Generation of Lymph Node-fat Pad Chimeras for the Study of Lymph Node Stromal Cell Origin

The stroma is a key component of the lymph node structure and function. However, little is known about its origin, exact cellular composition and the ...

University of Birmingham

University of Edinburgh

Tumor cells often escape immunosurveillance by down-regulating MHC class I molecule expression. For human Vgamma9Vdelta2 T cells, a major peripheral blood T cell subset with broad antitumor reactivity, this down-regulation can affect signals transmitted by both the inhibitory and the activating MHC class I and Ib-specific NK receptors (NKRs) that these lymphocytes frequently express. To assess the overall impact of MHC down-regulation on Vgamma9Vdelta2 T cell activation, we used stable beta(2)-microglobulin knockdown to generate tumor cells with a approximately 10-fold down-modulation of all MHC class I molecules. This down-modulation had little effect on T cell proliferation or cytokine production, but modified tumor cell killing efficiency. Ab-blocking studies identified ILT2 as an important inhibitor of tumor cell killing by Vgamma9Vdelta2 T cells. Down-modulation of MHC class I and Ib molecules severely reduced ILT2 inhibitory signaling, but still allowed signaling by activating CD94-based receptors. It also unveiled a frequent enhancing effect of NKG2D on tumor killing by Vgamma9Vdelta2 T cells. Current models suggest that activating NKRs have less affinity for their MHC ligands than homologous inhibitory NKRs. Our results show that, despite this, activating NKRs recognizing MHC class I molecules play an important role in the increased killing by Vgamma9Vdelta2 T cells of tumor cells with down-regulated MHC class I molecule expression, and suggest that these T cells will best lyse tumor cells combining MHC class I molecule expression down-regulation with up-regulated NKG2D ligand expression.

Complex interplay of activating and inhibitory signals received by Vgamma9Vdelta2 T cells revealed by target cell beta2-microglobulin knockdown.

&#x53D1;&#x5C55;&#x7684;&#x8F85;&#x52A9;&#x7684;&#x6DCB;&#x5DF4;&#x5668;&#x5B98;&#xFF0C;&#x5982;&#x6DCB;&#x5DF4;&#x7ED3; &#xFF08;LNs&#xFF09;&#xFF0C;&#x5728;&#x80DA;&#x80CE;&#x6DCB;&#x5DF4;&#x7EC4;&#x7EC7;&#x8BF1;&#x5BFC;&#x56E0;&#x5B50; (LTi) &#x7EC6;&#x80DE;&#x548C;&#x95F4;&#x8D28;&#x7EC6;&#x80DE;&#x4E4B;&#x95F4;&#x7684;&#x76F8;&#x4E92;&#x64CD;&#x4F5C;&#x7684;&#x7ED3;&#x679C;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x6700;&#x521D;&#x7684;&#x4E8B;&#x4EF6;&#x53D1;&#x751F;&#x4E4B;&#x524D;&#x8FDB;&#x884C; LTi &#x57FA;&#x8D28;&#x7EC6;&#x80DE;&#x76F8;&#x58F0;&#x4EA4;&#x4E92;&#x8BF1;&#x5BFC; LN &#x8BBE;&#x5907;&#x516C;&#x53F8;&#x5F62;&#x6210;&#x4E0D;&#x5145;&#x5206;&#x7684;&#x9610;&#x8FF0;&#x3002;&#x5728;&#x6B64;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x5C55;&#x793A;&#x5728;&#x5C0F;&#x9F20;&#x80DA;&#x80CE;&#x7684;&#x8179;&#x80A1;&#x6C9F; LN &#x8BBE;&#x5907;&#x516C;&#x53F8;&#x5F00;&#x53D1;&#x4ECE;&#x5468;&#x56F4;&#x7684;&#x6DCB;&#x5DF4;&#x56CA;&#x3001; &#x7403;&#x5F62;&#x7ED3;&#x6784;&#x82BD;&#x4ECE;&#x9759;&#x8109;&#x7684;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x7684;&#x95F4;&#x8D28;&#x7EC6;&#x80DE;&#x3002;&#x4F7F;&#x7528;&#x8179;&#x80A1;&#x6C9F;&#x548C;&#x80A0;&#x7CFB;&#x819C; LNs (mLNs)&#xFF0C;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x7684;&#x57FA;&#x8D28;&#x7EC6;&#x80DE;&#x652F;&#x6301;&#x4E24;&#x6B65;&#x6210;&#x719F;&#x6A21;&#x578B;&#x7684;&#x8BC1;&#x636E;&#xFF1A; &#x7B2C;&#x4E00;&#xFF0C;ICAM-1(-)VCAM-1(-) &#x95F4;&#x5145;&#x8D28;&#x4F53;&#x7EC6;&#x80DE;&#x6210;&#x4E3A;&#x8FDB;&#x7A0B;&#x72EC;&#x7ACB;&#x7684; LTi &#x7EC6;&#x80DE;&#x548C;&#x6DCB;&#x5DF4; &beta; &#x53D7;&#x4F53; (LTbetaR) &#x7684;&#x4FE1;&#x53F7;&#x8F6C;&#x5BFC;&#x4E2D;&#x7684;&#x5355;&#x5143;&#x683C;&#xFF0C;ICAM-1(int)VCAM-1(int)&#x3002;&#x7B2C;&#x4E8C;&#x6B65;&#x6D89;&#x53CA; ICAM-1(int)VCAM-1(int) &#x7EC6;&#x80DE;&#x5411; ICAM-1(high)VCAM-1(high) &#x9ECF;&#x819C;&#x6C49;&#x8BED;&#x7EC6;&#x80DE;&#x7C98;&#x9644; molecule-1(+) &#x7BA1;&#x7406;&#x5668;&#x7EC6;&#x80DE;&#x7684;&#x6210;&#x719F;&#x548C;&#x53D6;&#x51B3;&#x4E8E; LTi &#x7EC6;&#x80DE;&#x548C; LTbetaR&#x3002;&#x53E6;&#x5916; alphaLTbetaR &#x6FC0;&#x52A8;&#x5230; LN &#x673A;&#x5173;&#x6587;&#x5316;&#x662F;&#x4E0D;&#x8DB3;&#x4EE5;&#x4FC3;&#x4F7F; ICAM-1(int)VCAM-1(int) &#x7EC6;&#x80DE;&#x5230;&#x6210;&#x719F;&#x3002;LtbetaR(-/-) &#x80DA;&#x80CE;&#xFF0C;&#x4E24;&#x8005;&#x8179;&#x80A1;&#x6C9F;&#x548C;&#x63D0;&#x4F9B;&#x5DE8;&#x989D;&#x57FA;&#x8D28;&#x7EC6;&#x80DE;&#x663E;&#x793A;&#x5728; ICAM-1(int)VCAM-1(int) &#x9636;&#x6BB5;&#xFF0C;&#x4E00;&#x4E2A;&#x5757;&#xFF0C;&#x4E0E;&#x8179;&#x80A1;&#x6C9F; LNs&#xFF0C;mLNs &#x575A;&#x6301;&#x8F83;&#x957F;&#x548C;&#x6240;&#x5305;&#x542B;&#x7684; LTi &#x7EC6;&#x80DE;&#xFF0C;&#x6301;&#x7EED;&#x7684;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x7684;&#x767D;&#x7EC6;&#x80DE;&#x4ECB;&#x7D20;-7&#xFF0C;Cxcl13&#xFF0C;&#x4E0E;&#x76F8;&#x5173;&#x548C;&#xFF0C;&#x7A0B;&#x5EA6;&#x8F83;&#x8F7B;&#xFF0C;Ccl21.&#x91C7;&#x53D6;&#x5728;&#x4E00;&#x8D77;&#xFF0C;&#x8FD9;&#x4E9B;&#x7ED3;&#x679C;&#x7A81;&#x51FA;&#x663E;&#x793A;&#x4FE1;&#x53F7;&#x548C;&#x7EC6;&#x80DE;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#xFF0C;&#x8BF1;&#x4F7F;&#x57FA;&#x8D28;&#x7EC6;&#x80DE;&#x7684;&#x6210;&#x719F;&#xFF0C;&#x6700;&#x7EC8;&#x5BFC;&#x81F4;&#x6DCB;&#x5DF4;&#x7EC4;&#x7EC7;&#x5F62;&#x6210;&#x7684;&#x91CD;&#x8981;&#x6027;&#x3002;

