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Capturing resting T cells: the perils of PLL

Nature Immunology volume 19, pages 203–205 (2018)Cite this article

To the editor — Full understanding of lymphocyte activation will require thorough characterization of the ‘resting’ state and how it changes. Surfaces coated with the cationic homopolymer poly-l-lysine (PLL) are widely used for total internal reflection fluorescence (TIRF) imaging of the organization of surface proteins on resting lymphocytes1,2,3,4,5 because PLL is assumed to be inert. Here we found that PLL initiated T cell signaling and profoundly altered the activity of membrane proteins such as the T cell antigen receptor (TCR). Therefore, the emerging idea that receptors and signaling proteins cluster by default1,2,3,4,5, which has been based mostly on studies of lymphocytes interacting with PLL-coated surfaces, needs reconsideration.

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Fig. 1: PLL induces strong T cell signaling and profoundly alters TCR diffusional activity and organization.

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Acknowledgements

Supported by a Royal Society University Research Fellowship (UF120277 to S.F.L.) and Research Professorship (RP150066 to D.K.); the EPSRC (EP/L027631/1 to A.P.,); the Wellcome Trust (098274/Z/12/Z to S.J.D., and WT101609MA to R.A.F.); PA Cephalosporin Fund (C.E.); the Wolfson Imaging Centre Oxford (funded by the Wolfson Foundation and Wellcome Trust; 104924/14/Z/14); the Micron Advanced BioImaging Unit (Wellcome Trust Strategic Award 091911); the Medical Research Council (MC_UU_12010/Unit Programmes G0902418 and MC_UU_12025); an MRC/BBSRC/EPSRC award (MR/K01577X/1); and a Marie Skłodowska-Curie Intra-European grant (707348 to I.U.).

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  1. These authors contributed equally: Ana Mafalda Santos, Aleks Ponjavic, Marco Fritzsche and Ricardo A. Fernandes.

Authors and Affiliations

  1. Radcliffe Department of Medicine, University of Oxford, Oxford, UK

    Ana Mafalda Santos, Marco Fritzsche, Ricardo A. Fernandes, Jorge Bernardino de la Serna, Martin J. Wilcock, Falk Schneider, Iztok Urbančič, Consuelo Anzilotti, Meike Aßmann, Simon J. Davis & Christian Eggeling

  2. MRC Human Immunology Unit, John Radcliffe Hospital, University of Oxford, Oxford, UK

    Ana Mafalda Santos, Marco Fritzsche, Ricardo A. Fernandes, Jorge Bernardino de la Serna, Martin J. Wilcock, Falk Schneider, Iztok Urbančič, Consuelo Anzilotti, Meike Aßmann, Richard J. Cornall, Simon J. Davis & Christian Eggeling

  3. Department of Chemistry, University of Cambridge, Cambridge, UK

    Aleks Ponjavic, James McColl, Kristina A. Ganzinger, David Klenerman & Steven F. Lee

  4. Kennedy Institute of Rheumatology, University of Oxford, Oxford, UK

    Marco Fritzsche, David Depoil & Michael L. Dustin

  5. Nuffield Department of Clinical Medicine, University of Oxford, Oxford, UK

    Richard J. Cornall

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Contributions

A.M.S., A.P., M.F., R.A.F., J.B.S., M.J.W, F.S., I.U., J.M., C.A., K.A.G., M.A. and D.D. performed experiments; A.M.S., A.P., M.F., R.A.F., R.J.C, M.L.D., D.K., S.J.D., C.E. and S.F.L. drafted and/or edited the manuscript; D.K., S.J.D., C.E. and S.F.L. conceived of the study; and M.L.D., S.J.D., C.E. and S.F.L. supervised the experiments.

Corresponding authors

Correspondence to Christian Eggeling or Steven F. Lee.

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The authors declare no competing financial interests.

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Santos, A.M., Ponjavic, A., Fritzsche, M. et al. Capturing resting T cells: the perils of PLL. Nat Immunol 19, 203–205 (2018). https://doi.org/10.1038/s41590-018-0048-8

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