The Experts below are selected from a list of 300 Experts worldwide ranked by ideXlab platform
Irene Söderhäll - One of the best experts on this subject based on the ideXlab platform.
-
a comparative global proteomic analysis of the Hematopoietic lineages in the crustacean pacifastacus leniusculus
Developmental and Comparative Immunology, 2019Co-Authors: Irene Söderhäll, Kingkamon JunkunloAbstract:In crustaceans as in other arthropods, the circulating hemocytes are vital for protecting the animal against attacking microorganisms. As many hemocytes are destroyed early during an infection, new hemocytes must fast get in place to prevent disperse of a pathogenic microbe, In order to understand the Hematopoietic process in more detail we here report a complete proteomic analysis from purified cell types from the APC of the Hematopoietic Tissue, via the remaining parts of the HPT to the mature semigranular and granular hemocytes. Several possible cell type specific proteins are detected and new putative biomarkers within the crayfish Hematopoietic lineage that can be used to increase the understanding of how the differentiation process is regulated is described.
-
transfection of crayfish Hematopoietic Tissue cells
Developmental and Comparative Immunology, 2018Co-Authors: Hong Shi, Irene Söderhäll, Lingwei Ruan, Kenneth SoderhallAbstract:Transfection is a powerful tool useful for studying gene function. Establishing transfection methods that enable highly efficient DNA uptake has become increasingly important. The crayfish Hematopoietic Tissue (Hpt) cell cultures have been proven to be suitable for studies on immunity and cell differentiation in crustaceans including shrimps, but no efficient gene transfer and expression method is available for these cells. Here we report a novel and highly efficient DNA transfection system based on electroporation. This method depends on a recombinant plasmid with the promoter from white spot syndrome virus immediate-early gene wsv249. This plasmid could be introduced into primary cells and efficiently express foreign genes by electroporation. By optimizing different electroporation parameters, more than 30% transfection efficiency could be achieved with the relative viability of cells around 50%. This is the first report of gene introduction to crayfish Hpt cells and will be useful for the expanding our research on crustacean immunity.
-
Recent advances in crayfish Hematopoietic stem cell culture: a model for studies of hemocyte differentiation and immunity
Cytotechnology, 2013Co-Authors: Irene SöderhällAbstract:Hematopoiesis is the process by which blood cells (hemocytes) mature and subsequently enter the circulation and we have developed a new technique to culture the Hematopoietic progenitor cells in vitro. The reason for the successful culture was the isolation of a plasma protein that turned out to be a novel cytokine, astakine 1 (Ast1) containing a domain present in several vertebrates, so-called prokineticins. Now we have detected several astakines from other invertebrate species. Depending on our discovery of the cytokine Ast1 we have an opportunity to study in detail the differentiation of cells in the Hematopoietic Tissue of a crustacean, a Tissue of evolutionary interest for studies of the connection between the vascular system and the nervous system. We have been able to isolate the entire Hematopoietic Tissue and for the first time detected a link between this Tissue and the brain. We have further localized a proliferation center in the Tissue and characterized its different parts. We have also used this system to isolate a new Hematopoietic factor CHF that is important in the crossroad between apoptosis and hemocyte differentiation. Our technique for culture of crayfish Hematopoietic stem cells provides a simple tool for studying the mechanism of hematopoiesis, but also enables detailed studies of immune defense reactions. Further, the culture system has been used for studies of viral defense and the system is suitable for gene silencing which allows functional characterization of different molecules involved in host defense as well as in hemocyte differentiation.
