Dynamic imaging of chemokine-dependent cd8+ t cell help for cd8+ t cell responses

ABSTRACT Naive T lymphocytes move efficiently in lymphoid tissues while scanning dendritic cells in search of cognate complexes of peptide in major histocompatibility molecules. However, T

cell migration ceases after recognition of cognate antigen. We show here that during the initiation of antigen-specific CD8+ T cell responses, naive CD8+ polyclonal T cells

'preferentially' interacted in an antigen-independent way with mature dendritic cells competent to present antigen to antigen-specific CD8+ T cells. These antigen-independent

interactions required expression of the chemokine receptor CCR5 on polyclonal T cells and increased the efficiency of the induction of naive, low-precursor-frequency CD8+ T cell responses.

Thus, antigen-specific CD8+ T cells favor the priming of naive CD8+ T cells by promoting the CCR5-dependent recruitment of polyclonal CD8+ T cells to mature dendritic cells. Access through

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(1998). CAS  PubMed  Google Scholar  Download references ACKNOWLEDGEMENTS We thank C. Reis e Sousa (Cancer Research UK) for C57BL/6 OT-I _Rag2__−/−_ TCR-transgenic mice; and P. Guermonprez

for discussions. Supported by the Institut National de la Santé et de la Recherche Médicale, Centre National de la Recherche Scientifique, Ligue de Lutte Contre le Cancer, Association de la

Recherche contre le Cancer (A.B.), Institut Curie, European Community DC-Thera (LSBH-CT-2004-512074; “Dendritic cells for novel immunotherapies”), European Community CancerImmunotherapy

(LSHC-CT-2006-518234; “Cancer Immunology and Immunotherapy”), Ecole Normale Supérieure (A.S.) and a Marie Curie Intra-European Fellowship (A.K.N.). AUTHOR INFORMATION Author notes *

Stéphanie Hugues and Alix Scholer: These authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Institut Curie, Centre de Recherche, Paris, F-75248, France Stéphanie Hugues, 

Alix Scholer, Alexandre Boissonnas, Alexander Nussbaum, Sebastian Amigorena & Luc Fetler * Institut National de la Santé et de la Recherche Médical U653, Immunité et Cancer, Institut

Curie, Paris, F-75248, France Stéphanie Hugues, Alix Scholer, Alexandre Boissonnas, Alexander Nussbaum & Sebastian Amigorena * Institut National de la Santé et de la Recherche Médical

U543, Laboratoire d'Immunologie Cellulaire, Hôpital Pitié-Salpétrière, Paris, F-75013, France Christophe Combadière * Centre National de la Recherche Scientifique UMR 168, Laboratoire

Physico-Chimie Curie, Institut Curie, Paris, F-75248, France Luc Fetler Authors * Stéphanie Hugues View author publications You can also search for this author inPubMed Google Scholar * Alix

Scholer View author publications You can also search for this author inPubMed Google Scholar * Alexandre Boissonnas View author publications You can also search for this author inPubMed 

Google Scholar * Alexander Nussbaum View author publications You can also search for this author inPubMed Google Scholar * Christophe Combadière View author publications You can also search

for this author inPubMed Google Scholar * Sebastian Amigorena View author publications You can also search for this author inPubMed Google Scholar * Luc Fetler View author publications You

can also search for this author inPubMed Google Scholar CONTRIBUTIONS S.H., A.S. and L.F., design, immunobiology and imaging experiments, analysis, interpretation, coordination and writing;

A.B., help with experiments and interpretation; A.N., gp33-expressing B16 melanoma cells; C.C., CCR5-deficient mice; and S.A., design, interpretation, coordination and writing. CORRESPONDING

AUTHORS Correspondence to Sebastian Amigorena or Luc Fetler. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION

SUPPLEMENTARY TEXT AND FIGURES Supplementary Figures 1–6 (PDF 1191 kb) SUPPLEMENTARY MOVIE 1 Polyclonal CD8+ T (poly-T) cells (green, cytoplasmic labelling) and OVA-specific CD8+ T (OVA-T)

cells (red) were injected i.v. into mice that were immunized 4 h later with anti-DEC205-OVA conjugates plus anti-CD40. Two-photon imaging was performed 18 h later on draining LNs, after

labelling of LN resident DCs (green, membrane labelling). Time of imaging: 30 min. Playback speed: 200×. Size: 175×175μm. (AVI 1833 kb) SUPPLEMENTARY MOVIE 2 Polyclonal CD8+ T (poly-T) cells

