Glycan analysis of monoclonal antibodies secreted in deposition disorders indicates that subsets of plasma cells differentially process IgG glycans
Glycan analysis of monoclonal antibodies secreted in deposition disorders indicates that subsets of plasma cells differentially process IgG glycans
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DOI：
10.1002/art.22171
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发表时间：
2006-11-01
影响因子：
--
通讯作者：
Rudd, Pauline A.
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文献类型：
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作者：
Omtvedt, Lone A.;Royle, Louise;Rudd, Pauline A.
目标。比较类风湿关节炎（RA）患者多克隆血清IgG重链的糖基化与同一患者重链沉积病（HCDD）发作期间的单克隆血清IgG重链的糖基化，以确定糖基化过程是否对B细胞亚群具有特异性。用HiTrap蛋白G柱纯化血清IgG。免疫球蛋白在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳凝胶上运行，从凝胶带中分离IgG聚糖并荧光标记。采用正相高效液相色谱法和液相色谱-电喷雾电离-质谱法对多糖进行了分析。分析了1例血清阴性RA和HCDD患者血清免疫球蛋白的糖基化。主要的免疫球蛋白是截断的糖基化γ - 3重链，也存在少量的多克隆IgG。多糖谱显示，单克隆γ - 3重链含有完全半乳糖基化的双天线聚糖，与健康对照的IgG3相比，其焦点明显减少，但唾液酸含量增加。相比之下，多克隆IgG表现出ra样的特征，主要是聚焦的双链聚糖和低水平的半乳糖基化。同一患者在疾病缓解期间获得的血清IgG的聚糖谱与典型的RA谱相似。这些数据表明，不同类型的B细胞处理一组特定的IgG糖型。
Objective. To compare the glycosylation of polyclonal serum IgG heavy chains in a patient with rheumatoid arthritis (RA) with that of monoclonal serum IgG heavy chains in the same patient during an episode of heavy-chain deposition disease (HCDD), to establish whether glycosylation processing is specific for subsets of B cells.Methods. Serum IgG was purified using a HiTrap protein G column. Immunoglobulins were run on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, and IgG glycans were isolated from gel bands and fluorescently labeled. Glycans were analyzed by normal-phase high-performance liquid chromatography and by liquid chromatography-electrospray ionization-mass spectrometry.Results. The glycosylation of serum immunoglobulins from a patient with seronegative RA and HCDD was analyzed. The predominant immunoglobulin was a truncated glycosylated gamma 3 heavy chain, and a small amount of polyclonal IgG was also present. The glycan profile showed that the monoclonal gamma 3 heavy chain contained fully galactosylated biantennary glycans with significantly less fucose but more sialic acid than in IgG3 from healthy controls. In contrast, the polyclonal IgG showed an RA-like profile, with a predominance of fucosylated biantennary glycans and low levels of galactosylation. The glycan profile of serum IgG obtained from the same patient during disease remission resembled a typical RA profile.Conclusion. These data indicate that different types of B cells process a particular set of IgG glycoforms.