orthotopic-hind-limb-transplantation-in-the-mouse

Orthotopic Hind Limb Transplantation in the Mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

Innsbruck Medical University

&#x8F7B;&#x5EA6;&#x76AE;&#x80A4;&#x62D2;&#x7EDD;&#x662F;&#x91CD;&#x5EFA;&#x79FB;&#x690D;&#x4E2D;&#x4E00;&#x4E2A;&#x5E38;&#x89C1;&#x7684;&#x73B0;&#x8C61;&#x3002;&#x6765;&#x7167;&#x4EAE;&#x8FD9;&#x708E;&#x75C7;&#x53CD;&#x5E94;&#x7684;&#x4F5C;&#x7528;&#x6211;&#x4EEC;&#x8C03;&#x67E5;&#x6807;&#x5FD7;&#x7269;&#x7EC6;&#x80DE;&#x548C;&#x6297;&#x4F53;&#x4ECB;&#x5BFC;&#x7684;&#x62D2;&#x7EDD;&#x3001; &#x7C98;&#x9644;&#x5206;&#x5B50;&#x548C;&#x5BB9;&#x5FCD;&#x6807;&#x8BB0;&#x3002;&#x56DB;&#x5341;&#x4E03;&#x4E2A;&#x76AE;&#x80A4;&#x7A7F;&#x523A;&#x6D3B;&#x68C0; &#xFF08;&#x62D2;&#x7EDD;&#x4E00;&#x7EA7;&#xFF09; &#x4EBA;&#x7C7B;&#x7684;&#x624B;&#x7684;&#x514D;&#x75AB;&#x7EC4;&#x7EC7;&#x5316;&#x5B66;&#x65B9;&#x6CD5;&#x8FDB;&#x884C;&#x8C03;&#x67E5;&#x540C;&#x79CD;&#x5F02;&#x4F53;&#x79FB;&#x690D; &#xFF08;CD3&#x3001; CD4&#x3001; CD8&#x3001; CD20&#x3001; CD68&#x3001; C4d&#x3001; &#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x529F;&#x80FD;&#x76F8;&#x5173;&#x6297;&#x539F;-1&#x3001; &#x7EC6;&#x80DE;&#x95F4;&#x7C98;&#x9644;&#x5206;&#x5B50;-1&#x3001; E-&#x9009;&#x62E9;&#x7D20;&#x3001; P-&#x9009;&#x62E9;&#x7D20;&#x3001; &#x9ECF;&#x86CB;&#x767D;&#x3001; HLA-DR&#x3001; &#x4F9D;&#x5F97;&#x548C; Foxp3)&#x3002;&#x8868;&#x8FBE;&#x5F0F;&#x662F;&#x5728;&#x79FB;&#x690D;&#x540E;&#x7684;&#x65F6;&#x95F4;&#x9605;&#x8BFB;&#x3002;&#x6E17;&#x900F;&#x4E3B;&#x8981;&#x7531; CD3 + T &#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x7EC4;&#x6210;&#x3002;&#x5176;&#x4E2D;&#xFF0C;CD8 + &#x7EC6;&#x80DE;&#x6709;&#x6BD4; CD4 + &#x7EC6;&#x80DE;&#x66F4;&#x4E3A;&#x663E;&#x8457;&#x3002;CD20 + B &#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x88AB;&#x7A00;&#x758F;&#x548C; CD68 + &#x5DE8;&#x566C;&#x7EC6;&#x80DE;&#x88AB;&#x53D1;&#x73B0;&#x5728;&#x4E00;&#x4E9B;&#xFF0C;&#x4F46;&#x4E0D;&#x662F;&#x6240;&#x6709;&#x6837;&#x672C; (&#x5927;&#x7EA6; 10%&#x7684;&#x6E17;&#x900F;&#xFF09;&#x3002;CD4/CD8 &#x6BD4;&#x4F8B;&#x589E;&#x52A0;&#x540E;&#x7684;&#x7B2C;&#x4E00;&#x5E74;&#x3002;&#x5728;&#x8FDF;&#x4E8E; 1 &#x5E74;&#x7684;&#x65F6;&#x95F4;&#x70B9;&#xFF0C;C4d &#x67D3;&#x8272;&#x662F;&#x6837;&#x54C1;&#x4E2D;&#x4E3B;&#x8981;&#x662F;&#x79EF;&#x6781;&#x7684;&#x3002;&#x7C98;&#x9644;&#x5206;&#x5B50; LFA 1&#x3001; &#x7EC6;&#x80DE;&#x95F4;&#x7C98;&#x9644;&#x5206;&#x5B50; 1&#x3001; E-&#x9009;&#x62E9;&#x7D20;&#xFF0C;&#x53D1;&#x73B0;&#x4E0A;&#x8C03;&#xFF0C;P-&#x9009;&#x62E9;&#x7D20;&#xFF0C;&#x548C;&#x9ECF;&#x86CB;&#x767D;&#x548C; P-&#x9009;&#x62E9;&#x7D20;&#xFF0C;&#x8868;&#x8FBE;&#x589E;&#x52A0;&#x4E0E;&#x79FB;&#x690D;&#x540E;&#x7684;&#x65F6;&#x95F4;&#x3002;&#x4F9D;&#x5F97;&#x8868;&#x8FBE;&#x6700;&#x5F3A;&#x5728; 3 &#x4E2A;&#x6708;-1 &#x5E74;&#x6BDB;&#x56CA;&#x800C;&#x4E14;&#x89C2;&#x5BDF;&#x66F4;&#x591A; Foxp3 + &#x7EC6;&#x80DE;&#x5728;&#x7A0D;&#x540E;&#x7684;&#x65F6;&#x95F4;&#x70B9;&#x7684;&#x503E;&#x5411;&#x3002;&#x8F7B;&#x5EA6;&#x76AE;&#x80A4;&#x62D2;&#x7EDD;&#x540E;&#x624B;&#x79FB;&#x690D;&#x5E26;&#x6765;&#x4E86; T &#x7EC6;&#x80DE;&#x4E3A;&#x4E3B;&#x76AE;&#x80A4;&#x7EC6;&#x80DE;&#x6E17;&#x900F;&#x548C;&#x7C98;&#x9644;&#x5206;&#x5B50;&#x7684;&#x8868;&#x8FBE;&#x4E0A;&#x8C03;&#x3002;&#x540E;&#x5E74;&#x4E4B;&#x4E00;&#x4ECD;&#x7136;&#x662F;&#x9610;&#x660E; C4d &#x8868;&#x8FBE;&#x7684;&#x4F5C;&#x7528;&#x3002;

