Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, 7700, South Africa.
Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, 7700, South Africa.
Fish Shellfish Immunol. 2020 Apr;99:130-143. doi: 10.1016/j.fsi.2020.02.007. Epub 2020 Feb 8.
The South African abalone Haliotis midae is a commercially important species farmed at high densities in land-based aquaculture systems. Disease outbreaks have had a severe financial impact on the abalone industry yet the molecular mechanisms underlying the immune response of H. midae remain obscure. In this study, a comparative shotgun proteomics approach using iTRAQ coupled with LC-MS/MS was employed to investigate H. midae proteome changes in response to Vibrio anguillarum challenge. A total of 118 non-redundant, unique haemocyte proteins were identified and quantified, with 16 proteins significantly regulated. Hierarchical clustering and pathway analysis uncovered a coordinated response dominated by calcium and cAMP signalling via activation of MAPK cascades. Early up-regulated biological processes involve phagocytosis, nitric oxide production and ATP-synthesis, whilst down-regulated responses were predominantly involved in the regulation of apoptosis. The late up-regulated response involved protein kinase activity and detoxification processes. Expression of selected proteins was validated by Western blot. A putative allograft inflammatory factor-1 protein was further selected to establish its functional molecular role in haemocytes. Confocal imaging revealed that allograft inflammatory factor-1 regulates phagocytosis via a functional interaction with filamentous actin. This is the first time a high-throughput proteomics approach has been used to investigate the immune response of H. midae.
南非鲍 Haliotis midae 是一种商业上重要的物种，在陆基水产养殖系统中高密度养殖。疾病爆发对鲍鱼产业造成了严重的财务影响，但 H. midae 免疫反应的分子机制仍不清楚。在这项研究中，采用 iTRAQ 与 LC-MS/MS 相结合的比较 shotgun 蛋白质组学方法，研究了 H. midae 对鳗弧菌挑战的蛋白质组变化。共鉴定和定量了 118 个非冗余、独特的血细胞蛋白，其中 16 个蛋白显著调节。层次聚类和途径分析揭示了一个以钙和 cAMP 信号为主导的协调反应，通过 MAPK 级联的激活。早期上调的生物学过程涉及吞噬作用、一氧化氮产生和 ATP 合成，而下调的反应主要涉及细胞凋亡的调节。晚期上调的反应涉及蛋白激酶活性和解毒过程。通过 Western blot 验证了选定蛋白的表达。进一步选择了一种假定的同种异体炎症因子-1 蛋白，以确定其在血细胞中的功能分子作用。共聚焦成像显示同种异体炎症因子-1 通过与丝状肌动蛋白的功能相互作用调节吞噬作用。这是首次使用高通量蛋白质组学方法研究 H. midae 的免疫反应。
