Long‐term humoral and cellular immunity induced by a single immunization with replication‐defective adenovirus recombinant vector

Abstract

This study examines the suitability of replication-defective adenovirus vectors for engineering recombinant vaccines. The immunological abilities and limitations of E1-deleted adenoviruses containing the lacZ gene (Ad-beta-gal) were investigated by examining the humoral and cellular immune responses to the beta-galactosidase protein. BALB/c mice (H-2d) were given in a single injection of recombinant adenovirus. The cytotoxic T lymphocyte (CTL) response of spleen cells was evaluated. Recognized target cells were H-2d-derived tumor cells transfected by the lac Z gene, or incubated with the 876-884 beta-galactosidase peptide known to be restricted by the Ld molecule of the major histocompatibility complex. A long-lasting beta-galactosidase-specific cytotoxic T cell response was obtained. By contrast, CTL from mice immunized with the Ld-restricted peptide were less specific for the endogenous epitope presented by the transfectants expressing beta-galactosidase. Ad-beta-gal-immunized mice were also protected against an intra-cerebral challenge with a recombinant vaccinia virus expressing the lac-Z gene. These results suggest that Ad-beta-gal-induced CTL have protective abilities in vivo. The induction of beta-galactosidase-specific T helper lymphocytes and humoral IgG responses were also examined. A proliferative response occurred only late after immunization and the primed T lymphocytes produced interleukin-2, but no interleukin-4. A humoral IgG response to the beta-galactosidase protein was detected 15-30 days after a single immunization and remained stable for 6 months without boosting. Lastly, we followed the evolution of the immune response over the course of successive immunizations. The magnitude and kinetics of the cellular and humoral responses were similar to those obtained after a single immunization. Consistent with these observations, an adenovirus-specific neutralizing antibody response was detected as early as the second immunization. Thus, a single immunization with a replication-defective adenovirus recombinant vector induces long-lasting humoral and cellular immune responses specific to the transgene product.

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

“…Adenovirus has been known to induce specific host inflammatory responses in healthy people and animals, even after a single administration, 12 although adenovirus vectors with reduced immunogenicity and inflammatory potential are being developed.…”

“…[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.…”

Adenovirus-mediated gene transfer in dog prostate: a preclinical study of a relevant model system for gene therapy of human prostatic cancer

“…29 ± 31 However, in¯ammatory cells are also directly responsible for the transient expression of the transgene; in addition, the setting of an adenovirus-speci®c humoral immune response makes adenoviral readministrations dif®cult. Two major factors that are potentially responsible for the transient transgene expression and the in¯am-mation mediated by AMGT are the transgene itself 21,32,33 and the viral antigens. 28,34 ± 38 To optimize the persistence Figure 5 Detection by ELISA of adenovirus-speci®c IgM in the serum one and seven days after the intraprostatic delivery of AdRSVbgal.…”

“…21 Sera were incubated at 56 C for 30 min to inactivate complement. Serial dilutions of dog sera (1:40 ± 1:1280) in MEM supplemented with 10% FCS were incubated with 5610 5 pfu of AdRSVbgal for 1 h at 37 C. Each mixture was then used to infect 293 cells plated in a 96-well dish (5610 4 cells/well).…”