Muehlbauer Stefan M, Evering Teresa H, Bonuccelli Gloria, Squires Raynal C, Ashton Anthony W, Porcelli Steven A, Lisanti Michael P, Brojatsch Jürgen
Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Cell Cycle. 2007 Mar 15;6(6):758-66. doi: 10.4161/cc.6.6.3991. Epub 2007 Mar 7.
Murine macrophages have been classified as either susceptible or nonsusceptible to killing by anthrax lethal toxin (LT) depending upon genetic background. While considered resistant to LT killing, we found that bone marrow-derived macrophages (BMMs) from DBA/2, AKR, and C57BL/6 mice were slowly killed by apoptosis following LT exposure. LT killing was not restricted to in vitro assays, as splenic macrophages were also depleted in LT-injected C57BL/6 mice. Human macrophages, also considered LT resistant, similarly underwent slow apoptosis in response to LT challenge. In contrast, LT triggered rapid necrosis and broad protein release in BMMs derived from BALB/c and C3H/HeJ, but not C57BL/6 mice. Released proteins included processed interleukin-18, confirming reports of inflammasome and caspase-1 activation in LT-mediated necrosis in macrophages. Complete inhibition of caspase-1 activity was required to block LT-mediated necrosis. Strikingly, minimal residual caspase-1 activity was sufficient to trigger significant necrosis in LT-treated macrophages, indicating the toxicity of caspase-1 in this process. IL-18 release does not trigger cytolysis, as IL-18 is released late and only from LT-treated macrophages undergoing membrane perturbation. We propose that caspase-1-mediated macrophage necrosis is the source of the cytokine storm and rapid disease progression reported in LT-treated BALB/c mice.
根据遗传背景,小鼠巨噬细胞已被分类为对炭疽致死毒素(LT)杀伤敏感或不敏感。虽然通常认为对LT杀伤具有抗性,但我们发现,来自DBA/2、AKR和C57BL/6小鼠的骨髓来源巨噬细胞(BMM)在暴露于LT后会通过凋亡缓慢死亡。LT杀伤并不局限于体外试验,因为在注射LT的C57BL/6小鼠中,脾巨噬细胞也会减少。同样被认为对LT具有抗性的人类巨噬细胞,在受到LT刺激时也会类似地经历缓慢凋亡。相比之下,LT会在源自BALB/c和C3H/HeJ而非C5BL/6小鼠的BMM中引发快速坏死和广泛的蛋白质释放。释放的蛋白质包括加工后的白细胞介素-18,这证实了巨噬细胞中LT介导的坏死过程中炎性小体和半胱天冬酶-1激活的报道。需要完全抑制半胱天冬酶-1的活性来阻断LT介导的坏死。引人注目的是,最小的残余半胱天冬酶-1活性就足以在LT处理的巨噬细胞中引发显著坏死,表明半胱天冬酶-1在此过程中的毒性。IL-18的释放不会引发细胞溶解,因为IL-18释放较晚,且仅从经历膜扰动的LT处理巨噬细胞中释放。我们认为,半胱天冬酶-1介导的巨噬细胞坏死是LT处理的BALB/c小鼠中细胞因子风暴和疾病快速进展的来源。