-
invertebrate hematopoiesis an anterior proliferation center as a link between the Hematopoietic Tissue and the brain
Stem Cells and Development, 2012Co-Authors: Chadanat Noonin, Kenneth Soderhall, Pikul Jiravanichpaisal, Xionghui Lin, Irene SöderhällAbstract:During evolution, the innate and adaptive immune systems were developed to protect organisms from non-self substances. The innate immune system is phylogenetically more ancient and is present in most multicellular organisms, whereas adaptive responses are restricted to vertebrates. Arthropods lack the blood cells of the lymphoid lineage and oxygen-carrying erythrocytes, making them suitable model animals for studying the regulation of the blood cells of the innate immune system. Many crustaceans have a long life span and need to continuously synthesize blood cells, in contrast to many insects. The Hematopoietic Tissue (HPT) of Pacifastacus leniusculus provides a simple model for studying hematopoiesis, because the Tissue can be isolated, and the proliferation of stem cells and their differentiation can be studied both in vivo and in vitro. Here, we demonstrate new findings of a physical link between the HPT and the brain. Actively proliferating cells were localized to an anterior proliferation center (APC) in the anterior part of the Tissue near the area linking the HPT to the brain, whereas more differentiated cells were detected in the posterior part. The central areas of HPT expand in response to lipopolysaccharide-induced blood loss. Cells isolated from the APC divide rapidly and form cell clusters in vitro; conversely, the cells from the remaining HPT form monolayers, and they can be induced to differentiate in vitro. Our findings offer an opportunity to learn more about invertebrate hematopoiesis and its connection to the central nervous system, thereby obtaining new information about the evolution of different blood and nerve cell lineages.
-
transglutaminase activity in the Hematopoietic Tissue of a crustacean pacifastacus leniusculus importance in hemocyte homeostasis
BMC Immunology, 2008Co-Authors: Kenneth Soderhall, Irene SöderhällAbstract:Background Transglutaminases (TGases) form a group of enzymes that have many different substrates and among the most well known are fibrin for Factor XIIIa and the clotting protein in crustaceans. We also found that TGase is an abundant protein in the Hematopoietic Tissue (Hpt) cells of crayfish and hence we have studied the possible function of this enzyme in hematopoiesis.
Kenneth Soderhall - One of the best experts on this subject based on the ideXlab platform.
-
transfection of crayfish Hematopoietic Tissue cells
Developmental and Comparative Immunology, 2018Co-Authors: Hong Shi, Irene Söderhäll, Lingwei Ruan, Kenneth SoderhallAbstract:Transfection is a powerful tool useful for studying gene function. Establishing transfection methods that enable highly efficient DNA uptake has become increasingly important. The crayfish Hematopoietic Tissue (Hpt) cell cultures have been proven to be suitable for studies on immunity and cell differentiation in crustaceans including shrimps, but no efficient gene transfer and expression method is available for these cells. Here we report a novel and highly efficient DNA transfection system based on electroporation. This method depends on a recombinant plasmid with the promoter from white spot syndrome virus immediate-early gene wsv249. This plasmid could be introduced into primary cells and efficiently express foreign genes by electroporation. By optimizing different electroporation parameters, more than 30% transfection efficiency could be achieved with the relative viability of cells around 50%. This is the first report of gene introduction to crayfish Hpt cells and will be useful for the expanding our research on crustacean immunity.
-
invertebrate hematopoiesis an anterior proliferation center as a link between the Hematopoietic Tissue and the brain
Stem Cells and Development, 2012Co-Authors: Chadanat Noonin, Kenneth Soderhall, Pikul Jiravanichpaisal, Xionghui Lin, Irene SöderhällAbstract:During evolution, the innate and adaptive immune systems were developed to protect organisms from non-self substances. The innate immune system is phylogenetically more ancient and is present in most multicellular organisms, whereas adaptive responses are restricted to vertebrates. Arthropods lack the blood cells of the lymphoid lineage and oxygen-carrying erythrocytes, making them suitable model animals for studying the regulation of the blood cells of the innate immune system. Many crustaceans have a long life span and need to continuously synthesize blood cells, in contrast to many insects. The Hematopoietic Tissue (HPT) of Pacifastacus leniusculus provides a simple model for studying hematopoiesis, because the Tissue can be isolated, and the proliferation of stem cells and their differentiation can be studied both in vivo and in vitro. Here, we demonstrate new findings of a physical link between the HPT and the brain. Actively proliferating cells were localized to an anterior proliferation center (APC) in the anterior part of the Tissue near the area linking the HPT to the brain, whereas more differentiated cells were detected in the posterior part. The central areas of HPT expand in response to lipopolysaccharide-induced blood loss. Cells isolated from the APC divide rapidly and form cell clusters in vitro; conversely, the cells from the remaining HPT form monolayers, and they can be induced to differentiate in vitro. Our findings offer an opportunity to learn more about invertebrate hematopoiesis and its connection to the central nervous system, thereby obtaining new information about the evolution of different blood and nerve cell lineages.
