Glucocorticoid hormone differentially modulates the in vitro expansion and cytokine profile of thymic and splenic Treg cells

被引:9
|
作者
Pap, Ramona [1 ,2 ]
Ugor, Emese [1 ]
Litvai, Timea [1 ]
Prenek, Lilla [1 ]
Najbauer, Jozsef [1 ]
Nemeth, Peter [1 ]
Berki, Tfinea [1 ]
机构
[1] Univ Pecs, Med Sch, Clin Ctr, Dept Immunol & Biotechnol, H-7624 Pecs, Hungary
[2] Univ Pecs, Fac Pharm, Dept Pharmaceut Biol, H-7624 Pecs, Hungary
关键词
Dexamethasone; Treg; Foxp3; Glucocorticoid hormone; IL-10; TGF beta; REGULATORY T-CELLS; TGF-BETA; CUTTING EDGE; FOXP3; IL-2; INTERLEUKIN-2; DEXAMETHASONE; MECHANISMS; GENERATION; CONVERSION;
D O I
10.1016/j.imbio.2018.12.002
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Objective: Functional disturbances in regulatory T cells (Treg) have been described in autoimmune diseases, and their potential therapeutic use is intensively studied. Our goal was to investigate the influence of glucocorticoid hormone on the in vitro differentiation of Treg cells from thymic and splenic CD4(+) T cells under different conditions to establish methods for generating stable and functionally suppressive iTregs for future use in adoptive transfer experiments. Methods: Thymic and splenic CD4(+) T lymphocytes were isolated from 3 to 4 week-old control and in vivo dexamethasone (DX) pretreated BALB/c mice using magnetic bead negative selection, followed by CD25 positive selection. The cells were cultured with anti-CD3/CD28 beads and IL-2 in the presence or absence of TGF beta and/or DX for 3-6 days. Multiparametric flow cytometry was performed using CD4, CD25, CD8, TGF beta (LAP) cell surface and Foxp3, IL-4, IL-10, IL-17 and IFN gamma intracellular staining. Quantitative RT-PCR was performed to measure IL-10, TGF beta cytokine and Foxp3 mRNA levels. Results: Differentiation of thymus-derived CD4(+) cells in vitro into iTreg cells was most effective (24-25%) when anti-CD3/CD28 beads, IL-2, and TGF beta were present. Splenic CD4(+) T cell expansion under same conditions resulted in a higher (44-45%) iTreg cell ratio that further increased (up to 50% Treg) in the presence of DX. Elevated immunosuppressive cytokine (IL-10 and TGF beta) production by iTregs could be measured both at protein and mRNA levels without elevation of Th1/Th2 or Th17 cytokine production. We got the highest iTreg ratio (74%) and TGF beta production when CD4(+) CD25(+) splenic T cells were stimulated in the presence of TGF beta. In vivo 4 days DX pretreatment resulted in enhanced in vitro expansion and Foxp3 expression of thymus-derived iTregs and decreased differentiation of spleen-derived iTreg cells. In these Tregs the relative expression of IL-10 mRNA significantly decreased under all in vitro stimulation conditions, while TGFP mRNA level did not change. Conclusion: DX promotes the expansion of thymic and splenic Treg cells, and enhances Foxp3(+) expression and the production of immunosuppressive cytokines IL-10 and TGF beta in vitro. In vivo pretreatment of mice with DX inhibited the immunosuppressive cytokine production of in vitro differentiated Treg cells. We hypothesize that patients receiving GC therapy may need special attention prior to in vitro expansion and transplantation of Treg cells.
引用
收藏
页码:285 / 295
页数:11
相关论文
共 50 条
  • [31] Chronic ethanol exposure differentially affects the migratory capacity of hepatic compared to splenic dendritic cells in vitro and in vivo
    Lau, Audrey H.
    Colvin, Bridget L.
    Thomson, Angus W.
    ALCOHOL, 2006, 39 (02) : 115 - 115
  • [32] SUPPRESSION OF DNA-SYNTHESIS IN HEPATOMA-CELLS EXPOSED TO GLUCOCORTICOID HORMONE IN-VITRO
    LOEB, JN
    BOREK, C
    YEUNG, LL
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1973, 70 (12) : 3852 - 3856
  • [33] Glucocorticoid hormone treatment enhances the cytokine production of regulatory T cells by upregulation of Foxp3 expression
    Ugor, Emese
    Prenek, Lilla
    Pap, Ramona
    Berta, Gergely
    Ernszt, David
    Najbauer, Jozsef
    Nemeth, Peter
    Boldizsar, Ferenc
    Berki, Timea
    IMMUNOBIOLOGY, 2018, 223 (4-5) : 422 - 431
  • [34] In vitro processing of MDS, AML, and CML cells significantly alters the cytokine mRNA profile of the cells.
    Gao, XZ
    Chopra, H
    Bi, S
    Preisler, HD
    BLOOD, 1996, 88 (10) : 328 - 328
  • [35] Nerve growth factor stimulates proliferation, adhesion and thymopoietic cytokine expression in mouse thymic epithelial cells in vitro
    Lee, Hee-Woo
    Na, Yong-Jin
    Jung, Pil-Ku
    Kim, Mi-Na
    Kim, Sung-Min
    Chung, Joo-Seop
    Kim, Bong-Seon
    Kim, Jae-Bong
    Moon, Jeon Ok
    Yoon, Sik
    REGULATORY PEPTIDES, 2008, 147 (1-3) : 72 - 81
  • [36] Donor Treg expansion by liposomal α-galactosylceramide modulates Tfh cells and prevents sclerodermatous chronic graft-versus-host disease
    Sugiura, Hiroyuki
    Matsuoka, Ken-ichi
    Fukumi, Takuya
    Sumii, Yuichi
    Kondo, Takumi
    Ikegawa, Shuntaro
    Meguri, Yusuke
    Iwamoto, Miki
    Sando, Yasuhisa
    Nakamura, Makoto
    Toji, Tomohiro
    Ishii, Yasuyuki
    Maeda, Yoshinobu
    IMMUNITY INFLAMMATION AND DISEASE, 2021, 9 (03) : 721 - 733
  • [37] CYTOKINE REQUIREMENTS FOR OPTIMUM IN-VITRO EXPANSION OF CFU FROM PURIFIED CANINE PROGENITOR CELLS
    SCHUMM, M
    NEUNER, E
    GUNTHER, W
    KREMMER, E
    VOGL, C
    ELLWART, J
    MAILHAMMER, R
    WILMANNS, W
    THIERFELDER, S
    KOLB, HJ
    EXPERIMENTAL HEMATOLOGY, 1995, 23 (08) : 875 - 875
  • [38] C1q Differentially Modulates Phagocytosis and Cytokine Responses during Ingestion of Apoptotic Cells by Human Monocytes, Macrophages, and Dendritic Cells
    Fraser, Deborah A.
    Laust, Amanda K.
    Nelson, Edward L.
    Tenner, Andrea J.
    JOURNAL OF IMMUNOLOGY, 2009, 183 (10): : 6175 - 6185
  • [40] Ovarian cancer-cell glucocorticoid receptor activity modulates cytokine secretion promoting infiltration of immunosuppressive cells into the tumor microenvironment
    Taya, Manisha
    LeBlanc, Eleanore
    Bennett, Lynda
    Conzen, Suzanne D.
    CANCER RESEARCH, 2023, 83 (07)