Purification and characterization of silkworm hemocytes by flow cytometry
Purification and characterization of silkworm hemocytes by flow cytometry
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DOI：
10.1016/j.dci.2008.09.005
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发表时间：
2009-01-01
影响因子：
2.9
通讯作者：
Kamimura, Manabu
中科院分区：
文献类型：
--
作者：
Nakahara, Yuichi;Shimura, Sachiko;Kamimura, Manabu
家蚕的血细胞功能已得到充分研究，然而，由于缺乏适当的分离方法，对每个血细胞亚群的详细分析受到阻碍。在这里，我们使用一系列流式细胞术分析，用各种分子探针（例如碘化丙啶、绿色荧光蛋白、单克隆抗体和荧光凝集素）来表征蚕血细胞。其中，使用碘化丙啶的分离是最简单的，并且为血细胞亚群的分离提供了最可靠的结果。然后从这些分选的群体中合成 cDNA，并通过 RT-PCR 检查子集特异性基因表达。粒细胞、浆细胞和类细胞表达不同类别的免疫基因，表明它们在家蚕免疫中具有多种作用。相比之下，球状红细胞对免疫的贡献没有被记录，因为它们未能表达大多数基因。因此，球状红细胞的功能可能与其他三个血细胞亚群的功能不同。 (C) 2008 Elsevier Ltd. 保留所有权利。
Hemocyte functions are well-investigated in the silkworm, Bombyx mori, however, detailed analysis of each hemocyte subset has been hampered by the lack of appropriate separation method. Here we use an array of flow cytometric analyses to characterize silkworm hemocytes with various molecular probes, such as propidium iodide, green fluorescence protein, monoclonal antibodies, and fluorescent lectins. of these, separation using propidium iodide was the simplest and provided most reliable results for the isolation of the hemocyte subsets. cDNAs were then synthesized from these sorted populations and subset-specific gene expression was examined by RT-PCR. Granulocytes, plasmatocytes, and oenocytoids expressed different classes of immune genes, suggesting that they have multiple roles in silkworm immunity. In contrast, a contribution of spherulocytes to immunity was not documented in that they failed to express most of the genes. The functions of spherulocytes are thus likely to be distinct from those of the other three hemocyte subsets. (C) 2008 Elsevier Ltd. All rights reserved.