The xenoantibody response and immunoglobulin gene expression profile of cynomolgus monkeys transplanted with hDAF-transgenic porcine hearts.

摘要: Abstract:  Background:  Recent work has indicated a role for anti-Galα1-3Gal (Gal) and anti-non-Gal xenoantibodies in the primate humoral rejection response against human-decay accelerating factor (hDAF) transgenic pig organs. Our laboratory shown that anti-porcine xenograft antibodies humans non-human primates are encoded by small number of germline IgVH progenitors. In this study, we extended our analysis to identify genes encoding immunosuppressed cynomolgus monkeys (Macaca fascicularis) transplanted with hDAF-transgenic organs. Methods:  Three underwent heterotopic heart transplantation hDAF porcine xenografts. Two three animals were given GAS914, poly-l-lysine derivative bind anti-Gal neutralize them. One animal rejected its at post-operative day (POD) 39; second POD 78. The third monkey was euthanized on 36 but not rejected. Peripheral blood leukocytes (PBL) serum obtained from each before multiple time points after transplantation. We analyzed immune enzyme-linked immunosorbent assay (ELISA) confirm whether or induced graft placement. Immunoglobulin heavy-chain gene (VH) cDNA libraries then produced screened. generated soluble single-chain (scFv) establish binding specificity cloned immunoglobulin genes. Results:  Despite immunosuppression, which included use polymer two their hearts showed elevated levels cytotoxic anti-pig red cell (RBC) aortic endothelial (PAEC) antibodies. did reject decline anti-RBC, anti-PAEC, when compared pre-transplant levels. A VH3 family high level sequence similarity an allele VH3–11, designated VH3–11cyno, expressed GAS914 whose until IgM IgG directed N-acetyl lactosamine (a precursor Gal epitope) also animal. scFv new determine antibody could carbohydrate, demonstrated protein capable blocking human xenoantibody oligosaccharide, as had previously been VH3–11 scFv. Conclusions:  DAF-transgenic organs into induce responses mediated both xenoantibodies. Anti-non-Gal treated GAS914. Antibodies carbohydrate absence treatment. remained beating 78 days increased usage progenitor is structurally related, distinct IGHV311. This binds does rapid early 8 post-transplantation.