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Blood pressure regulation by CD4+ lymphocytes expressing choline acetyltransferase

Nature Biotechnology volume 34, pages 1066–1071 (2016)Cite this article

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Abstract

Blood pressure regulation is known to be maintained by a neuro-endocrine circuit, but whether immune cells contribute to blood pressure homeostasis has not been determined. We previously showed that CD4+ T lymphocytes that express choline acetyltransferase (ChAT), which catalyzes the synthesis of the vasorelaxant acetylcholine, relay neural signals1. Here we show that these CD4+CD44hiCD62Llo T helper cells by gene expression are a distinct T-cell population defined by ChAT (CD4 TChAT). Mice lacking ChAT expression in CD4+ cells have elevated arterial blood pressure, compared to littermate controls. Jurkat T cells overexpressing ChAT (JTChAT) decreased blood pressure when infused into mice. Co-incubation of JTChAT and endothelial cells increased endothelial cell levels of phosphorylated endothelial nitric oxide synthase, and of nitrates and nitrites in conditioned media, indicating increased release of the potent vasorelaxant nitric oxide. The isolation and characterization of CD4 TChAT cells will enable analysis of the role of these cells in hypotension and hypertension, and may suggest novel therapeutic strategies by targeting cell-mediated vasorelaxation.

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Figure 1: Gene expression analysis reveals differences between CD4+CD44hiCD62Llo ChAT-eGFP+ and ChAT-eGFP− T cells. (a) RNA from the ChAT-eGFP+ and ChAT-eGFP− subsets of CD4+CD44hiCD62Llo T cells was analyzed by Affymetrix Gene ST 1.
Figure 2: Increased blood pressure in mice with genetic ablation of choline acetyltransferase+ CD4+ cells.
Figure 3: Mean arterial blood pressure (MAP) after infusion of JTChAT lymphocytes in wild-type C57Bl/6 mice.

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Acknowledgements

We thank the flow cytometry facility at the Feinstein Institute, the laboratory of C. Benoist at Harvard Medical School for assistance, and G.K. Hansson for comments. We thank C. James Kirkpatrick (Institute of Pathology, Johannes-Gutenberg University, Mainz, Germany), who kindly provided PMEC-ST1.6R cells. This work benefitted from the data assembled by the ImmGen consortium. This work was supported by the following grants: NIH, NIGMS RO1 GM57226 to K.J.T., NIGMS RO1 GM098446 to H.Y. and NIGMS RO1GM089807 to K.J.T. and V.A.P., VR 2013-3003 to A.A., NIAID R01 029690 to T.L.R., Canadian Institutes for Health Research FRN 119339 to P.H.B., and Knut and Alice Wallenberg's Foundation 2014.0212, The Swedish Heart-Lung Foundation 20150767 and Svenska Läkaresällskapet to P.S.O. Jurkat cells (originally obtained from ATCC) were a gift from C. Chu, The Feinstein Institute for Medical Research. eNOS-deficient mice were provided by J. Lundberg and E. Weitzberg, Karolinska Institutet, Stockholm, Sweden.

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Authors and Affiliations

  1. Department of Medicine, Center for Bioelectronic Medicine, Center for Molecular Medicine, Solna, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden

    Peder S Olofsson

  2. Laboratory of Biomedical Science, The Feinstein Institute for Medical Research, Manhasset, New York, USA

    Peder S Olofsson, Benjamin E Steinberg, William M Hanes, Sangeeta S Chavan, Huan Yang, Valentin A Pavlov & Kevin J Tracey

  3. The Campbell Family Institute for Breast Cancer Research, University Health Network, Toronto, Ontario, Canada

    Benjamin E Steinberg, Maureen A Cox & Tak W Mak

  4. Division of Cardiology, Peter Munk Cardiac Centre, University Health Network, Toronto, Ontario, Canada

    Roozbeh Sobbi & Peter H Backx

  5. Center for Heart and Lung Research, The Feinstein Institute for Medical Research, Manhasset, New York, USA

    Mohamed N Ahmed, Shu Fang Liu & Edmund J Miller

  6. Robert S. Boas Center for Genomics and Human Genetics, Feinstein Institute for Medical Research, Manhasset, New York, USA

    Michaela Oswald & Peter K Gregersen

  7. Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden

    Ferenc Szekeres, Cecilia Lövdahl & Anders Arner

  8. School of Health and Education, University of Skövde, Skövde, Sweden

    Ferenc Szekeres

  9. Department of Medicine, Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden

    Andrea Introini & Kristina Broliden

  10. Center for Oncology and Cell Biology, The Feinstein Institute for Medical Research, Manhasset, New York, USA

    Nichol E Holodick & Thomas L Rothstein

  11. Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden

    Ulf Andersson & Kevin J Tracey

  12. The Center for Autoimmune and Musculoskeletal Diseases, The Feinstein Institute for Medical Research, Manhasset, New York, USA

    Betty Diamond

  13. Department of Biology, York University, Toronto, Ontario, Canada

    Peter H Backx

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Contributions

P.S.O. and K.J.T. conceived the study, planned and performed experiments, analyzed data and wrote the manuscript. U.A., B.D. and T.W.M. planned experiments and edited the manuscript. B.E.S., W.M.H. and M.A.C. planned and performed experiments and analyzed data. M.O. and P.K.G. planned experiments, analyzed gene expression and edited the manuscript. R.S. and P.H.B. planned experiments, measured murine blood pressure, performed telemetry and echocardiography, and analyzed data. M.N.A. and E.J.M. planned, performed and analyzed experiments with endothelial cells in vitro and murine blood pressure in response to cell infusion. F.S. and A.A. planned, performed and analyzed experiments on murine blood pressure in response to cell infusion. A.I., V.A.P., K.B., H.Y., S.S.C., S.F.L., C.L., T.L.R. and N.E.H. performed experiments and analyzed data.

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Correspondence to Peder S Olofsson.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–18 and Supplementary Tables 1 and 5 (PDF 1855 kb)

Supplementary Table 2

Included samples (XLSX 21 kb)

Supplementary Table 3

ChAT expr values (XLS 38 kb)

Supplementary Table 4

Subsets from the Immgen dataset included in the principal component analysis of splenic immune cell subsets (XLS 26 kb)

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Olofsson, P., Steinberg, B., Sobbi, R. et al. Blood pressure regulation by CD4+ lymphocytes expressing choline acetyltransferase. Nat Biotechnol 34, 1066–1071 (2016). https://doi.org/10.1038/nbt.3663

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