&#x4E2A;&#x4F53;&#x53D1;&#x80B2;&#x95F4;&#x8D28;&#x7EC4;&#x7EC7;&#x8005;&#x7EC6;&#x80DE;&#x6DCB;&#x5DF4;&#x7ED3;&#x53D1;&#x5C55;&#x8FC7;&#x7A0B;&#x3002;

The development of secondary lymphoid organs, such as lymph nodes (LNs), in the embryo results from the reciprocal action between lymphoid tissue inducer (LTi) cells and stromal cells. However, the initial events inducing LN anlagen formation before the LTi stromal cells cross-talk interactions take place are not fully elucidated. In this study, we show that the inguinal LN anlagen in mouse embryos developed from mesenchymal cells surrounding the lymph sacs, spherical structures of endothelial cells that bud from veins. Using inguinal and mesenteric LNs (mLNs), we provide evidence supporting a two-step maturation model for stromal cells: first, ICAM-1(-)VCAM-1(-) mesenchymal precursor cells become ICAM-1(int)VCAM-1(int) cells, in a process independent of LTi cells and lymphotoxin beta receptor (LTbetaR) signaling. The second step involves the maturation of ICAM-1(int)VCAM-1(int) cells to ICAM-1(high)VCAM-1(high) mucosal addressin cell adhesion molecule-1(+) organizer cells and depends on both LTi cells and LTbetaR. Addition of alphaLTbetaR agonist to LN organ cultures was sufficient to induce ICAM-1(int)VCAM-1(int) cells to mature. In LtbetaR(-/-) embryos, both inguinal and mLN stromal cells showed a block at the ICAM-1(int)VCAM-1(int) stage, and, contrary to inguinal LNs, mLNs persist longer and contained LTi cells, which correlated with the sustained gene expression of Il-7, Cxcl13, and, to a lesser degree, Ccl21. Taken together, these results highlight the importance of the signals and cellular interactions that induce the maturation of stromal cells and ultimately lead to the formation of lymphoid tissues.

Ontogeny of stromal organizer cells during lymph node development.

