Development of attenuated herpes simplex virus vector for analysis of neurological functions.

我们开发了一种减毒的HSV-1活载体βH1，在神经系统中表达β-半乳糖苷酶（β-gal）。皮下接种可导致接种部位感觉神经节神经元β-gal活性表达至少45天。βH1也足以通过肌肉注射将外源基因传递到大鼠双侧脊髓前角的运动神经元中，并且β-gal在90%的双侧脊髓前角运动神经元中持续表达至少182天。对HSV免疫的存在不妨碍转基因在肌内接种大鼠前角运动神经元中的传递。此外，将βH1接种到大鼠右侧尾状壳核中，不仅可以在接种部位的神经元中有效表达β-gal活性，还可以在接种部位的突起区域表达β-gal活性，该区域的扩散受抗hsv药物更昔洛韦（ganciclovir）的调节。βH1来源于小鼠减毒的HSV-1 HF毒株，在中枢神经系统（CNS），特别是脊髓运动神经元中表达β-gal活性，但不引起组织破坏或炎症。它将成为家族性肌萎缩侧索硬化症和中枢神经系统疾病基因治疗的广泛潜在载体

We have developed a live attenuated HSV-1 vector, βH1, expressing β-galactosidase (β-gal) in the nervous system. Its subcutaneous inoculation led the expression of β-gal activity in neurons of the sensory ganglia innervating the inoculation site for at least 45 days. βH1 was also sufficient to deliver the foreign gene into motor neurons in the bilateral anterior horn of the spinal cord of rats by an intramuscular inoculation and β-gal expression continued for at least 182 days in 90% of motor neurons in the bilateral anterior horn of spinal cord in a wide range. The presence of immunity to HSV was not an obstacle to deliver the transgene in anterior horn motor neurons in the intramuscularly inoculated rats. Furthermore, βH1 inoculation into the right caudate putamen of rats was efficient in expressing β-gal activity not only in the neurons of the inoculation site but also in the area projecting to the inoculation site, where this spread of area was regulated by an anti-HSV agent, ganciclovir. βH1 is originated from HSV-1 HF strain that is attenuated to mice, and expresses β-gal activity in the central nervous system (CNS), especially motor neurons in spinal cord without causing tissue destruction or inflammation. It would be a broad potential vector for gene therapy for familial amyotrophic lateral sclerosis and CNS diseases