Trifilo Matthew J, Ying Ge, Teng Chao, Oldstone Michael B A
Viral-Immunobiology Laboratory, Molecular and Integrative Neurosciences Department, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037, USA.
Virology. 2007 Aug 15;365(1):136-43. doi: 10.1016/j.virol.2007.03.032. Epub 2007 Apr 23.
To study the pathogenesis of chronic wasting disease (CWD) in deer and elk, transgenic (tg) mice were generated that expressed the prion protein (PrP) of deer containing a glycine at amino acid (aa) 96 and a serine at aa 225 under transcriptional control of the murine PrP promoter. This construct was introduced into murine PrP-deficient mice. As anticipated, neither non-tg mice nor PrP ko mice were susceptible when inoculated intracerebrally (i.c.) or orally with CWD brain material (scrapie pool from six mule deer) and followed for 600+ days (dpi). Deer PrP tg mice were not susceptible to i.c. inoculation with murine scrapie. In contrast, a fatal neurologic disease occurred accompanied by conversion of deer PrPsen to PrPres by western blot and immunohistochemistry after either i.c. inoculation with CWD brain into two lines of tg mice studied (312+32 dpi [mean+2 standard errors] for the heterozygous tg line 33, 275+46 dpi for the heterozygous tg line 39 and 210 dpi for the homozygous tg line 33) or after oral inoculation (381+55 dpi for the homozygous tg line 33 and 370+26 dpi for the homozygous tg line 39). Kinetically, following oral inoculation of CWD brain, PrPres was observed by day 200 when mice were clinically healthy in the posterior surface of the dorsum of the tongue primarily in serous and mucous glands, in the intestines, in large cells at the splenic marginal zone that anatomically resembled follicular dendritic cells and macrophages and in the olfactory bulb and brain stem but did not occur in the cerebellum, cerebral cortex or hippocampus or in hearts, lungs and livers of infected mice. After 350 days when mice become clinically ill the cerebellum, cerebral cortex and hippocampus became positive for PrPres and displayed massive spongiosis, neuronal drop out, gliosis and florid plaques.
为研究鹿和麋鹿慢性消耗病(CWD)的发病机制,构建了转基因(tg)小鼠,其在小鼠朊病毒蛋白(PrP)启动子的转录控制下,表达在氨基酸(aa)96位含有甘氨酸且在aa 225位含有丝氨酸的鹿朊病毒蛋白(PrP)。将该构建体导入小鼠PrP基因缺陷型小鼠。不出所料,当脑内(i.c.)或经口接种CWD脑材料(来自六只骡鹿的羊瘙痒病混合样本)并观察600多天(dpi)时,非转基因小鼠和PrP基因敲除小鼠均不易感。鹿PrP转基因小鼠对脑内接种小鼠羊瘙痒病不易感。相比之下,在研究的两系转基因小鼠中,脑内接种CWD脑材料后(杂合转基因系33为312 + 32 dpi [平均值 + 2个标准误],杂合转基因系39为275 + 46 dpi,纯合转基因系33为210 dpi)或经口接种后(纯合转基因系33为381 + 55 dpi,纯合转基因系39为370 + 26 dpi),均出现致命性神经疾病,同时通过蛋白质印迹法和免疫组织化学法检测到鹿PrPsen转化为PrPres。从动力学角度来看,经口接种CWD脑后,在第200天时,当小鼠临床健康时,在舌背后面主要在浆液腺和黏液腺、肠道、脾边缘区形态上类似滤泡树突状细胞和巨噬细胞的大细胞、嗅球和脑干中观察到PrPres,但在小脑、大脑皮层或海马体以及感染小鼠的心脏、肺和肝脏中未出现。350天后当小鼠出现临床疾病时,小脑、大脑皮层和海马体的PrPres呈阳性,并出现大量海绵状变性、神经元脱失、胶质细胞增生和明显的斑块。