Le d&#xE9;veloppement des organes lympho&#xEF;des secondaires, tels que les ganglions lymphatiques (SLN), dans les r&#xE9;sultats d&#x26;#39;embryons &#xE0; partir de l&#x26;#39;action r&#xE9;ciproque entre les tissus lympho&#xEF;des inducteurs (LTI) et les cellules stromales. Cependant, les &#xE9;v&#xE9;nements initiaux induisant LN &#xE9;bauches de formation avant que les cellules stromales LTi diaphonie interactions ont lieu ne sont pas compl&#xE8;tement &#xE9;lucid&#xE9;s. Dans cette &#xE9;tude, nous montrons que l&#x26;#39;&#xE9;bauche LN inguinale chez les embryons de souris d&#xE9;velopp&#xE9; &#xE0; partir de cellules m&#xE9;senchymateuses entourant les sacs lymphatiques, des structures sph&#xE9;riques de cellules endoth&#xE9;liales qui bourgeonnent &#xE0; partir des veines. Utilisation de ganglions inguinaux et m&#xE9;sent&#xE9;riques (mLNs), nous en apporter la preuve d&#x26;#39;un mod&#xE8;le de maturation en deux &#xE9;tapes pour les cellules stromales: d&#x26;#39;abord, ICAM-1 (-) VCAM-1 (-) des cellules pr&#xE9;curseurs m&#xE9;senchymateuses deviennent ICAM-1 (int) de VCAM-1 (int) des cellules, dans un processus ind&#xE9;pendant de cellules LTi et des r&#xE9;cepteurs b&#xEA;ta lymphotoxine (LTbetaR) de signalisation. La deuxi&#xE8;me &#xE9;tape consiste &#xE0; la maturation de l&#x26;#39;ICAM-1 (int) VCAM-1 (int), les cellules &#xE0; ICAM-1 (&#xE9;lev&#xE9;) de VCAM-1 (&#xE9;lev&#xE9;) des muqueuses cellules addressin cell adhesion organisateur mol&#xE9;cule-1 (+) et d&#xE9;pend &#xE0; la fois LTi et les cellules LTbetaR. Ajout d&#x26;#39;un agoniste alphaLTbetaR aux cultures d&#x26;#39;organes LN &#xE9;tait suffisante pour induire ICAM-1 (int) VCAM-1 (int), les cellules &#xE0; maturit&#xE9;. En LtbetaR (- / -) des embryons, les deux cellules stromales inguinaux et MLN a montr&#xE9; un bloc &#xE0; l&#x26;#39;ICAM-1 (int) VCAM-1 (int) stade, et, contrairement &#xE0; LN inguinales, mLNs persister plus longtemps et contenaient des cellules LTi, qui corr&#xE9;l&#xE9;e avec l&#x26;#39;expression g&#xE9;nique soutenue d&#x26;#39;IL-7, CXCL13, et, &#xE0; un degr&#xE9; moindre, CCL21. Pris ensemble, ces r&#xE9;sultats mettent en &#xE9;vidence l&#x26;#39;importance des signaux et des interactions cellulaires qui induisent la maturation des cellules stromales et finalement conduire &#xE0; la formation de tissus lympho&#xEF;des.

Ontogen&#xE8;se des cellules stromales de l&#x26;#39;organiseur durant le d&#xE9;veloppement des ganglions lymphatiques

El desarrollo de los &#xF3;rganos linfoides secundarios, tales como los ganglios linf&#xE1;ticos (LNS), en los resultados de embriones a partir de la acci&#xF3;n rec&#xED;proca entre inductoras de tejido linfoide (LTI) y c&#xE9;lulas del estroma. Sin embargo, los primeros eventos que inducen la formaci&#xF3;n de LN esbozos antes de que las c&#xE9;lulas del estroma LTi diafon&#xED;a interacciones tienen lugar no est&#xE1;n completamente dilucidados. En este estudio, mostramos que los esbozos LN inguinal en embriones de rat&#xF3;n desarrollado a partir de las c&#xE9;lulas mesenquimales que rodean los sacos linf&#xE1;ticos, estructuras esf&#xE9;ricas de c&#xE9;lulas endoteliales, que brotan de las venas. Usando LNs inguinales y mesent&#xE9;ricos (mLNs), proporcionamos evidencia que apoya un modelo de maduraci&#xF3;n de dos pasos para las c&#xE9;lulas del estroma: primero, la ICAM-1 (-) VCAM-1 (-) c&#xE9;lulas mesenquimales precursoras convertido ICAM-1 (int) VCAM-1 (int) c&#xE9;lulas, en un proceso independiente de las c&#xE9;lulas del receptor de LTI y linfotoxina beta (LTbetaR) de se&#xF1;alizaci&#xF3;n. El segundo paso consiste en la maduraci&#xF3;n de ICAM-1 (int) VCAM-1 (int) de las c&#xE9;lulas a la ICAM-1 (alto) VCAM-1 (alto) la mucosa adhesi&#xF3;n celular las c&#xE9;lulas del organizador mol&#xE9;cula-1 (+) y depende tanto de LTi c&#xE9;lulas y LTbetaR. La adici&#xF3;n de agonista alphaLTbetaR a cultivos de &#xF3;rganos LN era suficiente para inducir la ICAM-1 (int) VCAM-1 (int) c&#xE9;lulas para madurar. En LtbetaR (- / -) embriones, tanto las c&#xE9;lulas del estroma inguinales y MLN mostr&#xF3; un bloque en el ICAM-1 (int) VCAM-1 (int) el escenario, y, al contrario de LN inguinal, persisten mLNs m&#xE1;s largo y contiene c&#xE9;lulas LTI, que correlacionada con la expresi&#xF3;n g&#xE9;nica sostenida de IL-7, CXCL13, y, en menor grado, CCL21. En conjunto, estos resultados ponen de manifiesto la importancia de las se&#xF1;ales y las interacciones celulares que inducen la maduraci&#xF3;n de las c&#xE9;lulas del estroma y en &#xFA;ltima instancia conducir a la formaci&#xF3;n de los tejidos linfoides.

