{"id":{"repo_id":"freiburg-diss","oai_identifier":"oai:freidok.uni-freiburg.de:1948"},"canonical_url":"https://search.dev.ndltd.org/etd/freiburg-diss/oai:freidok.uni-freiburg.de:1948","repository":{"repo_id":"freiburg-diss","name":"University of Freiburg","base_url":"https://freidok.uni-freiburg.de/oai/oai2.php"},"display":{"title":"A comparison of gene expression in tolerated and rejected islet grafts by gene microarray analysis","abstract":"Background. It has recently been demonstrated that donor-specific tolerance to MHC-matched minor-antigen-mismatched islet allografts in diabetic NOD mice could be induced by simultaneous islet and bone marrow transplantation preventing both graft rejection and recurrence of autoimmunity. Despite the presence of tolerance and good graft function mononuclear cell infiltration surrounding the islet was found in tolerated grafts. To elucidate the differences between these mononuclear infiltrates in the presence of tolerance and destructive mononuclear infiltrates during graft rejection as well as their influence on intragraft gene expression, tolerated islet grafts from the described model and rejected islet grafts from untreated animals were subjected to gene expression studies. <br>Methods. Gene expression analysis in tolerated and rejected islet grafts was performed by using Affymetrix Murine U74A oligonucleotide arrays. Real-time PCR and RNase protection assay on selected genes were performed to confirm the results of microarray analysis. <br>Results. Of over 12,000 genes studied, 57 genes were expressed at consistently higher levels in tolerated islet grafts, and 524 genes in rejected islet grafts. Genes from a variety of different functional clusters were found to be different between rejected and tolerated grafts. In the rejected islet grafts, a number of T cell surface markers (e.g., CD3, CD8) and of cytotoxicity-related genes (e.g., granzyme B and Fas ligand) were highly expressed. Also in the rejected grafts, a number of cytokines and chemokines and their receptors (e.g., IL-1b, IFN-g, MIG, RANTES) were found at high levels. The differential expression of selected genes found by microarray analysis was also confirmed by real-time PCR and RNase protection assay. <br>Conclusion. In our studies, infiltrating mononuclear cells in rejected and tolerated islet grafts showed differential expression in a number of genes belonging to different functional clusters. Gene microarray analysis can be used to detect gene expression differences representative of the biologic mechanisms of tolerance and rejection and thereby provide us a new tool for the discovery of genes that can be used in immune monitoring and as target for new medical therapies for the treatment of rejection.","abstract_html":"Background. It has recently been demonstrated that donor-specific tolerance to MHC-matched minor-antigen-mismatched islet allografts in diabetic NOD mice could be induced by simultaneous islet and bone marrow transplantation preventing both graft rejection and recurrence of autoimmunity. Despite the presence of tolerance and good graft function mononuclear cell infiltration surrounding the islet was found in tolerated grafts. To elucidate the differences between these mononuclear infiltrates in the presence of tolerance and destructive mononuclear infiltrates during graft rejection as well as their influence on intragraft gene expression, tolerated islet grafts from the described model and rejected islet grafts from untreated animals were subjected to gene expression studies. &lt;br&gt;Methods. Gene expression analysis in tolerated and rejected islet grafts was performed by using Affymetrix Murine U74A oligonucleotide arrays. Real-time PCR and RNase protection assay on selected genes were performed to confirm the results of microarray analysis. &lt;br&gt;Results. Of over 12,000 genes studied, 57 genes were expressed at consistently higher levels in tolerated islet grafts, and 524 genes in rejected islet grafts. Genes from a variety of different functional clusters were found to be different between rejected and tolerated grafts. In the rejected islet grafts, a number of T cell surface markers (e.g., CD3, CD8) and of cytotoxicity-related genes (e.g., granzyme B and