(green, cytoplasmic labelling) were injected i.v. into mice that were immunized 4 h later with anti-DEC205-OVA conjugates plus anti-CD40. 2-photon imaging was performed 18 h later on

draining LNs, after labelling of LN resident DC (green, membrane labelling). Time of imaging: 30 min. Playback speed: 200×. Size: 280×280μm. (AVI 1551 kb) SUPPLEMENTARY MOVIE 3 Time–lapse

video–microscopy (acceleration 52×; 65×65 μm) showing transient interactions between polyclonal CD8+ T cells and bone marrow-derived DC previously incubated with OVA peptide (5μM) and LPS

for 18h. (AVI 1050 kb) SUPPLEMENTARY MOVIE 4 Time–lapse video-microscopy (acceleration 52×; 65×65 μm) showing long–lasting interactions between naive OVA–specific CD8+ T cells and bone

marrow-derived DC previously incubated with OVA peptide (5μM) and LPS for 18h. (AVI 1410 kb) SUPPLEMENTARY MOVIE 5 Time-lapse video-microscopy (acceleration 52×; 65×65 μm) showing

long-lasting interactions of both polyclonal CD8+ T cells (poly-T, green) and OVA–specific CD8+ T cells (OVA–T, red) with bone marrow-derived DC previously incubated with OVA peptide (5μM)

and LPS for 18h. (AVI 1757 kb) SUPPLEMENTARY MOVIE 6 Time-lapse video-microscopy (acceleration 52×; 65×65 μm) showing long-lasting interactions of only OVA–specific CD8+ T cells (OVA–T, red)

and Pertussis toxin pre-treated polyclonal CD8+ T cells (PTX treated poly–T, green) with bone marrow-derived DC previously incubated with OVA peptide (5μM) and LPS for 18h. (AVI 1757 kb)

SUPPLEMENTARY MOVIE 7 Time–lapse video-microscopy (acceleration 52×; 65×65 μm) of CFSE-labeled OVA–specific CD8+ T cells (OVA–T, green) and polyclonal CD8+ T cells (poly–T, unlabeled)

interacting with OVA–loaded bone marrow derived DC (OVA-DC, unlabeled) versus CMTMR–labeled unloaded bone marrow–derived DC (0-DC, red). (AVI 4387 kb) SUPPLEMENTARY MOVIE 8 CMTMR–labeled

polyclonal CD8+ T (poly–T) cells (blue) were injected i.v. into mice that were injected s.c. with a mixture (1:1) of GFP+ OVA–pulsed DC (red) and CFP+ unloaded DC (green). Two-photon imaging

was performed 18 h later on draining LNs. The numbers of contacts between poly-T cells and OVA-DC (red squares) or 0-DC (green squared) were quantified and incremented each time a new

contact appears. Time of imaging: 30 min. Playback speed: 200×. Size: 230×230μm. (AVI 2181 kb) SUPPLEMENTARY MOVIE 9 CMTMR–labeled polyclonal CD8+ T (poly–T) cells (blue) and OVA–specific

CD8+ T cells (unlabeled) were injected i.v. into mice that were injected s.c. with a mixture (1:1) of GFP+ OVA–pulsed DC (red) and CFP+ unloaded DC (green). Two–photon imaging was performed

18 h later on draining LNs. The numbers of contacts between poly–T cells and OVA–DC (red squares) or 0–DC (green squared) were quantified and incremented each time a new contact appears.

Time of imaging: 30 min. Playback speed: 200×. Size: 230×230μm. (AVI 5184 kb) SUPPLEMENTARY MOVIE 10 CMTMR–labeled CCR5–deficient polyclonal CD8+ T (poly–T) cells (blue) and OVA–specific

CD8+ T cells (unlabeled) were injected i.v. into mice that were injected s.c. with a mixture (1:1) of GFP+ OVA–pulsed DC (red) and CFP+ unloaded DC (green). Two–photon imaging was performed

18 h later on draining LNs. The numbers of contacts between poly–T cells and OVA–DC (red squares) or 0–DC (green squared) were quantified and incremented each time a new contact appears.

Time of imaging: 25 min. Playback speed: 200×. Size: 230×230μm. (AVI 1895 kb) SUPPLEMENTARY MOVIE 11 CMTMR–labeled polyclonal CD8+ T (poly–T) cells (blue) and GFP+ OVA–specific CD8+ T cells

(green) were injected i.v. into mice that were injected s.c. with CFP+ OVA–pulsed DC (red). Two–photon imaging was performed 18 h later on draining lymph nodes. Time of imaging: 15 min.

Playback speed: 200×. Size: 110×110μm. (AVI 533 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Hugues, S., Scholer, A., Boissonnas, A. _et al._

Dynamic imaging of chemokine-dependent CD8+ T cell help for CD8+ T cell responses. _Nat Immunol_ 8, 921–930 (2007). https://doi.org/10.1038/ni1495 Download citation * Received: 08 March 2007

* Accepted: 25 June 2007 * Published: 29 July 2007 * Issue Date: September 2007 * DOI: https://doi.org/10.1038/ni1495 SHARE THIS ARTICLE Anyone you share the following link with will be

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