ABSTRACT:The function of Toll pathway defense against bacterial infection has been well 37 established in shrimp, however how this pathway responds to viral infection is still largely 38 unknown. In this study, we report the Toll4-Dorsal-AMPs cascade restricts the white spot 39 syndrome virus (WSSV) infection of shrimp. A total of nine Tolls from Litopenaeus 40 vannamei namely Toll1-9 are identified, and RNAi screening in vivo reveals the Toll4 is 41 important for shrimp to oppose WSSV infection. Knockdown of Toll4 results in elevated viral 42 loads and renders shrimp more susceptible to WSSV. Furthermore, Toll4 could be a one of 43 upstream pattern recognition receptor (PRR) to detect WSSV, and thereby leading to nuclear 44 translocation and phosphorylation of Dorsal, the known NF-κB transcription factor of the 45 canonical Toll pathway. More importantly, silencing of Toll4 and Dorsal contributes to 46 impaired expression of a specific set of antimicrobial peptides (AMPs) such as 47anti-LPS-factor (ALF) and lysozyme (LYZ) family, which exert potent anti-WSSV activity.
48Two AMPs of ALF1 and LYZ1 as representatives are demonstrated to have the ability to 49 interact with several WSSV structural proteins. Taken together, we therefore identify the 50 Toll4-Dorsal pathway mediates strong resistance to WSSV infection by inducing some 51 specific AMPs.
53Author summary:The TLR pathway mediated antiviral immune response is well identified 54 in mammals, yet, Toll pathway governing this protection in invertebrates remains unknown.
55In the present study, we uncover that a shrimp Toll4 from a total of nine Tolls in L. vannamei 56 confers resistance to WSSV thought inducing the NF-κB transcription factor Dorsal to 57 inspiring the production of some antimicrobial peptides (AMPs) with antiviral activity. The 58 anti-LPS-factor (ALF) and lysozyme (LYZ) family are identified as the Toll4-Dorsal pathway 59 targeted genes with the ability to interact with viral structural proteins in response to WSSV 60 infection. These results suggest that the Toll receptor induces the expression of AMPs with 61 antiviral activity could be a general antiviral mechanism in invertebrates and Toll pathway 62 established antiviral defense could be conserved during evolution.63 KEYWORDS: Litopenaeus vannamei; Toll4-Dorsal pathway; Antiviral defense; WSSV; 64 antimicrobial peptides (AMPs) 65 66 67Multicellular organisms have evolved the ability to protect themselves from a wide 68 variety of pathogens including virus. In invertebrates including shrimps that lacking 69 immunoglobulin-based adaptive immune system, this protection is thus provided through the 70 action of an innate immune system. The innate immune response is generally initiated via the 71 detection of pathogen-associated molecular patterns (PAMPs), some evolutionarily conserved 72 structures or motifs shared by broad classes of invading organisms, by a wide diversity of 73 host pattern recognition receptors (PRRs) (1). One important class of PRRs is the Toll 74 receptor superfamily, ...