真核系统表达的猪α干扰素(PoIFN-α)发酵液,经离心、收集上清液、透析过滤处理,通过FPLC-HisTrap HP Kit亲和层析系统分离纯化,最后以SDS-PAGE银染法检验。结果表明,在不同pH和NaCl浓度情况下,发酵液经亲和层析系统分离纯化后均可以得...真核系统表达的猪α干扰素(PoIFN-α)发酵液,经离心、收集上清液、透析过滤处理,通过FPLC-HisTrap HP Kit亲和层析系统分离纯化,最后以SDS-PAGE银染法检验。结果表明,在不同pH和NaCl浓度情况下,发酵液经亲和层析系统分离纯化后均可以得到较为单一的目的蛋白组分,而在pH8.2,NaCl0mmol/L的情况下得到的目的蛋白更纯,且非特异性吸附也较少。展开更多
Interferon, a natural protein that is produced by a variety of cells during viral infection, activates the transcription of multiple functional genes in cells, regulates synergy among various signaling pathways, and m...Interferon, a natural protein that is produced by a variety of cells during viral infection, activates the transcription of multiple functional genes in cells, regulates synergy among various signaling pathways, and mediates many biological functions such as antiviral activity, immune regulation, and cell growth. However, clinical research on interferon in livestock is lacking. In this study, recombinant porcine interferon(PoIFNa) was used as an adjuvant, in combination with inactivated influenza virus, to vaccinate 6-week-old pigs via nasal infusion. The transcription of target genes was then monitored and the functions of PoIFNa were determined with respect to the activation of mucosal immunity. We found that a combination of low-dose PoIFNa and inactivated influenza virus could significantly up-regulate the expression of immunoregulatory cytokines such as IL-2, IL-18, IFN-c, IL-6, and IL-10 by real-time PCR, suggesting the induction of a strong mucosal innate immune response after administration. In addition, low-dose PoIFNa can significant enhancing the transcription of genes encoding homing factors including CCR9 and CCR10(P \ 0.001), thereby resulting in the induction of higher levels of HA-specific antibodies(P \ 0.05), which can be determined by ELISA and IFA. Post-immunization challenges with H1 N1 virus demonstrated that PoIFNa, combined with inactivated influenza virus, could alleviate clinical signs in pigs during the early stages of viral infection. These studies reveal low-dose PoIFNa as a potential mucosal adjuvant for influenza virus in pigs.展开更多
文摘真核系统表达的猪α干扰素(PoIFN-α)发酵液,经离心、收集上清液、透析过滤处理,通过FPLC-HisTrap HP Kit亲和层析系统分离纯化,最后以SDS-PAGE银染法检验。结果表明,在不同pH和NaCl浓度情况下,发酵液经亲和层析系统分离纯化后均可以得到较为单一的目的蛋白组分,而在pH8.2,NaCl0mmol/L的情况下得到的目的蛋白更纯,且非特异性吸附也较少。
基金supported by Grants from the National Key R&D Programme of China (2017YFD051105)the National Natural Science Foundation of China (31630079)+2 种基金the National Science and Technology Major Project (2018ZX10101004)the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB29010000)supported by Youth Innovation Promotion Association of CAS (2019)
文摘Interferon, a natural protein that is produced by a variety of cells during viral infection, activates the transcription of multiple functional genes in cells, regulates synergy among various signaling pathways, and mediates many biological functions such as antiviral activity, immune regulation, and cell growth. However, clinical research on interferon in livestock is lacking. In this study, recombinant porcine interferon(PoIFNa) was used as an adjuvant, in combination with inactivated influenza virus, to vaccinate 6-week-old pigs via nasal infusion. The transcription of target genes was then monitored and the functions of PoIFNa were determined with respect to the activation of mucosal immunity. We found that a combination of low-dose PoIFNa and inactivated influenza virus could significantly up-regulate the expression of immunoregulatory cytokines such as IL-2, IL-18, IFN-c, IL-6, and IL-10 by real-time PCR, suggesting the induction of a strong mucosal innate immune response after administration. In addition, low-dose PoIFNa can significant enhancing the transcription of genes encoding homing factors including CCR9 and CCR10(P \ 0.001), thereby resulting in the induction of higher levels of HA-specific antibodies(P \ 0.05), which can be determined by ELISA and IFA. Post-immunization challenges with H1 N1 virus demonstrated that PoIFNa, combined with inactivated influenza virus, could alleviate clinical signs in pigs during the early stages of viral infection. These studies reveal low-dose PoIFNa as a potential mucosal adjuvant for influenza virus in pigs.