&#x8F7B;&#x5EA6;&#x7684;&#x6392;&#x65A5;&#x53CD;&#x5E94;&#x7684;&#x75C5;&#x7406;&#x8868;&#x5F81; (&#x804C;&#x7CFB;) &#x5728;&#x4EBA;&#x7C7B;&#x624B;&#x540C;&#x79CD;&#x5F02;&#x4F53;&#x79FB;&#x690D;&#x7684;&#x76AE;&#x80A4;&#x6D3B;&#x68C0;&#x3002;

Mild skin rejection is a common observation in reconstructive transplantation. To enlighten the role of this inflammatory reaction we investigated markers for cellular and antibody mediated rejection, adhesion molecules and tolerance markers. Forty-seven skin biopsies (rejection grade I) of human hand allografts were investigated by immunohistochemistry (CD3, CD4, CD8, CD20, CD68, C4d, LFA-1, ICAM-1, E-selectin, P-selectin, VE-cadherin, HLA-DR, IDO, and Foxp3). Expression was read with respect to time after transplant. The infiltrate was mainly comprised of CD3+T-lymphocytes. Among these, CD8+cells were more prominent than CD4+cells. CD20+B-lymphocytes were sparse and CD68+macrophages were found in some, but not all samples (approximately 10% of the infiltrate). The CD4/CD8-ratio was increased after the first year. C4d staining was mainly positive in samples at time-points later than 1 year. Adhesion molecules LFA-1, ICAM-1, E-selectin, P-selectin, and VE-cadherin were found upregulated, and for P-selectin, expression increased with time after transplant. IDO expression was strongest at 3 months-1 year post-transplant and a tendency toward more Foxp3+ cells at later time points was observed. Mild skin rejection after hand transplantation presents with a T-cell dominated dermal cell infiltrate and upregulation of adhesion molecules. The role of C4d expression after year one remains to be elucidated.

Histopathologic characterization of mild rejection (grade I) in skin biopsies of human hand allografts.

Rejet l&#xE9;g&#xE8;re de la peau est une observation courante dans reconstructive transplantation. Pour &#xE9;clairer le r&#xF4;le de cette r&#xE9;action inflammatoire, nous avons &#xE9;tudi&#xE9; des marqueurs de rejet &#xE0; m&#xE9;diation cellulaire et des anticorps, des mol&#xE9;cules d&#x26;#39;adh&#xE9;sion et des marqueurs de tol&#xE9;rance. Quarante-sept biopsies de la peau (grade rejet I) d&#x26;#39;allogreffes main humaine a &#xE9;t&#xE9; &#xE9;tudi&#xE9;e par immunohistochimie (CD3, CD4, CD8, CD20, CD68, C4D, LFA-1, ICAM-1, E-s&#xE9;lectine, P-s&#xE9;lectine, VE-cadh&#xE9;rine , HLA-DR, IDO, et Foxp3). Expression a &#xE9;t&#xE9; lu par rapport au temps apr&#xE8;s la transplantation. L&#x26;#39;infiltrat est compos&#xE9; principalement de CD3 + des lymphocytes T. Parmi ceux-ci, des cellules CD8 + &#xE9;taient plus important que les cellules CD4 +. CD20 + des lymphocytes B &#xE9;taient rares et les macrophages CD68 + ont &#xE9;t&#xE9; trouv&#xE9;s dans certains cas, mais pas tous les &#xE9;chantillons (environ 10% de l&#x26;#39;infiltrat). Le CD4/CD8-ratio a augment&#xE9; apr&#xE8;s la premi&#xE8;re ann&#xE9;e. Coloration C4d &#xE9;tait essentiellement positive dans les &#xE9;chantillons au moment des points au plus tard 1 an. Les mol&#xE9;cules d&#x26;#39;adh&#xE9;sion LFA-1, ICAM-1, E-s&#xE9;lectine, P-s&#xE9;lectine, et VE-cadh&#xE9;rine ont &#xE9;t&#xE9; trouv&#xE9;s r&#xE9;gul&#xE9;s &#xE0; la hausse, et pour la P-s&#xE9;lectine, une expression accrue avec le temps apr&#xE8;s la transplantation. IDO expression la plus forte &#xE9;tait de 3 mois &#xE0; 1 an post-transplantation et une tendance vers une plus grande Foxp3 + cellules &#xE0; points dans le temps plus tard, a &#xE9;t&#xE9; observ&#xE9;e. Rejet l&#xE9;g&#xE8;re de la peau apr&#xE8;s la transplantation main pr&#xE9;sente avec un T-cell cellules dermiques domin&#xE9;e infiltrer et la r&#xE9;gulation positive de mol&#xE9;cules d&#x26;#39;adh&#xE9;sion. Le r&#xF4;le de l&#x26;#39;expression C4d apr&#xE8;s la premi&#xE8;re ann&#xE9;e reste &#xE0; &#xEA;tre &#xE9;lucid&#xE9;.

