Department of Bioengineering, University of Texas at Arlington, 500 UTA Blvd., Arlington, TX, 76019, USA.
Sci Rep. 2021 Jan 8;11(1):87. doi: 10.1038/s41598-020-79862-2.
Subtypes of B cell non-Hodgkin's lymphomas, including follicular lymphomas, have shown a unique high oligomannose presentation on their immunoglobulins that will interact with natural receptors of the innate immunity, reportedly causing stimulation and proliferation. From deep sequencing of the variable heavy and light chain sequences of follicular lymphoma involved tissue sections, we identified the consensus variable sequences possessing glycosylation sites at the complementarity determining region. Using this information, we developed a cell line, referred to here as BZ, which displays the consensus variable segments as part of a surface antibody (IgM) and confirmed its presentation of high oligomannose on the heavy chain both in vitro and in vivo. An mCherry expressing variant provided a reporter cell line displaying the high oligomannose surface biomarker while affording clear fluorescent signals for FACS screening as well as for fluorescent in vivo imaging of ectopic xenograft tumors. In developing this reporter cell line that displays the biomarker glycan of follicular lymphoma, we provide a tool that may be used for future screening and validation of receptive moieties for selectively binding high oligomannose for development of targeted diagnostics or therapeutics to such B cell malignancies that display this unique glycan.
B 细胞非霍奇金淋巴瘤的亚型，包括滤泡性淋巴瘤，其免疫球蛋白上表现出独特的高甘露寡糖，据报道，这些高甘露寡糖会与先天免疫的天然受体相互作用，从而导致刺激和增殖。通过对滤泡性淋巴瘤受累组织切片中重链和轻链可变区序列的深度测序，我们确定了具有补体决定区糖基化位点的共识可变序列。利用这些信息，我们开发了一种细胞系，称为 BZ，其表面抗体（IgM）部分展示了共识可变片段，并在体外和体内证实了其重链上高甘露寡糖的表达。一个表达 mCherry 的变体提供了一个报告细胞系，该细胞系显示出高甘露寡糖表面生物标志物，同时为 FACS 筛选以及异位异种移植肿瘤的荧光体内成像提供了清晰的荧光信号。在开发这种显示滤泡性淋巴瘤生物标志物聚糖的报告细胞系时，我们提供了一种工具，可用于未来对选择性结合高甘露寡糖的受体进行筛选和验证，以开发针对显示这种独特聚糖的 B 细胞恶性肿瘤的靶向诊断或治疗方法。
