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Molecular cloning of S almo salar T oll‐like receptors ( TLR 1, TLR 22, TLR 5M and TLR 5S) and expression analysis in SHK ‐1 cells during Piscirickettsia salmonis infection
Author(s) -
Salazar C,
Haussmann D,
Kausel G,
Figueroa J
Publication year - 2016
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/jfd.12354
Subject(s) - biology , immune system , innate immune system , salmo , pathogen , receptor , in vitro , microbiology and biotechnology , cloning (programming) , fish <actinopterygii> , immunology , genetics , fishery , computer science , programming language
In fish, the innate immune system is the primary response against infection. Toll‐like receptors (TLRs) recognize pathogens through pathogen‐associated molecular patterns (PAMPs), and some target molecules of TLRs are homologous between fish and mammals. Piscirickettsia salmonis is one of the main pathogens affecting the salmon industry in Chile. Better knowledge of mechanisms underlying its invasive capacity and recognition of target cells is crucial for vaccine development. Therefore, Salmo salar L. TLR1, TLR22, membrane TLR5M and soluble TLR5S sequences were cloned, and expression kinetics were analysed by RT‐qPCR in salmon head kidney cells (SHK‐1) infected with three different P. salmonis preparations: alive, formaldehyde treated, extract. Clearly, all analysed TLRs were expressed and transcription level changes were revealed at 2 hpi, 12 or 16 hpi and 24 hpi depending on P. salmonis infection scheme. Increased IL1‐beta expression confirmed TLR pathway response. Furthermore, significant expression modulations of several members of the TLR pathway in this in vitro model suggest that P. salmonis extract rather than formaldehyde‐inactivated bacteria might strengthen the salmon immune system.