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Article | Published: 01 February 2016

CD4+ T cell anergy prevents


autoimmunity and generates
regulatory T cell precursors
Lokesh A Kalekar, Shirdi E Schmiel, Sarada L Nandiwada, Wing Y Lam, Laura O Barsness, Na Zhang, Gretta L Stritesky,
Deepali Malhotra, Kristen E Pauken, Jonathan L Linehan, M Gerard O'Sullivan, Brian T Fife, Kristin A Hogquist, Marc K
Jenkins & Daniel L Mueller

Nature Immunology volume 17, pages 304–314 (2016) | Download Citation

Abstract
The role of anergy, an acquired state of T cell functional unresponsiveness, in natural peripheral tolerance remains unclear. In
this study, we found that anergy was selectively induced in fetal antigen–specific maternal CD4+ T cells during pregnancy. A
naturally occurring subpopulation of anergic polyclonal CD4+ T cells, enriched for self antigen–specific T cell antigen
receptors, was also present in healthy hosts. Neuropilin-1 expression in anergic conventional CD4+ T cells was associated
with hypomethylation of genes related to thymic regulatory T cells (Treg cells), and this correlated with their ability to
differentiate into Foxp3+ Treg cells that suppressed immunopathology. Thus, our data suggest that not only is anergy induction
important in preventing autoimmunity but also it generates the precursors for peripheral Treg cell differentiation.

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Acknowledgements
We thank J.A. Bluestone (University of California, San Francisco) for spleen and lymph node cells from Foxp3-Cre-GFP ×
R26-YFP mice and discussions; D. Mathis and C. Benoist (Harvard Medical School) and the Institut de Genetique et de
Biologie Moleculaire et Cellulaire (Strasbourg, France) for B6.g7 mice and KRN B6 mice; A. Rudensky (Memorial Sloan-
Kettering Cancer Center) for B6 Foxp3DTR knock-in mice; S.S. Way (University of Cincinnati) for B6 Foxp3GFP and Foxp3DTR
CD45.1 mice; J.A. Bluestone (University of San Francisco) for cells from the spleen and all lymph nodes of Foxp3-Cre-GFP ×
R26-YFP mice; S.C. Jameson, M. Mescher and M.A. Farrar for discussions; P.J. Titcombe for technical support; and N. Shah,
T. Martin and J. Motl for assistance in cell sorting. Supported by the Rheumatology Research Foundation (Within Our Reach:
Finding a Cure for Rheumatoid Arthritis campaign grant to D.L.M.) and the US National Institutes of Health (01 AI35296 to
D.L.M., B.T.F., K.A.H. and M.K.J.).

Author information
Affiliations

Department of Medicine, University of Minnesota


Medical School, Minneapolis, Minnesota, USA.
Lokesh A Kalekar, Shirdi E Schmiel, Sarada L Nandiwada, Wing Y Lam, Laura O Barsness, Na Zhang, Kristen E Pauken,
Brian T Fife & Daniel L Mueller

The Center for Immunology, University of Minnesota


Medical School, Minneapolis, Minnesota, USA.
Lokesh A Kalekar, Shirdi E Schmiel, Sarada L Nandiwada, Wing Y Lam, Laura O Barsness, Na Zhang, Gretta L Stritesky,
Deepali Malhotra, Kristen E Pauken, Jonathan L Linehan, Brian T Fife, Kristin A Hogquist, Marc K Jenkins & Daniel L Mueller

Department of Laboratory Medicine and Pathology,


University of Minnesota Medical School, Minneapolis,

https://www.nature.com/articles/ni.3331 7/12
5/6/2019 CD4+ T cell anergy prevents autoimmunity and generates regulatory T cell precursors | Nature Immunology

Minnesota, USA.
Gretta L Stritesky & Kristin A Hogquist

Department of Microbiology, University of Minnesota


Medical School, Minneapolis, Minnesota, USA.
Deepali Malhotra, Jonathan L Linehan & Marc K Jenkins

The Comparative Pathology Core at the Masonic Cancer


Center, University of Minnesota Medical School,
Minneapolis, Minnesota, USA.
M Gerard O'Sullivan

Contributions
L.A.K. and D.L.M. designed the experiments and analyzed the data; L.A.K. performed most of the experiments; S.E.S., S.L.N.,
W.Y.L., L.O.B., N.Z., G.L.S., D.M., K.E.P. and J.L.L. performed experiments or provided technical help; M.G.O.S. scored
histology slides; B.T.F., K.A.H. and M.K.J. provided scientific input; D.L.M. conceived of the study and directed the research;
and L.A.K. and D.L.M. wrote the manuscript.

Competing interests
The authors declare no competing financial interests.

Corresponding author
Correspondence to Daniel L Mueller.

Integrated supplementary information


Supplementary figures

1.
https://www.nature.com/articles/ni.3331 8/12
5/6/2019 CD4+ T cell anergy prevents autoimmunity and generates regulatory T cell precursors | Nature Immunology

A Foxp3−CD44hiCD73hiFR4hi anergic compartment


shows enrichment for autoreactive insulin-specific
CD4+ polyclonal T cells.

2.
KRN CD4+ T cells cause arthritis in lymphopenic Tcra−/−
mice following reversal of anergy.

3.
Gating strategy and purity after sorting.

4.
Experimental design for Figure 5.

5.
Reversal of anergy in polyclonal CD4+ T cells results in
autoantibody production after ablation of newly
generated Treg cells.

6.
Quantification and frequency of Treg cells recovered in
models of arthritis and colitis from polyclonal anergic
cells.

https://www.nature.com/articles/ni.3331 9/12
5/6/2019 CD4+ T cell anergy prevents autoimmunity and generates regulatory T cell precursors | Nature Immunology

7.
Formerly Foxp3-expressing cells make up a very small
fraction of anergic cells.

Supplementary information
PDF files

1.
Supplementary Text and Figures
Supplementary Figures 1–7 and Supplementary Table 1

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Publication history

Received

25 August 2015

Accepted

20 October 2015

Published
https://www.nature.com/articles/ni.3331 10/12
5/6/2019 CD4+ T cell anergy prevents autoimmunity and generates regulatory T cell precursors | Nature Immunology

01 February 2016

DOI
https://doi.org/10.1038/ni.3331

Subjects
AutoimmunityRegulatory T cells

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