Anand V, Chirmule N, Fersh M, Maguire A M, Bennett J
Department of Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia 19104-2689, USA.
Hum Gene Ther. 2000 Feb 10;11(3):449-57. doi: 10.1089/10430340050015914.
Subretinal delivery of recombinant adeno-associated virus (rAAV) results in a systemic humoral response in the adult immunocompetent mouse. We characterized this response and determined whether it is possible to readminister rAAV to the subretinal space despite the presence of antibodies to the vector. A systemic humoral response to rAAV capsid proteins was induced by either unilateral subretinal injection or by intradermal administration of 1 x 10(9) infectious units of rAAV carrying the cDNA encoding green fluorescent protein (GFP), rAAV-GFP. Experiments were performed in cohorts of adult C57BL/6 mice. Assessment of systemic humoral response to viral capsid protein was performed through enzyme-linked immunoabsorbent assay (ELISA) and infection inhibition studies of serum samples 3 weeks after virus delivery. The rAAV-GFP virus was readministered by subretinal injection. GFP expression after subretinal administration was evaluated ophthalmoscopically throughout the course of the experiment and histologically at the termination of the experiment. We observed significant systemic humoral responses to viral capsid protein after subretinal delivery of rAAV. Intradermal injection resulted in a larger humoral response (with a higher percentage of neutralizing antibodies) than subretinal injection. Additional transduction events were observed after readministration of rAAV despite the presence of strong humoral response to the vector.
在成年具有免疫活性的小鼠中，重组腺相关病毒（rAAV）的视网膜下递送会引发全身性体液反应。我们对这种反应进行了表征，并确定在存在针对载体的抗体的情况下，是否有可能再次向视网膜下间隙注射rAAV。通过单侧视网膜下注射或皮内注射1×10⁹感染单位携带编码绿色荧光蛋白（GFP）的cDNA的rAAV（rAAV-GFP），诱导对rAAV衣壳蛋白的全身性体液反应。实验在成年C57BL/6小鼠群体中进行。在病毒递送3周后，通过酶联免疫吸附测定（ELISA）和血清样本的感染抑制研究，评估对病毒衣壳蛋白的全身性体液反应。通过视网膜下注射再次给予rAAV-GFP病毒。在整个实验过程中通过检眼镜检查评估视网膜下给药后的GFP表达，并在实验结束时进行组织学评估。我们观察到rAAV视网膜下递送后对病毒衣壳蛋白有显著的全身性体液反应。皮内注射比视网膜下注射产生更大的体液反应（具有更高百分比的中和抗体）。尽管对载体存在强烈的体液反应，但在再次给予rAAV后仍观察到额外的转导事件。