Caract&#xE9;risation histologique du rejet l&#xE9;g&#xE8;re (grade I) dans des biopsies cutan&#xE9;es de allogreffes main de l&#x26;#39;homme

Milde Haut Ablehnung ist eine gemeinsame Beobachtung im rekonstruktive Transplantation. Die Rolle dieser entz&uuml;ndlichen Reaktion zu erleuchten, die wir untersuchten Marker f&uuml;r Mobilfunk und Antik&ouml;rper vermittelte Ablehnung, Adh&auml;sionsmolek&uuml;le und Toleranzmarken. Siebenundvierzig Haut Biopsien (Ablehnung Grad I) von Menschenhand Herztransplanate durch Immunhistochemie untersucht wurden (CD3, CD4, CD8, CD20, CD68, C4d, LFA-1, ICAM-1, E-selectin, P-selectin, VE-Cadherin HLA-DR, IDO und Foxp3). Ausdruck wurde in Bezug auf die Zeit nach der Transplantation gelesen. CD3 + T-Lymphozyten umfasste haupts&auml;chlich den Infiltraten. Unter diesen waren CD8 + Zellen mehr im Vordergrund als CD4 + Zellen. CD20 + B-Lymphozyten waren sp&auml;rlich und CD68 + Makrophagen fanden sich in einigen, aber nicht alle Proben (etwa 10 % der Infiltrate). Das CD4/CD8-Verh&auml;ltnis wurde nach dem ersten Jahr erh&ouml;ht. C4d F&auml;rbung war haupts&auml;chlich positiven Proben zu Zeitpunkten, sp&auml;ter als 1 Jahr. Adh&auml;sionsmolek&uuml;le LFA-1, ICAM-1, E-selectin, P-selectin und VE-Cadherin herraufreguliert gefunden wurden, und f&uuml;r P-selectin, erh&ouml;hte Ausdruck mit Zeit nach der Transplantation. IDO Ausdruck war st&auml;rkste bei 3 Monate-1 Jahr-Post-transplant und eine Tendenz zu mehr Foxp3 + Zellen zu sp&auml;teren Zeitpunkten beobachtet wurde. Milde Haut Ablehnung nach Hand-Transplantation mit einer T-Zelle pr&auml;sentiert dominiert dermal Zelle infiltrieren und Hochregulation der Adh&auml;sionsmolek&uuml;le. Die Rolle der C4d Ausdruck nach Jahr bleibt eine gekl&auml;rt werden.

Histopathologische Charakterisierung von milden Ablehnung (Grad I) in Haut Biopsien von Menschenhand Herztransplanate.

Pelle lieve rifiuto &egrave; un&#39;osservazione comune nel trapianto ricostruttivo. Per illuminare il ruolo di questa reazione infiammatoria abbiamo studiato marcatori per cellulari e anticorpo mediata rifiuto, molecole di adesione e gli indicatori di tolleranza. Quarantasette pelle biopsie (rifiuto di grado I) della mano umana allograft sono stati investigati dall&#39;immunoistochimica (CD3, CD4, CD8, CD20, CD68, C4d, LFA-1, ICAM-1, E-selectina, P-Selectina, VE-caderina, HLA-DR, IDO e Foxp3). Espressione &egrave; stata letta rispetto al tempo dopo il trapianto. L&#39;infiltrato era composto principalmente da CD3 + T-linfociti. Tra questi, CD8 + cellule erano pi&ugrave; prominenti di CD4 + cellule. CD20 + B-linfociti erano sparse e CD68 + macrofagi sono stati trovati in alcuni, ma non tutti i campioni (circa il 10% dell&#39;infiltrato). Dopo il primo anno &egrave; stato aumentato il rapporto CD4/CD8. C4D colorazione era principalmente positiva nei campioni a tempo-punti entro 1 anno. Molecole di adesione LFA-1, ICAM-1, E-selectina, sono stati trovati sovraregolati P-Selectina e VE-caderina, e per la P-Selectina, espressione aumentata con il momento dopo il trapianto. IDO espressione era pi&ugrave; forte a post-transplant di 3 mesi-1 anno ed &egrave; stata osservata una tendenza verso pi&ugrave; Foxp3 + cellule in punti temporali successive. Rigetto lieve della pelle dopo trapianto di mano si presenta con una T-cellula dominato infiltrato cellulare dermico e sovraregolazione di molecole di adesione. Il ruolo dell&#39;espressione C4d dopo anno uno rimane per essere chiarita.

