Yanay Ofer, Brzezinski Margaret, Christensen Jeffrey, Liggitt Denny, Dale David C, Osborne William R A
Department of Pediatrics, University of Washington, Seattle, WA 98195, USA.
Hum Gene Ther. 2006 Apr;17(4):464-9. doi: 10.1089/hum.2006.17.464.
Cyclic neutropenia occurs in humans and gray collie dogs, is characterized by recurrent neutropenia, and is treated by daily injections of recombinant granulocyte colony-stimulating factor (G-CSF). After showing that canine recombinant G-CSF increased neutrophil counts in an affected dog, we administered intramuscularly 2 x 10(9) infectious units (IU) of a lentiviral vector encoding canine G-CSF cDNA. Elevated, therapeutic neutrophil production was obtained for nearly 18 months. Lentiviral vector treatment provided a mean neutrophil count of 29,230 +/- 12,930 cells/microl, which was significantly increased over both the pretreatment value (5,240 +/- 4,800 cells/microl; p < 0.0001) and the neutrophil count during G-CSF administration (17,820 +/- 11,100 cells/microl; p < 0.0001). By systemic infusion of recombinant G-CSF to normal dogs we estimated that 2 x 10(9) IU of lentivirus delivered 3.5 microg of G-CSF per kilogram per day. After lentiviral vector treatment the gray collie gained weight, showed no clinical signs of infection and fever, and no longer needed housing in a pathogen-free environment. Genomic DNA harvested from muscle at the injection sites was positive for provirus, whereas gonad, lung, spleen, heart, liver, kidney, and noninjected muscle samples were negative. These studies show that an adult animal is responsive long-term to lentivirus-mediated G-CSF delivery, suggesting this approach may be applied for treatment of adult patients with cyclic and other neutropenias.
周期性中性粒细胞减少症发生于人类和灰色柯利犬,其特征为反复出现中性粒细胞减少,可通过每日注射重组粒细胞集落刺激因子(G-CSF)进行治疗。在证实犬重组G-CSF可使患病犬的中性粒细胞计数增加后,我们肌肉注射了2×10⁹感染单位(IU)编码犬G-CSF cDNA的慢病毒载体。获得了近18个月的升高的、治疗性中性粒细胞生成。慢病毒载体治疗使中性粒细胞平均计数达到29,230±12,930个/微升,与预处理值(5,240±4,800个/微升;p<0.0001)和G-CSF给药期间的中性粒细胞计数(17,820±11,100个/微升;p<0.0001)相比均显著增加。通过向正常犬全身输注重组G-CSF,我们估计2×10⁹IU的慢病毒每天每千克可递送3.5微克G-CSF。慢病毒载体治疗后,灰色柯利犬体重增加,未出现感染和发热的临床症状,且不再需要饲养在无病原体环境中。从注射部位肌肉采集的基因组DNA检测到前病毒呈阳性,而性腺、肺、脾、心、肝、肾和未注射肌肉的样本均为阴性。这些研究表明,成年动物对慢病毒介导的G-CSF递送具有长期反应性,提示这种方法可能适用于治疗患有周期性和其他中性粒细胞减少症的成年患者。