Intratracheal gene delivery with adenoviral vector induces elevated systemic IgG and mucosal IgA antibodies to adenovirus and beta-galactosidase.
Intratracheal gene delivery with adenoviral vector induces elevated systemic IgG and mucosal IgA antibodies to adenovirus and beta-galactosidase.
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使用腺病毒载体进行气管内基因递送可诱导针对腺病毒和 β-半乳糖苷酶的全身 IgG 和粘膜 IgA 抗体升高。
DOI：
10.1089/hum.1995.6.7-895
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Pascual,DW
中科院分区：
文献类型：
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作者：
VanGinkel,FW;Liu,C;Simecka,JW;Dong,JY;Greenway,T;Frizzell,RA;Kiyono,H;McGhee,JR;Pascual,DW
使用腺病毒载体重复基因递送至肺上皮细胞的一个主要问题是诱导对载体的免疫应答，从而阻碍有效的基因转导。为了评估对腺病毒载体的免疫应答，重复的肠内（i.t.）使用携带lacZ基因的复制缺陷型腺病毒5（Ade 5）载体在CD-1小鼠中进行基因给药，并与通过常规鼻内（i. n.）和腹膜内（i. p.）免疫途径。评价血清IgG、伊加和IgM抗体对腺病毒载体和β-半乳糖苷酶（β-Gal）的反应动力学。通过i.t.给予两到三剂腺病毒载体，即，或腹膜内途径导致血清IgG滴度超过1：200，000，而血清IgM和伊加被中度诱导。主要鼠IgG亚类的分析确定为IgG 2带IgG 2a。为了确定该抗体应答的定位，采用ELISP 0 T测定。从肺、下呼吸道淋巴结（LRLN）、鼻道（NP）和脾中分离淋巴细胞。对于i.t.和i.n.-在给予Ade 5和β-Gal的小鼠中，对Ade 5和β-Gal的最高伊加斑点形成细胞（SFC）应答位于NP和肺中。肺和LRLN均显示Ade 5和β-Gal的IgG SFC数量升高（比脾IgG SFC应答高4至12倍）。这一证据表明，肺和相关淋巴组织是血清抗体的来源。血清抗体的进一步分析表明，和信息技术-给药组产生了最大的Ade 5中和滴度，表明重复基因转移的有效性降低部分是由于循环中和抗体。因此，重复的i.t.滴注将刺激对载体的局部和全身抗体应答。
One major concern about using adenoviral vectors for repetitive gene delivery to lung epithelial cells is the induction of an immune response to the vector, thus, impeding effective gene transduction. To assess the immune response to the adenoviral vector, repetitive intratracheal (i.t.) gene dosing was performed in CD-1 mice using the replication-deficient adenovirus 5 (Ade5) vector carrying thelacZgene, and compared to the antibody responses induced by conventional intranasal (i.n.) and intraperitoneal (i.p.) routes of immunization. Kinetics of serum IgG, IgA, and IgM antibody responses to the adenoviral vector and toβ-galactosidase (β-Gal) were evaluated. Two or three adenoviral vector doses given by i.t., i.n., or i.p. routes resulted in serum IgG titers in excess of 1:200,000, whereas serum IgM and IgA were moderately induced. Analysis of the predominant murine IgG subclass was determined to be IgG2band IgG2a. To determine the localization of this antibody response, the ELISPOT assay was employed. Lymphocytes were isolated from the lung, the lower respiratory lymph nodes (LRLN), the nasal passages (NP), and the spleen. For i.t- and i.n.-administered mice, the highest IgA spot-forming cell (SFC) response to Ade5 andβ-Gal was located in the NP and in the lung. Both the lung and the LRLN showed elevated numbers of IgG SFCs (4- to 12-fold greater than splenic IgG SFC response) for Ade5 andβ-Gal. This evidence suggests that the lung and associated lymphoid tissues were the source for serum antibodies. Further analysis of serum antibodies showed that the i.p.- and i.t.-administered groups yielded the greatest neutralization titers to Ade5, suggesting that the reduced effectiveness of repetitive gene transfer is in part due to circulating neutralizing antibodies. Thus, repetitive i.t. instillation will stimulate a localized and systemic antibody response to the vector.