Caratterizzazione istopatologica di rigetto lieve (grado I) in biopsie cutanee del allograft mano umana.

&#x8EFD;&#x5EA6;&#x306E;&#x76AE;&#x819A;&#x9664;&#x53BB;&#x306F;&#x3001;&#x518D;&#x5EFA;&#x79FB;&#x690D;&#x306B;&#x304A;&#x3051;&#x308B;&#x4E00;&#x822C;&#x7684;&#x306A;&#x89B3;&#x6E2C;&#x3067;&#x3042;&#x308B;&#x3002;&#x3053;&#x306E;&#x708E;&#x75C7;&#x53CD;&#x5FDC;&#x306E;&#x5F79;&#x5272;&#x3092;&#x5553;&#x767A;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3001;&#x6211;&#x3005;&#x306F;&#x3001;&#x7D30;&#x80DE;&#x3084;&#x6297;&#x4F53;&#x3092;&#x4ECB;&#x3057;&#x305F;&#x62D2;&#x7D76;&#x53CD;&#x5FDC;&#x3001;&#x63A5;&#x7740;&#x5206;&#x5B50;&#x3068;&#x8010;&#x6027;&#x30DE;&#x30FC;&#x30AB;&#x30FC;&#x306E;&#x30DE;&#x30FC;&#x30AB;&#x30FC;&#x3092;&#x8ABF;&#x3079;&#x305F;&#x3002;&#x4EBA;&#x9593;&#x306E;&#x624B;&#x540C;&#x7A2E;&#x79FB;&#x690D;&#x7247;&#x306E;&#x56DB;&#x5341;&#x4E03;&#x4EBA;&#x306E;&#x76AE;&#x819A;&#x751F;&#x691C;&#xFF08;&#x62D2;&#x5426;&#x30B0;&#x30EC;&#x30FC;&#x30C9;I&#xFF09;&#x306F;&#x3001;&#x514D;&#x75AB;&#x7D44;&#x7E54;&#x5316;&#x5B66;&#xFF08;CD3&#x3001;CD4&#x3001;CD8&#x3001;CD20&#x3001;CD68&#x3001;C4D&#x3001;LFA-1&#x3001;ICAM-1&#x3001;E-&#x30BB;&#x30EC;&#x30AF;&#x30C1;&#x30F3;&#x3001;P-&#x30BB;&#x30EC;&#x30AF;&#x30C1;&#x30F3;&#x3001;VE-&#x30AB;&#x30C9;&#x30D8;&#x30EA;&#x30F3;&#x306B;&#x3088;&#x308A;&#x8ABF;&#x3079;&#x305F;&#x3001;HLA-DR&#x3001;IDO&#x3001;&#x3068;Foxp3&#xFF09;&#x3002;&#x5F0F;&#x306F;&#x3001;&#x79FB;&#x690D;&#x5F8C;&#x306E;&#x6642;&#x9593;&#x3092;&#x57FA;&#x6E96;&#x306B;&#x3057;&#x3066;&#x8AAD;&#x307F;&#x8FBC;&#x307E;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x6F5C;&#x5165;&#x306F;&#x3001;&#x4E3B;&#x306B;CD3 + T&#x30EA;&#x30F3;&#x30D1;&#x7403;&#x3067;&#x69CB;&#x6210;&#x3055;&#x308C;&#x3066;&#x3044;&#x305F;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x3046;&#x3061;&#x3001;CD8 +&#x7D30;&#x80DE;&#x306F;&#x3001;CD4 +&#x7D30;&#x80DE;&#x3088;&#x308A;&#x3082;&#x9855;&#x8457;&#x3067;&#x3042;&#x3063;&#x305F;&#x3002; CD20 + B&#x30EA;&#x30F3;&#x30D1;&#x7403;&#x306F;&#x758E;&#x3067;&#x3042;&#x3063;&#x305F;&#x3068;CD68 +&#x30DE;&#x30AF;&#x30ED;&#x30D5;&#x30A1;&#x30FC;&#x30B8;&#x306F;&#x3001;&#x3059;&#x3079;&#x3066;&#x306E;&#x30B5;&#x30F3;&#x30D7;&#x30EB;&#xFF08;&#x6F5C;&#x5165;&#x306E;&#x7D04;10&#xFF05;&#xFF09;&#x304C;&#x4E00;&#x90E8;&#x306B;&#x3042;&#x308B;&#x304C;&#x3001;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002; CD4/CD8-ratio&#x306F;&#x3001;&#x6700;&#x521D;&#x306E;&#x5E74;&#x5F8C;&#x306B;&#x5897;&#x52A0;&#x3057;&#x305F;&#x3002; C4D&#x67D3;&#x8272;&#x306F;&#x5F8C;1&#x5E74;&#x4EE5;&#x4E0A;&#x306E;&#x30DD;&#x30A4;&#x30F3;&#x30C8;&#x6642;&#x306B;&#x30B5;&#x30F3;&#x30D7;&#x30EB;&#x3067;&#x4E3B;&#x306B;&#x967D;&#x6027;&#x3067;&#x3042;&#x3063;&#x305F;&#x3002;&#x63A5;&#x7740;&#x5206;&#x5B50;LFA-1&#x3001;ICAM-1&#x3001;E-&#x30BB;&#x30EC;&#x30AF;&#x30C1;&#x30F3;&#x3001;P-&#x30BB;&#x30EC;&#x30AF;&#x30C1;&#x30F3;&#x3001;&#x304A;&#x3088;&#x3073;VE-&#x30AB;&#x30C9;&#x30D8;&#x30EA;&#x30F3;&#x3092;&#x30A2;&#x30C3;&#x30D7;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30C8;&#x3055;&#x308C;&#x767A;&#x898B;&#x3055;&#x308C;&#x3001;P-&#x30BB;&#x30EC;&#x30AF;&#x30C1;&#x30F3;&#x3001;&#x767A;&#x73FE;&#x306E;&#x305F;&#x3081;&#x306B;&#x79FB;&#x690D;&#x5F8C;&#x6642;&#x9593;&#x306E;&#x7D4C;&#x904E;&#x3068;&#x3068;&#x3082;&#x306B;&#x5897;&#x52A0;&#x3057;&#x305F;&#x3002; IDO&#x306E;&#x767A;&#x73FE;&#x306F;&#x3001;3&#x30AB;&#x6708;1&#x5E74;&#x5F8C;&#x306E;&#x79FB;&#x690D;&#x3068;Foxp3&#x306E;&#x50BE;&#x5411;&#x3067;&#x306F;&#x6700;&#x5F37;&#x3060;&#x3063;&#x305F;+&#x5F8C;&#x306E;&#x6642;&#x70B9;&#x3067;&#x7D30;&#x80DE;&#x304C;&#x89B3;&#x5BDF;&#x3055;&#x308C;&#x305F;&#x3002;&#x624B;&#x306E;&#x79FB;&#x690D;&#x5F8C;&#x306E;&#x8EFD;&#x5EA6;&#x306E;&#x76AE;&#x819A;&#x62D2;&#x7D76;&#x306F;T&#x7D30;&#x80DE;&#x306B;&#x652F;&#x914D;&#x76AE;&#x819A;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x3059;&#x308B;&#x3068;&#x63A5;&#x7740;&#x5206;&#x5B50;&#x306E;&#x30A2;&#x30C3;&#x30D7;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002; 1&#x5E74;&#x5F8C;&#x306E;C4D&#x8868;&#x73FE;&#x306E;&#x5F79;&#x5272;&#x304C;&#x89E3;&#x660E;&#x3055;&#x308C;&#x3066;&#x3044;&#x306A;&#x3044;&#x3002;