Ontogenia de las c&#xE9;lulas del estroma Organizador durante el desarrollo del ganglio linf&#xE1;tico

Die Entwicklung der sekund&auml;ren lymphatischen Organe wie Lymphknoten (LNs), im Embryo ergibt sich aus der Wechselwirkung zwischen Lymphgewebe Induktor (LTi) und stromal Zellen. Jedoch sind die urspr&uuml;nglichen Ereignisse induzierende LN Anlagen Bildung, bevor die LTi stromal Zellen Nebensprechen Interaktionen stattfinden nicht vollst&auml;ndig aufgekl&auml;rt. In dieser Studie zeigen wir, dass die Bikinizone LN-Anlagen in Maus-Embryonen entwickelt von mesenchymalen Zellen, die rund um die Lymphe T&uuml;ten, kugelf&ouml;rmige Strukturen der Endothelzellen, die Knospe von Venen. Dank inguinale und mesenterica LNs (mLNs) bieten wir Nachweise f&uuml;r einen zweistufigen Reifung Modell f&uuml;r stromal Zellen: Erstens wurde ICAM-1(-)VCAM-1(-) Mesenchymale Vorl&auml;uferzellen ICAM-1(int)VCAM-1(int) Zellen, in einem Prozess unabh&auml;ngig von LTi-Zellen und Lymphotoxin Beta-Rezeptor (LTbetaR)-Signalisierung. Der zweite Schritt beinhaltet die Reifung der ICAM-1(int)VCAM-1(int) Zellen zu ICAM-1(high)VCAM-1(high) Schleimhaut addressin Zelle Adh&auml;sion molecule-1(+) Veranstalter Zellen und h&auml;ngt von LTi-Zellen und LTbetaR. Zusatz von AlphaLTbetaR Agonist LN Orgel Kulturen konnten ICAM-1(int)VCAM-1(int) Zellen ausreifen zu induzieren. LtbetaR(-/-) Embryonen, beide inguinale und mLN stromal Zellen zeigten einen Block im ICAM-1(int)VCAM-1(int) Stadium, und, entgegen inguinale LNs, mLNs bleiben l&auml;ngere und enthaltene LTi-Zellen, die mit der anhaltenden Genexpression von Il-7, Cxcl13, korreliert und, in geringerem Ma&szlig;e, Ccl21. Taken zusammen, diese Ergebnisse unterstreichen die Bedeutung der Signale und zellulare Wechselwirkungen, die induzieren der Reifung der stromal Zellen und f&uuml;hren letztlich zur Bildung der lymphatischen Gewebe.

Ontogenese der stromal Veranstalter Zellen w&auml;hrend der Entwicklung der Lymphknoten.

Lo sviluppo di organi linfoidi secondari, quali i linfonodi (LNs), nell&#39;embrione risultati dall&#39;azione reciproca tra tessuto linfoide dell&#39;induttore (LTi) e cellule stromale. Tuttavia, gli eventi iniziali che inducono la formazione anlagen LN prima avvengono le interazioni cross-talk di cellule stromali LTi non sono completamente chiariti. In questo studio, mostriamo che la anlagen LN inguinali in embrioni di topo sviluppato da cellule mesenchimali che circonda il sacs linfonodi, strutture sferiche delle cellule endoteliali che bud da vene. Utilizzando inguinale e mesenterici LNs (mln), forniamo le prove a sostegno di un modello di maturazione in due fasi per le cellule stromale: in primo luogo, le cellule mesenchimali precursore ICAM-1(-)VCAM-1(-) diventano cellule ICAM-1(int)VCAM-1(int), in un processo indipendente di cellule LTi e segnalazione di lymphotoxin beta del recettore (LTbetaR). Il secondo passo riguarda la maturazione delle cellule ICAM-1(int)VCAM-1(int) a ICAM-1(high)VCAM-1(high) addressin cella adesione molecule-1(+) Organizzatore cellule della mucosa e dipende sia LTi cellule e LTbetaR. Aggiunta di alphaLTbetaR agonista a culture organo LN era sufficiente per indurre le cellule ICAM-1(int)VCAM-1(int) a maturare. In LtbetaR(-/-) gli embrioni, sia inguinale e cellule stromali mLN ha mostrato un blocco nella fase ICAM-1(int)VCAM-1(int) e, contrariamente a LNs inguinale, mln persistere pi&ugrave; a lunghe e conteneva cellule di LTi correlato con l&#39;espressione del gene sostenuta di Il-7, Cxcl13, e, in misura minore, Ccl21. preso insieme, questi risultati sottolineano l&#39;importanza dei segnali e interazioni cellulari che inducono la maturazione delle cellule stromale e infine conducono alla formazione di tessuti linfoidi.

Ontogenesi delle cellule stromali organizzatore durante lo sviluppo del linfonodo.

