Reduzierung der Immunogenität des Herzens durch gentechnische Veränderungen während der ex vivo Herzperfusion
Katharina Schmalkuche (Hannover / DE), Tamina Rother (Hannover / DE), Jonathan Burgmann (Hannover / DE), Henrike Voß (Hannover / DE), Arjang Ruhparwar (Hannover / DE), Jan-Dieter Schmitto (Hannover / DE), Rainer Blasczyk (Hannover / DE), Constanca Ferreira de Figueiredo (Hannover / DE)
Lentiviral vectors encoding for shRNAs targeting β2-microglobulin sh(β2m) and class II transactivator (shCIITA) were delivered to the heart during two hours of normothermic ex-vivo heart perfusion (EVHP). Hearts perfused with lentiviral vectors encoding for non‐specific shRNA (shNS) or non-transduced (non-TD) hearts served as controls. After perfusion different cardiac cell types wer isolated, culture in vitro and evaluated for the expression of the reporter gene in luminescence assays and the SLA expression by real-time PCR and flow cytometry analyses.
Transduction efficiency was extremely high in all types of cells in the heart tissue including endothelial cells, fibroblasts and cardiomyocytes. Compared to shNS transduced hearts, silenced hearts showed up to 95% decreased SLA class I and up to 65% SLA class II-DQ expression, including the vascular endothelium as the major interface between donor and recipient. Histological analyses, perfusate troponin T (non-TD: 0.4±0.1 vs. TD: 0.2±0.1 ng/ml) and LDH levels (Abs A490-A690: non-TD: 0.961±0.1 vs. TD: 0.860±0.1) indicated no additional cell injury or tissue damage caused by lentiviral vector transduction in comparison to non-TD hearts after normothermic EVHP. Additionally, pro-inflammatory cytokine secretion signatures did not significantly differ between shβ2m/shCIITA and shNS transduced hearts.
This study clearly shows the potential of heart genetic engineering and its use in creating immunological graft invisibility. It is very likely that this technology will open up a completely new direction in overcoming organ rejection..Auf unserem Internetauftritt verwenden wir Cookies. Bei Cookies handelt es sich um kleine (Text-)Dateien, die auf Ihrem Endgerät (z.B. Smartphone, Notebook, Tablet, PC) angelegt und gespeichert werden. Einige dieser Cookies sind technisch notwendig um die Webseite zu betreiben, andere Cookies dienen dazu die Funktionalität der Webseite zu erweitern oder zu Marketingzwecken. Abgesehen von den technisch notwendigen Cookies, steht es Ihnen frei Cookies beim Besuch unserer Webseite zuzulassen oder nicht.