Zebrafish Kidney Phagocytes Utilize Macropinocytosis and Ca2+-Dependent Endocytic Mechanisms

Background The innate immune response constitutes the first line of defense against invading pathogens and consists of a variety of immune defense mechanisms including active endocytosis by macrophages and granulocytes. Endocytosis can be used as a reliable measure of selective and non-selective mec...

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Bibliographic Details
Published in:PloS one 2009-02, Vol.4 (2), p.e4314
Main Authors: Hohn, Claudia, Lee, Sang-Ryul, Pinchuk, Lesya M., Petrie-Hanson, Lora
Format: Article
Language:English
Subjects:
Animal tissues
Assaying
Calcium
Cytometry
Cytoskeleton
Danio rerio
Dextran
Edwardsiella
Edwardsiella ictaluri
Endocytosis
Epidemiology
Fish
Ictalurus punctatus
Immune response
Immune system
Immunology
Immunology/Immune Response
Immunology/Immunity to Infections
Immunology/Innate Immunity
Immunology/Leukocyte Activation
Infectious Diseases/Bacterial Infections
Innate immunity
Internalization
Kidneys
Leukocytes (granulocytic)
Leukocytes (neutrophilic)
Lymphocytes
Macrophages
Mannose
Monocytes
Phagocytes
Pimephales promelas
Veterinary colleges
Zebrafish
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Summary:Background The innate immune response constitutes the first line of defense against invading pathogens and consists of a variety of immune defense mechanisms including active endocytosis by macrophages and granulocytes. Endocytosis can be used as a reliable measure of selective and non-selective mechanisms of antigen uptake in the early phase of an immune response. Numerous assays have been developed to measure this response in a variety of mammalian and fish species. The small size of the zebrafish has prevented the large-scale collection of monocytes/macrophages and granulocytes for these endocytic assays. Methodology/Principal Findings Pooled zebrafish kidney hematopoietic tissues were used as a source of phagocytic cells for flow-cytometry based endocytic assays. FITC-Dextran, Lucifer Yellow and FITC-Edwardsiella ictaluri were used to evaluate selective and non-selective mechanisms of uptake in zebrafish phagocytes. Conclusions/Significance Zebrafish kidney phagocytes characterized as monocytes/macrophages, neutrophils and lymphocytes utilize macropinocytosis and Ca2+-dependant endocytosis mechanisms of antigen uptake. These cells do not appear to utilize a mannose receptor. Heat-killed Edwardsiella ictaluri induces cytoskeletal interactions for internalization in zebrafish kidney monocytes/macrophages and granulocytes. The proposed method is easy to implement and should prove especially useful in immunological, toxicological and epidemiological research.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0004314