Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, China.
Front Immunol. 2023 Sep 1;14:1200297. doi: 10.3389/fimmu.2023.1200297. eCollection 2023.
African swine fever (ASF) is an infectious disease caused by African swine fever virus (ASFV) that is highly contagious and has an extremely high mortality rate (infected by virulent strains) among domestic and wild pigs, causing huge economic losses to the pig industry globally. In this study, SDS-PAGE gel bands hybridized with ASFV whole virus protein combined with ASFV-convalescent and ASFV-positive pig serum were identified by mass spectrometry. Six antigens were detected by positive serum reaction bands, and eight antigens were detected in ASFV-convalescent serum. In combination with previous literature reports and proteins corresponding to MHC-II presenting peptides screened from ASFV-positive pig urine conducted in our lab, seven candidate antigens, including KP177R (p22), K78R (p10), CP204L (p30), E183L (p54), B602L (B602L), EP402R-N (CD2V-N) and F317L (F317L), were selected. Subunit-Group 1 was prepared by mixing above-mentioned seven ASFV recombinant proteins with MONTANIDETM1313 VG N mucosal adjuvant and immunizing pigs intranasally and intramuscularly. Subunit-Group 2 was prepared by mixing four ASFV recombinant proteins (p22, p54, CD2V-N1, B602L) with Montanide ISA 51 VG adjuvant and immunizing pigs by intramuscular injection. Anticoagulated whole blood, serum, and oral fluid were collected during immunization for flow cytometry, serum IgG as well as secretory sIgA antibody secretion, and cytokine expression testing to conduct a comprehensive immunogenicity assessment. Both immunogen groups can effectively stimulate the host to produce ideal humoral, mucosal, and cellular immune responses, providing a theoretical basis for subsequent functional studies, such as immunogens challenge protection and elucidation of the pathogenic mechanism of ASFV.
非洲猪瘟（ASF）是由非洲猪瘟病毒（ASFV）引起的一种传染病，具有高度传染性，在国内外猪群中死亡率极高（感染强毒株），给全球养猪业造成巨大经济损失。本研究采用 SDS-PAGE 凝胶条带与 ASFV 全病毒蛋白杂交，再经质谱鉴定。阳性血清反应带检测到 6 种抗原，ASFV 恢复期血清中检测到 8 种抗原。结合文献报道和本实验室前期筛选的 ASFV 阳性猪尿液中 MHC-II 呈递肽对应的蛋白，选择了 7 种候选抗原，包括 KP177R（p22）、K78R（p10）、CP204L（p30）、E183L（p54）、B602L（B602L）、EP402R-N（CD2V-N）和 F317L（F317L）。将上述 7 种 ASFV 重组蛋白与 MONTANIDETM1313 VG N 黏膜佐剂混合制备亚单位 1 组，通过鼻内和肌肉注射免疫猪。将 4 种 ASFV 重组蛋白（p22、p54、CD2V-N1、B602L）与 Montanide ISA 51 VG 佐剂混合制备亚单位 2 组，通过肌肉注射免疫猪。免疫过程中采集抗凝全血、血清和唾液，进行流式细胞术、血清 IgG 和分泌性 sIgA 抗体分泌以及细胞因子表达检测，全面评估免疫原性。两组免疫原都能有效地刺激宿主产生理想的体液、黏膜和细胞免疫应答，为后续免疫原的功能研究（如免疫原攻毒保护和 ASF 发病机制的阐明）提供了理论依据。
