Identification of barramundi (Lates calcarifer) DC-SCRIPT, a specific molecular marker for dendritic cells in fish

Zoccola, Emmanuelle, Delamare-Deboutteville, Jerome and Barnes, Andrew C. (2015) Identification of barramundi (Lates calcarifer) DC-SCRIPT, a specific molecular marker for dendritic cells in fish. PLoS ONE, 10 7: 1-25. doi:10.1371/journal.pone.0132687


Author Zoccola, Emmanuelle
Delamare-Deboutteville, Jerome
Barnes, Andrew C.
Title Identification of barramundi (Lates calcarifer) DC-SCRIPT, a specific molecular marker for dendritic cells in fish
Formatted title
Identification of barramundi (Lates calcarifer) DC-SCRIPT, a specific molecular marker for dendritic cells in fish
Journal name PLoS ONE   Check publisher's open access policy
ISSN 1932-6203
Publication date 2015-07-14
Year available 2015
Sub-type Article (original research)
DOI 10.1371/journal.pone.0132687
Open Access Status DOI
Volume 10
Issue 7
Start page 1
End page 25
Total pages 25
Place of publication San Francisco, CA United States
Publisher Public Library of Science
Language eng
Formatted abstract
Antigen presentation is a critical step bridging innate immune recognition and specific immune memory. In mammals, the process is orchestrated by dendritic cells (DCs) in the lymphatic system, which initiate clonal proliferation of antigen-specific lymphocytes. However, fish lack a classical lymphatic system and there are currently no cellular markers for DCs in fish, thus antigen-presentation in fish is poorly understood. Recently, antigen-presenting cells similar in structure and function to mammalian DCs were identified in various fish, including rainbow trout (Oncorhynchus mykiss) and zebrafish (Danio rerio). The present study aimed to identify a potential molecular marker for DCs in fish and therefore targeted DC-SCRIPT, a well-conserved zinc finger protein that is preferentially expressed in all sub-types of human DCs. Putative dendritic cells were obtained in culture by maturation of spleen and pronephros-derived monocytes. DC-SCRIPT was identified in barramundi by homology using RACE PCR and genome walking. Specific expression of DC-SCRIPT was detected in barramundi cells by Stellaris mRNA FISH, in combination with MHCII expression when exposed to bacterial derived peptidoglycan, suggesting the presence of DCs in L. calcarifer. Moreover, morphological identification was achieved by light microscopy of cytospins prepared from these cultures. The cultured cells were morphologically similar to mammalian and trout DCs. Migration assays determined that these cells have the ability to move towards pathogens and pathogen associated molecular patterns, with a preference for peptidoglycans over lipopolysaccharides. The cells were also strongly phagocytic, engulfing bacteria and rapidly breaking them down. Barramundi DCs induced significant proliferation of responder populations of T-lymphocytes, supporting their role as antigen presenting cells. DC-SCRIPT expression in head kidney was higher 6 and 24 h following intraperitoneal challenge with peptidoglycan and lipopolysaccharide and declined after 3 days relative to PBS-injected controls. Relative expression was also lower in the spleen at 3 days post challenge but increased again at 7 days. As DC-SCRIPT is a constitutively expressed nuclear receptor, independent of immune activation, this may indicate initial migration of immature DCs from head kidney and spleen to the injection site, followed by return to the spleen for maturation and antigen presentation. DC-SCRIPT may be a valuable tool in the investigation of antigen presentation in fish and facilitate optimisation of vaccines and adjuvants for aquaculture.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
School of Biological Sciences Publications
 
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