Voth Daniel E, Hamm Elaine E, Nguyen Lan G, Tucker Amy E, Salles Isabelle I, Ortiz-Leduc William, Ballard Jimmy D
Department of Microbiology and Immunology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Cell Microbiol. 2005 Aug;7(8):1139-49. doi: 10.1111/j.1462-5822.2005.00539.x.
Oedema factor (OF) and protective antigen (PA) are secreted by Bacillus anthracis, and their binary combination yields oedema toxin (OT). Following PA-mediated delivery to the cytosol, OF functions as an adenylate cyclase generating high levels of cAMP. To assess OT as a possible cause of tissue damage and cell death, a novel approach was developed, which utilized a developing zebrafish embryo model to study toxin activity. Zebrafish embryos incubated with OT exhibited marked necrosis of the liver, cranium and gastrointestinal tract, as well as reduced swim bladder inflation. The OT-treated embryos survived after all stages of development but succumbed to the toxin within 7 days. Additional analysis of specific cell lines, including macrophage and non-macrophage, showed OT-induced cell death is cell type-specific. There was no discernible correlation between levels of OF-generated cAMP and cell death. Depending on the type of cell analysed, cell death could be detected in low levels of cAMP, and, conversely, cell survival was observed in one cell line in which high levels of cAMP were found following treatment with OT. Collectively, these data suggest OT is cytotoxic in a cell-dependent manner and may contribute to disease through direct cell killing leading to tissue necrosis.
水肿因子(OF)和保护性抗原(PA)由炭疽芽孢杆菌分泌,二者二元组合产生水肿毒素(OT)。在PA介导下转运至胞质溶胶后,OF作为一种腺苷酸环化酶发挥作用,产生高水平的环磷酸腺苷(cAMP)。为了评估OT是否可能是组织损伤和细胞死亡的原因,开发了一种新方法,利用发育中的斑马鱼胚胎模型来研究毒素活性。用OT孵育的斑马鱼胚胎表现出肝脏、颅骨和胃肠道明显坏死,以及鳔充气减少。经OT处理的胚胎在所有发育阶段后均存活,但在7天内死于毒素。对包括巨噬细胞和非巨噬细胞在内的特定细胞系的进一步分析表明,OT诱导的细胞死亡具有细胞类型特异性。OF产生的cAMP水平与细胞死亡之间没有明显的相关性。根据所分析的细胞类型,在低水平的cAMP中可检测到细胞死亡,相反,在用OT处理后发现高水平cAMP的一个细胞系中观察到细胞存活。总体而言,这些数据表明OT以细胞依赖性方式具有细胞毒性,并可能通过直接导致组织坏死的细胞杀伤作用促成疾病。