Classification of Hemocytes from Four Crustaceans and Cross-Reactivity of Their Antisera (original) (raw)
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Journal of Invertebrate Pathology, 2012
Monoclonal antibodies (MAbs) to hemocytes of mud crab, Scylla serrata, were produced by immunizing mice with formalin-fixed hemocytes. Of the six MAbs produced, two (MAb 1D11 and MAb 1A2) reacted specifically with hemocyte proteins in western blot. MAb 1A2 showed strong immunofluorescent reaction with granular hemocytes and a weak reaction with semigranular cells. However, hyaline cells did not show any reaction. The MAbs also showed strong cross-reactivity with the hemocytes of different crab species but not with other crustaceans. The MAbs produced can be used as a marker for granular hemocytes of crabs.
Crustacean haemocytes and haematopoiesis
Aquaculture, 2000
Crustacean haemocytes play important roles in the host immune response including recognition, phagocytosis, melanization, cytotoxicity and cell-cell communication. Classification of the haemocyte types in decapod crustaceans is based mainly on the presence of cytoplasmic granules into hyaline cells, semigranular cells, and granular cells. Each cell type is active in defence reactions, for example; in crayfish, the hyaline cells are chiefly involved in phagocytosis, the semigranular cells are the cells active in encapsulation, while the granular cells participate in Ž . storage and release of the prophenoloxidase proPO system and cytotoxicity. The haematopoietic tissue has been described in several crustacean decapod species and shown to be the haemocyteproducing organ. Tentative stem cells have been shown to be present in this tissue. Using in situ hybridization, we demonstrated that proPO is not present in the haematopoietic tissue of crayfish which suggests that protein expression is different between circulating haemocytes and the cells in the haematopoietic tissue. q
First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea, Decapoda)
European Journal of Histochemistry, 2010
For the first time, a morphological study of haemocytes from the crab Carcinus aestuarii was carried out by means of light microscopy and differing cytochemical assays. Analysis of haemocyte size frequency distribution (performed by means of a Coulter Counter) revealed the presence of two distinct haemocyte fractions in C. aestuarii haemolymph, depending on cell size. The first fraction was of about 3-5 µm in diameter and 30-50 fL in volume, the second was of about 6-12 µm in diameter and over 200 fL in volume. Mean cell diameter and volume were 8.20±1.7 µm and 272.30±143.5 fL, respectively. Haemocytes observed under light microscope were distinguished in three cell types: granulocytes (28%; 11.94±1.43 µm in diameter) with evident cytoplasmic granules, semigranulocytes (27%; 12.38±1.76 µm in diameter) with less granules than granulocytes, and hyalinocytes (44%; 7.88±1.6 µm in diameter) without granules. In addition, a peculiar cell type was occasionally found (about 1%): it was 25-30 µm in diameter and had a great vacuole and a peripheral cytoplasm with granules. Granulocyte and semigranulocyte granules stained in vivo with Neutral Red, indicating that they were lysosomes. Giemsa's dye confirmed that granulocytes and semigranulocytes were larger than hyalinocytes. Pappenheim's panoptical staining and Ehrlich's triacid mixture allowed to distinguish granule-containing cells (including semigranulocytes) in acidophils (64%), basophils (35%) and neutrophils (1%). Hyalinocytes showed always a basophilic cytoplasm. Haemocytes were positive to the PAS reaction for carbohydrates, even if cytoplasm carbohydrate distribution varied among cell types. Lastly, lipids were found on cell membrane and in cytoplasm of all haemocyte types in the form of black spots produced after Sudan Black B staining. The morphological characterisation of C. aestuarii haemocytes by light microscopy was necessary before performing both ultrastructural and functional studies of circulating cells.
Fish & Shellfish Immunology, 1995
In order to ensure shrimp aquaculture production, immunology has become a priority in terms of disease control and prevention. Research must be devoted to the identification of anti-infectious effectors at the cellular and molecular levels. In this review, recent results obtained in the shrimp Penaeus japonicus are presented and discussed with reference to the Crustacea and other arthropods. Various cellular and plasma haemolymph components were characterised both antigenically and functionally. Haemocyte subpopulations, separated by isopycnic centrifugation on Percoll were discriminated using two monoclonal antibodies, 40E2 and 40El0. The granular cells were recognised by the antibody 40E2 specific for a plasmatic protein. By comparison, the hyaline and semi-granular cells were recognised by the antibody 40El0, which immunoprecipitated a plasmatic protein identified as an agglutinin. Moreover, an a2-macroglobulin-like protein present both in plasma and in haemocytes was antigenically characterized in P. japonicus as well as a plasma clotting factor. Functional studies have been undertaken by considering antimicrobial effectors and haemocyte-mediated oxidative defence mechanisms studied by chemiluminescence. Prospects on findings in shrimp immunology are discussed with reference to their application to zoosanitary prophylaxis and disease prevention. ~$)
Fish & Shellfish Immunology, 1995
In order to ensure shrimp aquaculture production, immunology has become a priority in terms of disease control and prevention. Research must be devoted to the identification of anti-infectious effectors at the cellular and molecular levels. In this review, recent results obtained in the shrimp Penaeus japonicus are presented and discussed with reference to the Crustacea and other arthropods. Various cellular and plasma haemolymph components were characterised both antigenically and functionally. Haemocyte subpopulations, separated by isopycnic centrifugation on Percoll were discriminated using two monoclonal antibodies, 40E2 and 40E10. The granular cells were recognised by the antibody 40E2 specific for a plasmatic protein. By comparison, the hyaline and semi-granular cells were recognised by the antibody 40E10, which immunoprecipitated a plasmatic protein identified as an agglutinin. Moreover, an α2-macroglobulin-like protein present both in plasma and in haemocytes was antigenically characterized in P. japonicus as well as a plasma clotting factor. Functional studies have been undertaken by considering antimicrobial effectors and haemocyte-mediated oxidative defence mechanisms studied by chemiluminescence. Prospects on findings in shrimp immunology are discussed with reference to their application to zoosanitary prophylaxis and disease prevention.
