Production and characterisation of a monoclonal antibody against the thymocytes of the sea bassDicentrarchus labrax(L.) (Teleostea, Percicthydae)

Scapigliati, Giuseppe; Mazzini, Massimo; Mastrolia, Lucia; Romano, Nicla; Abelli, Luigi

doi:10.1006/fsim.1995.0039

Cited by 79 publications

(96 citation statements)

References 27 publications

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“…In immunoblotting of reduced membrane proteins, E6 MAb recognized a polypeptide of 166.2 kDa. This finding is in accordance with earlier reports that MAbs against thymocytes recognize high molecular weight polypeptides in teleosts (Scapigliati et al 1995;Rombout et al 1997;Beelen et al 2004;Chaudhary et al 2012). However, thymocyte-specific molecules of such high molecular weight have not been identified in mammalian thymocytes.…”

Section: Discussionsupporting

confidence: 94%

“…Flow cytometry revealed that 89.7 % of the gated cells presumed to be lymphocytes were found to be positive by E6 MAb. In earlier studies, percentage of cells reported to be positive by MAbs against thymocytes was 74.9 % in sea bass (Scapigliati et al 1995), 76.1 % in yellowtail (Nishimura et al 1995), 73 % in channel catfish (Passer et al 1996); 87.5 % in hybrid surubim (Beelen et al 2004) and 77.7 % in catla (Chaudhary et al 2012). However, Rombout et al (1997) reported that WCL9 MAb reacted with only 30-50 % of thymocytes and this MAb probably detected only a subpopulation of thymic lymphocytes.…”

Section: Discussionmentioning

confidence: 93%

“…There are only a few MAbs that have been described as being specific to fish thymocytes; DLT 15 for the sea bass Dicentrarchus labrax (Scapigliati et al 1995), CfT1 for channel catfish Ictalurus punctatus (Passer et al 1996), WCL9 for carp Cyprinus carpio (Rombout et al 1997), PST33 for surubim catfish Pseudoplatystoma corruscans (Beelen et al 2004) and B8 MAb against thymocytes of Indian major carp, Catla catla (Chaudhary et al 2012). In most of earlier studies, the immunizing antigens were of thymic origin such as unmodified thymocytes (Secombes et al 1983;Yamaguchi et al 1996;Nishimura et al 1997), paraformaldehyde-fixed thymocytes (Scapigliati et al 1995), thymocytes plus Ignegative cells from PBL (Passer et al 1996) or thymocyte plasma membrane (Rombout et al 1997), plasma membrane preparations (Rombout et al 1998), and paraformaldehyde-fixed nylon wool enriched thymus MNCs (Chaudhary et al 2012).…”

Section: Discussionmentioning

confidence: 99%

“…In most of earlier studies, the immunizing antigens were of thymic origin such as unmodified thymocytes (Secombes et al 1983;Yamaguchi et al 1996;Nishimura et al 1997), paraformaldehyde-fixed thymocytes (Scapigliati et al 1995), thymocytes plus Ignegative cells from PBL (Passer et al 1996) or thymocyte plasma membrane (Rombout et al 1997), plasma membrane preparations (Rombout et al 1998), and paraformaldehyde-fixed nylon wool enriched thymus MNCs (Chaudhary et al 2012). Some of these MAbs cross-reacted with other leukocyte populations (Secombes et al 1983;Yamaguchi et al 1996;Nishimura et al 1997) and, consequently, cannot be considered specific markers for T-cells.…”

Section: Discussionmentioning

confidence: 99%

“…Monoclonal antibodies have been raised against T lymphocytes of sea bass (Scapigliati et al 1995), yellowtail (Nishimura et al 1995), channel catfish (Passer et al 1996), common carp (Secombes et al 1983;Yamaguchi et al 1996;Rombout et al 1997) hybrid surubim catfish (Beelen et al 2004) and Catla catla (Chaudhary et al 2012). In most of the cases, thymocytes either live or fixed or membrane lysates prepared from thymocytes have been used as antigen to immunize the mice and produce MAbs.…”

