Adenovirus capsid-display of the retro-oriented human complement inhibitor DAF reduces Ad vector–triggered immune responses in vitro and in vivo

Seregin, Sergey S.; Aldhamen, Yasser A.; Appledorn, Daniel M.; Hartman, Zachary C.; Schuldt, Nathaniel J.; Scott, Jeannine M.; Godbehere, Sarah; Jiang, Haixiang; Frank, Michael M.; Amalfitano, Andrea

doi:10.1182/blood-2010-03-276949

Cited by 38 publications

(62 citation statements)

References 46 publications

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“…To this end, we directly quantified the abundance of cytokines and chemokines in the plasma of mice treated with Ad5-VCA0956 using a multiplexed assay system at 6 and 24 h.p.i. Consistent with prior studies showing that the adenovirus vector stimulates the secretion of proinflammatory cytokines and chemokines (27,28), we observed 9 cytokines and chemokines that were modestly induced in the Ad5-null-treated mice compared to the naive mice (Ͼ3-fold) (IFN-␥, MCP-1, G-CSF, MIP-1␣, IL-6, MIP-1␤, IL12p40, KC, and RANTES), and these differences were greatest at the 6-hour time point (Fig. 6).…”

Section: Synthesis Of C-di-gmp In Vivosupporting

confidence: 76%

“…The transfection efficiency was determined to be 18.2%, which was obtained by transfecting HeLa cells with a plasmid containing green fluorescent protein (GFP) under CMV promoter control and measuring the percentage of GFP-positive cells using flow cytometry. The infection efficiency of HeLa cells was determined to be 82.2% by infecting HeLa cells with Ad5-gfp (28) and quantifying the percentage of GFP-positive cells using flow cytometry.…”

Section: Methodsmentioning

confidence: 99%

“…Recombination, viral propagation of the Ad5 vectors, and subsequent virus characterization were performed as previously described (27,28). Viral particle (vp) numbers were determined by optical density measurement at 260 nm and validated as previously described (29).…”

Section: Methodsmentioning

confidence: 99%

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Stimulation of Innate Immunity byIn VivoCyclic di-GMP Synthesis Using Adenovirus

Koestler

Seregin

Rastall

et al. 2014

Clin. Vaccine Immunol.

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The bacterial second messenger cyclic di-GMP (c-di-GMP) stimulates inflammation by initiating innate immune cell recruitment and triggering the release of proinflammatory cytokines and chemokines. These properties make c-di-GMP a promising candidate for use as a vaccine adjuvant, and numerous studies have demonstrated that administration of purified c-di-GMP with different antigens increases protection against infection in animal models. Here, we have developed a novel approach to produce c-di-GMP inside host cells as an adjuvant to exploit a host-pathogen interaction and initiate an innate immune response. We have demonstrated that c-di-GMP can be synthesized in vivo by transducing a diguanylate cyclase (DGC) gene into mammalian cells using an adenovirus serotype 5 (Ad5) vector. Expression of DGC led to the production of c-di-GMP in vitro and in vivo, and this was able to alter proinflammatory gene expression in murine tissues and increase the secretion of numerous cytokines and chemokines when administered to animals. Furthermore, coexpression of DGC modestly increased T-cell responses to a Clostridium difficile antigen expressed from an adenovirus vaccine, although no significant differences in antibody titers were observed. This adenovirus c-di-GMP delivery system offers a novel method to administer c-di-GMP as an adjuvant to stimulate innate immunity during vaccination.

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Section: Synthesis Of C-di-gmp In Vivosupporting

confidence: 76%

Section: Methodsmentioning

confidence: 99%

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Stimulation of Innate Immunity byIn VivoCyclic di-GMP Synthesis Using Adenovirus

Koestler

Seregin

Rastall

et al. 2014

Clin. Vaccine Immunol.