Fas ligand) were highly expressed. Also in the rejected grafts, a number of cytokines and chemokines and their receptors (e.g., IL-1b, IFN-g, MIG, RANTES) were found at high levels. The differential expression of selected genes found by microarray analysis was also confirmed by real-time PCR and RNase protection assay. &lt;br&gt;Conclusion. In our studies, infiltrating mononuclear cells in rejected and tolerated islet grafts showed differential expression in a number of genes belonging to different functional clusters. Gene microarray analysis can be used to detect gene expression differences representative of the biologic mechanisms of tolerance and rejection and thereby provide us a new tool for the discovery of genes that can be used in immune monitoring and as target for new medical therapies for the treatment of rejection.","abstract_has_math":false,"creators":["Berg, Tobias"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Reincke, Martin"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":null,"date_issued":"","date_published":null,"updated_at":"2026-07-24T02:22:33Z","subjects":["Transplantation immunology","islet transplantation","microarray","immune tolerance","chemokines"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://freidok.uni-freiburg.de/data/1948","outbound_label":"Repository record","outbound_source":"source_url"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Reincke, Martin"]},{"key":"dc:creator","label":"Author","values":["Berg, Tobias"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:type","label":"Dc Type","values":["DoctoralThesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Transplantation immunology","islet transplantation","microarray","immune tolerance","chemokines"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Background. It has recently been demonstrated that donor-specific tolerance to MHC-matched minor-antigen-mismatched islet allografts in diabetic NOD mice could be induced by simultaneous islet and bone marrow transplantation preventing both graft rejection and recurrence of autoimmunity. Despite the presence of tolerance and good graft function mononuclear cell infiltration surrounding the islet was found in tolerated grafts. To elucidate the differences between these mononuclear infiltrates in the presence of tolerance and destructive mononuclear infiltrates during graft rejection as well as their influence on intragraft gene expression, tolerated islet grafts from the described model and rejected islet grafts from untreated animals were subjected to gene expression studies. <br>Methods. Gene expression analysis in tolerated and rejected islet grafts was performed by using Affymetrix Murine U74A oligonucleotide arrays. Real-time PCR and RNase protection assay on selected genes were performed to confirm the results of microarray analysis. <br>Results. Of over 12,000 genes studied, 57 genes were expressed at consistently higher levels in tolerated islet grafts, and 524 genes in rejected islet grafts. Genes from a variety of different functional clusters were found to be different between rejected and tolerated grafts. In the rejected islet grafts, a number of T cell surface markers (e.g., CD3, CD8) and of cytotoxicity-related genes (e.g., granzyme B and Fas ligand) were highly expressed. Also in the rejected grafts, a number of cytokines and chemokines and their receptors (e.g., IL-1b, IFN-g, MIG, RANTES) were found at high levels. The differential expression of selected genes found by microarray analysis was also confirmed by real-time PCR and RNase protection assay. <br>Conclusion. In our studies, infiltrating mononuclear cells in rejected and tolerated islet grafts showed differential expression in a number of genes belonging to different functional clusters. Gene microarray analysis can be used to detect gene expression differences representative of the biologic mechanisms of tolerance and rejection and thereby provide us a new tool for the discovery of genes that can be used in immune monitoring and as target for new medical therapies for the treatment of rejection.","In jüngster Zeit konnte gezeigt werden, dass es durch gleichzeitige Knochenmark- und Inselzelltransplantation möglich ist, in diabetischen NOD-Mäusen