BACKGROUND: Brain death impairs donor organ quality and accelerates immune responses after transplantation. Detailed aspects of immune activation following brain death remain unclear. We have established a mouse model and investigated the immediate consequences of brain death and anesthesia on immune responses. METHODS: C57JBl/6 mice (n=6/group) were anesthetized with isoflurane (ISF) or ketamine/xylazine (KX); subsequently, animals underwent brain death induction and were followed for 3h under continuous ventilation. Blood pressure was monitored continuously and animals were resuscitated with normal saline to achieve normotension. Immune activation in brain dead animals was analyzed by IFNγ-ELispot, MLR, and flow-cytometry. Sham-operated and naïve animals served as controls. RESULTS: Blood pressure remained stable in both BD/KX and BD/ISF animals during the 3h observation time. Brain death was linked to systemic immune activation: IFNγ-expression of splenocytes and lymphocyte proliferation rates was significantly elevated subsequent to brain death (p<0.02, <0.01); T-cell activation markers CD28 and CD69 had increased in brain dead animals (p<0.03, <0.02). Isoflurane treatment in sham controls throughout the observation period (3.5h) revealed anesthesia associated IFNγ-expression and lymphocyte activation which were not observed when animals were treated with ketamine/xylazine (p<0.04, <0.009). CONCLUSIONS: This study reports on a reproducible and hemodynamically stable brain death mouse model. Hemodynamic stability was not impacted through either isoflurane or ketamine/xylazine induction. Of clinical relevance, prolonged anesthesia with isoflurane had been linked to pro-inflammatory cytokine activation. Brain death caused systemic immune activation in organ donors.
BACKGROUND:Brain death impairsdonor organ quality and accelerates immune responses after transplantation. Detailed aspects of immune activation following brain death remain unclear. We have established a mouse model and investigated the immediate consequences of brain death and anesthesia on immune responses. METHODS: C57JBl/6 mice (n=6/group) were anesthetized with isoflurane (ISF) or ketamine/xylazine (KX); subsequently, animals underwent brain death induction and were followed for 3h under continuous ventilation. Blood pressure was monitored continuously and animals were resuscitated with normal saline to achieve normotension. Immune activation in brain dead animals was analyzed by IFNγ-ELispot, MLR, and flow-cytometry. Sham-operated and naïve animals served as controls. RESULTS: Blood pressure remained stable in both BD/KX and BD/ISF animals during the 3h observation time. Brain death was linked to systemic immune activation: IFNγ-expression of splenocytes and lymphocyte proliferation rates was significantly elevated subsequent to brain death (p<0.02, <0.01); T-cell activation markers CD28 and CD69 had increased in brain dead animals (p<0.03, <0.02). Isoflurane treatment in sham controls throughout the observation period (3.5h) revealed anesthesia associated IFNγ-expression and lymphocyte activation which were not observed when animals were treated with ketamine/xylazine (p<0.04, <0.009). CONCLUSIONS: This study reports on a reproducible and hemodynamically stable brain deathmouse model. Hemodynamic stability was not impacted through either isoflurane or ketamine/xylazine induction. Of clinical relevance, prolonged anesthesia with isoflurane had been linked to pro-inflammatory cytokine activation. Brain death caused systemic immune activation in organ donors.
Authors: Marcus Buggert; Son Nguyen; Gonzalo Salgado-Montes de Oca; Bertram Bengsch; Samuel Darko; Amy Ransier; Emily R Roberts; Daniel Del Alcazar; Irene Bukh Brody; Laura A Vella; Lalit Beura; Sathi Wijeyesinghe; Ramin S Herati; Perla M Del Rio Estrada; Yuria Ablanedo-Terrazas; Leticia Kuri-Cervantes; Alberto Sada Japp; Sasikanth Manne; Shant Vartanian; Austin Huffman; Johan K Sandberg; Emma Gostick; Gregory Nadolski; Guido Silvestri; David H Canaday; David A Price; Constantinos Petrovas; Laura F Su; Golnaz Vahedi; Yoav Dori; Ian Frank; Maxim G Itkin; E John Wherry; Steven G Deeks; Ali Naji; Gustavo Reyes-Terán; David Masopust; Daniel C Douek; Michael R Betts Journal: Sci Immunol Date: 2018-06-01
Authors: R Oberhuber; P Ritschl; C Fabritius; A-V Nguyen; M Hermann; P Obrist; E R Werner; M Maglione; B Flörchinger; S Ebner; T Resch; J Pratschke; K Kotsch Journal: Am J Transplant Date: 2015-06-23 Impact factor: 8.086