNG ADULT;

EID: 84871445044     PISSN: 00221759     EISSN: 18727905     Source Type: Journal    
DOI: 10.1016/j.jim.2012.09.014     Document Type: Article

References (24)

1. Baecher-Allan C.M., Hafler D.A. Functional analysis of highly defined, FACS-isolated populations of human regulatory CD4+ CD25+ T cells. Clin. Immunol. 2005, 117:192. (discussion 193).
2. Baron U., Floess S., Wieczorek G., Baumann K., Grutzkau A., Dong J., Thiel A., Boeld T.J., Hoffmann P., Edinger M., Turbachova I., Hamann A., Olek S., Huehn J. DNA demethylation in the human FOXP3 locus discriminates regulatory T cells from activated FOXP3(+) conventional T cells. Eur. J. Immunol. 2007, 37:2378.
3. Chen L., Cohen A.C., Lewis D.B. Impaired allogeneic activation and T-helper 1 differentiation of human cord blood naive CD4 T cells. Biol. Blood Marrow Transplant. 2006, 12:160.
4. d'Hennezel E., Piccirillo C.A. Analysis of human FOXP3+ Treg cells phenotype and function. Methods Mol. Biol. 2011, 707:199.
5. d'Hennezel E., Yurchenko E., Sgouroudis E., Hay V., Piccirillo C.A. Single-cell analysis of the human T regulatory population uncovers functional heterogeneity and instability within FOXP3+ cells. J. Immunol. 2011, 186:6788.
6. Floess S., Freyer J., Siewert C., Baron U., Olek S., Polansky J., Schlawe K., Chang H.D., Bopp T., Schmitt E., Klein-Hessling S., Serfling E., Hamann A., Huehn J. Epigenetic control of the foxp3 locus in regulatory T cells. PLoS Biol. 2007, 5:e38.
7. Godfrey W.R., Spoden D.J., Ge Y.G., Baker S.R., Liu B., Levine B.L., June C.H., Blazar B.R., Porter S.B. Cord blood CD4(+)CD25(+)-derived T regulatory cell lines express FoxP3 protein and manifest potent suppressor function. Blood 2005, 105:750.
8. Grant J., Bourcier K., Wallace S., Pan D., Conway A., Seyfert-Margolis V., Wallace P.K. Validated protocol for FoxP3 reveals increased expression in type 1 diabetes patients. Cytometry B Clin. Cytom. 2009, 76:69.
9. Liu J., Lluis A., Illi S., Layland L., Olek S., von Mutius E., Schaub B. T regulatory cells in cord blood-FOXP3 demethylation as reliable quantitative marker. PLoS One 2010, 5:e13267.
10. Lundgren A., Stromberg E., Sjoling A., Lindholm C., Enarsson K., Edebo A., Johnsson E., Suri-Payer E., Larsson P., Rudin A., Svennerholm A.M., Lundin B.S. Mucosal FOXP3-expressing CD4+ CD25high regulatory T cells in Helicobacter pylori-infected patients. Infect. Immun. 2005, 73:523.
11. Ly N.P., Ruiz-Perez B., McLoughlin R.M., Visness C.M., Wallace P.K., Cruikshank W.W., Tzianabos A.O., O'Connor G.T., Gold D.R., Gern J.E. Characterization of regulatory T cells in urban newborns. Clin. Mol. Allergy 2009, 7:8.
12. Manroe B.L., Weinberg A.G., Rosenfeld C.R., Browne R. The neonatal blood count in health and disease. I. Reference values for neutrophilic cells. J. Pediatr. 1979, 95:89.
13. McClymont S.A., Putnam A.L., Lee M.R., Esensten J.H., Liu W., Hulme M.A., Hoffmuller U., Baron U., Olek S., Bluestone J.A., Brusko T.M. Plasticity of human regulatory T cells in healthy subjects and patients with type 1 diabetes. J. Immunol. 2011, 186:3918.
14. Miyara M., Yoshioka Y., Kitoh A., Shima T., Wing K., Niwa A., Parizot C., Taflin C., Heike T., Valeyre D., Mathian A., Nakahata T., Yamaguchi T., Nomura T., Ono M., Amoura Z., Gorochov G., Sakaguchi S. Functional delineation and differentiation dynamics of human CD4+ T cells expressing the FoxP3 transcription factor. Immunity 2009, 30:899.
15. Mold J.E., Michaelsson J., Burt T.D., Muench M.O., Beckerman K.P., Busch M.P., Lee T.H., Nixon D.F., McCune J.M. Maternal alloantigens promote the development of tolerogenic fetal regulatory T cells in utero. Science 2008, 322:1562.
16. Polansky J.K., Kretschmer K., Freyer J., Floess S., Garbe A., Baron U., Olek S., Hamann A., von Boehmer H., Huehn J. DNA methylation controls Foxp3 gene expression. Eur. J. Immunol. 2008, 38:1654.
17. Schaub B., Liu J., Schleich I., Hoppler S., Sattler C., von Mutius E. Impairment of T helper and T regulatory cell responses at birth. Allergy 2008, 63:1438.
18. Sehouli J., Loddenkemper C., Cornu T., Schwachula T., Hoffmuller U., Grutzkau A., Lohneis P., Dickhaus T., Grone J., Kruschewski M., Mustea A., Turbachova I., Baron U., Olek S. Epigenetic quantification of tumor-infiltrating T-lymphocytes. Epigenetics 2011, 6:236.
19. Seroogy C.M., Soares L., Ranheim E.A., Su L., Holness C., Bloom D., Fathman C.G. The gene related to anergy in lymphocytes, an E3 ubiquitin ligase, is necessary for anergy induction in CD4 T cells. J. Immunol. 2004, 173:79.
20. Steinborn A., Engst M., Haensch G.M., Mahnke K., Schmitt E., Meuer S., Sohn C. Small for gestational age (SGA) neonates show reduced suppressive activity of their regulatory T cells. Clin. Immunol. 2010, 134:188.
21. Takahashi T., Kuniyasu Y., Toda M., Sakaguchi N., Itoh M., Iwata M., Shimizu J., Sakaguchi S. Immunologic self-tolerance maintained by CD25+ CD4+ naturally anergic and suppressive T cells: induction of autoimmune disease by breaking their anergic/suppressive state. Int. Immunol. 1998, 10:1969.
22. Tran D.Q., Ramsey H., Shevach E.M. Induction of FOXP3 expression in naive human CD4+ FOXP3 T cells by T-cell receptor stimulation is transforming growth factor-beta dependent but does not confer a regulatory phenotype. Blood 2007, 110:2983.
23. Weinberg A.G., Rosenfeld C.R., Manroe B.L., Browne R. Neonatal blood cell count in health and disease. II. Values for lymphocytes, monocytes, and eosinophils. J. Pediatr. 1985, 106:462.
24. Wieczorek G., Asemissen A., Model F., Turbachova I., Floess S., Liebenberg V., Baron U., Stauch D., Kotsch K., Pratschke J., Hamann A., Loddenkemper C., Stein H., Volk H.D., Hoffmuller U., Grutzkau A., Mustea A., Huehn J., Scheibenbogen C., Olek S. Quantitative DNA methylation analysis of FOXP3 as a new method for counting regulatory T cells in peripheral blood and solid tissue. Cancer Res. 2009, 69:599.