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Adenovirus-mediated expression of human CD55 or CD59 protects adult porcine islets from complement-mediated cell lysis by human serum
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Oncology, Radiology and Clinical Immunology, Clinical Immunology.
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2003 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 75, no 5, 697-702 p.Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Protection against complement activation may reduce acute islet damage in pig-to-human islet xenotransplantation. Expression of the human complement regulatory proteins decay-accelerating factor (DAF, CD55) or CD59 was induced on intact adult porcine islets (APIs) by adenoviral transduction. The functional capacity of the transgenes was examined in vitro after exposure to fresh human serum.

METHODS: Intact APIs were transduced with adenoviral vectors Ad.hDAF or Ad.hCD59 or a control vector. After 3 days, the islets were trypsin dissociated to a single-cell suspension. A cytotoxicity assay was performed in which the islet cells were incubated with human complement active AB serum. Flow cytometry and immunohistochemistry were used to evaluate transgene expression.

RESULTS: APIs could be transduced to express hDAF or hCD59. Flow cytometry analysis of islet single cells revealed that only a fraction of the cells expressed the transgene; immunohistochemical staining of transduced islets demonstrated that mainly cells located in the periphery of the islets were expressing the protein. Cells from nontransduced islets or islets expressing the control protein were sensitive to lysis in human sera (66+/-4.0% and 73+/-3.7% cytotoxicity, respectively). Single cells from islets transduced with hDAF and hCD59 were partially protected from lysis. Islet cells expressing hCD59 were slightly less sensitive to lysis (33+/-3.3%) than cells expressing hDAF (45+/-3.5%).

CONCLUSIONS: These data show that intact pig islets can be transduced to express human regulators of complement activation on the surface and that pig islet cells expressing hDAF or hCD59 are less sensitive to complement-mediated lysis.

Place, publisher, year, edition, pages
2003. Vol. 75, no 5, 697-702 p.
National Category
Medical and Health Sciences
URN: urn:nbn:se:uu:diva-91264DOI: 10.1097/01.TP.0000053249.39753.D6PubMedID: 12640312OAI: oai:DiVA.org:uu-91264DiVA: diva2:163935
Available from: 2004-01-15 Created: 2004-01-15 Last updated: 2010-10-11Bibliographically approved
In thesis
1. Islet Xenotransplantation: An Experimental Study of Barriers to Clinical Transplantation
Open this publication in new window or tab >>Islet Xenotransplantation: An Experimental Study of Barriers to Clinical Transplantation
2004 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Xenotransplantation av Langerhanska öar : Experimentiella studier av hinder för klinisk tillämpning
Abstract [en]

In the field of transplantation, the increasing deficit of human donors have lead to an interest in animals as an alternative source of organs and tissues.

Different in vitro systems and rodent models of xenotransplantation were used to examine the most significant barriers that have to be overcome, before isolated islets of Langerhans from pigs can be used as a cure for insulin-dependent diabetes mellitus in humans.

In clinical transplantation, islets are infused into the liver through the portal vein. During this procedure the islets are susceptible to harmful innate reactions triggered in blood. Adenoviral vectors generating transgenic expression of human complement regulatory proteins were evaluated in pig islets and shown to confer protection against acute complement-mediated damage.

Transplanted islets escaping this immediate destruction will be targets of a cellular immune response. Using a new mouse model of islet xenograft rejection, it was demonstrated that macrophages, effector cells in the rejection, were part of an MHC-restricted xenospecific immune response mediated by T cells. In a strain of knockout mice it was further shown that this process can proceed in the absence of an important signalling system, mediated by Toll-like receptors, between cells in innate and adaptive immunity. These findings illustrate some of the mechanistic differences compared to cellular islet allograft rejection which partly explain why immunosuppressive drugs used in clinical allotransplantation is not sufficient for preventing xenograft rejection.

Porcine endogenous retroviruses (PERV) remain a safety concern in xenotransplantation. Characterization of PERV in pig islets indicated that virus expression is low in vitro but increases during the immediate time period following transplantation. This suggests that antiviral therapies administered at the time of transplantation could be used for preventing the risk of PERV transmission after xenotransplantation.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2004. 55 p.
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 0282-7476 ; 1317
Medicine, xenotransplantation, islets of Langerhans, rejection, diabetes, porcine endogenous retroviruses, quantitative RT-PCR, cytokines, animal models, Medicin
National Category
Dermatology and Venereal Diseases
urn:nbn:se:uu:diva-3942 (URN)91-554-5850-5 (ISBN)
Public defence
2004-02-06, Rudbecksalen, Rudbecklaboratoriet, Dag Hammarskölds väg 20, Uppsala, 09:15
Available from: 2004-01-15 Created: 2004-01-15Bibliographically approved

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