Effects of human TFPI and CD47 expression and selectin and integrin inhibition during GalTKO.hCD46 pig lung perfusion with human blood.
CD47
TFPI
integrin
lung xenotransplantation
selectin
Journal
Xenotransplantation
ISSN: 1399-3089
Titre abrégé: Xenotransplantation
Pays: Denmark
ID NLM: 9438793
Informations de publication
Date de publication:
03 2022
03 2022
Historique:
revised:
30
10
2021
received:
12
09
2021
accepted:
17
12
2021
pubmed:
3
3
2022
medline:
12
4
2022
entrez:
2
3
2022
Statut:
ppublish
Résumé
Loss of barrier function when GalTKO.hCD46 porcine lungs are perfused with human blood is associated with coagulation pathway dysregulation, innate immune system activation, and rapid sequestration of human formed blood elements. Here, we evaluate whether genetic expression of human tissue factor pathway inhibitor (hTFPI) and human CD47 (hCD47), alone or with combined selectin and integrin adhesion pathway inhibitors, delays GalTKO.hCD46 porcine lung injury or modulates neutrophil and platelet sequestration. In a well-established paired ex vivo lung perfusion model, GalTKO.hCD46.hTFPI.hCD47 transgenic porcine lungs (hTFPI.hCD47, n = 7) were compared to GalTKO.hCD46 lungs (reference, n = 5). All lung donor pigs were treated with a thromboxane synthase inhibitor, anti-histamine, and anti-GPIb integrin-blocking Fab, and were pre-treated with Desmopressin. In both genotypes, one lung of each pair was additionally treated with PSGL-1 and GMI-1271 (P- and E-selectin) and IB4 (CD11b/18 integrin) adhesion inhibitors (n = 6 hTFPI.hCD47, n = 3 reference). All except for two reference lungs did not fail within 480 min when experiments were electively terminated. Selectin and integrin adhesion inhibitors moderately attenuated initial pulmonary vascular resistance (PVR) elevation in hTFPI.hCD47 lungs. Neutrophil sequestration was significantly delayed during the early time points following reperfusion and terminal platelet activation was attenuated in association with lungs expressing hTFPI.hCD47, but additional adhesion pathway inhibitors did not show further effects with either lung genotype. Expression of hTFPI.hCD47 on porcine lung may be useful as part of an integrated strategy to prevent neutrophil adhesion and platelet activation that are associated with xenograft injury. Additionally, targeting canonical selectin and integrin adhesion pathways reduced PVR elevation associated with hTFPI.hCD47 expression, but did not significantly attenuate neutrophil or platelet sequestration. We conclude that other adhesive mechanisms mediate the residual sequestration of human formed blood elements to pig endothelium that occurs even in the context of the multiple genetic modifications and drug treatments tested here.
Sections du résumé
BACKGROUND
Loss of barrier function when GalTKO.hCD46 porcine lungs are perfused with human blood is associated with coagulation pathway dysregulation, innate immune system activation, and rapid sequestration of human formed blood elements. Here, we evaluate whether genetic expression of human tissue factor pathway inhibitor (hTFPI) and human CD47 (hCD47), alone or with combined selectin and integrin adhesion pathway inhibitors, delays GalTKO.hCD46 porcine lung injury or modulates neutrophil and platelet sequestration.
METHODS
In a well-established paired ex vivo lung perfusion model, GalTKO.hCD46.hTFPI.hCD47 transgenic porcine lungs (hTFPI.hCD47, n = 7) were compared to GalTKO.hCD46 lungs (reference, n = 5). All lung donor pigs were treated with a thromboxane synthase inhibitor, anti-histamine, and anti-GPIb integrin-blocking Fab, and were pre-treated with Desmopressin. In both genotypes, one lung of each pair was additionally treated with PSGL-1 and GMI-1271 (P- and E-selectin) and IB4 (CD11b/18 integrin) adhesion inhibitors (n = 6 hTFPI.hCD47, n = 3 reference).
RESULTS
All except for two reference lungs did not fail within 480 min when experiments were electively terminated. Selectin and integrin adhesion inhibitors moderately attenuated initial pulmonary vascular resistance (PVR) elevation in hTFPI.hCD47 lungs. Neutrophil sequestration was significantly delayed during the early time points following reperfusion and terminal platelet activation was attenuated in association with lungs expressing hTFPI.hCD47, but additional adhesion pathway inhibitors did not show further effects with either lung genotype.
CONCLUSION
Expression of hTFPI.hCD47 on porcine lung may be useful as part of an integrated strategy to prevent neutrophil adhesion and platelet activation that are associated with xenograft injury. Additionally, targeting canonical selectin and integrin adhesion pathways reduced PVR elevation associated with hTFPI.hCD47 expression, but did not significantly attenuate neutrophil or platelet sequestration. We conclude that other adhesive mechanisms mediate the residual sequestration of human formed blood elements to pig endothelium that occurs even in the context of the multiple genetic modifications and drug treatments tested here.