&#x3053;&#x306E;&#x3088;&#x3046;&#x306A;&#x30EA;&#x30F3;&#x30D1;&#x7D44;&#x7E54;&#x8A98;&#x5C0E;&#xFF08;LTI&#xFF09;&#x7D30;&#x80DE;&#x3068;&#x30B9;&#x30C8;&#x30ED;&#x30FC;&#x30DE;&#x7D30;&#x80DE;&#x3068;&#x306E;&#x9593;&#x306E;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x304B;&#x3089;&#x80DA;&#x306E;&#x7D50;&#x679C;&#x3067;&#x30EA;&#x30F3;&#x30D1;&#x7BC0;&#xFF08;LNS&#xFF09;&#x3068;&#x3057;&#x3066;&#x3001;&#x4E8C;&#x6B21;&#x30EA;&#x30F3;&#x30D1;&#x5668;&#x5B98;&#x306E;&#x958B;&#x767A;&#x3002;&#x3057;&#x304B;&#x3057;&#x3001;LTI&#x30B9;&#x30C8;&#x30ED;&#x30FC;&#x30DE;&#x7D30;&#x80DE;&#x306E;&#x30AF;&#x30ED;&#x30B9;&#x30C8;&#x30FC;&#x30AF;&#x306E;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x304C;&#x884C;&#x308F;&#x308C;&#x308B;&#x524D;&#x306B;&#x3001;LN&#x539F;&#x57FA;&#x5F62;&#x6210;&#x3092;&#x8A98;&#x5C0E;&#x3059;&#x308B;&#x6700;&#x521D;&#x306E;&#x30A4;&#x30D9;&#x30F3;&#x30C8;&#x306F;&#x5B8C;&#x5168;&#x306B;&#x306F;&#x89E3;&#x660E;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x305B;&#x3093;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;&#x3001;&#x30DE;&#x30A6;&#x30B9;&#x80DA;&#x306B;&#x304A;&#x3051;&#x308B;&#x9F20;&#x5F84;&#x90E8;&#x306E;LN&#x539F;&#x57FA;&#x304C;&#x30EA;&#x30F3;&#x30D1;&#x56A2;&#x5468;&#x56F2;&#x306E;&#x9593;&#x8449;&#x7CFB;&#x7D30;&#x80DE;&#x3001;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x305D;&#x306E;&#x82BD;&#x306E;&#x7403;&#x72B6;&#x69CB;&#x9020;&#x304B;&#x3089;&#x958B;&#x767A;&#x3055;&#x308C;&#x305F;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x308B;&#x3002; &#xFF08; - &#xFF09;VCAM-1&#xFF08; - &#xFF09;&#x9593;&#x8449;&#x7CFB;&#x524D;&#x99C6;&#x7D30;&#x80DE;&#x306F;&#x3001;ICAM-1&#xFF08;int&#xFF09;&#x306F;VCAM-1&#x3068;&#x306A;&#x3063;&#x3066;&#x3001;&#x6700;&#x521D;&#x306B;ICAM-1&#xFF1A;&#x9F20;&#x5F84;&#x90E8;&#x304A;&#x3088;&#x3073;&#x8178;&#x9593;&#x819C;LNS&#xFF08;mLNs&#xFF09;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x3001;&#x6211;&#x3005;&#x306F;&#x9593;&#x8CEA;&#x7D30;&#x80DE;&#x306E;2&#x3064;&#x306E;&#x6BB5;&#x968E;&#x306E;&#x6210;&#x719F;&#x30E2;&#x30C7;&#x30EB;&#x3092;&#x652F;&#x6301;&#x3059;&#x308B;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;LTI&#x7D30;&#x80DE;&#x304A;&#x3088;&#x3073;&#x30EA;&#x30F3;&#x30DB;&#x30C8;&#x30AD;&#x30B7;&#x30F3;&#x3B2;&#x53D7;&#x5BB9;&#x4F53;&#xFF08;LTbetaR&#xFF09;&#x30B7;&#x30B0;&#x30CA;&#x30EA;&#x30F3;&#x30B0;&#x306E;&#x72EC;&#x7ACB;&#x3057;&#x305F;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306E;&#xFF08;int&#xFF09;&#x306E;&#x7D30;&#x80DE;&#x3002;&#x7B2C;&#x4E8C;&#x6BB5;&#x968E;&#x306F;&#x3001;ICAM-1&#xFF08;&#x9AD8;&#xFF09;VCAM-1&#xFF08;&#x9AD8;&#xFF09;&#x7C98;&#x819C;&#x30A2;&#x30C9;&#x30EC;&#x30B7;&#x30F3;&#x7D30;&#x80DE;&#x63A5;&#x7740;&#x5206;&#x5B50;1&#xFF08;+&#xFF09;&#x30AA;&#x30FC;&#x30AC;&#x30CA;&#x30A4;&#x30B6;&#x30FC;&#x7D30;&#x80DE;&#x306B;ICAM-1&#xFF08;int&#xFF09;&#x306F;VCAM-1&#xFF08;int&#xFF09;&#x306E;&#x7D30;&#x80DE;&#x306E;&#x6210;&#x719F;&#x3092;&#x4F34;&#x3044;&#x3001;LTI&#x306E;&#x4E21;&#x65B9;&#x306B;&#x4F9D;&#x5B58;&#x7D30;&#x80DE;&#x3068;LTbetaR&#x3002; LN&#x306E;&#x5668;&#x5B98;&#x57F9;&#x990A;&#x306B;alphaLTbetaR&#x30A2;&#x30B4;&#x30CB;&#x30B9;&#x30C8;&#x306E;&#x6DFB;&#x52A0;&#x306F;&#x3001;&#x6210;&#x719F;&#x3057;&#x3066;ICAM-1&#xFF08;int&#xFF09;&#x306F;VCAM-1&#xFF08;int&#xFF09;&#x306E;&#x7D30;&#x80DE;&#x3092;&#x8A98;&#x5C0E;&#x3059;&#x308B;&#x306E;&#x306B;&#x5341;&#x5206;&#x3067;&#x3042;&#x3063;&#x305F;&#x3002; &#xFF08; -  /  - &#xFF09;LtbetaR&#x3067;&#x306F;&#x80DA;&#x306F;&#x3001;&#x4E21;&#x65B9;&#x306E;&#x9F20;&#x5F84;&#x90E8;&#x304A;&#x3088;&#x3073;MLN&#x30B9;&#x30C8;&#x30ED;&#x30FC;&#x30DE;&#x7D30;&#x80DE;&#x306F;&#x3001;ICAM-1&#x306E;&#x30D6;&#x30ED;&#x30C3;&#x30AF;&#x3092;&#x793A;&#x3057;&#x305F;&#xFF08;int&#xFF09;&#x3092;&#x9F20;&#x5F84;LNS&#x306B;&#x53CD;&#x3057;&#x3066;VCAM-1&#xFF08;int&#xFF09;&#x306E;&#x30B9;&#x30C6;&#x30FC;&#x30B8;&#x3001;&#x305D;&#x3057;&#x3066;&#x3001;mLNs&#x304C;&#x9577;&#x304F;&#x6301;&#x7D9A;&#x3057;&#x3001;LTI&#x7D30;&#x80DE;&#x3092;&#x542B;&#x3093;&#x3067;&#x3001;&#x3069;&#x306E;CXCL13&#x306F;&#x3001;IL-7&#x306E;&#x6301;&#x7D9A;&#x7684;&#x306A;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x3068;&#x306E;&#x76F8;&#x95A2;&#x3001;&#x304A;&#x3088;&#x3073;&#x3001;&#x7A0B;&#x5EA6;&#x306F;&#x4F4E;&#x3044;&#x3001;CCL21&#x306B;&#x3002;&#x4E00;&#x7DD2;&#x306B;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x7D50;&#x679C;&#x306F;&#x3001;&#x30B9;&#x30C8;&#x30ED;&#x30FC;&#x30DE;&#x7D30;&#x80DE;&#x306E;&#x6210;&#x719F;&#x3092;&#x8A98;&#x5C0E;&#x3057;&#x3001;&#x6700;&#x7D42;&#x7684;&#x306B;&#x306F;&#x30EA;&#x30F3;&#x30D1;&#x7D44;&#x7E54;&#x306E;&#x5F62;&#x6210;&#x306B;&#x3064;&#x306A;&#x304C;&#x308B;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x3068;&#x7D30;&#x80DE;&#x9593;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306E;&#x91CD;&#x8981;&#x6027;&#x3092;&#x5F37;&#x8ABF;&#x3002;

