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Characterization of monoclonal antibodies to haemocyte types of the shrimp, Fenneropenaeus chinensis

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[Five monoclonal antibodies (MAbs) (1A7, 1C6, 1E4, 1G8, 2C3) against haemocytes of Fenneropenaeus chinensis (Osbeck, 1765) were produced by immunizing Balb/c mice, followed by indirect immunofluorescence assay tests (IIFAT). These showed specificity for more than one haemocyte type and for various haemocyte components of F. chinensis. Using Wright's staining and IIFAT with haemocyte monolayers prepared by sucrose gradient centrifugation, MAb 1A7, 1C6 reacted with membranes of hyalinocytes and the cytoplasm of semi-granulocytes, respectively; MAb 1E4 reacted with membranes and the cytoplasm of hyalinocytes, and with membranes of semi-granulocytes; MAb 1G8 reacted with haemocyte granules; and MAb 2C3 reacted with surface membranes of all haemocyte types. Western-blotting of haemocytes and haemocyte membrane analysis demonstrated that MAb 1G8 recognized an antigen of 96 kDa in haemocyte; that MAb 1E4 reacted with antigens of 88 kDa and 79 kDa in the haemocyte membrane; and that MAb 2C3 reacted specifically to a single membrane protein band with an approximate molecular weight of 153 kDa; however, there was no protein band detected by either MAbs 1A7, or 1C6. Cinq anticorps monoclonaux (MAbs) (1A7, 1C6, 1E4, 1G8, 2C3) contre les hémocytes de Fenneropeneaus chinensis (Osbeck, 1765) ont été produits par immunization de souris Balb/c, et contrôlés par un test en immunofluorescence indirecte (IIFAT). Ils ont présenté une spécificité pour plusieurs types d'hémocytes et pour divers composants d'hémocytes de F. chinensis. En utilisant la coloration de Wright et IIFAT avec une monocouche d'hémocytes preparée par centrifugation à gradient de concentration en glucose, MAb 1A7, 1C6 ont réagit respectivement avec les membranes des hémocytes hyalins et le cytoplasme des semi-granuleux; MAb 1E4 a réagit avec les membranes et le cytoplasme des hémocytes granuleux; et MAb 2C3 a réagit avec les membranes de tous les types d'hémocytes. Les résultats du Western-blot des hémocytes et de l'analyse de leur membrane ont démontré que MAb 1G8 reconnaît un antigène de 96 kDa dans les hémocytes; que MAb 1E4 a réagit avec des antigènes de 88 kDa et 79 kDa au niveau de la membrane des hémocytes; et que MAb 2C3 a réagit spécifiquement à une bande protéique, d'une poids moléculaire approximatif de 153 kDa; cependant aucune bande protéique n'a été détectée par MABs 1A7 ou 1C6., Five monoclonal antibodies (MAbs) (1A7, 1C6, 1E4, 1G8, 2C3) against haemocytes of Fenneropenaeus chinensis (Osbeck, 1765) were produced by immunizing Balb/c mice, followed by indirect immunofluorescence assay tests (IIFAT). These showed specificity for more than one haemocyte type and for various haemocyte components of F. chinensis. Using Wright's staining and IIFAT with haemocyte monolayers prepared by sucrose gradient centrifugation, MAb 1A7, 1C6 reacted with membranes of hyalinocytes and the cytoplasm of semi-granulocytes, respectively; MAb 1E4 reacted with membranes and the cytoplasm of hyalinocytes, and with membranes of semi-granulocytes; MAb 1G8 reacted with haemocyte granules; and MAb 2C3 reacted with surface membranes of all haemocyte types. Western-blotting of haemocytes and haemocyte membrane analysis demonstrated that MAb 1G8 recognized an antigen of 96 kDa in haemocyte; that MAb 1E4 reacted with antigens of 88 kDa and 79 kDa in the haemocyte membrane; and that MAb 2C3 reacted specifically to a single membrane protein band with an approximate molecular weight of 153 kDa; however, there was no protein band detected by either MAbs 1A7, or 1C6. Cinq anticorps monoclonaux (MAbs) (1A7, 1C6, 1E4, 1G8, 2C3) contre les hémocytes de Fenneropeneaus chinensis (Osbeck, 1765) ont été produits par immunization de souris Balb/c, et contrôlés par un test en immunofluorescence indirecte (IIFAT). Ils ont présenté une spécificité pour plusieurs types d'hémocytes et pour divers composants d'hémocytes de F. chinensis. En utilisant la coloration de Wright et IIFAT avec une monocouche d'hémocytes preparée par centrifugation à gradient de concentration en glucose, MAb 1A7, 1C6 ont réagit respectivement avec les membranes des hémocytes hyalins et le cytoplasme des semi-granuleux; MAb 1E4 a réagit avec les membranes et le cytoplasme des hémocytes granuleux; et MAb 2C3 a réagit avec les membranes de tous les types d'hémocytes. Les résultats du Western-blot des hémocytes et de l'analyse de leur membrane ont démontré que MAb 1G8 reconnaît un antigène de 96 kDa dans les hémocytes; que MAb 1E4 a réagit avec des antigènes de 88 kDa et 79 kDa au niveau de la membrane des hémocytes; et que MAb 2C3 a réagit spécifiquement à une bande protéique, d'une poids moléculaire approximatif de 153 kDa; cependant aucune bande protéique n'a été détectée par MABs 1A7 ou 1C6.]

Affiliations: 1: Laboratory of Pathology and Immunology of Aquatic Animals, LMMEC, Ocean University of China, Qingdao 266003, China

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/content/journals/10.1163/156854008x354993
2008-09-01
2016-12-07

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