PDF(Pubmed)
Abstract：
BF/C2 is a crucial molecule in the coagulation complement cascade pathway and plays a significant role in the immune response of grass carp through the classical, alternative, and lectin pathways during GCRV infection. In vivo experiments demonstrated that the mRNA expression levels of BF/C2 (A, B) in grass carp positively correlated with GCRV viral replication at various stages of infection. Excessive inflammation leading to death coincided with peak levels of BF/C2 (A, B) mRNA expression and GCRV viral replication. Correspondingly, BF/C2 (A, B) recombinant protein, CIK cells and GCRV co-incubation experiments yielded similar findings. Therefore, 3 h (incubation period) and 9 h (death period) were selected as critical points for this study. Transcriptome sequencing analysis revealed significant differences in the expression of BF/C2A and BF/C2B during different stages of CIK infection with GCRV and compared to the blank control group (PBS). Specifically, the BF/C2A_3 and BF/C2A_9 groups exhibited 2729 and 2228 differentially expressed genes (DEGs), respectively, with 1436 upregulated and 1293 downregulated in the former, and 1324 upregulated and 904 downregulated in the latter. The BF/C2B_3 and BF/C2B_9 groups showed 2303 and 1547 DEGs, respectively, with 1368 upregulated and 935 downregulated in the former, and 818 upregulated and 729 downregulated in the latter. KEGG functional enrichment analysis of these DEGs identified shared pathways between BF/C2A and PBS groups at 3 and 9 h, including the C-type lectin receptor signaling pathway, protein processing in the endoplasmic reticulum, Toll-like receptor signaling pathway, Salmonella infection, apoptosis, tight junction, and adipocytokine signaling pathway. Additionally, the BF/C2B groups at 3 and 9 h shared pathways related to protein processing in the endoplasmic reticulum, glycolysis/gluconeogenesis, and biosynthesis of amino acids. The mRNA levels of these DEGs were validated in cellular models, confirming consistency with the sequencing results. In addition, the mRNA expression levels of these candidate genes (mapk1, il1b, rela, nfkbiab, akt3a, hyou1, hsp90b1, dnajc3a et al.) in the head kidney, kidney, liver and spleen of grass carp immune tissue were significantly different from those of the control group by BF/C2 (A, B) protein injection in vivo. These candidate genes play an important role in the response of BF/C2 (A, B) to GCRV infection and it also further confirmed that BF/C2 (A, B) of grass carp plays an important role in coping with GCRV infection.
摘要：
BF/C2是凝血补体级联途径中的关键分子,在草鱼免疫应答中发挥重要作用,另类，和GCRV感染期间的凝集素途径。体内实验证明，BF/C2的mRNA表达水平（A，B）在草鱼中，在感染的各个阶段与GCRV病毒复制呈正相关。导致死亡的过度炎症与BF/C2的峰值水平相吻合（A，B)mRNA表达和GCRV病毒复制。相应地,BF/C2(A,B)重组蛋白，CIK细胞和GCRV共孵育实验产生了类似的发现。因此,选择3小时(潜伏期)和9小时(死亡期)作为本研究的临界点。转录组测序分析显示，在CIK感染GCRV的不同阶段，BF/C2A和BF/C2B的表达与空白对照组（PBS）相比存在显着差异。具体来说,BF/C2A_3和BF/C2A_9组表现出2729和2228个差异表达基因(DEGs),分别,前者为1436上调，1293下调，在后者中，1324个上调，904个下调。BF/C2B_3和BF/C2B_9组显示2303和1547DEG，分别,前者有1368个上调，935个下调，在后者中，818个上调，729个下调。这些DEGG的KEGG功能富集分析在3和9小时确定了BF/C2A和PBS组之间的共享途径，包括C型凝集素受体信号通路，内质网中的蛋白质加工，Toll样受体信号通路,沙门氏菌感染,凋亡，紧密连接,和脂肪细胞因子信号通路。此外,BF/C2B组在3和9小时共享与内质网中蛋白质加工相关的途径，糖酵解/糖异生，和氨基酸的生物合成。这些DEGs的mRNA水平在细胞模型中得到验证，确认与测序结果的一致性。此外,这些候选基因的mRNA表达水平(mapk1，il1b，rela,nfkbiab,akt3a,hyou1，hsp90b1，dnajc3a等。)在头肾中，肾,通过BF/C2，草鱼免疫组织的肝脏和脾脏与对照组的显着差异（A，B)体内蛋白质注射。这些候选基因在BF/C2（A，B)对GCRV的感染也进一步证实了BF/C2(A,B)的草鱼在应对GCRV感染中起着重要作用。
