Department of Pathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, New York, United States of America.
PLoS One. 2020 Jul 1;15(7):e0235143. doi: 10.1371/journal.pone.0235143. eCollection 2020.
To clarify the significance of quantitative analyses of amyloid proteins in clinical practice and in research relating to systemic amyloidoses, we applied mass spectrometry-based quantification by isotope-labeled cell-free products (MS-QBIC) to formalin-fixed, paraffin-embedded (FFPE) tissues. The technique was applied to amyloid tissues collected by laser microdissection of Congo red-stained lesions of FFPE specimens. Twelve of 13 amyloid precursor proteins were successfully quantified, including serum amyloid A (SAA), transthyretin (TTR), immunoglobulin kappa light chain (IGK), immunoglobulin lambda light chain (IGL), beta-2-microglobulin (B2M), apolipoprotein (Apo) A1, Apo A4, Apo E, lysozyme, Apo A2, gelsolin, and fibrinogen alpha chain; leukocyte cell-derived chemotaxin-2 was not detected. The quantification of SAA, TTR, IGK, IGL, and B2M confirmed the responsible proteins, even when the immunohistochemical results were not decisive. Considerable amounts of Apo A1, Apo A4, and Apo E were deposited in parallel amounts with the responsible proteins. Quantification of amyloid protein by MS-QBIC is feasible and useful for the classification of and research on systemic amyloidoses.
为了阐明定量分析淀粉样蛋白在临床实践和系统性淀粉样变性相关研究中的意义，我们应用基于同位素标记细胞游离产物的质谱定量（MS-QBIC）方法对福尔马林固定、石蜡包埋（FFPE）组织进行分析。该技术应用于刚果红染色的 FFPE 标本激光微切割获得的淀粉样组织。成功定量了 13 种淀粉样前体蛋白中的 12 种，包括血清淀粉样蛋白 A（SAA）、转甲状腺素蛋白（TTR）、免疫球蛋白 k 轻链（IGK）、免疫球蛋白 lambda 轻链（IGL）、β2-微球蛋白（B2M）、载脂蛋白（Apo）A1、Apo A4、Apo E、溶菌酶、Apo A2、凝胶蛋白和纤维蛋白原 alpha 链；白细胞衍生趋化因子-2 未被检测到。SAA、TTR、IGK、IGL 和 B2M 的定量结果证实了负责蛋白的存在，即使免疫组化结果不具有决定性。大量的 Apo A1、Apo A4 和 Apo E 与负责蛋白以相同的量沉积。MS-QBIC 对淀粉样蛋白的定量分析对于系统性淀粉样变性的分类和研究是可行且有用的。