Toleranz gegenüber MHC-identischen Inseltransplantaten zu induzieren und so sowohl eine Abstoßung als auch ein Wiederauftreten der Autoimmunreaktion zu verhindern. Trotz vorhandener Toleranz und guter Transplantatfunktion zeigte sich in diesen Transplantaten eine Infiltration durch mononukleäre Zellen, die die Inseln umgaben, ohne sie in ihrer Struktur oder Funktion zu schädigen. Um die Unterschiede zwischen diesen toleranzassoziierten Infiltraten und den destruktiven Infiltraten während Abstoßung sowie ihren Einfluß auf die Genexpression in den Inseltransplantaten zu ermitteln, wurden vergleichende Genexpressionsstudien mit tolerierten Transplantaten aus dem beschriebenen Toleranzinduktionsmodell und abgestoßenen Transplantaten von unbehandelten Tieren durchgeführt. <br>Das Genexpressionsmuster in tolerierten und abgestoßenen Inseltransplantaten wurde mittels Affymetrix U74A-Oligonukleotidarrays verglichen. Die Ergebnisse dieser Versuche wurde mittels konventionellen Methoden der Genexpressionsanalyse (quantitative realtime PCR, RNase-Protektions-Assay) bestätigt. <br>Von mehr als 12.000 untersuchten Genen wurden 57 Gene vermehrt in tolerierten Inseltransplantaten exprimiert und 524 in abgestoßenen Inseltransplantaten. Tolerierte und abgestoßene Inseltransplantate unterscheiden sich dabei in der Expression von Genen unterschiedlicher funktioneller Gruppen. In abgestoßenen Inseltransplantaten fand sich eine höhere Expression einer Anzahl von Oberflächenmarkern von T-Lymphozyten (z. B. CD3, CD8) und von Genen, die bei ihren zytotoxischen Eigenschaften eine Rolle spielen (z. B. Granzyme B, Fas Ligand). Ebenso fand sich in den abgestoßenen Transplantaten eine gesteigerte Expression einer Anzahl an Zytokinen und Chemokinen sowie ihrer Rezeptoren (z. B. IL-1b, IFN-g, MIG, RANTES). Die unterschiedliche Expression von einigen ausgewählten Genen, die mittels Microarray-Analyse ermittelt wurde, konnte mittels quantitativer realtime-PCR und RNase-Protektions-Assay bestätigt werden. Die Ergebnisse dieser Versuche legen nahe, dass sich die mononukleären Zellen, die im Kontext einer ablaufenden Abstoßung und die bei Toleranz ein Transplantat infiltieren, im Expressionsmuster der Gene unterschiedlicher funktioneller Gruppen unterscheiden. Die Microarray-Analyse mit Oligonukleotidarrays kann genutzt werden, um die Genexpressionsunterschiede zu ermitteln, die repräsentativ für die biologischen Mechanismen von Toleranz und Abstoßung sind. Auf diese Weise kann sie somit Zielgene für die Entwicklung von Methoden zur Überwachung der Immunantwort und Ansatzpunkte für neue Medikamente liefern."]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["A comparison of gene expression in tolerated and rejected islet grafts by gene microarray analysis","Vergleich der Genexpression in tolerierten und abgestossenen Inseltransplantaten mit LS-Mikroarray-Aalyse"]}]}],"canonical_facts":{"dc:contributor":["Reincke, Martin"],"dc:creator":["Berg, Tobias"],"dc:description.abstract":["Background. It has recently been demonstrated that donor-specific tolerance to MHC-matched minor-antigen-mismatched islet allografts in diabetic NOD mice could be induced by simultaneous islet and bone marrow transplantation preventing both graft rejection and recurrence of autoimmunity. Despite the presence of tolerance and good graft function mononuclear cell infiltration surrounding the islet was found in tolerated grafts. To elucidate the differences between these mononuclear infiltrates in the presence of tolerance and destructive mononuclear infiltrates during graft rejection as well as their influence on intragraft gene expression, tolerated islet grafts from the described model and rejected islet grafts from untreated animals were subjected to gene expression studies. <br>Methods. Gene expression analysis in tolerated and rejected islet grafts was performed by using Affymetrix Murine U74A oligonucleotide arrays. Real-time PCR and RNase protection assay on selected genes were performed to confirm the results of microarray analysis. <br>Results. Of over 12,000 genes studied, 57 genes were expressed at consistently higher levels in tolerated islet grafts, and 524 genes in rejected islet grafts. Genes from a variety of different functional