&#x4EBA;&#x9593;&#x306E;&#x624B;&#x540C;&#x7A2E;&#x79FB;&#x690D;&#x306E;&#x76AE;&#x819A;&#x751F;&#x691C;&#x3067;&#x306F;&#x8EFD;&#x5EA6;&#x306E;&#x62D2;&#x7D76;&#x53CD;&#x5FDC;&#x306E;&#x75C5;&#x7406;&#x7D44;&#x7E54;&#x5B66;&#x7684;&#x7279;&#x6027;&#xFF08;I&#x30B0;&#x30EC;&#x30FC;&#x30C9;&#xFF09;

&#xAC00;&#xBCBC;&#xC6B4; &#xD53C;&#xBD80; &#xAC70;&#xBD80; &#xC7AC;&#xAC74; &#xC774;&#xC2DD;&#xC5D0; &#xACF5;&#xD1B5;&#xC801;&#xC778; &#xAD00;&#xCC30; &#xC774;&#xB2E4;. &#xACC4;&#xBABD;&#xC774; &#xC5FC;&#xC99D; &#xBC18;&#xC751;&#xC758; &#xC5ED;&#xD560;&#xC744; &#xC6B0;&#xB9AC;&#xAC00; &#xC138;&#xD3EC;&#xC758; &#xD45C;&#xC2DD; &#xC870;&#xC0AC; &#xD558; &#xACE0; &#xD56D; &#xCCB4; &#xC911;&#xC7AC; &#xAC70;&#xBD80;, &#xC811;&#xCC29; &#xBD84;&#xC790; &#xBC0F; &#xACF5;&#xCC28; &#xB9C8;&#xCEE4;. 47 &#xD53C;&#xBD80; &#xC0DD; &#xAC80; (&#xAC70;&#xBD80; &#xD559;&#xB144;) &#xC778;&#xAC04;&#xC758; &#xC190;&#xC758; allografts &#xAE30;&#xC220;&#xC5D0; &#xC758;&#xD574; &#xC870;&#xC0AC; &#xB418;&#xC5C8;&#xB2E4; (CD3, CD4, CD8, CD20, CD68, C4d, LFA-1, ICAM-1 E selectin P selectin, VE cadherin, HLA-DR, IDO, &#xADF8;&#xB9AC;&#xACE0; Foxp3). &#xC2DD; &#xC774;&#xC2DD; &#xD6C4; &#xC2DC;&#xAC04;&#xC5D0; &#xAD00;&#xD558;&#xC5EC; &#xC77D; &#xC5C8;&#xB294; &#xB370;. CD3 + T &#xC138;&#xD3EC;&#xB294; &#xCE68;&#xD22C; &#xC8FC;&#xB85C; &#xAD6C;&#xC131; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC774; &#xC911;, CD8 + &#xC138;&#xD3EC; CD4 + &#xC138;&#xD3EC; &#xBCF4;&#xB2E4; &#xB354; &#xC720;&#xBA85; &#xD588;&#xB2E4;. CD20 + B &#xC138;&#xD3EC; &#xC2A4;&#xD30C;&#xC2A4; &#xB418;&#xC5C8;&#xACE0; CD68 + &#xB300; &#xC2DD; &#xC138;&#xD3EC;&#xB294; &#xBAA8;&#xB4E0; &#xC0D8;&#xD50C; (&#xC57D; 10%&#xB294; &#xCE68;&#xD22C;) &#xD558;&#xC9C0;&#xB9CC; &#xC77C;&#xBD80;&#xC5D0;&#xC11C; &#xBC1C;&#xACAC; &#xB410;&#xB2E4;. CD4/CD8-&#xBE44;&#xB294; &#xCCAB;&#xBC88;&#xC9F8; &#xB144; &#xD6C4;&#xC5D0; &#xC99D;&#xAC00; &#xD588;&#xB2E4;. C4d &#xC5BC;&#xB8E9;&#xC740; &#xC2DC;&#xAC04; &#xD3EC;&#xC778;&#xD2B8; 1 &#xB144; &#xB098;&#xC911;&#xC5D0; &#xC0D8;&#xD50C;&#xC5D0;&#xC11C; &#xC8FC;&#xB85C; &#xAE0D;&#xC815;&#xC801; &#xC774;&#xC5C8;&#xB2E4;. &#xC811;&#xCC29; &#xBD84;&#xC790; LFA-1, ICAM-1, &#xC804;&#xC790; selectin P selectin &#xADF8;&#xB9AC;&#xACE0; VE cadherin upregulated, &#xBC1C;&#xACAC; &#xD558; &#xACE0; P selectin &#xC704;&#xD55C; &#xC2DD; &#xC99D;&#xAC00; &#xC774;&#xC2DD; &#xD6C4; &#xC2DC;&#xAC04;&#xC774;. &#xC774; &#xC2DD; 3 &#xAC1C;&#xC6D4;-1 &#xB144; post-transplant&#xC5D0;&#xC11C; &#xAC00;&#xC7A5; &#xAC15;&#xD55C; &#xB418;&#xC5C8;&#xACE0; &#xB098;&#xC911; &#xC2DC;&#xAC04; &#xC9C0;&#xC810;&#xC5D0;&#xC11C; &#xB354; &#xB9CE;&#xC740; Foxp3 + &#xC140; &#xACBD;&#xD5A5;&#xC744; &#xAD00;&#xCC30; &#xD588;&#xB2E4;. &#xAC00;&#xBCBC;&#xC6B4; &#xD53C;&#xBD80; &#xC81C;&#xAC70; &#xD6C4; &#xC190;&#xC73C;&#xB85C; &#xC774;&#xC2DD; &#xC120;&#xBB3C; T &#xC138;&#xD3EC;&#xC640; &#xD53C;&#xBD80; &#xC138;&#xD3EC;&#xC758; &#xCE68;&#xD22C; &#xC811;&#xCC29; &#xBD84;&#xC790;&#xC758; upregulation &#xC9C0;&#xBC30;. 1 &#xB144; &#xD6C4; C4d &#xD45C;&#xD604;&#xC758; &#xC5ED;&#xD560; &#xD558;&#xB098; &#xD574;&#xBA85; &#xB420; &#xC788;&#xB2E4;.

&#xAC00;&#xBCBC;&#xC6B4; &#xAC70;&#xC808;&#xC758; &#xBCD1;&#xB9AC;&#xD559; &#xD2B9;&#xC131;&#xD654; (&#xC804; &#xD559;&#xB144;) &#xC778;&#xAC04;&#xC758; &#xC190; allografts&#xC758; &#xD53C;&#xBD80; biopsies&#xC5D0;.

