Subretinal delivery of adeno-associated virus serotype 2 results in minimal immune responses that allow repeat vector administration in immunocompetent mice.
Subretinal delivery of adeno-associated virus serotype 2 results in minimal immune responses that allow repeat vector administration in immunocompetent mice.
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DOI：
10.1002/jgm.1327
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发表时间：
2009-06
影响因子：
3.5
通讯作者：
Ali, Robin R.
中科院分区：
文献类型：
--
作者：
Barker, Susie E.;Broderick, Cathryn A.;Robbie, Scott J.;Duran, Yanai;Natkunarajah, Mythili;Buch, Prateek;Balaggan, Kamaljit S.;MacLaren, Robert E.;Bainbridge, James W. B.;Smith, Alexander J.;Ali, Robin R.
腺相关病毒血清型2(AAV2)载体在眼部基因转移中显示出良好的应用前景。然而，有效的长期治疗的一个潜在障碍是针对载体或转基因产品的免疫反应的发展。我们评估了单次和重复注射AAV2后小鼠的细胞和体液反应，并检测了它们对RPE65和绿色荧光蛋白转基因表达的影响。在视网膜下注射载体后，在与AAV2共培养的刺激下，引流淋巴结中的脾细胞和T细胞表现出最小的激活。在任何服用AAV2的小鼠的眼液中都没有检测到中和抗体(NAB)，在接受较低剂量的小鼠的血清中也没有检测到。接受较高剂量载体注射的小鼠的血清中存在一定比例的NaB。此外，治疗组和对照组小鼠的血清或眼液中针对RPE65蛋白或AAV2衣壳的免疫球蛋白滴度没有差异。组织学检查显示，与未注射的眼睛相比，没有视网膜毒性或白细胞渗透的证据。小剂量AAV.hRPE65.hRPE65对RPE65−/−小鼠的双眼重复注射可导致转基因表达和功能挽救，但再次注射大剂量AAV2可提高第二只眼的NAB滴度和可变的转基因表达。这些在小鼠身上获得的数据表明，在视网膜下注射AAV2后，对AAV2的免疫反应是剂量依赖的。低剂量AAV2在眼内的耐受性良好，免疫反应最小，重复给药后转基因表达是可以实现的。较高的剂量会导致NABS的表达，从而降低重复给药的效果。版权所有©2009 John Wiley&Sons，Ltd.
Adeno-associated virus serotype 2 (AAV2) vectors show considerable promise for ocular gene transfer. However, one potential barrier to efficacious long-term therapy is the development of immune responses against the vector or transgene product. We evaluated cellular and humoural responses in mice following both single and repeated subretinal administration of AAV2, and examined their effects on RPE65 and green fluorescent protein transgene expression. Following subretinal administration of vector, splenocytes and T-cells from draining lymph nodes showed minimal activation following stimulation by co-culture with AAV2. Neutralizing antibodies (NAbs) were not detected in the ocular fluids of any mice receiving AAV2 or in the serum of mice receiving a lower dose. NAbs were present in the serum of a proportion of mice receiving a higher dose of the vector. Furthermore, no differences in immunoglobulin titre in serum or ocular fluids against RPE65 protein or AAV2 capsid between treated and control mice were detected. Histological examination showed no evidence of retinal toxicity or leukocyte infiltration compared to uninjected eyes. Repeat administration of low-dose AAV.hRPE65.hRPE65 to both eyes of RPE65−/− mice resulted in transgene expression and functional rescue, but re-administration of high-dose AAV2 resulted in boosted NAb titres and variable transgene expression in the second injected eye. These data, which were obtained in mice, suggest that, following subretinal injection, immune responses to AAV2 are dose-dependent. Low-dose AAV2 is well tolerated in the eye, with minimal immune responses, and transgene expression after repeat administration of vector is achievable. Higher doses lead to the expression of NAbs that reduce the efficacy of repeated vector administration. Copyright © 2009 John Wiley & Sons, Ltd.
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影响因子：
20.3
作者：
Jiang, Haiyan;Couto, Linda B.;Pierce, Glenn F.
通讯作者：
Pierce, Glenn F.
影响因子：
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Camelo, S;Kezic, J;McMenamin, PG
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McMenamin, PG