N-linked glycosylation modulates the immunogenicity of recombinant human factor VIII in hemophilia A mice.
N-linked glycosylation modulates the immunogenicity of recombinant human factor VIII in hemophilia A mice.
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DOI：
10.3324/haematol.2018.188219
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发表时间：
2018-11
期刊：
影响因子：
10.1
通讯作者：
Lillicrap D
中科院分区：
文献类型：
--
作者：
Lai JD;Swystun LL;Cartier D;Nesbitt K;Zhang C;Hough C;Dennis JW;Lillicrap D
对因子 VIII 的免疫反应仍然是治疗严重甲型血友病的最大并发症。最近的流行病学证据强调，幼仓鼠肾细胞产生的重组因子 VIII 比中国仓鼠卵巢细胞产生的因子 VIII 更具免疫原性。假设糖基化差异会影响这些合成浓缩物的免疫原性。在两种 A 型血友病小鼠模型中，与中国仓鼠卵巢细胞衍生的因子 VIII 相比，幼仓鼠肾细胞衍生的因子 VIII 引发了更强的免疫反应。此外，幼仓鼠肾细胞中产生的因子 VIII 表现出加速从循环中清除，与冯·维勒布兰德因子无关。总 N 连接聚糖的凝集素和质谱分析揭示了高甘露糖聚糖、唾液酸化和聚糖位点占用的差异。因子 VIII 去唾液酸化不影响与小鼠脾细胞或树突状细胞的结合，也不影响表面共刺激分子的表达。然而，我们确实观察到幼仓鼠肾源性因子 VIII 在幼年 A 型血友病小鼠中特异性的免疫球蛋白 M 水平升高。去 N-糖基化增强了免疫球蛋白 M 结合，表明 N-聚糖占据掩盖了表位。在健康个体中也观察到幼仓鼠肾源性因子 VIII 特异性的免疫球蛋白 M 和免疫球蛋白 G 水平升高，并且去 N-糖基化增加了免疫球蛋白 G 的结合。总的来说，我们的数据表明，幼仓鼠肾细胞产生的因子 VIII 比中国仓鼠卵巢细胞产生的因子 VIII 更具免疫原性，并且 N 连接糖基化位点的不完全占据导致免疫球蛋白 M 和免疫球蛋白 G 因子 VIII 免疫复合物的形成，从而有助于增强这些小鼠模型的清除率和免疫原性。 血友病A.
Immune responses to factor VIII remain the greatest complication in the treatment of severe hemophilia A. Recent epidemiological evidence has highlighted that recombinant factor VIII produced in baby hamster kidney cells is more immunogenic than factor VIII produced in Chinese hamster ovary cells. Glycosylation differences have been hypothesized to influence the immunogenicity of these synthetic concentrates. In two hemophilia A mouse models, baby hamster kidney cell-derived factor VIII elicited a stronger immune response compared to Chinese hamster ovary cell-derived factor VIII. Furthermore, factor VIII produced in baby hamster kidney cells exhibited accelerated clearance from circulation independent of von Willebrand factor. Lectin and mass spectrometry analysis of total N-linked glycans revealed differences in high-mannose glycans, sialylation, and the occupancy of glycan sites. Factor VIII desialylation did not influence binding to murine splenocytes or dendritic cells, nor surface co-stimulatory molecule expression. We did, however, observe increased levels of immunoglobulin M specific to baby hamster kidney-derived factor VIII in naïve hemophilia A mice. De-N-glycosylation enhanced immunoglobulin M binding, suggesting that N-glycan occupancy masks epitopes. Elevated levels of immunoglobulin M and immunoglobulin G specific to baby hamster kidney-derived factor VIII were also observed in healthy individuals, and de-N-glycosylation increased immunoglobulin G binding. Collectively, our data suggest that factor VIII produced in baby hamster kidney cells is more immunogenic than that produced in Chinese hamster ovary cells, and that incomplete occupancy of N-linked glycosylation sites leads to the formation of immunoglobulin M- and immunoglobulin G-factor VIII immune complexes that contribute to the enhanced clearance and immunogenicity in these mouse models of hemophilia A.