-
transglutaminase activity in the Hematopoietic Tissue of a crustacean pacifastacus leniusculus importance in hemocyte homeostasis
BMC Immunology, 2008Co-Authors: Kenneth Soderhall, Irene SöderhällAbstract:Background Transglutaminases (TGases) form a group of enzymes that have many different substrates and among the most well known are fibrin for Factor XIIIa and the clotting protein in crustaceans. We also found that TGase is an abundant protein in the Hematopoietic Tissue (Hpt) cells of crayfish and hence we have studied the possible function of this enzyme in hematopoiesis.
-
transglutaminase activity in the Hematopoietic Tissue of a crustacean pacifastacus leniusculus importance in hemocyte homeostasis
BMC Immunology, 2008Co-Authors: Xionghui Lin, Kenneth Soderhall, Irene SöderhällAbstract:Transglutaminases (TGases) form a group of enzymes that have many different substrates and among the most well known are fibrin for Factor XIIIa and the clotting protein in crustaceans. We also found that TGase is an abundant protein in the Hematopoietic Tissue (Hpt) cells of crayfish and hence we have studied the possible function of this enzyme in hematopoiesis. TGase is one of the most abundant proteins in the Hpt and its mRNA expression as well as enzyme activity is very high in the Hpt cells, lesser in the semi-granular hemocytes and very low in the granular cells. In cultured Hematopoietic Tissues, high activity was present in cells in the centre of the Tissue, whereas cells migrating out of the Tissue had very low TGase activity. RNAi experiments using dsRNA for TGase completely knocked down the transcript and as a result the cell morphology was changed and the cells started to spread intensely. If astakine, a cytokine directly involved in hematopoiesis, was added the cells started to spread and adopt a morphology similar to that observed after RNAi of TGase. Astakine had no effect on TGase expression, but after a prolonged incubation for one week with this invertebrate cytokine, TGase activity inside and outside the cells was completely lost. Thus it seems as if astakine addition to the Hpt cells and RNAi of TGase in the cell culture will lead to the same results, i.e. loss of TGase activity in the cells and they start to differentiate and spread. The results of this study suggest that TGase is important for keeping the Hpt cells in an undifferentiated stage inside the Hematopoietic Tissue and if expression of TGase mRNA is blocked the cells start to differentiate and spread. This shows a new function for transglutaminase in preventing Hematopoietic stem cells from starting to differentiate and migrate into the hemolymph, whereas their proliferation is unaffected. Astakine is also important for the hematopoiesis, since it induces hemocyte synthesis in the Hpt but now we also show that it in some unknown way participates in the differentiation of the Hpt cells.
-
cell mediated immunity in arthropods hematopoiesis coagulation melanization and opsonization
Immunobiology, 2006Co-Authors: Pikul Jiravanichpaisal, Ok Luel Lee, Kenneth SoderhallAbstract:The functions of hemocytes in innate immune response are reviewed with emphasized on their roles in coagulation, melanization and opsonization. Also the ways in which hemocytes are produced in and released from Hematopoietic Tissue are discussed.
Feng Yang - One of the best experts on this subject based on the ideXlab platform.
-
white spot syndrome virus enters crayfish Hematopoietic Tissue cells via clathrin mediated endocytosis
Virology, 2015Co-Authors: Jiajun Huang, Feng YangAbstract:White spot syndrome virus (WSSV) is a major pathogen of aquacultured shrimp. However, the mechanism of its entry remains poorly understood. In this study, by analyzing the internalization of WSSV using crayfish Hematopoietic Tissue (HPT) cells, we showed that WSSV virions were engulfed by cell membrane invaginations sharing the features of clathrin-coated pits and then internalized into coated cytoplasmic vesicles. Further investigation indicated that WSSV internalization was significantly inhibited by chlorpromazine (CPZ) but not genistein. The internalized virions were colocalized with endogenous clathrin as well as transferrin which undergoes clathrin-dependent uptake. Preventing endosome acidification by ammonium chloride (NH4Cl) or chloroquine (CQ) dramatically reduced WSSV entry as well. Moreover, disturbance of dynamin activity or depletion of membrane cholesterol also blocked WSSV uptake. These data indicate that WSSV enters crayfish HPT cells via clathrin-mediated endocytosis in a pH-dependent manner, and membrane cholesterol as well as dynamin is critical for efficient viral entry.