Section: Introductionmentioning

confidence: 99%

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Development and characterization of a monoclonal antibody against the putative T cells of Labeo rohita

et al. 2015

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In this study, we have described the development and characterization of monoclonal antibodies (MAbs) directed against thymocytes of rohu, Labeo rohita. MAbs were obtained by immunizing BALB/c mice with freshly isolated and nylon wool column enriched mononuclear cells of thymus. Positive clones against thymocytes were screened by cellular ELISA. The hybridoma showing strong reactivity with nylon wool enriched mononuclear cells, and non-reactivity with a rohu thymus macrophage cell line and rohu serum was selected and subjected to single cell cloning by limiting dilution. The MAbs secreted by a positive clone were designated as E6 MAb. Western blotting of reduced protein from enriched thymocytes showed that E6 reacted with a 166.2 kDa polypeptide and belongs to the IgG1 subclass. Flow cytometric analysis of gated lymphocytes, revealed that the percentage of E6 positive (E6?) cells in thymus (n = 5, 720.4 ± 79.70 g) was 89.7 %. Similarly, the percentage of E6? cells in kidney, spleen and blood (n = 5) was 6.71, 1.71 and 1.88 %, respectively. In indirect immunoperoxidase test, E6? cells appeared to be lymphoid cells with a high nucleus to cytoplasmic ratio and were densely packed in the central region of thymus whereas, a few cells were found to be positive in kidney and spleen sections. E6 MAb also reacted with a small population of lymphocytes in blood smear. This MAb appears to be a suitable marker for T lymphocytes and can be a valuable tool in studying immune response and ontogeny of L. rohita immune system.

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Section: Discussionsupporting

confidence: 94%

Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development and characterization of a monoclonal antibody against the putative T cells of Labeo rohita

et al. 2015

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Blood cells of sea bass (Dicentrarchus labrax l.). Flow cytometric and microscopic studies

2000

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Studies of fish blood cells made to date presented numerous problems derived from both the nomenclature and the techniques used. A combination of quantitative and morphological methods is needed if the classification of fish blood cells is to advance from it present provisional state. The aim of the present paper was first to isolate sea bass blood cell populations by flow cytometry and second to characterize then microscopically. Blood cell populations from sea bass (Dicentrarchus labrax L.) were isolated according to their FSC (size) and SSC (granularity) properties by flow cytometry. The isolated populations were then processed for light and transmission and scanning electron microscopic characterization. Sea bass blood leukocytes isolated by flow cytometry consisted of two main cell subpopulations. Subsequent microscopic study of these cells revealed that the first subpopu-lation was composed of small cells (3-5 µm) of low granularity and consisted of thrombocytes and lymphocytes whereas, the second subpopulation was formed of 6-9 µm sized cells of high granularity consisting of granulocytes and monocyte/macrophages. The combined use of flow cytometry and electron microscopy makes it possible to characterize the different cell types present in sea bass peripheral blood with a high degree of certainty. Although sea bass basically follows the common vertebrate hematological pattern, significant modifications such as the presence of circulating immature erythrocytes, plasma cells and monocyte/macrophages and different forms of thrombocytes can be established with respect to this pattern. Anat

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Immunodetection of Lymphocyte Subpopulations Involved in Allograft Rejection in a Teleost,Dicentrarchus labrax(L.)

Abelli

Baldassini

Mastrolia

et al. 1999

Cellular Immunology

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Production and characterisation of a monoclonal antibody against the thymocytes of the sea bassDicentrarchus labrax(L.) (Teleostea, Percicthydae)

Cited by 79 publications

References 27 publications

Development and characterization of a monoclonal antibody against the putative T cells of Labeo rohita

Development and characterization of a monoclonal antibody against the putative T cells of Labeo rohita

Blood cells of sea bass (Dicentrarchus labrax l.). Flow cytometric and microscopic studies

Immunodetection of Lymphocyte Subpopulations Involved in Allograft Rejection in a Teleost,Dicentrarchus labrax(L.)

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