First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea
2013
For the first time, a morphological study of haemocytes from the crab Carcinus aestuarii was carried out by means of light microscopy and differing cytochemical assays. Analysis of haemocyte size frequency distribution (performed by means of a Coulter Counter) revealed the presence of two distinct haemocyte fractions in C. aestuarii haemolymph, depending on cell size. The first fraction was of about 3-5 µm in diameter and 30-50 fL in volume, the second was of about 6-12 µm in diameter and over 200 fL in volume. Mean cell diameter and volume were 8.20±1.7 µm and 272.30±143.5 fL, respectively. Haemocytes
Characterisation of shrimp haemocytes and plasma components by monoclonal antibodies
Journal of Cell Science, 1995
Various haemolymph components of the shrimp Penaeus japonicus were identified and characterised by monoclonal antibodies. Three groups of monoclonal antibodies were raised. Their reactivity to haemocyte types and/or secreted molecules was determined by immunofluorescence and the molecular masses of the antigens were analysed by western-blotting. A 170 kDa protein, in reducing conditions, was recognized by four panhaemocytic monoclonal antibodies from group 1. This protein was present both in the plasma and in the haemocytes from which it appears to be secreted. The shrimp haemocytes were separated by isopycnic centrifugation on a Percoll gradient and the different subpopulations were antigenically analysed using the two monoclonal antibodies, 40E2-2A and 40E10-2B, from group 2. The granular cells were labelled by 40E2-2A which was specific for a protein of 142 kDa also present in plasma. By comparison, the 40E10-2B monoclonal antibody was assumed to be the marker for small hyaline a...
Cellular Immunity in Crustaceans and the proPO System
The molecular mechanism of cellular immunity in arthropods has until recently been largely unknown, but with the d~.velopment of a technique to isolate and handle the different blood cell types of crustaceans and with the purification of several proteins associated with the so-called proPO system of freshwater crayfish the processes have now begun to be better understood. In this article Mats ff ohansson and Kenneth Srderh~ill discuss the function of the proPO system in cellular immune reactions in crustaceans and in particular the role of a protein with a molecular mass of 76 kDa, which has been shown to be involved in the communication between the different blood cell types of crayfish.
Pakistan Veterinary Journal, 2013
The Litopenaeus vannamei is a prime aquaculture species and has a worth market value all over the world. In this study, we investigated the genesis and morphology of hematopoietic tissue (HPT) and types of hemocyte based on its morphology and cytochemical feature using histological and histochemical methods. The results revealed that HPT in L. vannamei is not visible histologically in mysis and early two post larvae (P 1 & P 2), and emerges in P 3 which locates in epigastric region. The HPT showed a continuous propagation of cells in successive post larval stages. Four types of cells can be identified in the HPT from adult specimen and mitotic activity is visible in the HPT. The hemocytes have been differentiated into five types and some of these types are co-related with the HPT cells. The cytochemical studies suggested that type I and II cells of hemocytes are PAS positive while scanty presence of prophenoloxidase was observed in type I cells. Similarly the type I cells are Sudan Black B positive and rest of the cells showed weak activity against lipid detective stain. This is the first effort towards genesis of HPT and its relation with circulating hemocytes in L. vannamei.
Comparative biochemistry and physiology. B. Comparative biochemistry, 1993
1. LP1, a high density lipoprotein common to both sexes of the penaeid shrimp Penaeus semisulcatus was isolated as the upper yellow-brown layer resulting from ultracentrifugation of male hemolymph. The hemolymph solution was adjusted to a hydrated density of 1.2 g/ml. A similiar HDL fraction was isolated from the hemolymph of vitellogenic females containing, in addition to the LPI, the female-specific lipoprotein vitellogenin (Vg). 2. Hemocyanin (Hcy), the oxygen carrier of crustaceans was isolated as pellet, resulting from the same ultracentrifugation. Hcy was identified by its blue color, its copper content and its characteristic absorbance spectrum. It is the most abundant hemolymph protein, and is composed of two major subunits of 85 and 95 kD. 3. Polyclonal antibodies were raised in rabbits against Hcy and LPI. The antisera were found to be specific and no crossreaction was observed between them and an anti-Vg/Vt serum prepared during earlier studies. All three antisera proved to be suitable research tools for the identification of these three physiologically important proteins (Vg, LP1, Hcy).