&#x30EA;&#x30F3;&#x30D1;&#x7BC0;&#x306E;&#x958B;&#x767A;&#x4E2D;&#x306B;&#x9593;&#x8CEA;&#x30AA;&#x30FC;&#x30AC;&#x30CA;&#x30A4;&#x30B6;&#x30FC;&#x7D30;&#x80DE;&#x306E;&#x500B;&#x4F53;&#x767A;&#x751F;

2 &#xCC28; &#xB9BC;&#xD504; &#xAE30;&#xAD00; &#xB9BC;&#xD504;&#xC808; (LNs), &#xB9BC;&#xD504; &#xC870;&#xC9C1; &#xC720;&#xB3C4; (LTi) &#xC140;&#xACFC; stromal &#xC138;&#xD3EC; &#xAC04;&#xC758; &#xC0C1;&#xD638; &#xD589;&#xB3D9;&#xC5D0;&#xC11C; &#xBC30;&#xC544; &#xACB0;&#xACFC; &#xB4F1;&#xC758; &#xAC1C;&#xBC1C;. &#xADF8;&#xB7EC;&#xB098;, LTi stromal &#xC138;&#xD3EC; &#xD06C;&#xB85C;&#xC2A4; &#xD1A0;&#xD06C; &#xC0C1;&#xD638; &#xC791;&#xC6A9; &#xC790;&#xB9AC;&#xB97C; &#xCC28;&#xC9C0;&#xD560; &#xD558;&#xAE30; &#xC804;&#xC5D0; LN anlagen &#xD615;&#xC131;&#xC744; &#xC720;&#xB3C4; &#xD558;&#xB294; &#xCD08;&#xAE30; &#xC774;&#xBCA4;&#xD2B8; &#xC644;&#xBCBD; &#xD558; &#xAC8C; &#xD574;&#xBA85; &#xD558;&#xC9C0;&#xB294;. &#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC;&#xB294; &#xB9C8;&#xC6B0;&#xC2A4; &#xBC30;&#xC544;&#xC5D0;&#xC11C; &#xC0AC; &#xD0C0; &#xAD6C;&#xB2C8; LN anlagen &#xC8FC;&#xBCC0; &#xB9BC;&#xD504; &#xB0AD;, &#xAD6C;&#xD615; &#xAD6C;&#xC870; &#xC815; &#xB9E5;&#xC5D0;&#xC11C; &#xBC84;&#xB4DC; &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC758; &#xAC04; &#xC5FD; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xAC1C;&#xBC1C;&#xC744; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC; stromal cells 2 &#xB2E8;&#xACC4; &#xC131;&#xC219; &#xBAA8;&#xB378;&#xC744; &#xC9C0; &#xC6D0;&#xD558;&#xB294; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xC0AC; &#xD0C0; &#xAD6C;&#xB2C8;&#xC640; mesenteric LNs (mLNs)&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC;: &#xCCAB;&#xC9F8;, ICAM-1(-)VCAM-1(-) &#xC5FD; &#xC804;&#xC870; &#xC138;&#xD3EC;&#xAC00; ICAM-1(int)VCAM-1(int) &#xC140; LTi &#xC140; &#xBC0F; lymphotoxin &#xBCA0;&#xD0C0; &#xC218;&#xC6A9; &#xCCB4; (LTbetaR) &#xC2E0;&#xD638; &#xD504;&#xB85C;&#xC138;&#xC2A4; &#xAD00;&#xACC4; &#xC5C6;&#xC774;. &#xB450; &#xBC88;&#xC9F8; &#xB2E8;&#xACC4; ICAM-1(high)VCAM-1(high) &#xC810; &#xB9C9; addressin &#xC138;&#xD3EC; &#xC811;&#xCC29; molecule-1(+) &#xC8FC;&#xCD5C;&#xC790; &#xC138;&#xD3EC; ICAM-1(int)VCAM-1(int) &#xC138;&#xD3EC;&#xC758; &#xC131;&#xC219;&#xC744; &#xD3EC;&#xD568; &#xD558; &#xACE0; LTi &#xC140; &#xBC0F; Ltbetar&#xC5D0; &#xB530;&#xB77C; &#xB2EC;&#xB77C; &#xC9D1;&#xB2C8;&#xB2E4;. AlphaLTbetaR &#xAE38; &#xD56D; &#xC81C; LN &#xC7A5;&#xAE30; &#xBB38;&#xD654;&#xC5D0;&#xC758; &#xCD94;&#xAC00; &#xC131;&#xC219; ICAM-1(int)VCAM-1(int) &#xC140;&#xC744; &#xC720;&#xBC1C; &#xD558;&#xAE30;&#xC5D0; &#xCDA9;&#xBD84; &#xD588;&#xB2E4;. LtbetaR(-/-) &#xBC30;&#xC544; &#xB458; &#xB2E4; &#xC0AC; &#xD0C0; &#xAD6C;&#xB2C8;&#xC5D0; &#xACE0; mLN stromal &#xC138;&#xD3EC; ICAM-1(int)VCAM-1(int) &#xB2E8;&#xACC4;&#xC5D0;&#xC11C; &#xBE14;&#xB85D;, &#xC0AC; &#xD0C0; &#xAD6C;&#xB2C8; LNs &#xBC18;&#xB300;&#xB85C; Mlns&#xC640; &#xC77C;&#xB9AC;&#xB178;&#xC774;-7, Cxcl13, &#xC9C0;&#xC18D;&#xC801;&#xC778;&#xB41C; &#xC720;&#xC804;&#xC790; &#xBC1C;&#xD604; &#xC0C1;&#xAD00; &#xD558;&#xB294; &#xB354; &#xC774;&#xC0C1; &#xD558; &#xACE0; &#xD3EC;&#xD568; &#xB41C; LTi &#xC140; &#xC720;&#xC9C0; &#xC8FC;&#xBA70;, Ccl21. &#xD568;&#xAED8; &#xCC0D;&#xC740; &#xB0AE;&#xC740; &#xC815;&#xB3C4;&#xB85C;&#xC774; &#xACB0;&#xACFC; &#xC2E0;&#xD638; stromal &#xC138;&#xD3EC;&#xC758; &#xC131;&#xC219; &#xC720;&#xB3C4; &#xD558; &#xACE0; &#xAD81;&#xADF9;&#xC801;&#xC73C;&#xB85C; &#xB9BC;&#xD504; &#xC870;&#xC9C1;&#xC758; &#xD615;&#xC131;&#xC73C;&#xB85C; &#xC774;&#xC5B4;&#xC9C8; &#xC138;&#xD3EC; &#xC0C1;&#xD638; &#xC791;&#xC6A9;&#xC758; &#xC911;&#xC694;&#xC131;&#xC744; &#xAC15;&#xC870; &#xD569;&#xB2C8;&#xB2E4;.

Ontogeny stromal &#xC8FC;&#xCD5C;&#xC790; &#xC138;&#xD3EC; &#xB9BC;&#xD504; &#xB178;&#xB4DC; &#xAC1C;&#xBC1C; &#xD558;&#xB294; &#xB3D9;&#xC548;.

NF-κB1-dependent signaling directs the development of CD4(+) Th2 cells during allergic airway inflammation and protective responses to helminth infection. Here, we show that IL-4 and IL-13 production is NF-κB1-dependent in mouse OVA-specific CD4(+) (OTII) T cells responding to alum-precipitated OVA (alumOVA) immunization. More surprisingly, we found that NF-κB1 deficiency in OTII cells also selectively impairs their CXCR5 induction by alumOVA without affecting upregulation of BCL6, IL-21, OX40 and CXCR4 mRNA and PD-1 protein. This results in functional impairment of follicular helper T cells. Thus, fewer germinal center B cells develop in LN responses to alumOVA in T-cell-deficient mice reconstituted with NF-κB1(-/-) OTII cells as opposed to NF-κB1(+/+) OTII cells, while plasma cell numbers are comparable. Unlike CXCR5 induction in CD4(+) T cells, NF-κB1-deficient recirculating follicular B cells are shown to express normal levels of CXCR5. The selective effects of NF-κB1-deficiency on Th2 and follicular helper T cell induction do not appear to be due to altered expression of the Th2-associated transcription factors - GATA-3, c-Maf and Ikaros. Altogether, these results suggest that NF-κB1 regulates the expression of CXCR5 on CD4(+) T cells primed in vivo, and thus selectively controls the T-cell-dependent germinal center component of B-cell response to alumOVA.

Selective effects of NF-κB1 deficiency in CD4⁺ T cells on Th2 and TFh induction by alum-precipitated protein vaccines.

Although in vitro IL-4 directs CD4 T cells to produce T helper 2 (Th2)-cytokines, these cytokines can be induced in vivo in the absence of IL-4-signalling. Thus, mechanism(s), different from the in vitro pathway for Th2-induction, contribute to in vivo Th2-differentiation. The pathway for in vivo IL-4-independent Th2-differentiation has yet to be characterized.

Helios is associated with CD4 T cells differentiating to T helper 2 and follicular helper T cells in vivo independently of Foxp3 expression.