Identifiants
pubmed: 35234315
doi: 10.1111/xen.12725
pmc: PMC10207735
mid: NIHMS1894639
doi:
Substances chimiques
CD47 Antigen
0
CD47 protein, human
0
Integrins
0
Lipoproteins
0
Selectins
0
lipoprotein-associated coagulation inhibitor
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e12725Subventions
Organisme : NIAID NIH HHS
ID : U01 AI066335
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI090959
Pays : United States
Informations de copyright
© 2022 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Références
Methods Mol Biol. 2012;885:105-23
pubmed: 22565993
J Leukoc Biol. 2011 Aug;90(2):313-21
pubmed: 21551251
Transplantation. 2006 Apr 27;81(8):1157-64
pubmed: 16641602
Transplantation. 2012 Apr 15;93(7):686-92
pubmed: 22391577
Crit Care Med. 2000 Jul;28(7):2507-14
pubmed: 10921586
Curr Opin Organ Transplant. 2012 Apr;17(2):148-54
pubmed: 22327911
Xenotransplantation. 2014 Sep-Oct;21(5):397-419
pubmed: 25176336
Transplantation. 2018 Mar;102(3):406-416
pubmed: 28968355
Cell Mol Biol (Noisy-le-grand). 2015 Feb 28;61(1):1-6
pubmed: 25817339
J Biol Chem. 2000 Oct 13;275(41):31715-21
pubmed: 10922378
Curr Opin Organ Transplant. 2018 Dec;23(6):621-627
pubmed: 30234737
Thromb Haemost. 2017 Jun 2;117(6):1171-1181
pubmed: 28300869
Xenotransplantation. 2014 Sep-Oct;21(5):454-64
pubmed: 25130043
Int J Surg. 2015 Nov;23(Pt B):247-254
pubmed: 26278663
J Immunol. 2006 Jan 1;176(1):616-24
pubmed: 16365457
Xenotransplantation. 2018 Sep;25(5):e12391
pubmed: 29527745
Vet Immunol Immunopathol. 1994 Oct;43(1-3):185-91
pubmed: 7856051
Arterioscler Thromb Vasc Biol. 2016 Sep;36(9):1809-20
pubmed: 27417582
Xenotransplantation. 2011 Mar-Apr;18(2):94-107
pubmed: 21496117
Curr Opin Hematol. 2017 May;24(3):265-273
pubmed: 28178038
Expert Opin Biol Ther. 2008 Jan;8(1):1-4
pubmed: 18081532
Transplantation. 1999 Sep 27;68(6):832-9
pubmed: 10515384
Xenotransplantation. 2017 Mar;24(2):
pubmed: 28303661
Xenotransplantation. 2008 Feb;15(1):27-35
pubmed: 18333911
Curr Opin Organ Transplant. 2018 Dec;23(6):615-620
pubmed: 30300328
Transplantation. 2002 Dec 15;74(11):1596-603
pubmed: 12490794
Blood. 2011 Dec 22;118(26):6743-51
pubmed: 22021370
Blood. 1995 Oct 1;86(7):2760-6
pubmed: 7545469
Xenotransplantation. 2008 May-Jun;15(3):191-7
pubmed: 18611227
Xenotransplantation. 2016 May;23(3):222-236
pubmed: 27188532
Transpl Immunol. 2009 Jun;21(2):81-6
pubmed: 19427901
Transplant Proc. 1996 Apr;28(2):625
pubmed: 8623309
Xenotransplantation. 2021 Nov;28(6):e12712
pubmed: 34657336
Am J Transplant. 2008 Dec;8(12):2516-26
pubmed: 19032222
Anticancer Res. 2005 Mar-Apr;25(2A):757-64
pubmed: 15868907
Am J Transplant. 2014 May;14(5):1084-95
pubmed: 24698431
J Immunol. 2005 Feb 15;174(4):2004-11
pubmed: 15699129
J Appl Physiol (1985). 2001 Jun;90(6):2257-68
pubmed: 11356791
J Heart Lung Transplant. 2013 Nov;32(11):1123-30
pubmed: 23932853
Kidney Int. 2014 Feb;85(2):265-75
pubmed: 24088952
Front Immunol. 2020 Apr 15;11:622
pubmed: 32351506
Methods Mol Biol. 2020;2110:173-196
pubmed: 32002909
Cell Mol Immunol. 2011 Jul;8(4):285-8
pubmed: 21258362
Am J Respir Crit Care Med. 1998 Mar;157(3 Pt 1):938-49
pubmed: 9517615
Xenotransplantation. 2017 May;24(3):
pubmed: 28393401
Xenotransplantation. 2018 Mar;25(2):e12381
pubmed: 29359469
Nat Commun. 2016 Apr 05;7:11138
pubmed: 27045379
Agents Actions. 1985 Dec;17(2):150-2
pubmed: 4096304
Xenotransplantation. 2014 Sep-Oct;21(5):420-30
pubmed: 25176471
Circulation. 1999 Mar 16;99(10):1363-9
pubmed: 10077522
Thromb Haemost. 1997 Dec;78(6):1488-94
pubmed: 9423800
Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):5062-6
pubmed: 17360380
Am J Transplant. 2022 Jan;22(1):28-45
pubmed: 34424601
Cardiovasc Res. 1999 Jan;41(1):65-76
pubmed: 10325954
Xenotransplantation. 2012 Jul-Aug;19(4):256-64
pubmed: 22909139
Xenotransplantation. 2017 Mar;24(2):
pubmed: 28258595
Xenotransplantation. 2012 May-Jun;19(3):144-58
pubmed: 22702466
Am J Pathol. 2001 May;158(5):1809-19
pubmed: 11337379
J Immunol. 1989 Apr 1;142(7):2352-8
pubmed: 2538507
Xenotransplantation. 2014 Mar-Apr;21(2):99-114
pubmed: 25268248
Thromb Res. 2014 Mar;133(3):477-80
pubmed: 24393660
Nat Biotechnol. 2002 Mar;20(3):251-5
pubmed: 11875425
Am J Transplant. 2014 Feb;14(2):488-9
pubmed: 24330419
Transplantation. 2005 Sep 27;80(6):828-35
pubmed: 16210972
J Pharmacol Exp Ther. 2001 Aug;298(2):658-64
pubmed: 11454928