Expansion and de novo generation of potentially therapeutic regulatory T cells in patients with autoimmune hepatitis

Maria Serena Longhi, Francesca Meda, Pengyun Wang, Marianne Samyn, Giorgina Mieli-Vergani, Diego Vergani, Yun Ma

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

CD4+CD25+ regulatory T cells (T-regs) are central to the maintenance of immune tolerance and represent an immune intervention candidate in autoimmune hepatitis (AIH), a condition characterized by impaired T-reg number and function. We investigated whether T-regs can be expanded from the existing CD4+CD25+ T cell pool and generated de novo from CD4+CD25- T cells in AIH patients and healthy controls. Purified CD4+CD25+ and CD4+CD25- T cells from 24 patients with type 1 AIH and 22 healthy controls were cultured for up to 5 weeks with anti-CD3/anti-CD28 T cell expander and high-dose interleukin-2 (IL-2). Cell phenotypes, suppressor ability, forkhead winged/helix transcription factor box P3 (FOXP3) gene, and protein expression were assessed weekly by cytofluorimetry, proliferation assay, real-time polymerase chain reaction (PCR), and immunoblot. During culture, the number of CD4+CD25+ T cells derived from the existing T-reg pool (expanded T-regs) and generated de novo from CD4+CD25- T cells (newly generated T-regs) increased constantly up to week 4 in both healthy controls and, to a lesser extent, in AIH patients. Expanded T-regs retained conventional T-reg phenotype and, compared with baseline, demonstrated more vigorous suppressive function and increased FOXP3 gene and protein expression. Newly generated T-regs not only acquired T-reg phenotype but underwent greater growth and were more resistant to apoptosis than expanded T-regs. Their suppressive function augmented throughout culture, reaching a peak at week 4, preceded by a peak FOXP3 gene and protein expression at week 2. Suppressor function and FOXP3 expression of both expanded and newly generated T-regs were higher in normal controls than in AIH patients. Conclusion: Functionally enhanced T-regs can be expanded and generated de novo in patients with AIH. This finding may assist in reconstituting impaired immune regulation and restoring peripheral tolerance through T-reg infusion in this condition.

Original languageEnglish
Pages (from-to)581-591
Number of pages11
JournalHepatology
Volume47
Issue number2
DOIs
Publication statusPublished - Feb 2008

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Autoimmune Hepatitis
Regulatory T-Lymphocytes
T-Lymphocytes
Phenotype
Gene Expression
Winged-Helix Transcription Factors
Therapeutics
Peripheral Tolerance
Forkhead Transcription Factors
Immune Tolerance
Proteins
Interleukin-2
Real-Time Polymerase Chain Reaction
Maintenance
Apoptosis
Growth

ASJC Scopus subject areas

  • Hepatology

Cite this

Expansion and de novo generation of potentially therapeutic regulatory T cells in patients with autoimmune hepatitis. / Longhi, Maria Serena; Meda, Francesca; Wang, Pengyun; Samyn, Marianne; Mieli-Vergani, Giorgina; Vergani, Diego; Ma, Yun.

In: Hepatology, Vol. 47, No. 2, 02.2008, p. 581-591.

Research output: Contribution to journalArticle

Longhi, Maria Serena ; Meda, Francesca ; Wang, Pengyun ; Samyn, Marianne ; Mieli-Vergani, Giorgina ; Vergani, Diego ; Ma, Yun. / Expansion and de novo generation of potentially therapeutic regulatory T cells in patients with autoimmune hepatitis. In: Hepatology. 2008 ; Vol. 47, No. 2. pp. 581-591.
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