腺相关病毒（AAV）在动物实验中的应用（二）
三、肝脏
AAV2、AAV5、AAV7 和 AAV8，其中 AAV7 和 AAV8 的效率是 AAV2 的 10-100 倍。注射部位常选择门静脉、外周静脉和尾静脉。注射病毒滴度一般选择 8×10^10-2×10^12(GC/kg)（人、灵长类）；1×10^11 GC/只鼠，稀释成 50-100 μl 进行尾静脉注射；注射 2 周之后可以检测。下图为肝脏注射的一些情况：
Fig 6. In vivo vector specificity analysis
四、心脏和动脉
对心脏和动脉而言，常用血清型有：AAV1、AAV6、AAV8 和 AAV9 。其中 AAV2、AAV5、AAV7 也可以使用，但是表达比较慢，3 个月后表达量才能与 AAV1、AAV6 表达量相当。所有血清型中 AAV8 和 AAV9 最有效，2 个月左右其表达量达到最高。
常见的注射部位为冠状动脉、主动脉、心肌和尾静脉。注射病毒滴度为 1.9×10^11-10^12 GC/ml，稀释成 100 μl- 250 μl（大鼠剂量稍高于小鼠）注射；但是对于心脏原点注射而言，常采用 10^10 GC/ml 病毒滴度，稀释成 20 μl 使用。检测时间：一般在2周之后可以检测。下图为注射的一些情况：
Biodistribution of LacZ expression and vector genomes 4 weeks after direct injection of 5 ×1011 vg of AAV8- and AAV9-CB-LacZ into the rat heart., heart; Li, liver; Lu, lung; Br, brain; Te, testis; Ki, kidney; Sp, spleen; St, stomach; Ga, gastrocnemius.
HUMAN GENE THERAPY 19:1359–1368
五、肺脏
肺脏常用血清型有 AAV1、AAV2、AAV5、AAV6 和 AAV9。由于肺脏表面唾液酸受体分布比较多，因此 AAV5 型比较常用；相比之下 AAV2 和 AAV6 转导效率相当。常见的注射方式有滴鼻、雾化吸入或气管内注射，注射病毒滴度为 10^12 GC/ml ，稀释成 3 μl，注射完之后 2 周可以检测表达情况。下图为注射的一些情况：
Histochemical detection of AP expression in mouse lungs 1 month after vector exposure.
JOURNAL OF VIROLOGY, Feb. 2000, p. 1524–1532
六、肾脏
肾脏常用血清型有 AAV2、AAV8 和 AAV9。注射病毒的滴度一般选择 1-5×10^11 GC/ml，稀释成 50-200 μl 注射。常见的注射部位有肾动脉、肾静脉和左髂脉、腹主动脉，注射完 2 周之后可以检测表达情况。下图为注射的一些情况：
In vivo long-term transduction by AAV serotype 2 vector into the rat kidney.
Nephron Exp Nephrol 2004;96 :e119–e126
七、肌肉
肌肉组织常用血清型有 AAV2、AAV8 和 AAV9。其中 AAV8、AAV9 最常用，心肌、胰腺和肾上腺也是如此。骨骼肌分为快速和慢速肌纤维，AAV2 常作用于慢速肌纤维，AAV6 则对二者都有效。对不同年龄阶段的鼠，注射剂量有所不同：幼鼠选择的病毒量为 7×10^10-1.2×10^11 GC/g；成年鼠选择的病毒量为 3.5 ×10^11-7×10^12 GC/只；但是低剂量的 AAV9 不能到达所有心肌细胞。下图为注射的一些情况：
Transduction of muscle fibersby different AAV serotypes.
J Gene Med 2005; 7: 442–451.
Representative photomicrographs of sections of skeletal muscle
MOLECULAR THERAPY Vol. 14, No. 1, July 2006
八、肠
肠组织常用血清型有 AAV1、AAV2 和 AAV5。常见的注射方式有口服、灌肠、腹腔注射、肠系膜动脉注射。一般选择注射滴度 1×10^11 GC/ml ,稀释成 100 μl（PBS或者稀释液稀释）使用，注射 2 周后可以检测。
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