Abstract FR:

The participation of innate immunity mediated by natural autoreactive an polyreactive IgM antibodies is shown in recent developpements of ischemia reperfusion injury. The purpose of this thesis was to settle an ex-vivo model to confirm this theory. Lymphoblastic cells lines (LCLs) were established with infection by Eptein-Barr Virus (EBV) associated with unmethylated single-stranded DNA motifs (CpG oligonucleotides) as an antigenic stimulus. For each subtype of B Cell (Transitional, Mature naive and Memory), the effect of immortalization on phenotypic markers was studied in flowcytometry. The production of immunoglobulin was estimated by ELISA as well as the Polyreactivity of these antibodies. Autoreactivity was appreciating by an ana HEP-2 test. On the other hand, an ex-vivo normothermic reperfusion device was built using a modified (oxygenation an temperature control) perfusion pulsatile machine (MOX 100 or RM3). All 4 subtypes of B-cells from human peripheral blood were transformed into actively proliferating lymphoblastoid cell lines. He effect of transformation were relevant for 8 out of 16 studied phenotype markers: CD5, CD10, CD23, CD24, CD27, CD38, IgD, IgM. Immunoglobulin secretion : IgM, IgA or IgG can be secreted, but no IgD. The type of Ig secreted differs according to the original subtype of B-Cell. There is a very strong and coherent link between Ig membrane expression and Ig secretion. Some LCLs were secreting IgM type autoreactive antibodies. Ther were no IgA or IgG type autreactive antibodies. Some LCLs were secreting IgM type polyreactive antibodies. We succeed in building a ex-vivo normothermic reperfusion kidney device with an active diuresis of 6 hours. We succeed in building a ex-vivo normothermic reperfusion kidney device with an active diuresis of 6 hours in order to study Innate immunity mediated by Natural IgM during ischemia reperfusion injury. Mass production of autoreactive an polyreactive antibodies for this model is possible.