El rechazo leve en la piel es un dato caracter&#xED;stico de la reconstrucci&#xF3;n del trasplante. Para ilustrar el papel de esta reacci&#xF3;n inflamatoria que investig&#xF3; los marcadores del rechazo mediado por celular y de anticuerpos, mol&#xE9;culas de adhesi&#xF3;n y marcadores de tolerancia. Cuarenta y siete biopsias de la piel (grado de rechazo I) de aloinjertos humanos de la mano fueron investigados por inmunohistoqu&#xED;mica (CD3, CD4, CD8, CD20, CD68, C4D, LFA-1, ICAM-1, E-selectina, P-selectina, VE-cadherina , HLA-DR, IDO, y Foxp3). Expresi&#xF3;n se ley&#xF3; con respecto al tiempo despu&#xE9;s del trasplante. El infiltrado estaba compuesta principalmente de CD3 + T-linfocitos. Entre ellas, las c&#xE9;lulas CD8 + fueron m&#xE1;s prominente que las c&#xE9;lulas CD4 +. CD20 + linfocitos B eran escasas y macr&#xF3;fagos CD68 + se encontraron en algunos, pero no todas las muestras (aproximadamente el 10% del infiltrado). El CD4/CD8-ratio se increment&#xF3; despu&#xE9;s del primer a&#xF1;o. Tinci&#xF3;n C4d fue principalmente positiva en las muestras en el momento de los puntos m&#xE1;s tardar el 1 a&#xF1;o. Las mol&#xE9;culas de adhesi&#xF3;n LFA-1, ICAM-1, E-selectina, P-selectina, y VE-cadherina se encontraron upregulated, y de la P-selectina, la expresi&#xF3;n aument&#xF3; con el tiempo despu&#xE9;s del trasplante. IDO la expresi&#xF3;n m&#xE1;s fuerte fue de 3 meses a 1 a&#xF1;o post-trasplante y una tendencia hacia una mayor Foxp3 + en las c&#xE9;lulas en los puntos de tiempo posteriores se observ&#xF3;. El rechazo leve en la piel despu&#xE9;s del trasplante de la mano se presenta con una c&#xE9;lula T de c&#xE9;lulas dominada infiltrado d&#xE9;rmico y la sobrerregulaci&#xF3;n de las mol&#xE9;culas de adhesi&#xF3;n. El papel de la expresi&#xF3;n C4d despu&#xE9;s de un a&#xF1;o est&#xE1; todav&#xED;a por dilucidar.

Caracterizaci&#xF3;n histopatol&#xF3;gica del rechazo leve (grado I) en biopsias de piel de los aloinjertos de mano Humanos

Reconstructive transplantation represents a bona fide option for select patients with devastating tissue loss, which could better restore the appearance, anatomy, and function than any other conventional treatment currently available. Despite favorable outcomes, broad clinical application of reconstructive transplantation is limited by the potential side effects of chronic multidrug immunosuppression. Thus, any reconstructive measures to improve these non-life-threatening conditions must address a delicate balance of risks and benefits. Today, several exciting novel therapeutic strategies, such as the implementation of cellular therapies including bone marrow or stem cells that integrate the concepts of immune regulation with those of nerve regeneration, are on the horizon. The development of reliable and reproducible small and large animal models is essential for the study of the unique immunological and biological aspects of vascularized composite allografts and to translate such novel immunoregulatory and tolerance-inducing strategies and therapeutic concepts from the bench to bedside. This review provides an overview of the multitude of small and large animal models that have been particularly designed for basic and translational research related to reconstructive transplantation.

Animal models for basic and translational research in reconstructive transplantation.

Vascularized composite allotransplantation (VCA) is an effective treatment option for patients suffering from limb loss or severe disfigurement. However, postoperative courses of VCA recipients have been complicated by skin rejection, and long-term immunosuppression remains a necessity for allograft survival. To widen the scope of this quality-of-life improving procedure minimization of immunosuppression in order to limit risks and side effects is needed. In some aspects, the molecular mechanisms and dynamics of skin allograft rejection seem similar to inflammatory skin conditions. T cells are key players in skin rejection and are recruited to the skin via activation of adhesion molecules, cytokines, and chemokines. Blocking these molecules has not only shown success in the treatment of inflammatory dermatoses, but also prolonged graft survival in various models of solid organ transplantation. In addition to T cell recruitment, ectopic lymphoid structures within the allograft associated with chronic rejection in solid organ transplantation might contribute to the strong alloimmune response towards the skin. Selectively targeting the molecules involved offers exciting novel therapeutic options in the prevention and treatment of skin rejection after VCA.

Mechanisms and mediators of inflammation: potential models for skin rejection and targeted therapy in vascularized composite allotransplantation.

Kv1.3-channels are critically involved in activation and function of effector memory T cells. Blocking Kv1.3-channels was investigated for its effect on skin rejection in a rat limb-transplantation-model. Animals received the Kv1.3-blocker correolide C systemically or locally as intra-graft-treatment in combination with tacrolimus. Systemic (intraperitoneal) administration of correolide C resulted in slight, but significant prolongation of allograft survival compared with untreated and placebo treated controls. In 4/6 correolide C treated animals, histology showed an intact epidermis and a mild infiltrate by day 10. High correolide C plasma trough levels correlated with prolonged allograft survival. A decrease in CD4+ and CD8+ effector memory T cells was observed in allograft skin, peripheral blood and the spleen on day 5. When applied subcutaneously in combination with systemic tacrolimus (30Â days+/-anti-lymphocyte serum) detectable, but insignificant prolongation of graft survival was achieved. 2/5 animals showed an intact epidermis and a mild infiltrate until day 45. Tapering systemic tacrolimus and weaning on day 50 resulted in rejection by day 55, regardless of local correolide C treatment. Subcutaneous injection did not lead to systemic plasma levels. The Kv1.3-channel is a potential drug target worth exploring in more detail for immunosuppression in vascularized composite allotransplantation.