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“…In one example, the Ad vector tolerated capsid display of the human decay-accelerating factor (DAF), a natural complement inhibitor. Introduction and in-frame genetic fusion of the complement inhibitory portions of the DAF protein onto the C-terminus of the Ad capsid cement protein pIX have been previously confirmed by our group to be viable (Seregin et al, 2010a). These DAF-displaying Ad vectors were not only confirmed to retain the native Ad vectors' ability to transduce genes both in vitro and in vivo, but the unique Ad vectors were also confirmed to be significantly safer; this was validated primarily by the vectors having a minimized induction of typical Ad capsid-triggered innate immune responses, coupled with a reduced activation of immune effector cell types early after administration (Seregin et al, 2010a).…”

mentioning

confidence: 99%

Use of DAF-Displaying Adenovirus Vectors Reduces Induction of Transgene- and Vector-Specific Adaptive Immune Responses in Mice

et al. 2011

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Adenovirus (Ad)-based vectors are attractive candidates for a variety of gene-transfer applications. In this study, we found that decay-accelerating factor (DAF)-displaying Ads induce significantly decreased cellular immune responses to transgenes expressed from the vectors in both Ad5-naive and Ad5-immune mice. Specifically, we found a diminished ability of splenocytes to secrete interferon-g after recall exposure to multiple peptides derived from antigens expressed by DAF-displaying Ads. We also confirmed that DAF-displaying Ads induce decreased numbers of antigen-specific, CD8þ effector memory and central memory CD8 þ T cells, thereby uncovering a unique role of complement in modulating the induction of robust memory T-cell responses. We also confirmed that DAF-displaying Ads generate significantly reduced titers of Ad capsid-specific neutralizing antibodies after gene transfer in vivo. In conclusion, DAF-displaying Ad5-based vectors exhibit decreased induction of complement-dependent, innate immune responses, resulting in both an improved safety profile and a decreased propensity to induce humoral and cellular adaptive immune responses to Ad capsid proteins and Ad vector-expressed transgene products. This attractive combination of features will be beneficial in a variety of clinically relevant gene-transfer applications.

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“…Hoeben and colleagues (18,77) showed that pIX fused to a hyperstable antibody directed to ␤-galactosidase or a single-chain T-cell receptor targeted to MAGE-A1 antigen was efficiently incorporated into the Ad capsid and bound its ligand. Other research groups have used pIX to display complement-inhibiting polypeptides (64,65). Finally, C-terminal cysteines added to pIX have been used as a target for the chemical addition of targeting ligands bearing reactive thiol groups (15).…”

mentioning

confidence: 99%

Retargeting of Adenovirus Vectors through Genetic Fusion of a Single-Chain or Single-Domain Antibody to Capsid Protein IX

Poulin

Lanthier

Smith

et al. 2010

J Virol

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Adenovirus (Ad) vectors are the most commonly used system for gene therapy applications, due in part to their ability to infect a wide array of cell types and tissues. However, many therapies would benefit from the ability to target the Ad vector only to specific cells, such as tumor cells for cancer gene therapy. In this study, we investigated the utility of capsid protein IX (pIX) as a platform for the presentation of single-chain variable-fragment antibodies (scFv) and single-domain antibodies (sdAb) for virus retargeting. We show that scFv can be displayed on the capsid through genetic fusion to native pIX but that these molecules fail to retarget the virus, due to improper folding of the scFv. Redirecting expression of the fusion protein to the endoplasmic reticulum (ER) results in correct folding of the scFv and allows it to recognize its epitope; however, ER-targeted pIX-scFv was incorporated into the Ad capsid at a very low level which was not sufficient to retarget virus infection. In contrast, a pIX-sdAb construct was efficiently incorporated into the Ad capsid and enhanced virus infection of cells expressing the targeted receptor. Taken together, our data indicate that pIX is an effective platform for presentation of large targeting polypeptides on the surface of the virus capsid, but the nature of the ligand can significantly affect its association with virions.

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Adenovirus capsid-display of the retro-oriented human complement inhibitor DAF reduces Ad vector–triggered immune responses in vitro and in vivo

Cited by 38 publications

References 46 publications

Stimulation of Innate Immunity byIn VivoCyclic di-GMP Synthesis Using Adenovirus

Stimulation of Innate Immunity byIn VivoCyclic di-GMP Synthesis Using Adenovirus

Use of DAF-Displaying Adenovirus Vectors Reduces Induction of Transgene- and Vector-Specific Adaptive Immune Responses in Mice

Retargeting of Adenovirus Vectors through Genetic Fusion of a Single-Chain or Single-Domain Antibody to Capsid Protein IX

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