Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells

Denis-Mize, Kimberly; Dupuis, Marc; MacKichan, Mary Lee; Singh, Manmohan; Doe, Barbara; O’Hagan, Derek T.; Ulmer, Jeffrey B.; Donnelly, John; McDonald, Donald M.; Ott, Gary

doi:10.1038/sj.gt.3301347

Cited by 150 publications

(68 citation statements)

References 36 publications

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“…Substantially increased antibody titers may be a result of prolonged expression of antigen in situ, as was seen in mice (54). In vitro studies have shown that PLG microparticles protect plasmid DNA from nuclease digestion (12), thereby potentially prolonging the availability of the plasmid in vivo. This may be important, since the amount of antigen produced in situ by a DNA vaccine is very small (60).…”

Section: Discussionmentioning

confidence: 99%

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Enhanced Potency of Plasmid DNA Microparticle Human Immunodeficiency Virus Vaccines in Rhesus Macaques by Using a Priming-Boosting Regimen with Recombinant Proteins

Otten¹,

Schaefer²,

Doe³

et al. 2005

J Virol

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DNA vaccines have been used widely in experimental primate models of human immunodeficiency virus (HIV), but their effectiveness has been limited. In this study, we evaluated three technologies for increasing the potency of DNA vaccines in rhesus macaques. These included DNA encoding Sindbis virus RNA replicons (pSINCP), cationic poly(lactide-co-glycolide) (PLG) microparticles for DNA delivery, and recombinant protein boosting. The DNA-based pSINCP replicon vaccines encoding HIV Gag and Env were approximately equal in potency to human cytomegalovirus (CMV) promoter-driven conventional DNA vaccines (pCMV). The PLG microparticle DNA delivery system was particularly effective at enhancing antibody responses induced by both pCMV and pSINCP vaccines and had less effect on T cells. Recombinant Gag and Env protein boosting elicited rapid and strong recall responses, in some cases to levels exceeding those seen after DNA or DNA/PLG priming. Of note, Env protein boosting induced serum-neutralizing antibodies and increased frequencies of gamma interferon-producing CD4 T cells severalfold. Thus, PLG microparticles are an effective means of delivering DNA vaccines in nonhuman primates, as demonstrated for two different types of DNA vaccines encoding two different antigens, and are compatible for use with DNA prime-protein boost regimens.Both neutralizing antibodies and cell-mediated immunity (CMI) likely will be required to protect against viruses that can establish chronic infections, such as human immunodeficiency virus (HIV) and hepatitis C virus (HCV). Moreover, in animal models for HIV, both neutralizing antibodies against Env (9, 35-37) and cytotoxic T lymphocytes (CTL) that target multiple viral antigens (3-5, 17, 18, 21, 26, 30, 32, 38, 39, 45, 48, 51-53, 59) can contribute to protection through prevention of infection and clearance of virus-infected cells, respectively. Vaccines consisting of inactivated pathogens or recombinant proteins generally are not effective at inducing CTL and typically are used to induce protective antibodies. In contrast, viruses and intracellular bacteria can induce CTL responses, in part due to neoexpression of the antigens during infection.Plasmid DNA vaccines were born out of the need for inducing both antibody and CMI responses, including CTL, without the problems associated with live organism-based vaccines, such as potential reversion to virulence and antivector immunity that can limit boosting. Indeed, DNA vaccines that express antigens from strong viral promoters have been used to elicit protective antibodies and CMI in many animal models (14, 23). However, naked DNA vaccines, i.e., plasmid DNA in saline, have proven to be only modestly potent in humans, thereby limiting their utility. Many approaches have been explored to improve DNA vaccine potency, including better expression vectors, DNA formulation and delivery systems, adjuvants, and the use of booster vaccines.We developed an alternative DNA vector that launches a self-amplifying Sindbis virus (alphavirus) RNA replicon (1...

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Section: Discussionmentioning

confidence: 99%

“…PLG microparticle adsorption captures DNA with high efficiency but still allows for rapid release of the DNA from particles. The mode of action of the PLG delivery system may involve uptake and expression of DNA by APCs as well as prolonged expression of antigen by protecting plasmid DNA from nuclease digestion (12).…”

mentioning

confidence: 99%

Enhanced Potency of Plasmid DNA Microparticle Human Immunodeficiency Virus Vaccines in Rhesus Macaques by Using a Priming-Boosting Regimen with Recombinant Proteins

Otten¹,

Schaefer²,

Doe³

et al. 2005

J Virol

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“…The search for efficient and safe synthetic vaccine carrier systems has yielded various types of biodegradable anionic and cationic microparticles, which are able to induce noticeable immune responses in vivo when loaded with peptide or protein antigens (10,23,33) or plasmid DNA (11,34). The adequate stimulation of DCs is of key importance for the initiation of acquired and innate immune responses by these cells.…”

Section: Discussionmentioning

confidence: 99%

Composition and Surface Charge of DNA-Loaded Microparticles Determine Maturation and Cytokine Secretion in Human Dendritic Cells

