Stromal mesenteric lymph node cells are essential for the generation of gut-homing T cells in vivo Export

@article{citeulike:3417284, abstract = {T cells primed in the gut-draining mesenteric lymph nodes (mLN) are imprinted to express {alpha}4{beta}7-integrin and chemokine receptor CCR9, thereby enabling lymphocytes to migrate to the small intestine. In vitro activation by intestinal dendritic cells (DC) or addition of retinoic acid (RA) is sufficient to instruct expression of these gut-homing molecules. We report that in vivo stroma cells, but not DC, allow the mLN to induce the generation of gut tropism. Peripheral LN (pLN) transplanted into the gut mesenteries fail to support the generation of gut-homing T cells, even though gut-derived DC enter the transplants and prime T cells. DC that fail to induce {alpha}4{beta}7-integrin and CCR9 in vitro readily induce these factors in vivo upon injection into mLN afferent lymphatics. Moreover, uniquely mesenteric but not pLN stroma cells express high levels of RA-producing enzymes and support induction of CCR9 on activated T cells in vitro. These results demonstrate a hitherto unrecognized contribution of stromal cell delivered signals, including RA, on the imprinting of tissue tropism in vivo. 10.1084/jem.20080039}, author = {Hammerschmidt, Swantje I. and Ahrendt, Manuela and Bode, Ulrike and Wahl, Benjamin and Kremmer, Elisabeth and Forster, Reinhold and Pabst, Oliver}, citeulike-article-id = {3417284}, citeulike-linkout-0 = {http://dx.doi.org/10.1084/jem.20080039}, citeulike-linkout-1 = {http://jem.rupress.org/content/jem.20080039v1/.abstract}, citeulike-linkout-2 = {http://jem.rupress.org/content/jem.20080039v1/.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/18852290}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=18852290}, comment = {MADCAM-1 (a4b7 ligand) is expressed all along intestinal venules CCL25 (CCR9 ligand) Prox S.I > Ileum (low expression) > Colon (no expression) ---=note-separator=--- mLN were excised from WT mice and replaced by either pLN or mLN fragments isolated from EGFP mice. T cells acquire gut homing receptors when MLNs where replaced with MLNs but not when they were replaced with pLNs ---=note-separator=--- bm-1 mice carry a mutated H2-K molecule and cannot present Ova peptide in the context of MHC class I ---=note-separator=--- Intralymph node injection of DCs. BM-DCs (GM-CSF) and splenic DCs don't induce gut homing receptors in vitro but they do it when they are injected into MLNs. Experiment design DO11.10 transferred into WT recipient 1 day later BM-DCs i.l injected 3 d later cells are harvested and analyzed for a4b7,CCr9, CFSE analysis. ---=note-separator=--- CD45- gp38+ stroma cells express RALDH1 and RALDH3 MLNs CD103+DCs express RALDH2 (but not the CD103-) In the pLNs CD103- DCs express more RALDH2 than CD103+ counterparts (but the level is low when compared with MLN CD103+ DCs) ---=note-separator=--- ISOLATION OF STROMA CELLS LNs were digested at 37 C for 60 min with 0.1 mg/mL Liberase Blendzyme 2 (Roche) and 50 U/mL DNase I. Digested LN cell suspensions were cultured in DMEM/10\% FCS/PS for 10d. Medium was enchanged every 2nd day to remove non adherent cells. ---=note-separator=--- The numbers of CD103+ DCs in CCR7-/- are greatly reduced and the cells remaining are RALDH2 negative. RALDH 1,2 and 3 expression in the stroma is not affected (so the stroma does not get educated to produce RA by DCs that come from LP) BM DCs failed to induce CCR9 upregulation when injected into MLNs of CCR7 -/- animals but not when injected in the MLNs of WT mice Is the antigen presented by the BM-DCs or could be transferred to other DCs present in the MLNs? When they injected MHC II -/- or bm1 BM DCs there was not proliferation of the OTI or OTII cells so it must be direct presentation.}, day = {13}, doi = {10.1084/jem.20080039}, issn = {1540-9538}, journal = {J. Exp. Med.}, keywords = {ilfs, stroma}, month = {October}, pages = {jem.20080039+}, posted-at = {2009-11-12 00:05:47}, priority = {2}, title = {Stromal mesenteric lymph node cells are essential for the generation of gut-homing T cells in vivo}, url = {http://dx.doi.org/10.1084/jem.20080039}, year = {2008} }

