Park Gyu-Nam, Shin Jihye, Choe SeEun, Kim Ki-Sun, Kim Jae-Jo, Lim Seong-In, An Byung-Hyun, Hyun Bang-Hun, An Dong-Jun
Virus Disease Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
College of Veterinary Medicine, Seoul University, Gwanak-ro, Gwanak-gu, Seoul 08826, Republic of Korea.
Vaccines (Basel). 2023 Oct 21;11(10):1622. doi: 10.3390/vaccines11101622.
A chimeric (KD26_E2LOM) was prepared by inserting the E2 gene of the classical swine fever virus (CSFV) LOM strain into the backbone of the bovine viral diarrhea virus (BVDV) KD26 strain. KD26_E2LOM was obtained by transfecting the cDNA pACKD26_E2LOM into PK-15 cells. KD26_E2LOM chimeric proliferated to titers of 10 TCID/mL and 10 TCID/mL at 96 h post-inoculation into PK-15 cells or MDBK cells, respectively. It also reacted with antibodies specific for CSFV E2 and BVDV E, but not with an anti-BVDV E2 antibody. Piglets (55-60 days old) inoculated with a high dose (10 TCID/mL) of KD26_E2LOM produced high levels of CSFV E2 antibodies. In addition, no co-habiting pigs were infected with KD26_E2LOM; however, some inoculated pigs excreted the virus, and the virus was detected in some organs. When pregnant sows were inoculated during the first trimester (55-60 days) with a high dose (10 TCID/mL) of KD26_E2LOM, anti-CSFV E2 antibodies were produced at high levels; chimeric was detected in one fetus and in the ileum of one sow. When 5-day-old calves that did not consume colostrum received a high dose (10 TCID/mL) of KD26_E2LOM, one calf secreted the virus in both feces and nasal fluid on Day 2. A high dose of KD26_E2LOM does not induce specific clinical signs in most animals, does not spread from animal to animal, and generates CSFV E2 antibodies with DVIA functions. Therefore, chimeric KD26_E2LOM is a potential CSFV live marker vaccine.
通过将经典猪瘟病毒(CSFV)LOM株的E2基因插入牛病毒性腹泻病毒(BVDV)KD26株的主干中制备了一种嵌合体(KD26_E2LOM)。通过将cDNA pACKD26_E2LOM转染到PK - 15细胞中获得KD26_E2LOM。将KD26_E2LOM嵌合体分别接种到PK - 15细胞或MDBK细胞中,在接种后96小时,其增殖滴度分别达到10⁵ TCID₅₀/mL和10⁶ TCID₅₀/mL。它还与针对CSFV E2和BVDV E的特异性抗体发生反应,但不与抗BVDV E2抗体发生反应。用高剂量(10⁶ TCID₅₀/mL)的KD26_E2LOM接种55 - 60日龄仔猪,可产生高水平的CSFV E2抗体。此外,同居猪未被KD26_E2LOM感染;然而,一些接种猪排出了病毒,并且在一些器官中检测到了该病毒。当怀孕母猪在妊娠早期(55 - 60天)用高剂量(10⁶ TCID₅₀/mL)的KD26_E2LOM接种时,可产生高水平的抗CSFV E2抗体;在一头仔猪和一头母猪的回肠中检测到了嵌合体。当未摄入初乳的5日龄犊牛接受高剂量(10⁶ TCID₅₀/mL)的KD26_E2LOM时,一头犊牛在第2天粪便和鼻液中都分泌了病毒。高剂量的KD26_E2LOM在大多数动物中不诱导特定的临床症状,不发生动物间传播,并产生具有鉴别诊断(DVIA)功能的CSFV E2抗体。因此,嵌合体KD26_E2LOM是一种潜在的CSFV活标记疫苗。