Abstract
Foot-and-mouth disease virus (FMDV) is one of the most important animal pathogens in the world. FMDV naturally infects swine, cattle, and other cloven-hoofed animals. FMD is not adequately controlled by vaccination. An alternative strategy is to develop swine that are genetically resistant to infection. Here, we generated FMDV-specific shRNA transgenic cells targeting either nonstructural protein 2B or polymerase 3D of FMDV. The shRNA-positive transgenic cells displayed significantly lower viral production than that of the control cells after infection with FMDV (P < 0.05). Twenty-three transgenic cloned swine (TGCS) and nine non-transgenic cloned swine (Non-TGCS) were produced by somatic cell nuclear transfer (SCNT). In the FMDV challenge study, one TGCS was completely protected, no clinical signs, no viremia and no viral RNA in the tissues, no non-structural antibody response, another one TGCS swine recovered after showing clinical signs for two days, whereas all of the normal control swine (NS) and Non-TGCS developed typical clinical signs, viremia and viral RNA was determined in the tissues, the non-structural antibody was determined, and one Non-TGCS swine died. The viral RNA load in the blood and tissues of the TGCS was reduced in both challenge doses. These results indicated that the TGCS displayed resistance to the FMDV infection. Immune cells, including CD3+, CD4+, CD8+, CD21+, and CD172+ cells, and the production of IFN-γ were analyzed, there were no significant differences observed between the TGCS and NS or Non-TGCS, suggesting that the FMDV resistance may be mainly derived from the RNAi-based antiviral pathway. Our work provides a foundation for a breeding approach to preventing infectious disease in swine.
Highlights
Foot-and-mouth disease virus (FMDV) is one of the most important animal pathogens in the world
Fire et al described a process found in Caenorhabditis elegans[11], called RNA interference (RNAi), a naturally occurring process triggered by a double-stranded RNA or short hairpin RNA that can be processed into small interfering RNAs, interact with a protein complex, and cleave their complementary RNAs12, 13
Every swine was inoculated successfully with FMDV, one transgenic cloned swine was protected under 100 S ID50 FMDV challenge, no clinical signs, no viremia, and no viral RNA in the tissues, no non-structural antibody response, and one transgenic cloned swine had recuperated and was healthy after showing lesions for two days in the 10 SID50 FMDV challenge study, whereas all of the Non-TGCS and normal control swine (NS) developed clinical symptoms, viremia, and viral RNA was determined in the tissues, non-structural antibody was determined., and one Non-TGCS died
Summary
Foot-and-mouth disease virus (FMDV) is one of the most important animal pathogens in the world. FMDV naturally infects swine, cattle, and other cloven-hoofed animals. The shRNA-positive transgenic cells displayed significantly lower viral production than that of the control cells after infection with FMDV (P < 0.05). The viral RNA load in the blood and tissues of the TGCS was reduced in both challenge doses These results indicated that the TGCS displayed resistance to the FMDV infection. One novel alternative strategy to control FMDVs in livestock is to introduce novel genes that provide resistance to FMDVs. Fire et al described a process found in Caenorhabditis elegans[11], called RNA interference (RNAi), a naturally occurring process triggered by a double-stranded RNA (dsRNA) or short hairpin RNA (shRNA) that can be processed into small interfering RNAs (siRNAs), interact with a protein complex, and cleave their complementary RNAs12, 13. The antiviral RNAi pathway in mammalian cells has been reported[17,18]
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