L-selectin-negative CCR7 effector and memory CD8+ T cells enter reactive lymph nodes and kill dendritic cells

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Abstract

T lymphocytes lacking the lymph node–homing receptors L-selectin and CCR7 do not migrate to lymph nodes in the steady state. Instead, we found here that lymph nodes draining sites of mature dendritic cells or adjuvant inoculation recruited L-selectin-negative CCR7 effector and memory CD8+ T cells. This recruitment required CXCR3 expression on T cells and occurred through high endothelial venules in concert with lumenal expression of the CXCR3 ligand CXCL9. In reactive lymph nodes, recruited T cells established stable interactions with and killed antigen-bearing dendritic cells, limiting the ability of these dendritic cells to activate naive CD4+ and CD8+ T cells. The inducible recruitment of blood-borne effector and memory T cells to lymph nodes may represent a mechanism for terminating primary and limiting secondary immune responses.

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Figure 1: In vitro–generated L-selectin-negative CCR7 effector CD8+ T cells are excluded from resting lymph nodes but migrate to reactive lymph nodes.
Figure 2: Effector and effector memory CD8+ T cells generated by in vivo priming rapidly migrate to reactive lymph nodes.
Figure 3: Effector T cells roll and stick on HEVs of reactive lymph nodes.
Figure 4: CXCR3 is required for the migration of effector CD8+ T cells to reactive lymph nodes.
Figure 5: CXCL9 is transiently displayed on the lumenal surfaces of HEVs in reactive lymph nodes.
Figure 6: Effector and memory CD8+ T cells kill antigen-presenting DCs in reactive lymph nodes.
Figure 7: Priming of naive CD4+ and CD8+ T cells is inhibited in mice containing effector or memory CD8+ T cells.

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Acknowledgements

We thank L. Lefrancois (University of Connecticut) for the Listeria monocytogenes strain expressing OVA; P. Dellabona (DIBIT San Raffaele Research Institute), C. Gerard (Harvard Medical School), J. Kirberg (Max Planck Institute) and M. Lipp (Max Delbruck Center) for transgenic and knockout mice; M. Manz and M. Uguccioni for critical reading and comments; A. Almeida for discussions; and D. Jarrossay, E. Mira-Catò, L. Perlini and M. Convert for technical help. Supported by the Swiss National Science Foundation (31-101962 and 31-109832), the European Commission FP6 'Network of Excellence' initiative (N. LSHG-CT-2003-502935 MAIN, LSHB-CT-2004-512074 DC-THERA and LSHG-CT-2005-005203 MUGEN), the intramural program of the National Institute of Allergy and Infectious Diseases of the National Institutes of Health, the Cancer Research Institute (A.H.) and the Helmut Horten Foundation (for the Institute for Research in Biomedicine).

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Correspondence to Federica Sallusto.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Priming of naive CD4+ T cells is partially recovered in mice carrying effector or memory CD8+ T cells when MHC class I and class II peptides are provided on different DCs. (PDF 50 kb)

Supplementary Video 1

Dynamic intravital 2-photon imaging movies of prolonged T cell-DC interaction followed by DC cell body fragmentation. SIINFEKL pulsed-DCs (green) and control peptide pulsed-DCs (purple) were activated with LPS and co-injected in the footpad of recipient mice 24 hours prior to intravenous injection of in vitro-generated L-selectin effector OT-I cells (red). Intravital 2-photon microscopy imaging was performed on the draining popliteal lymph node of anesthetized recipient animals in the interfollicular region 60-160 μm below the capsule starting at 3 hours after adoptive transfer of T cells. An example of prolonged T cell interaction with SIINFEKL pulsed-DCs is shown (highlighted by dotted circle). These prolonged interactions are followed by fragmentation of the DC cell body over the subsequent several minutes. Control peptide pulsed-DCs (purple) are either ignored by or exhibit transient interaction with T cells, and did not exhibit any fragmentation during the same observation period. Clock (h:min:sec) denotes the time after i.v. OT-I cell transfer. Scale bar as indicated. Playback speed: 450X. Total time: 3 hrs 18 mins. (AVI 7938 kb)

Supplementary Video 2

Dynamic intravital 2-photon imaging movies of prolonged T cell-DC interaction followed by DC cell body fragmentation. SIINFEKL pulsed-DCs (green) and control peptide pulsed-DCs (purple) were activated with LPS and co-injected in the footpad of recipient mice 24 hours prior to intravenous injection of in vitro-generated L-selectin effector OT-I cells (red). Intravital 2-photon microscopy imaging was performed on the draining popliteal lymph node of anesthetized recipient animals in the interfollicular region 60-160 μm below the capsule starting at 3 hours after adoptive transfer of T cells. Shown is an example of prolonged T cell interaction with SIINFEKL pulsed-DCs followed by fragmentation of the DC cell body. Clock (h:min:sec) denotes the time after i.v. OT-I cell transfer. Scale bar as indicated. Playback speed: 450X. Total time: 1 hr 8 mins 45 secs (Movie 2). (AVI 1206 kb)

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Guarda, G., Hons, M., Soriano, S. et al. L-selectin-negative CCR7 effector and memory CD8+ T cells enter reactive lymph nodes and kill dendritic cells. Nat Immunol 8, 743–752 (2007) doi:10.1038/ni1469

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