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Development of a monoclonal antibody specific to granulocytes and its application for variation of granulocytes in scallop Chlamys farreri after acute viral necrobiotic virus (AVNV) infection

Author:
Lin, Tingting, Xing, Jing, Sheng, Xiuzhen, Tang, Xiaoqian, Zhan, Wenbin
Source:
Fish & shellfish immunology 2011 v.30 no.6 pp. 1348-1353
ISSN:
1050-4648
Subject:
Chlamys, Western blotting, cytoplasmic granules, enzyme-linked immunosorbent assay, epitopes, flow cytometry, fluorescent antibody technique, granulocytes, hemocytes, mice, monoclonal antibodies, scallops, viruses
Abstract:
A monoclonal antibody (MAb 6H7) specific to granulocytes of scallop Chlamys farreri was produced by immunising mice with separated granulocytes as an antigen. Characterised using a flow cytometric immunofluorescence assay, MAb 6H7 reacted to granulocytes by 87.1% of total positive haemocytes. At the ultrastructural level, MAb 6H7 demonstrated epitope in cytoplasmic granules of granulocytes. Western blotting analysis indicated that a peptide of 155 kDa was recognised by MAb 6H7. It was therefore used to investigate granulocyte variation in C. farreri after acute viral necrobiotic virus (AVNV) infection using an enzyme-linked immunosorbent assay. The result illustrated that granulocytes varied greatly by AVNV infection, and their amount significantly increased on day 1 post-injection, then decreased on days 2, 3 and 4, thereafter, rebounded and approached to a second peak on day 6, finally went down gradually to the control level on day 8.
Agid:
828750