高效薄层层析-病毒覆盖结合法比较细胞表面神经节苷脂与不同动物源性新城疫病毒结合特性

【目的】神经节苷脂是新城疫病毒入侵宿主细胞的受体,但不同动物源性的新城疫病毒利用受体的特性是否存在差异尚不明确。以鹅源新城疫病毒NA-1株和鸡源新城疫病毒F48E9株为研究对象,比较两株病毒受体结合特性的差异。【方法】提取鸡胚和鹅胚成纤维细胞新城疫病毒受体成分——神经节苷脂,用高效薄层层析法比较两种细胞所含神经节苷脂的类型和含量;通过高效薄层层析-病毒覆盖结合法比较两种病毒与不同细胞神经节苷脂的结合特性,最后通过病毒红细胞吸附抑制试验进一步验证神经节苷脂与病毒之间的相互作用关系。【结果】鸡胚和鹅胚成纤维细胞所含的神经节苷脂成分存在明显差异;高效薄层层析-病毒覆盖结合实验结果显示NA-1和F48E9与神经节苷脂的结合模式不同,NA-1主要与神经节苷脂GD1a结合,即包含双SAα2,3Gal末端的神经节苷脂,而F48E9可与更多类型的神经节苷脂结合,从单唾液酸到三唾液酸形式的神经节苷脂如GM1、GD1a、GD1b、GT1b等。【结论】鹅源新城疫病毒NA-1株和鸡源新城疫病毒F48E9株在入侵靶细胞时优先选择利用的受体不同。

Comparison of the binding characteristics of gangliosides with Newcastle Disease viruses of different animal species by High performance thin layer chromatography-virus overlay protein blot assay

Abstract: Objective] Gangliosides on the cells surface can act as the Newcastle disease virus receptors, but the differences of receptor specificities between Newcastle disease virus strains isolated from different avian species have not been determined. Accordingly, we attempted to determine the difference of binding specificity to gangliosides between two Newcastle disease virus NA-1 (goose-origin) and F48E9 (chicken-origin). Methods] Gangliosides were extracted from chicken embryo fibroblast cells (CEF) and goose embryo fibroblast cells (GEF). The extracted gangliosides were characterized and quantified by high performance thin-layer chromatography. We performed virus overlay assays and hemadsorption inhibition assays to assess the binding specificity of NA-1 and F48E9 to different gangliosides. Results] The results showed that ganglioside compositions of the CEF and GEF were different. In the thin-layer chromatographic binding assay, we analyzed the binding of the virus to different gangliosides, detecting that NA-1 mainly bound to GD1a, F48E9 mainly bound to GM1, GD1a, GD1b. Conclusion] In conclusion, our results suggest that two viruses used different receptors for entry into different target cells.