clusters were found to be different between rejected and tolerated grafts. In the rejected islet grafts, a number of T cell surface markers (e.g., CD3, CD8) and of cytotoxicity-related genes (e.g., granzyme B and Fas ligand) were highly expressed. Also in the rejected grafts, a number of cytokines and chemokines and their receptors (e.g., IL-1b, IFN-g, MIG, RANTES) were found at high levels. The differential expression of selected genes found by microarray analysis was also confirmed by real-time PCR and RNase protection assay. <br>Conclusion. In our studies, infiltrating mononuclear cells in rejected and tolerated islet grafts showed differential expression in a number of genes belonging to different functional clusters. Gene microarray analysis can be used to detect gene expression differences representative of the biologic mechanisms of tolerance and rejection and thereby provide us a new tool for the discovery of genes that can be used in immune monitoring and as target for new medical therapies for the treatment of rejection.","In jüngster Zeit konnte gezeigt werden, dass es durch gleichzeitige Knochenmark- und Inselzelltransplantation möglich ist, in diabetischen NOD-Mäusen Toleranz gegenüber MHC-identischen Inseltransplantaten zu induzieren und so sowohl eine Abstoßung als auch ein Wiederauftreten der Autoimmunreaktion zu verhindern. Trotz vorhandener Toleranz und guter Transplantatfunktion zeigte sich in diesen Transplantaten eine Infiltration durch mononukleäre Zellen, die die Inseln umgaben, ohne sie in ihrer Struktur oder Funktion zu schädigen. Um die Unterschiede zwischen diesen toleranzassoziierten Infiltraten und den destruktiven Infiltraten während Abstoßung sowie ihren Einfluß auf die Genexpression in den Inseltransplantaten zu ermitteln, wurden vergleichende Genexpressionsstudien mit tolerierten Transplantaten aus dem beschriebenen Toleranzinduktionsmodell und abgestoßenen Transplantaten von unbehandelten Tieren durchgeführt. <br>Das Genexpressionsmuster in tolerierten und abgestoßenen Inseltransplantaten wurde mittels Affymetrix U74A-Oligonukleotidarrays verglichen. Die Ergebnisse dieser Versuche wurde mittels konventionellen Methoden der Genexpressionsanalyse (quantitative realtime PCR, RNase-Protektions-Assay) bestätigt. <br>Von mehr als 12.000 untersuchten Genen wurden 57 Gene vermehrt in tolerierten Inseltransplantaten exprimiert und 524 in abgestoßenen Inseltransplantaten. Tolerierte und abgestoßene Inseltransplantate unterscheiden sich dabei in der Expression von Genen unterschiedlicher funktioneller Gruppen. In abgestoßenen Inseltransplantaten fand sich eine höhere Expression einer Anzahl von Oberflächenmarkern von T-Lymphozyten (z. B. CD3, CD8) und von Genen, die bei ihren zytotoxischen Eigenschaften eine Rolle spielen (z. B. Granzyme B, Fas Ligand). Ebenso fand sich in den abgestoßenen Transplantaten eine gesteigerte Expression einer Anzahl an Zytokinen und Chemokinen sowie ihrer Rezeptoren (z. B. IL-1b, IFN-g, MIG, RANTES). Die unterschiedliche Expression von einigen ausgewählten Genen, die mittels Microarray-Analyse ermittelt wurde, konnte mittels quantitativer realtime-PCR und RNase-Protektions-Assay bestätigt werden. Die Ergebnisse dieser Versuche legen nahe, dass sich die mononukleären Zellen, die im Kontext einer ablaufenden Abstoßung und die bei Toleranz ein Transplantat infiltieren, im Expressionsmuster der Gene unterschiedlicher funktioneller Gruppen unterscheiden. Die Microarray-Analyse mit Oligonukleotidarrays kann genutzt werden, um die Genexpressionsunterschiede zu ermitteln, die repräsentativ für die biologischen Mechanismen von Toleranz und Abstoßung sind. Auf diese Weise kann sie somit Zielgene für die Entwicklung von Methoden zur Überwachung der Immunantwort und Ansatzpunkte für neue Medikamente liefern."],"dc:format.medium":["application/pdf"],"dc:subject":["Transplantation immunology","islet transplantation","microarray","immune tolerance","chemokines"],"dc:title":["A comparison of gene expression in tolerated and rejected islet grafts by gene microarray analysis","Vergleich der Genexpression in tolerierten und abgestossenen Inseltransplantaten mit LS-Mikroarray-Aalyse"],"dc:type":["DoctoralThesis"]},"updated_at":"2026-07-24T02:22:33Z"}