Removal of single-site N-linked glycans on factor VIII alters binding of domain-specific monoclonal antibodies.
Removal of single-site N-linked glycans on factor VIII alters binding of domain-specific monoclonal antibodies.
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DOI：
10.1111/jth.15616
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Batsuli G
中科院分区：
文献类型：
--
作者：
Ito J;Baldwin WH;Cox C;Healey JF;Parker ET;Legan ER;Li R;Gill S;Batsuli G
一部分血友病A患者会产生中和抗体，称为糖蛋白因子VIII（FVIII）抑制剂。有多种风险因素会导致抑制剂形成的风险。然而，关于FVIII天冬酰胺（N）连接糖基化在FVIII免疫中的作用的知识有限。评价位点特异性N-连接聚糖去除对FVIII生化特性、小鼠骨髓源性树突状细胞（BMDC）的内吞作用和抗体应答的影响。制备了四种重组B结构域缺失（BDD）FVIII变体，其具有去除N-连接聚糖的单位点氨基酸取代。生产了经确认去除N-连接聚糖且糖基化特征与BDD FVIII相似的BDD FVIII-N41 G、FVIII-N239 A、FVIII-N1810 A和FVIII-N2118 A。与BDD FVIII相比，FVIII变体的凝血酶活化或血管性血友病因子结合无差异，但在所有变体中均观察到FVIII表达、活性和比活性降低。BDD FVIII-N41 G和FVIII-N1810 A降低了BMDC的摄取，但与BDD FVIII相比，免疫的血友病A小鼠中的抗体形成没有差异。12种结构域特异性FVIII MAb库中有一半与≥1种FVIII变体的结合显著降低，A1结构域MAb 2-116与FVIII-N239 A的结合降低50%。FVIII N-连接聚糖的修饰减少了BMDC对FVIII的内吞作用和结构域特异性FVIII MAb的结合，但未改变血友病A小鼠中的从头抗体产生，表明N-聚糖对抑制剂形成无显著影响。
A portion of individuals with hemophilia A develop neutralizing antibodies called inhibitors to glycoprotein factor VIII (FVIII). There are multiple risk factors that contribute to the risk of inhibitor formation. However, knowledge of the role of FVIII asparagine (N)-linked glycosylation in FVIII immunity is limited. To evaluate the effect of site-specific N-linked glycan removal on FVIII biochemical properties, endocytosis by murine bone marrow-derived dendritic cells (BMDCs), and antibody responses. Four recombinant B domain deleted (BDD) FVIII variants with single-site amino acid substitutions to remove N-linked glycans were produced for experimental assays. BDD FVIII-N41G, FVIII-N239A, FVIII-N1810A, and FVIII-N2118A with confirmed removal of N-linked glycans and similar glycosylation profiles to BDD FVIII were produced. There were no differences in thrombin activation or von Willebrand factor binding of FVIII variants compared to BDD FVIII, however, reduced FVIII expression, activity, and specific activity was observed with all variants. BDD FVIII-N41G and FVIII-N1810A had reduced uptake by BMDCs, but there were no differences in antibody development in immunized hemophilia A mice compared to BDD FVIII. Half of a repertoire of 12 domain-specific FVIII MAbs had significantly reduced binding to ≥1 FVIII variant with a 50% decrease in A1 domain MAb 2-116 binding to FVIII-N239A. Modifications of FVIII N-linked glycans reduced FVIII endocytosis by BMDCs and binding of domain-specific FVIII MAbs, but did not alter de novo antibody production in hemophilia A mice suggesting that N-glycans do not significantly contribute to inhibitor formation.
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影响因子：
7.5
作者：
Kannicht, Christoph;Ramstrom, Margareta;Sandberg, Helena
通讯作者：
Sandberg, Helena
影响因子：
10.4
作者：
Canis, K.;Anzengruber, J.;Malisauskas, M.
通讯作者：
Malisauskas, M.
影响因子：
3.9
作者：
Dimichele, D
通讯作者：
Dimichele, D
DOI：
10.1073/pnas.84.23.8573
发表时间：
1987-12-01
影响因子：
11.1
作者：
ARUFFO, A;SEED, B
通讯作者：
SEED, B
影响因子：
20.3
作者：
Eubanks, Joshua;Baldwin, W. Hunter;Meeks, Shannon L.
通讯作者：
Meeks, Shannon L.