Lipshutz G S, Flebbe-Rehwaldt L, Gaensler K M
Department of Surgery and, University of California San Francisco, San Francisco, California 94143, USA.
Mol Ther. 2000 Oct;2(4):374-80. doi: 10.1006/mthe.2000.0136.
Adenovirus-mediated gene delivery is limited by the induction of immune responses that produce toxicity and prevent reexpression. To determine whether adenoviral delivery in the preimmune fetus would produce tolerance, we assessed luciferase (luc) expression following sequential pre- and postnatal adenoviral-mediated gene delivery. Day 15 fetuses were injected intrahepatically with 1 x 10(7) pfu of an adenoviral-luc vector (Ad-luc). Following in utero injection, hepatic luc expression persisted 1 month postnatally. No humoral response to adenovirus or luc was detected. Adult mice, previously injected in utero, were reinjected intravenously with 5 x 10(8) pfu of Ad-luc at 3 months of age and again at 6 months with either 5 x 10(8) pfu of Ad-luc or cationic liposome-DNA complexes (CLDC). Following the first postnatal injection, animals injected in utero had levels of luc comparable to those of age-matched naive controls. However, both control and experimental animals subsequently developed antibodies to adenovirus and luc. No further expression was achieved with a second postnatal injection of Ad-luc or with delivery of CLDC-luc. These studies demonstrate that the delivery of adenoviral vectors in utero at E15 does not elicit an immune response. However, delivery of recombinant adenovirus postnatally results in brisk and limiting immune responses regardless of the in utero exposure.
腺病毒介导的基因递送受到免疫反应诱导的限制，这种免疫反应会产生毒性并阻止基因的重新表达。为了确定在免疫前胎儿中进行腺病毒递送是否会产生耐受性，我们评估了在出生前和出生后连续进行腺病毒介导的基因递送后荧光素酶（luc）的表达。在妊娠第15天的胎儿肝内注射1×10⁷ 个腺病毒-luc载体（Ad-luc）的空斑形成单位（pfu）。宫内注射后，肝内luc表达在出生后持续了1个月。未检测到对腺病毒或luc的体液反应。之前在宫内注射过的成年小鼠在3月龄时静脉内再次注射5×10⁸ pfu的Ad-luc，并在6月龄时再次注射5×10⁸ pfu的Ad-luc或阳离子脂质体-DNA复合物（CLDC）。出生后首次注射后，宫内注射的动物的luc水平与年龄匹配的未接触过的对照相当。然而，对照动物和实验动物随后都产生了针对腺病毒和luc的抗体。出生后第二次注射Ad-luc或递送CLDC-luc均未实现进一步的表达。这些研究表明，在胚胎第15天进行宫内腺病毒载体递送不会引发免疫反应。然而，无论宫内是否接触过，出生后递送重组腺病毒都会导致迅速且具有限制性的免疫反应。