Several tumor necrosis factor receptor (TNFR) family members activate both the classical and the alternative NF-κB pathways. However, how a single receptor engages these two distinct pathways is still poorly understood. Using lymphotoxin β receptor (LTβR) as a prototype, we showed that activation of the alternative, but not the classical, NF-κB pathway relied on internalization of the receptor. Further molecular analyses revealed a specific cytosolic region of LTβR essential for its internalization, TRAF3 recruitment, and p100 processing. Interestingly, we found that dynamin-dependent, but clathrin-independent, internalization of LTβR appeared to be required for the activation of the alternative, but not the classical, NF-κB pathway. In vivo, ligand-induced internalization of LTβR in mesenteric lymph node stromal cells correlated with induction of alternative NF-κB target genes. Thus, our data shed light on LTβR cellular trafficking as a process required for specific biological functions of NF-κB.

Induction of the alternative NF-κB pathway by lymphotoxin αβ (LTαβ) relies on internalization of LTβ receptor.

The C-type lectin receptor CLEC-2 signals through a pathway that is critically dependent on the tyrosine kinase Syk. We show that homozygous loss of either protein results in defects in brain vascular and lymphatic development, lung inflation, and perinatal lethality. Furthermore, we find that conditional deletion of Syk in the hematopoietic lineage, or conditional deletion of CLEC-2 or Syk in the megakaryocyte/platelet lineage, also causes defects in brain vascular and lymphatic development, although the mice are viable. In contrast, conditional deletion of Syk in other hematopoietic lineages had no effect on viability or brain vasculature and lymphatic development. We show that platelets, but not platelet releasate, modulate the migration and intercellular adhesion of lymphatic endothelial cells through a pathway that depends on CLEC-2 and Syk. These studies found that megakaryocyte/platelet expression of CLEC-2 and Syk is required for normal brain vasculature and lymphatic development and that platelet CLEC-2 and Syk directly modulate lymphatic endothelial cell behavior in vitro.

CLEC-2 and Syk in the megakaryocytic/platelet lineage are essential for development.

Tissue glucocorticoid (GC) levels are regulated by the GC-activating enzyme 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1). This enzyme is expressed in cells and tissues arising from mesenchymal stromal cells. Proinflammatory cytokines dramatically increase expression of 11β-HSD1 in stromal cells, an effect that has been implicated in inflammatory arthritis, osteoporosis, obesity, and myopathy. Additionally, GCs act synergistically with proinflammatory cytokines to further increase enzyme expression. The present study was undertaken to investigate the mechanisms underlying this regulation.

Inflammatory regulation of glucocorticoid metabolism in mesenchymal stromal cells.

The thymic medulla provides a specialized microenvironment for the negative selection of T cells, with the presence of autoimmune regulator (Aire)-expressing medullary thymic epithelial cells (mTECs) during the embryonic-neonatal period being both necessary and sufficient to establish long-lasting tolerance. Here we showed that emergence of the first cohorts of Aire(+) mTECs at this key developmental stage, prior to αβ T cell repertoire selection, was jointly directed by Rankl(+) lymphoid tissue inducer cells and invariant Vγ5(+) dendritic epidermal T cell (DETC) progenitors that are the first thymocytes to express the products of gene rearrangement. In turn, generation of Aire(+) mTECs then fostered Skint-1-dependent, but Aire-independent, DETC progenitor maturation and the emergence of an invariant DETC repertoire. Hence, our data attributed a functional importance to the temporal development of Vγ5(+) γδ T cells during thymus medulla formation for αβ T cell tolerance induction and demonstrated a Rank-mediated reciprocal link between DETC and Aire(+) mTEC maturation.

Rank signaling links the development of invariant γδ T cell progenitors and Aire(+) medullary epithelium.

Lymph node development during embryogenesis involves lymphotoxin-β receptor engagement and subsequent differentiation of a poorly defined population of mesenchymal cells into lymphoid tissue organizer cells. Here, we showed that embryonic mesenchymal cells with characteristics of adipocyte precursors present in the microenvironment of lymph nodes gave rise to lymph node organizer cells. Signaling through the lymphotoxin-β receptor controlled the fate of adipocyte precursor cells by blocking adipogenesis and instead promoting lymphoid tissue stromal cell differentiation. This effect involved activation of the NF-κB2-RelB signaling pathway and inhibition of the expression of the key adipogenic factors Pparγ and Cebpα. In vivo organogenesis assays show that embryonic and adult adipocyte precursor cells can migrate into newborn lymph nodes and differentiate into a variety of lymph node stromal cells. Thus, we propose that adipose tissues act as a source of lymphoid stroma for lymph nodes and other lymphoid structures associated with fat.

Cecile Benezech

School of Immunity and Infection,
IBR-MRC Centre for Immune Regulation,
College of Medical and Dental Sciences,
School of Immunity and Infection, IBR-MRC Centre for Immune Regulation, College of Medical and Dental Sciences

University of Birmingham

Generation of Lymph Node-fat Pad Chimeras for the Study of Lymph Node Stromal Cell Origin

Cecile Benezech

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}School of Immunity and Infection,IBR-MRC Centre for Immune Regulation,College of Medical and Dental Sciences,School of Immunity and Infection, IBR-MRC Centre for Immune Regulation, College of Medical and Dental Sciences

University of Birmingham

Generation of Lymph Node-fat Pad Chimeras for the Study of Lymph Node Stromal Cell Origin

School of Immunity and Infection,
IBR-MRC Centre for Immune Regulation,
College of Medical and Dental Sciences,
School of Immunity and Infection, IBR-MRC Centre for Immune Regulation, College of Medical and Dental Sciences