TY - JOUR ID - citeulike:3417284 L3 - citeulike-article-id:3417284 N2 - T cells primed in the gut-draining mesenteric lymph nodes (mLN) are imprinted to express {alpha}4{beta}7-integrin and chemokine receptor CCR9, thereby enabling lymphocytes to migrate to the small intestine. In vitro activation by intestinal dendritic cells (DC) or addition of retinoic acid (RA) is sufficient to instruct expression of these gut-homing molecules. We report that in vivo stroma cells, but not DC, allow the mLN to induce the generation of gut tropism. Peripheral LN (pLN) transplanted into the gut mesenteries fail to support the generation of gut-homing T cells, even though gut-derived DC enter the transplants and prime T cells. DC that fail to induce {alpha}4{beta}7-integrin and CCR9 in vitro readily induce these factors in vivo upon injection into mLN afferent lymphatics. Moreover, uniquely mesenteric but not pLN stroma cells express high levels of RA-producing enzymes and support induction of CCR9 on activated T cells in vitro. These results demonstrate a hitherto unrecognized contribution of stromal cell delivered signals, including RA, on the imprinting of tissue tropism in vivo. 10.1084/jem.20080039 JF - J. Exp. Med. SN - 1540-9538 TI - Stromal mesenteric lymph node cells are essential for the generation of gut-homing T cells in vivo SP - jem.20080039 KW - ilfs KW - stroma N1 - MADCAM-1 (a4b7 ligand) is expressed all along intestinal venules CCL25 (CCR9 ligand) Prox S.I > Ileum (low expression) > Colon (no expression) N1 - mLN were excised from WT mice and replaced by either pLN or mLN fragments isolated from EGFP mice. T cells acquire gut homing receptors when MLNs where replaced with MLNs but not when they were replaced with pLNs N1 - bm-1 mice carry a mutated H2-K molecule and cannot present Ova peptide in the context of MHC class I N1 - Intralymph node injection of DCs. BM-DCs (GM-CSF) and splenic DCs don't induce gut homing receptors in vitro but they do it when they are injected into MLNs. Experiment design DO11.10 transferred into WT recipient 1 day later BM-DCs i.l injected 3 d later cells are harvested and analyzed for a4b7,CCr9, CFSE analysis. N1 - CD45- gp38+ stroma cells express RALDH1 and RALDH3 MLNs CD103+DCs express RALDH2 (but not the CD103-) In the pLNs CD103- DCs express more RALDH2 than CD103+ counterparts (but the level is low when compared with MLN CD103+ DCs) N1 - ISOLATION OF STROMA CELLS LNs were digested at 37 C for 60 min with 0.1 mg/mL Liberase Blendzyme 2 (Roche) and 50 U/mL DNase I. Digested LN cell suspensions were cultured in DMEM/10% FCS/PS for 10d. Medium was enchanged every 2nd day to remove non adherent cells. N1 - The numbers of CD103+ DCs in CCR7-/- are greatly reduced and the cells remaining are RALDH2 negative. RALDH 1,2 and 3 expression in the stroma is not affected (so the stroma does not get educated to produce RA by DCs that come from LP) BM DCs failed to induce CCR9 upregulation when injected into MLNs of CCR7 -/- animals but not when injected in the MLNs of WT mice Is the antigen presented by the BM-DCs or could be transferred to other DCs present in the MLNs? When they injected MHC II -/- or bm1 BM DCs there was not proliferation of the OTI or OTII cells so it must be direct presentation. AU - Hammerschmidt, Swantje AU - Ahrendt, Manuela AU - Bode, Ulrike AU - Wahl, Benjamin AU - Kremmer, Elisabeth AU - Forster, Reinhold AU - Pabst, Oliver PY - 2008/10/13/ UR - http://dx.doi.org/10.1084/jem.20080039 ER -