-
crayfish Hematopoietic Tissue cells but not hemocytes are permissive for white spot syndrome virus replication
Fish & Shellfish Immunology, 2015Co-Authors: Jiajun Huang, Feng YangAbstract:Hemocytes are the major immune cells of crustaceans which are believed to be essential for the pathogenesis of white spot syndrome virus (WSSV) infection. Crayfish hemocytes and Hematopoietic Tissue (HPT) cells have been found to be susceptible to WSSV infection, but the procedure of WSSV infection to both cell types has not yet been carefully investigated. In this study, we analyzed the infection and proliferation of WSSV in crayfish hemocytes as well as HPT cells in detail through transmission electronic microscopy (TEM). The results showed that WSSV could enter both hemocytes and HPT cells through endocytosis, but the production of progeny virus was only achieved in HPT cells. Further investigation demonstrated that although WSSV could transcribe its genes in both cell types, viral genome replication and structural protein expression were unsuccessful in hemocytes, which may be responsible for the failure of progeny production. Therefore, we propose that both hemocytes and HPT cells are susceptible to WSSV infection but only HPT cells are permissive to WSSV replication. These findings will extend our knowledge of the interaction between WSSV and the host immune system.
Raquel Riquelme - One of the best experts on this subject based on the ideXlab platform.
-
generation of mouse zebrafish Hematopoietic Tissue chimeric embryos for hematopoiesis and host pathogen interaction studies
Disease Models & Mechanisms, 2018Co-Authors: Margarita Paradakusz, Cristina Penaranda, Elliott J Hagedorn, Anne E Clatworthy, Anil V Nair, Jonathan E Henninger, Christoph M Ernst, Brian Li, Raquel RiquelmeAbstract:ABSTRACT Xenografts of the Hematopoietic system are extremely useful as disease models and for translational research. Zebrafish xenografts have been widely used to monitor blood cancer cell dissemination and homing due to the optical clarity of embryos and larvae, which allow unrestricted in vivo visualization of migratory events. Here, we have developed a xenotransplantation technique that transiently generates hundreds of Hematopoietic Tissue chimeric embryos by transplanting murine bone marrow cells into zebrafish blastulae. In contrast to previous methods, this procedure allows mammalian cell integration into the fish developmental Hematopoietic program, which results in chimeric animals containing distinct phenotypes of murine blood cells in both circulation and the Hematopoietic niche. Murine cells in chimeric animals express antigens related to (i) Hematopoietic stem and progenitor cells, (ii) active cell proliferation and (iii) myeloid cell lineages. We verified the utility of this method by monitoring zebrafish chimeras during development using in vivo non-invasive imaging to show novel murine cell behaviors, such as homing to primitive and definitive Hematopoietic Tissues, dynamic Hematopoietic cell and Hematopoietic niche interactions, and response to bacterial infection. Overall, transplantation into the zebrafish blastula provides a useful method that simplifies the generation of numerous chimeric animals and expands the range of murine cell behaviors that can be studied in zebrafish chimeras. In addition, integration of murine cells into the host Hematopoietic system during development suggests highly conserved molecular mechanisms of hematopoiesis between zebrafish and mammals. This article has an associated First Person interview with the first author of the paper.
