用流式细胞仪测定扇贝血细胞吞噬活性

Application of flow Cytometry in the Measurement of Phagocytic Activity in Scallop Haemocytes

采用流式细胞仪技术对栉孔扇贝(Chlamysfarreri)和海湾扇贝(Argopectenirradians)血细胞吞噬活性进行了测定。通过对缓冲体系的筛选和比较,建立了扇贝血细胞吞噬活性的流式细胞仪检测方法,并应用该技术对两种扇贝血细胞的吞噬活性进行了比较研究。结果发现在TBS(0.05mol.L-1Tris-HCl,pH7.4;2%glucose;2%NaCl;20mmol.L-1EDTA)和PBS体系中,扇贝血细胞的吞噬活性几乎完全受到抑制,在CMPBS(0.1mol.L-1PBS,pH7.4;2%NaCl;2%glucose;1.0mmol.L-1CaCl2;0.5mmol.L-1MgCl2)体系中细胞吞噬活性略有升高,而在海水缓冲液中细胞吞噬活性最高,其中海湾扇贝和栉孔扇贝血细胞的吞噬率分别达到了26.73%和19.89%,且海湾扇贝血细胞的吞噬率显著高于栉孔扇贝(P<0.05)。研究结果为进一步探讨扇贝血细胞吞噬功能提供了方法依据。

Phagocytosis is considered as the predominant mechanism of internal defense in bivalve species. Flow-cytometric method for phagocytic activity of scallop haemocytes was established by screening four different buffers based on their inhibitive effects. The phagocytic activity was almost suppressed in TBS and PBS buffers, and slightly activated in CMPBS buffer with the introduction of Mg^2＋ and Ca^2＋ , whereas the capacity in FSW increased significantly compared to the other three buffers, which indicated that FSW was the preferable buffer for phagoeytosis assay. Moreover, FCM was employed to compare the phagocytic capacity of haemocytes between two kinds of scallops, Chlamys farreri and Argopecten irradians. The phagocytosis rate of Argopecten irradians （26.73%） was significantly higher than that of Chlamys farreri （ 19.89% ）. The results indicated this method was practicable and feasible to elucidate the phagocytic capacity and useful to study the immune defense mechanisms of bivalve.