Targeting the Kv1.3 potassium channel for immunosuppression in vascularized composite allotransplantation - a pilot study.

Wider application of vascularized composite allotransplantation (VCA) is limited by the need for chronic immunosuppression. Recent data suggest that the lymphatic system plays an important role in mediating rejection. This study used near-infrared (NIR) lymphography to describe lymphatic reconstitution in a rat VCA model. Syngeneic (Lewis-Lewis) and allogeneic (Brown Norway-Lewis) rat orthotopic hind limb transplants were performed without immunosuppression. Animals were imaged pre- and postoperatively using indocyanine green (ICG) lymphography. Images were collected using an NIR imaging system. Co-localization was achieved through use of an acrylic paint/hydrogen peroxide mixture. In all transplants, ICG first crossed graft suture lines on postoperative day (POD) 5. Clinical signs of rejection also appeared on POD 5 in allogeneic transplants, with most exhibiting Grade 3 rejection by POD 6. Injection of an acrylic paint/hydrogen peroxide mixture on POD 5 confirmed the existence of continuous lymphatic vessels crossing the suture line and draining into the inguinal lymph node. NIR lymphography is a minimally invasive imaging modality that can be used to study lymphatic vessels in a rat VCA model. In allogeneic transplants, lymphatic reconstitution correlated with clinical rejection. Lymphatic reconstitution may represent an early target for immunomodulation.

Near-infrared lymphography as a minimally invasive modality for imaging lymphatic reconstitution in a rat orthotopic hind limb transplantation model.

The effect of cold ischemia (CI) in vascularized composite allotransplantation is unknown. We herein assess tissue-specific damage, acceptable CI time, and the effect of preservation solutions in a syngenic rat hindlimb transplant model.

Histomorphometric evaluation of ischemia-reperfusion injury and the effect of preservation solutions histidine-tryptophan-ketoglutarate and University of Wisconsin in limb transplantation.

Broader clinical application of reconstructive hand and face transplantation is hindered by the need for lifelong immunosuppression for allograft maintenance. In this review, we summarize various cell-based approaches to tolerance induction currently under investigation in both clinical and pre-clinical models to alleviate the need for chronic immunosuppression. These include strategies to induce mixed hematopoietic chimerism, therapy with T and B regulatory cells, regulatory macrophages, tolerogenic dendritic cells, and mesenchymal stem cells. The vascularized, intragraft bone components inherent to reconstructive transplants serve as a continuous source of donor-derived hematopoietic cells, and make hand and face transplants uniquely well suited for cell-based approaches to tolerance that may ultimately tilt the risk-benefit balance for these life-changing, but not life-saving, procedures.

Exploring cell-based tolerance strategies for hand and face transplantation.

Many surgeries are complicated by the need to anastomose, or reconnect, micrometre-scale vessels. Although suturing remains the gold standard for anastomosing vessels, it is difficult to place sutures correctly through collapsed lumen, making the procedure prone to failure. Here, we report a multiphase transitioning peptide hydrogel that can be injected into the lumen of vessels to facilitate suturing. The peptide, which contains a photocaged glutamic acid, forms a solid-like gel in a syringe and can be shear-thin delivered to the lumen of collapsed vessels (where it distends the vessel) and the space between two vessels (where it is used to approximate the vessel ends). Suturing is performed directly through the gel. Light is used to initiate the final gel-sol phase transition that disrupts the hydrogel network, allowing the gel to be removed and blood flow to resume. This gel adds a new tool to the armamentarium for micro- and supermicrosurgical procedures.

Article	Year
Histopathologic characterization of mild rejection (grade I) in skin biopsies of human hand allografts. Transplant international : official journal of the European Society for Organ Transplantation\| PubMed ID: 21981770	2012
Animal models for basic and translational research in reconstructive transplantation. Birth defects research. Part C, Embryo today : reviews\| PubMed ID: 22457176	2012
Mechanisms and mediators of inflammation: potential models for skin rejection and targeted therapy in vascularized composite allotransplantation. Clinical & developmental immunology\| PubMed ID: 23049603	2012
Targeting the Kv1.3 potassium channel for immunosuppression in vascularized composite allotransplantation - a pilot study. Transplant international : official journal of the European Society for Organ Transplantation\| PubMed ID: 23489391	2013
Near-infrared lymphography as a minimally invasive modality for imaging lymphatic reconstitution in a rat orthotopic hind limb transplantation model. Transplant international : official journal of the European Society for Organ Transplantation\| PubMed ID: 23879384	2013
Histomorphometric evaluation of ischemia-reperfusion injury and the effect of preservation solutions histidine-tryptophan-ketoglutarate and University of Wisconsin in limb transplantation. Transplantation\| PubMed ID: 25073033	2014
Exploring cell-based tolerance strategies for hand and face transplantation. Expert review of clinical immunology\| PubMed ID: 26289376	2015
A multiphase transitioning peptide hydrogel for suturing ultrasmall vessels. Nature nanotechnology\| PubMed ID: 26524396	2016

Johanna Grahammer

Biography

JoVE Journal Publications

Other Publications