Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan.
Int J Mol Sci. 2020 May 21;21(10):3632. doi: 10.3390/ijms21103632.
Glycosphingolipids containing very-long-chain fatty acids (VLCFAs) regulate several immune responses, such as cytokine production, immune signaling, and antibody induction. We previously reported that stimulation with an inflammatory mediator, TNF-α, promotes the expression of glycosphingolipids in vascular endothelial cells. The major component is globotetraosylceramide containing VLCFAs (Gb4Cer-VLCFAs), but its role in inflammatory responses has not been fully investigated. In this study, the antibody-inducing properties of Gb4Cer-VLCFAs were analyzed using serum and hybridoma cells generated from Gb4Cer-VLCFA-immunized mice. The reactivity of serum antibodies against Gb4Cer indicated that immunization with Gb4Cer-VLCFAs immediately induced the production of anti-Gb4Cer antibodies. Over 81% of hybridomas generated from the splenocytes of an immunized mouse produced anti-Gb4Cer antibodies, a subset of which recognized an epitope shared by Gb4Cer and its precursor globotriaosylceramide (Gb3Cer). Further biochemical analyses of established monoclonal antibodies revealed that these antibodies included IgM and IgG3, which specifically react with Gb4Cer and Gb3Cer. These results indicate that immunization with Gb4Cer-VLCFAs can efficiently induce the production of anti-Gb4Cer and -Gb3Cer antibodies by B cells.
含有超长链脂肪酸 (VLCFA) 的糖鞘脂调节多种免疫反应，如细胞因子产生、免疫信号和抗体诱导。我们之前报道过，炎症介质 TNF-α 的刺激可促进血管内皮细胞中糖鞘脂的表达。主要成分是含有 VLCFA 的四糖苷神经酰胺 (Gb4Cer-VLCFAs)，但其在炎症反应中的作用尚未得到充分研究。在这项研究中，使用来自 Gb4Cer-VLCFA 免疫小鼠的血清和杂交瘤细胞分析了 Gb4Cer-VLCFAs 的抗体诱导特性。血清抗体对 Gb4Cer 的反应性表明，Gb4Cer-VLCFAs 的免疫接种立即诱导了抗 Gb4Cer 抗体的产生。从免疫小鼠的脾细胞中产生的超过 81%的杂交瘤产生了抗 Gb4Cer 抗体，其中一部分识别 Gb4Cer 和其前体神经节苷脂 (Gb3Cer) 共享的表位。对已建立的单克隆抗体的进一步生化分析表明，这些抗体包括 IgM 和 IgG3，它们特异性地与 Gb4Cer 和 Gb3Cer 反应。这些结果表明，Gb4Cer-VLCFAs 的免疫接种可以有效地诱导 B 细胞产生抗 Gb4Cer 和 -Gb3Cer 抗体。