-
generation of mouse zebrafish Hematopoietic Tissue chimeric embryos for hematopoiesis and host pathogen interaction studies
bioRxiv, 2017Co-Authors: Margarita Paradakusz, Cristina Penaranda, Elliott J Hagedorn, Anne E Clatworthy, Anil V Nair, Jonathan E Henninger, Christoph M Ernst, Brian Li, Raquel Riquelme, Humberto JijonAbstract:ABSTRACT Xenografts of the Hematopoietic system are extremely useful as disease models and for translational research. Zebrafish xenografts have been widely used to monitor blood cancer cell dissemination and homing due to the optical clarity of embryos and larvae, which allow unrestricted in vivo visualization of migratory events. To broaden the scope of xenotransplantation studies in zebrafish, we have developed a technique that transiently generates Hematopoietic Tissue chimeras by transplanting murine bone marrow cells into zebrafish blastulae. This procedure leads to mammalian cell integration into the fish developmental Hematopoietic program. Monitoring zebrafish chimeras at different time points post fertilization using in vivo time-lapse and confocal imaging showed murine cell co-localization with developing primitive and definitive Hematopoietic Tissues, intravasation into fish circulation, and dynamic Hematopoietic cell-vascular endothelial and Hematopoietic cell-niche interactions. Immunohistochemistry assays performed in chimeric animals showed that, after engraftment, murine cells expressed antigens related to i) Hematopoietic stem and progenitor cells, ii) active cell proliferation, and iii) myeloid cell lineages. Lastly, xenografted zebrafish larvae infected with Klebsiella pneumoniae showed murine immune cells trafficking to bacterial foci and interacting with bacterial cells. Overall, these results show that mammalian bone marrow cells xenografted in zebrafish integrate into the host Hematopoietic system revealing highly conserved molecular mechanisms of hematopoiesis between zebrafish and mammals. In addition, this procedure introduces a useful and simple method that improves and broadens the scope of Hematopoietic Tissue xenotransplantation studies in zebrafish.
Jiajun Huang - One of the best experts on this subject based on the ideXlab platform.
-
white spot syndrome virus enters crayfish Hematopoietic Tissue cells via clathrin mediated endocytosis
Virology, 2015Co-Authors: Jiajun Huang, Feng YangAbstract:White spot syndrome virus (WSSV) is a major pathogen of aquacultured shrimp. However, the mechanism of its entry remains poorly understood. In this study, by analyzing the internalization of WSSV using crayfish Hematopoietic Tissue (HPT) cells, we showed that WSSV virions were engulfed by cell membrane invaginations sharing the features of clathrin-coated pits and then internalized into coated cytoplasmic vesicles. Further investigation indicated that WSSV internalization was significantly inhibited by chlorpromazine (CPZ) but not genistein. The internalized virions were colocalized with endogenous clathrin as well as transferrin which undergoes clathrin-dependent uptake. Preventing endosome acidification by ammonium chloride (NH4Cl) or chloroquine (CQ) dramatically reduced WSSV entry as well. Moreover, disturbance of dynamin activity or depletion of membrane cholesterol also blocked WSSV uptake. These data indicate that WSSV enters crayfish HPT cells via clathrin-mediated endocytosis in a pH-dependent manner, and membrane cholesterol as well as dynamin is critical for efficient viral entry.
-
crayfish Hematopoietic Tissue cells but not hemocytes are permissive for white spot syndrome virus replication
Fish & Shellfish Immunology, 2015Co-Authors: Jiajun Huang, Feng YangAbstract:Hemocytes are the major immune cells of crustaceans which are believed to be essential for the pathogenesis of white spot syndrome virus (WSSV) infection. Crayfish hemocytes and Hematopoietic Tissue (HPT) cells have been found to be susceptible to WSSV infection, but the procedure of WSSV infection to both cell types has not yet been carefully investigated. In this study, we analyzed the infection and proliferation of WSSV in crayfish hemocytes as well as HPT cells in detail through transmission electronic microscopy (TEM). The results showed that WSSV could enter both hemocytes and HPT cells through endocytosis, but the production of progeny virus was only achieved in HPT cells. Further investigation demonstrated that although WSSV could transcribe its genes in both cell types, viral genome replication and structural protein expression were unsuccessful in hemocytes, which may be responsible for the failure of progeny production. Therefore, we propose that both hemocytes and HPT cells are susceptible to WSSV infection but only HPT cells are permissive to WSSV replication. These findings will extend our knowledge of the interaction between WSSV and the host immune system.