Induction of eye-derived tolerance does not depend on naturally occurring CD4+CD25+ T regulatory cells

Invest Ophthalmol Vis Sci. 2006 Mar;47(3):1047-55. doi: 10.1167/iovs.05-0110.

Abstract

Purpose: Regulatory CD4+ T cells (T regs) arise in the spleens of mice with anterior chamber-associated immune deviation (ACAID), an eye-derived tolerance evoked by injection of antigen into the ocular anterior chamber (AC). The current study was conducted to investigate the possibility that these T regs express CD25 and are derived from natural CD4+CD25+ T cells.

Methods: Naïve T cells from DO11.10 mice were activated in vitro by ovalbumin (OVA)-pulsed, TGFbeta-treated antigen-presenting cells (APCs), and the expression of CD25 assayed by flow cytometry. OVA-specific ACAID T regs were obtained from the spleens of DO11.10 mice with ACAID to OVA. Immunomagnetic enrichment was used to sort out CD4+CD25+, and CD4+CD25- ACAID T cells before they were injected into OVA-immunized mice or examined for mRNA expression of the regulatory T-cell transcription factor Foxp3. In addition, before AC injection of OVA, systemic depletion of CD25+ T cells was performed with injections of anti-IL-2 receptor antibody into the mice.

Results: OVA-specific T cells from DO11.10 mice expressed CD25 when exposed to OVA-pulsed, TGFbeta-treated APCs, even when the DO11.10 T cells were depleted of CD25+ cells before their in vitro stimulation. In addition, DH was suppressed in naïve mice that were injected with CD4+CD25+ or CD4+CD25- ACAID T cells. The CD4+CD25+, but not the CD4+CD25-, ACAID T regs expressed Foxp3. Finally, OVA induced ACAID in mice depleted of CD25+ cells.

Conclusions: Some of the CD4+ T regs of ACAID arise from CD25- precursors, and the induction of ACAID is not dependent on the presence of natural CD4+CD25+ T regs.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adoptive Transfer
  • Animals
  • Anterior Chamber / immunology
  • Antigen-Presenting Cells / immunology
  • CD4 Antigens / immunology*
  • Female
  • Flow Cytometry
  • Forkhead Transcription Factors / genetics
  • Hypersensitivity, Delayed / immunology
  • Immune Tolerance / physiology*
  • Lymphocyte Activation / immunology
  • Lymphocyte Depletion
  • Mice
  • Mice, Inbred BALB C
  • Mice, Transgenic
  • Ovalbumin
  • RNA, Messenger / metabolism
  • Receptors, Interleukin-2 / immunology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • T-Lymphocytes, Regulatory / immunology*
  • Transforming Growth Factor beta / pharmacology

Substances

  • CD4 Antigens
  • Forkhead Transcription Factors
  • Foxp3 protein, mouse
  • RNA, Messenger
  • Receptors, Interleukin-2
  • Transforming Growth Factor beta
  • Ovalbumin