Lazarte Jassy Mary S, Kim Young Rim, Lee Jung Seok, Chun Jin Hong, Kim Si Won, Jung Jae Wook, Kim Jaesung, Kayansamruaj Pattanapon, Thompson Kim D, Kim Hyeongsu, Jung Tae Sung
Laboratory of Aquatic Animal Diseases, Research Institute of Natural Science, College of Veterinary Medicine, Gyeongsang National University, 501-201, 501, Jinju-daero, Jinju-si, Gyeongsangnam-do 52828, Korea.
Center of Excellence in Aquatic Animal Health Management, Faculty of Fisheries, Kasetsart University, 50 Ngamwongwan Rd, Ladyao, Chatuchak, Bangkok 10900, Thailand.
Vaccines (Basel). 2021 Jan 16;9(1):55. doi: 10.3390/vaccines9010055.
The causative agent of acute hepatopancreatic necrosis disease (AHPND) is the bacterium, , which secretes toxins into the gastrointestinal tract of its host. toxins A and B (PirA/PirB) have been implicated in the pathogenesis of this disease, and are, therefore, the focus of studies developing treatments for AHPND. We previously produced recombinant antibodies based on the hagfish variable lymphocyte receptor B (VLRB) capable of neutralizing some viruses, suggesting that this type of antibody may have a potential application for treatment of AHPND. Here, recombinant PirA/PirB, produced using a bacterial expression system, were used as antigens to screen a hagfish VLRB cDNA library to obtain PirA/PirB-specific antibodies. A cell line secreting these antibodies was established by screening and cloning the DNA extracted from hagfish B cells. Supernatants collected from cells secreting the PirA/PirB antibodies were collected and concentrated, and used to passively immunize shrimp to neutralize the toxins PirA or PirB associated with AHPND. Briefly, 10 μg of PirA and PirB antibodies, 7C12 and 9G10, respectively, were mixed with the shrimp feed, and fed to shrimp for three days consecutive days prior to experimentally infecting the shrimp with (containing toxins A and B), and resulting mortalities recorded for six days. Results showed significantly higher level of survival in shrimp fed with the PirB-9G10 antibody (60%) compared to the group fed the PirA-7C12 antibody (3%) and the control group (0%). This suggests that VLRB antibodies may be a suitable alternative to immunoglobulin-based antibodies, as passive immunization treatments for effective management of AHPND outbreaks within shrimp farms.
急性肝胰腺坏死病(AHPND)的病原体是 细菌,它会向宿主的胃肠道分泌毒素。毒素A和B(PirA/PirB)与该病的发病机制有关,因此是开发AHPND治疗方法研究的重点。我们之前基于盲鳗可变淋巴细胞受体B(VLRB)制备了能够中和某些病毒的重组抗体,这表明这类抗体可能在AHPND治疗中具有潜在应用价值。在此,使用细菌表达系统生产的重组PirA/PirB被用作抗原,筛选盲鳗VLRB cDNA文库以获得PirA/PirB特异性抗体。通过筛选和克隆从盲鳗B细胞中提取的DNA,建立了分泌这些抗体的细胞系。收集并浓缩从分泌PirA/PirB抗体的细胞中收集的上清液,用于对虾进行被动免疫,以中和与AHPND相关的毒素PirA或PirB。简要地说,分别将10μg的PirA和PirB抗体7C12和9G10与虾饲料混合,在对虾进行实验性感染 (含有毒素A和B)之前连续三天喂给虾,并记录六天的死亡情况。结果显示,与喂食PirA - 7C12抗体的组(3%)和对照组(0%)相比,喂食PirB - 9G10抗体的虾的存活率显著更高(60%)。这表明VLRB抗体可能是基于免疫球蛋白的抗体的合适替代品,作为对虾养殖场内AHPND爆发进行有效管理的被动免疫治疗方法。