Intratracheal Gene Delivery with Adenoviral Vector Induces Elevated Systemic IgG and Mucosal IgA Antibodies to Adenovirus andβ-Galactosidase

Abstract

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 the lacZ gene, 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 to beta-galactosidase (beta-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 IgG2b and 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 and beta-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 and beta-Gal. This evidence suggests that the lung and associated lymphoid tissues were the source for serum antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)

Simian immunodeficiency virus-based lentivirus vector for retinal gene transfer: a preclinical safety study in adult rats

“…This hypothesis, however, does not seem to be likely, because the current construct of SIV is a self-inactivating (SIN) vector which lacks any promoter activity in its LTRs, 19 and theoretically expresses no virus-related genes except for the transgene driven by an exogenous promoter in the expression cassette. This finding may be supported by a few recent papers, including Doi et al, 25 who demonstrated that no retinal damage was seen in the eyes treated with empty HIV-based lentivirus vector. Another hypothesis is a host immune response against the transgene products, including b-galactosidase and GFP, which can evoke CTL and cytotoxic antibodies.…”

“…[26][27][28] Recent reports also suggested that the gradual reduction of GFP expression might be because of a host immune response against GFP itself when using the lentivirus vector. 22,25 However, these findings may not be sufficient to conclude these issues, because they used only one reporter gene (GFP). In contrast, we herein demonstrated that a loss of transgene expression was found in a dosedependent manner and the expression of both GFP and lacZ were not rejected when using low titer viruses.…”

Effects of bronchopulmonary inflammation induced by Pseudomonas aeruginosa on adenovirus-mediated gene transfer to airway epithelial cells in mice

“…[5][6][7][8][9][10] Several potential barriers to successful adenovirus-mediated gene transfer to the airways have been recognized which include the need for repeated delivery to maintain transgene expression, 11 the host inflammatory response to the vector, and specific immune responses to adenovirus vectors. [12][13][14] In vivo models used so far to conduct published studies of adenovirus-mediated gene transfer to the lungs have employed animals with uninfected, uninflamed lungs at the time of vector administration. [11][12][13][14][15][16][17][18][19][20] Given the prominence of the chronic infection and inflammation in the airways of CF patients, we examined the influence of chronic bronchopulmonary inflammation induced by P. aeruginosa on the delivery of transgenes to airway epithelium.…”

“…[12][13][14] In vivo models used so far to conduct published studies of adenovirus-mediated gene transfer to the lungs have employed animals with uninfected, uninflamed lungs at the time of vector administration. [11][12][13][14][15][16][17][18][19][20] Given the prominence of the chronic infection and inflammation in the airways of CF patients, we examined the influence of chronic bronchopulmonary inflammation induced by P. aeruginosa on the delivery of transgenes to airway epithelium. In the current studies, we tested the hypo-thesis that lung inflammation in mice induced by P. aeruginosa will be associated with a quantitative reduction in short-term, adenovirus-mediated reporter gene expression in airway epithelial cells.…”

Effect of fractalkine-Fc delivery in experimental lung metastasis using DNA/704 nanospheres

“…15 Indeed, the ultimate aim of this area of research was for the development of a cellular therapy for humans and the literature was divided on the long-term impact that MSCs have on tumor growth with both growth stimulatory and inhibitory effects described, 27 as well as on the risk of using adenoviral vector. 28,29 Non-viral methods could 10 Indeed, partially dependent on the level of gene transfer, the chemokine concentration at tumor site plays a critical role in inducing tumor regression. As the efficiencies of non-viral gene transfer methods are usually considered to be lower than those of cell-or viral-mediated methods, we can suppose that prolonged circulating half-life of FKN-Fc is required to compensate reduced efficiency of 704-mediated gene transfer.…”