Siddharthan Venkatraman, Wang Hong, Davies Christopher J, Hall Jeffery O, Morrey John D
Institute for Antiviral Research, Department of Animal, Dairy, and Veterinary Sciences, Utah State University, Logan, Utah, United States of America.
Center for Integrated BioSystems, Department of Animal, Dairy, and Veterinary Sciences, Utah State University, Logan, Utah, United States of America.
PLoS One. 2014 Sep 2;9(9):e106535. doi: 10.1371/journal.pone.0106535. eCollection 2014.
Evidence indicates that West Nile virus (WNV) employs Ca(2+) influx for its replication. Moreover, calcium buffer proteins, such as calbindin D28k (CB-D28k), may play an important role mitigating cellular destruction due to disease processes, and more specifically, in some neurological diseases. We addressed the hypothesis that CB-D28k inhibits WNV replication in cell culture and infected rodents. WNV envelope immunoreactivity (ir) was not readily co-localized with CB-D28k ir in WNV-infected Vero 76 or motor neuron-like NSC34 cells that were either stably or transiently transfected with plasmids coding for CB-D28k gene. This was confirmed in cultured cells fixed on glass coverslips and by flow cytometry. Moreover, WNV infectious titers were reduced in CB-D28k-transfected cells. As in cell culture studies, WNV env ir was not co-localized with CB-D28k ir in the cortex of an infected WNV hamster, or in the hippocampus of an infected mouse. Motor neurons in the spinal cord typically do not express CB-D28k and are susceptible to WNV infection. Yet, CB-D28k was detected in the surviving motor neurons after the initial phase of WNV infection in hamsters. These data suggested that induction of CB-D28k elicit a neuroprotective response to WNV infection.
有证据表明,西尼罗河病毒(WNV)利用钙离子内流进行复制。此外,钙缓冲蛋白,如钙结合蛋白D28k(CB-D28k),可能在减轻疾病过程导致的细胞破坏中发挥重要作用,更具体地说,在某些神经疾病中。我们探讨了CB-D28k在细胞培养和感染啮齿动物中抑制WNV复制的假说。在稳定或瞬时转染了编码CB-D28k基因的质粒的WNV感染的Vero 76细胞或运动神经元样NSC34细胞中,WNV包膜免疫反应性(ir)不易与CB-D28k ir共定位。这在固定在玻璃盖玻片上的培养细胞和通过流式细胞术中得到了证实。此外,在转染了CB-D28k的细胞中,WNV感染滴度降低。与细胞培养研究一样,在感染WNV的仓鼠皮质或感染小鼠的海马中,WNV env ir与CB-D28k ir不共定位。脊髓中的运动神经元通常不表达CB-D28k,易受WNV感染。然而,在仓鼠WNV感染初始阶段后,在存活的运动神经元中检测到了CB-D28k。这些数据表明,CB-D28k的诱导引发了对WNV感染的神经保护反应。