Little is known about the molecular mechanisms that determine the entry into the lymph node and intranodal positioning of lymph-derived cells. By injecting cells directly into afferent lymph vessels of popliteal lymph nodes, we demonstrate that lymph-derived T cells entered lymph-node parenchyma mainly from peripheral medullary sinuses, whereas dendritic cells (DCs) transmigrated through the floor of the subcapsular sinus on the afferent side. Transmigrating DCs induced local changes that allowed the concomitant entry of T cells at these sites. Signals mediated by the chemokine receptor CCR7 were absolutely required for the directional migration of both DCs and T cells into the T cell zone but were dispensable for the parenchymal entry of lymph-derived T cells and dendrite probing of DCs. Our findings provide insight into the molecular and structural requirements for the entry into lymph nodes and intranodal migration of lymph-derived cells of the immune system.
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We thank M. Herberg for animal care; E. Kremmer (Helmholtz Zentrum München) for antibodies; and G. Bernhardt for discussions and comments on the manuscript. Supported by Deutsche Forschungsgemeinschaft (SFB566-A14, SFB587-B3 and EXC62, 'Rebirth', to R.F.) and Boehringer Ingelheim Fonds (A.B.).
The authors declare no competing financial interests.
Supplementary Figures 1–5 (PDF 444 kb)
Ccr7-/- BMDCs do not reach the paracortical TCZ after intralymphatic application. (MOV 6411 kb)
Long-term two-photon microscopical imaging of intranodal DC migration behavior. (MOV 9414 kb)
Directional displacement and uropod formation of wt BMDCs is not reflected in Ccr7-/- BMDCs. (MOV 3477 kb)
Close-up view of cellular morphology during intranodal migration. (MOV 1190 kb)
Wt and Ccr7-/- BMDCs exhibit comparable dendrite movement and sweeping behavior. (MOV 2993 kb)
After i.l. application, primary skin-derived wt DC display an intranodal migration comparable to wt BMDCs. (MOV 9439 kb)
Transmigration of wt CD4 through lymphatic endothelium of the capsule-lining sinus floor. (MOV 6430 kb)
Transmigration of wt CD4 through medullar sinus endothelium. (MOV 1676 kb)
Upon exiting the capsule-lining sinus, wt CD4 preferably migrate within peripheral MC. (MOV 10025 kb)
Intralymphatically administered CD4 T cells enter LN parenchyma in a CCR7-dependent manner primarily within peripheral medullary areas. (MOV 3958 kb)
Immigration of DC allows CD4 T cells to enter the LN at SCS. (MOV 7426 kb)
LPS-treated CD4 cells fail to enter popLN via SCS. (MOV 1003 kb)
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