Beschreibung:
<jats:title>Abstract</jats:title><jats:sec><jats:title>Background</jats:title><jats:p>Some patients with acute or acute‐on‐chronic hepatic failure die before a suitable human liver allograft becomes available. Encouraging results have been achieved in such patients by the transplantation of human hepatocyte progenitor cells from fetal liver tissue. The aim of the study was to explore survival of hepatocytes from genetically engineered pigs after direct injection into the spleen and other selected sites in immunosuppressed baboons to monitor the immune response and the metabolic function and survival of the transplanted hepatocytes.</jats:p></jats:sec><jats:sec><jats:title>Methods</jats:title><jats:p>Baboons (n=3) were recipients of <jats:styled-content style="fixed-case">GTKO</jats:styled-content>/<jats:styled-content style="fixed-case">hCD</jats:styled-content>46 pig hepatocytes<jats:bold>.</jats:bold> All three baboons received anti‐thymocyte globulin (<jats:styled-content style="fixed-case">ATG</jats:styled-content>) induction and tapering methylprednisolone. Baboon 1 received maintenance immunosuppressive therapy with tacrolimus and rapamycin. Baboons 2 and 3 received an anti‐<jats:styled-content style="fixed-case">CD</jats:styled-content>40mAb/rapamycin‐based regimen that prevents sensitization to pig solid organ grafts. The baboons were euthanized 4 or 5 weeks after hepatocyte transplantation. The baboon immune response was monitored by the measurement of anti‐non‐Gal IgM and IgG antibodies (by flow cytometry) and <jats:styled-content style="fixed-case">CFSE</jats:styled-content>‐mixed lymphocyte reaction. Monitoring for hepatocyte survival and function was by (i) real‐time <jats:styled-content style="fixed-case">PCR</jats:styled-content> detection of porcine <jats:styled-content style="fixed-case">DNA</jats:styled-content>, (ii) real‐time <jats:styled-content style="fixed-case">PCR</jats:styled-content> for porcine gene expression, and (iii) pig serum albumin levels (by <jats:styled-content style="fixed-case">ELISA</jats:styled-content>). The sites of hepatocyte injection were examined microscopically.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Detection of porcine <jats:styled-content style="fixed-case">DNA</jats:styled-content> and porcine gene expression was minimal at all sites of hepatocyte injection. Serum levels of porcine albumen were very low—500‐1000‐fold lower than in baboons with orthotopic pig liver grafts, and approximately 5000‐fold lower than in healthy pigs. No hepatocytes or infiltrating immune cells were seen at any of the injection sites. Two baboons (Baboons 1 and 3) demonstrated a significant increase in anti‐pig IgM and an even greater increase in IgG, indicating sensitization to pig antigens.</jats:p></jats:sec><jats:sec><jats:title>Discussion and Conclusions</jats:title><jats:p>As a result of this disappointing experience, the following points need to be considered. (i) Were the isolated pig hepatocytes functionally viable? (ii) Are pig hepatocytes more immunogenic than pig hearts, kidneys, artery patch grafts, or islets? (iii) Does injection of pig cells (antigens) into the spleen and/or lymph nodes stimulate a greater immune response than when pig tissues are grafted at other sites? (iv) Did the presence of the recipient's intact liver prevent survival and proliferation of pig hepatocytes? (v) Is pig <jats:styled-content style="fixed-case">CD</jats:styled-content>47‐primate <jats:styled-content style="fixed-case">SIRP</jats:styled-content>‐α compatibility essential? In conclusion, the transplantation of genetically engineered pig hepatocytes into multiple sites in immunosuppressed baboons was associated with very early graft failure. Considerable further study is required before clinical trials should be undertaken.</jats:p></jats:sec>