Institute of Organic Chemistry, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
Department of Pathology, Oral Pathology Unit, Medical School and Clinical Center, Pécs University, Hungary.
Anal Cell Pathol (Amst). 2019 Aug 19;2019:6165140. doi: 10.1155/2019/6165140. eCollection 2019.
The aim of this study was to analyse the composition of amyloid mass and the plasmacytic infiltrate of localized amyloidosis of the upper aerodigestive tract.
Biopsy materials were studied by light microscopy, immunohistochemistry (IHC), and mRNA in situ hybridization (mRNA-ISH). The amyloid mass was also analysed with high-performance liquid chromatography mass spectrometry- (HPLC-MS-) based proteomics.
Nodular and diffuse forms of amyloid deposition were detected. IHC analysis revealed -light chain (LC) in two cases, -LC in one case. The remaining two were positive with both. Proteins, well known from other amyloidoses like amyloid A (AA), prealbumin/transthyretin (PA), apolipoprotein A-I (ApoAI), and amyloid P component (APC), and also keratin were found with variable intensities in the cases. HPLC-MS revealed dozens of proteins with both LCs in all the lesions but sometimes with surprisingly small intensities. mRNA-ISH analysis revealed identical and dominance and only one normal / cell ratio.
Cellular infiltrate and protein components in the amyloid showed congruent results in all but one case. The only exception with normal cell ratio and -dominant amyloid could be originated from the different protein-secreting activity of plasma cell clones. HPLC-MS analysis explored both LCs in all the amyloid in variable amount, but other proteins with much higher intensities like keratins, apolipoprotein A-IV (ApoAIV), were also detected. Proteins like AA, PA, ApoAI, and APC, previously known about amyloid-forming capability, also appeared. This indicates that localized amyloid in the upper aerodigestive tract is not a homogenous immunoglobulin mass but a mixture of proteins. The sometimes very low light chain intensities might also suggest that not all the localized amyloidosis cases of the upper aerodigestive tract are of convincingly AL type, and the analysis of the cellular infiltrate might indicate that not all are monoclonal.
本研究旨在分析上呼吸道局部淀粉样变中淀粉样物质的组成和浆细胞浸润。
通过光镜、免疫组化(IHC)和 mRNA 原位杂交(mRNA-ISH)研究活检材料。还通过基于高效液相色谱质谱(HPLC-MS)的蛋白质组学分析淀粉样物质。
检测到结节状和弥漫状淀粉样沉积。免疫组化分析显示,在两种情况下为轻链(LC),在一种情况下为 LC。其余两种均为阳性。在所有病例中,都发现了来自其他淀粉样变的蛋白质,如淀粉样蛋白 A(AA)、前白蛋白/转甲状腺素蛋白(PA)、载脂蛋白 A-I(ApoAI)和淀粉样蛋白 P 成分(APC),以及角蛋白,其强度不同。HPLC-MS 显示所有病变中都存在数十种带有两种 LC 的蛋白质,但有时强度很小。mRNA-ISH 分析显示,在所有情况下均存在相同的 和 优势,且正常/细胞比值仅为 1。
除了一个病例外,淀粉样物质的细胞浸润和蛋白质成分在所有病例中均一致。唯一的例外是正常细胞比值和 -优势淀粉样物质,可能源自浆细胞克隆的不同蛋白质分泌活性。HPLC-MS 分析以不同的量探索了所有淀粉样物质中的两种 LC,但也检测到了角蛋白、载脂蛋白 A-IV(ApoAIV)等其他蛋白,其强度要高得多。以前已知具有形成淀粉样能力的 AA、PA、ApoAI 和 APC 等蛋白质也出现了。这表明上呼吸道局部淀粉样变不是同质的免疫球蛋白物质,而是蛋白质混合物。有时 LC 强度非常低也可能表明并非所有上呼吸道局部淀粉样变病例都是可确信的 AL 型,细胞浸润分析可能表明并非所有病例都是单克隆的。