Application of monoclonal antibody against granulocytes of scallop Chlamys farreri on granulocytes occurrence at different developmental stages and antigenic cross-reactivity of granulocytes in five other bivalve species.
A monoclonal antibody (MAb) 6H7 raised specifically against granulocytes of scallop (Chlamys farreri) was employed to observe granulocyte occurrence successively in blastulae, gastrulae, trochophore larvae, D-shape larvae, umbo-veliger larvae and creeping larvae of C. farreri by immunohistochemistry assay contrasted with H&E stain using semi-thin sections. Moreover, the reactivity of the MAb with granulocytes of C. farreri, Bay scallop Argopecten irradians, Japanese scallop Patinopecten yessoensis, Blue mussel Mytilus edulis, Pacific oyster Crassostrea gigas and Manila clam Ruditapes philippinarum, was detected by immunofluorescence assay (IFA) with differential interference contrast and fluorescent microscopy and flow cytometric immunofluorescence assay (FCIFA). The results showed that positive signals were first observed at D-shape larval stage, about 28 h post fertilization, after that, umbo-veliger larvae exhibited the positive cells with a diameter of 3-5 μm distributed in velum, digestive gland and esophagus. Then in creeping larvae, the number of positive cells increased with average diameter of 5-7 μm, and widely distributed in foot, digestive gland, gills and adductor muscles. No positive signal was found in blastulae, gastrulae and trochophore larvae. The results of IFA and FCIFA showed MAb 6H7 reacted to granulocytes of C. farreri, A. irradians, P. yessoensis and C. gigas, and the positive percentage reactivity were 53 ± 2.5%, 15 ± 2.5%, 12 ± 2.1% and 19 ± 2.1%, respectively, however, no cross-reaction was detected in hemocytes of R. philippinarum and M. edulis.
Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, Qingdao 266003, PR China.
Jing
J
Xing
Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, Qingdao 266003, PR China.
Xiaoqian
X
Tang
Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, Qingdao 266003, PR China.
Yongqing
Y
Ni
Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, Qingdao 266003, PR China. Electronic address: wbzhan@ouc.edu.cn.
Wenbin
W
Zhan
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D005434:Flow Cytometry; D006098:Granulocytes / Q000276:immunology* / Q000648:ultrastructure; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D008857:Microscopy, Interference; D049897:Pectinidae / Q000276:immunology* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal
Chlamys farreri; Cross-reactivity; Granulocyte; Monoclonal antibody; Occurrence