Abstract

? Xenotransplantation is of interest due to the insufficient supply of human organs. The pig is considered a likely donor species for humans. However, there are major barriers to successful xenotransplantation. The first two barriers, hyperacute rejection and acute vascular rejection (AVR), are mediated primarily by humoral immunity. If hyperacute rejection is avoided, AVR will occur. Strategies to interfere with AVR include reducing humoral immunity or inducing tolerance in the recipient, and decreasing immunogenicity or inducing protection in the graft. Our goal is to investigate induction of protection in the donor organ from AVR and achieve accommodation. HYPOTHESIS: The cytokines IL-4 and IL-13 induce protection in pig vascular tissues from effectors of immunologic injury, including complement, induction of adhesion molecules, and apoptosis. Activation of specific cellular mechanisms result in protection from xenogeneic injury.
SPECIFIC AIMS. I: Characterize the activation mechanism and identify the specific genes that are responsible for protection in endothelial cells activated with IL-4/IL-13. IL-4-mediated phosphatidylinositol-3 kinase (PI-3K) and Akt kinase activation and post-translational modification of the pro-apoptotic protein Bad will be investigated. Selected genes will be tested to determine if they actually mediate protection, using RNA interference to block gene function. Protection by overexpressing the gene after recombinant adenoviral transduction of endothelial cells will also be used. II: Identify other candidate genes potentially involved in cytoprotection induced with IL-4. Highly focussed experiments will be performed using a pig cDNA microarray to identify genes that change expression after IL-4 treatment in the presence and absence of specific PI-3K inhibitors. III. Induction of protection by IL-4 in an artery model of pig-to-primate xenotransplantation. Protection will be induced by incubation of a pig artery with IL-4 and by gene transfer of IL-4, IL-13, or other specific identified genes into the arterial tissues. Protection of the artery from injury that normally occurs upon perfusion with human blood will be assessed. Accomplishing these goals will result in understanding mechanisms of protection in pig tissues and to what extent they contribute to successful xenotransplantation, and would facilitate understanding the pathophysiology of vascular diseases. ? ?

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL062195-05
Application #
6730650
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Massicot-Fisher, Judith
Project Start
1999-08-16
Project End
2007-03-31
Budget Start
2004-04-01
Budget End
2005-03-31
Support Year
5
Fiscal Year
2004
Total Cost
$297,000
Indirect Cost

  Institution

Name
University of Minnesota Twin Cities
Department
Surgery
Type
Schools of Medicine
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455

  Publications

Benson, Barbara A; Vercellotti, Gregory M; Dalmasso, Agustin P (2015) IL-4 and IL-13 induce protection from complement and melittin in endothelial cells despite initial loss of cytoplasmic proteins: membrane resealing impairs quantifying cytotoxicity with the lactate dehydrogenase permeability assay. Xenotransplantation 22:295-301
Dalmasso, Agustin P; Goldish, Daniel; Benson, Barbara A et al. (2014) Interleukin-4 induces up-regulation of endothelial cell claudin-5 through activation of FoxO1: role in protection from complement-mediated injury. J Biol Chem 289:838-47
Dalmasso, Agustin P (2012) On the intersections of basic and applied research in xenotransplantation. Xenotransplantation 19:137-43
Black, Sylvester M; Benson, Barbara A; Idossa, Damé et al. (2011) Protection of porcine endothelial cells against apoptosis with interleukin-4. Xenotransplantation 18:343-54
Black, Sylvester M; Schott, Megan E; Batdorf, Bjorn H et al. (2010) IL-4 induces protection of vascular endothelial cells against killing by complement and melittin through lipid biosynthesis. Eur J Immunol 40:803-12
Black, S M; Schott, M E; Benson, B A et al. (2008) Interleukin-4 induces lipogenesis in porcine endothelial cells, which in turn is critical for induction of protection against complement-mediated injury. Transplant Proc 40:638-40
Black, Sylvester M; Grehan, John F; Rivard, Andrew L et al. (2006) Porcine endothelial cells and iliac arteries transduced with AdenoIL-4 are intrinsically protected, through Akt activation, against immediate injury caused by human complement. J Immunol 177:7355-63
Grehan, John F; Levay-Young, Brett K; Fogelson, Jeremy L et al. (2005) IL-4 and IL-13 induce protection of porcine endothelial cells from killing by human complement and from apoptosis through activation of a phosphatidylinositide 3-kinase/Akt pathway. J Immunol 175:1903-10
Grehan, John F; Levay-Young, Brett K; Benson, Barbara A et al. (2005) Alpha Gal ligation of pig endothelial cells induces protection from complement and apoptosis independently of NF-kappa B and inflammatory changes. Am J Transplant 5:712-9
Grubbs, Brian C; Benson, Barbara A; Dalmasso, Agustin P (2003) Characteristics of CD59 up-regulation induced in porcine endothelial cells by alphaGal ligation and its association with protection from complement. Xenotransplantation 10:387-97

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