March 2012
Volume 53, Issue 14
ARVO Annual Meeting Abstract  |   March 2012
ACAID Tolerogenic APC Induce Two Types Of CD4+ Treg Cells By Two Different Mechanisms
Author Affiliations & Notes
  • Rose Mathew
    Immunology, Schepens Eye Research Institute/MEEI, Boston, Massachusetts
  • Joan Stein-Streilein
    Immunology, Schepens Eye Research Institute/MEEI, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  Rose Mathew, None; Joan Stein-Streilein, None
  • Footnotes
    Support  EY011983, EY016476
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 6311. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Rose Mathew, Joan Stein-Streilein; ACAID Tolerogenic APC Induce Two Types Of CD4+ Treg Cells By Two Different Mechanisms. Invest. Ophthalmol. Vis. Sci. 2012;53(14):6311.

      Download citation file:

      © ARVO (1962-2015); The Authors (2016-present)

  • Supplements

Purpose: : ACAID induction, in vivo or in vitro, generates three types of Tregs: efferent CD8+, afferent CD4+CD25+ and CD4+CD25-. Previously, we showed CD8+ Treg cells required F4/80 protein expression on the APC and iNKT cells for their development. The purpose of this study was to determine if the generation of ACAID induced afferent CD4+ Treg cells subpopulations were also dependent on the described ACAID mechanisms.

Methods: : ACAID tolerogenic APC (Tol APC) were generated from C57BL/6, PD-L1 KO, or F4/80 KO by overnight culture of peritoneal exudate cells (PEC) with TGFβ2 and Ovalbumin (OVA). Spleen cells from primed OVA tg (OTII) mice, Jα281 KO or F4/80 KO mice were depleted of natural T regs by incubation with CD25 antibody (PC61.5). The remaining natural T reg-depleted cells were co-cultured (5D) with OVA pulsed-Tol APC. Following culture the T cells were stained for select surface markers (CD4, CD25, LAP-1, FoxP3, PD-L1) and analysed by flow cytometry. The efferent suppressor function of the CD25+ or CD25- CD4+T cells was tested in a Local Adoptive Transfer (LAT) assay and afferent suppressor function was tested in a footpad assay and proliferation and assessed by bromodeoxyuridine (BrdU) incorporation into the draining lymph nodes.

Results: : We observed that post ACAID culture, there were two distinct populations of CD4+ T reg cells. ACAID induced CD4+CD25- Tregs were LAP-1+ FoxP3- and PD-L1lo and exhibited only afferent suppressor function. CD4+CD25+ Tregs were LAP-1- FoxP3+ and PD-L1hi and exhibited both afferent and efferent suppressor function. Moreover, ACAID induced CD4+CD25+ Tregs required PD-L1 and not F4/80KO and iNKT cells.

Conclusions: : Thus the ACAID Tol APC induces two populations of T reg cells by two different mechanisms. The CD4+CD25- Treg cells are dependent on F4/80 protein, iNKT cells, but not PD-L1 for their generation. The CD4+CD25+ Treg cells are generated by the activation of the TCR in the presence of TGFβ and PD-L1+ APC and supports the notion that PD-L1 is involved in the generation of inducible Treg cells within immune privileged sites where TGFβ is high.This research was supported in part by NIH grant: EY011983, EY016476

Keywords: ACAID • immune tolerance/privilege • immunomodulation/immunoregulation 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.