Jilek¹,

Ulrich²,

Merkle³

et al. 2004

Pharm Res

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Purpose. Biodegradable microparticles prepared from poly(lactide) (PLA) and poly(lactide-co-glycolide) (PLGA) have been shown to be promising carrier systems for vaccine delivery. Here, we have investigated the capacity of different PLA and PLGA microparticle formulations to induce stimulation of human blood monocyte-derived dendritic cells (DCs). Methods. Stimulation of human derived dendritic cells by plain microparticles were compared with microparticles loaded with plasmid DNA or double-stranded salmon DNA either by encapsulation or adsorption to the surface of cationic microparticles. Stimulation of DCs was monitored by the up-regulation of surface maturation markers CD83 and CD86 and the secretion of IL-12 and TNF-␣. Results. Slowly degrading PLA microparticles did not induce any detectable stimulation or activation of DCs. In contrast, fast degrading PLGA microparticles were able to influence DC maturation and cytokine secretion dependent on their surface charge. Anionic PLGA microparticles induced an up-regulation of CD83 and high TNF-␣ secretion, which was further enhanced up to the level of the potent stimulator lipopolysaccharide (LPS) when plasmid DNA was encapsulated. Moreover, the secretion of significant amounts of IL-12 was observed. Cationic PLGA microparticles induced an up-regulation of CD86 and moderate TNF-␣ secretion, but no IL-12 secretion, with no additional effects in the presence of plasmid DNA. Conclusions. The data suggest that the composition and charge of biodegradable DNA-loaded microparticles profoundly influences maturation and cytokine secretion in DCs. Thus, the individual formulation of microparticles used as a vaccine carrier system might considerably influence the profile of the immune response.

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“…To this end, we have developed a microparticle-based PLG formulation of DNA (25), based on the hypothesis that particles ϳ1 m in diameter would be readily internalized by phagocytic cells, such as immature dendritic cells (16), thereby facilitating nonspecific targeting. Indeed, this formulation is very potent in mice (25) and appears to function, at least in part, by facilitating DNA uptake by APCs (9). PLG has been used previously for delivery of small-molecule drugs (17), proteins (21,22), and DNA (8,15,18,27).…”

Section: Discussionmentioning

confidence: 99%

Induction of Potent Immune Responses by Cationic Microparticles with Adsorbed Human Immunodeficiency Virus DNA Vaccines

O’Hagan¹,

Singh²,

Ugozzoli³

et al. 2001

J Virol

172

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The effectiveness of cationic microparticles with adsorbed DNA at inducing immune responses was investigated in mice, guinea pigs, and rhesus macaques. Plasmid DNA vaccines encoding human immunodeficiency virus (HIV) Gag and Env adsorbed onto the surface of cationic poly(lactide-coglycolide) (PLG) microparticles were shown to be substantially more potent than corresponding naked DNA vaccines. In mice immunized with HIV gag DNA, adsorption onto PLG increased CD8 ؉ T-cell and antibody responses by ϳ100-and ϳ1,000-fold, respectively. In guinea pigs immunized with HIV env DNA adsorbed onto PLG, antibody responses showed a more rapid onset and achieved markedly higher enzyme-linked immunosorbent assay and neutralizing titers than in animals immunized with naked DNA. DNA vaccines have been shown to induce immune responses and protective immunity in many animal models of infectious disease (for a review, see reference 11). In mice, such responses can often be achieved with low doses (Ͻ1 g) of naked DNA. However, the immunogenicity of DNA vaccines in larger animals (e.g., guinea pigs, rabbits, and nonhuman primates) has been much lower than that observed in mice, even at higher doses of DNA. In human clinical trials, certain DNA vaccines have been shown to induce immune responses (5,19,30), but multiple immunizations of high doses of DNA were required. Therefore, in order to provide protective efficacy in humans, the potency of DNA vaccines needs to be increased. So far, it appears that DNA vaccines are more effective at priming T-cell responses than antibodies, as exemplified by induction of cytotoxic T lymphocytes (CTL) but no antibodies against malaria circumsporozoite protein in humans (30). Similarly, we show here that in mice, human immunodeficiency virus (HIV) gag DNA primed CD8 ϩ T-cell responses at doses of DNA 10-to 100-fold lower than that required for priming of antibody responses. Therefore, technologies aimed at increasing the potency of DNA vaccines need to be especially effective at boosting humoral responses.The technology described herein, formulation of DNA onto cationic poly(lactide-coglycolide) (PLG) microparticles, has been developed as a means to better target DNA to antigen-presenting cells (APCs). PLG microparticles are an attractive approach for vaccine delivery, since the polymer is biodegradable and biocompatible and has been used to develop several drug delivery systems (21). In addition, PLG microparticles have also been used for a number of years as delivery systems for entrapped vaccine antigens (24). More recently, PLG microparticles have been described as a delivery system for entrapped DNA vaccines (15,18). Nevertheless, recent observations have shown that DNA is damaged during microencapsulation, leading to a significant reduction in supercoiled DNA (2, 29). Moreover, the encapsulation efficiency is often low. Therefore, we developed a novel approach of adsorbing DNA onto the surface of PLG microparticles to avoid the problems associated with microencapsulation of DNA. This approach, ...

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Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells

Cited by 150 publications

References 36 publications

Enhanced Potency of Plasmid DNA Microparticle Human Immunodeficiency Virus Vaccines in Rhesus Macaques by Using a Priming-Boosting Regimen with Recombinant Proteins

Enhanced Potency of Plasmid DNA Microparticle Human Immunodeficiency Virus Vaccines in Rhesus Macaques by Using a Priming-Boosting Regimen with Recombinant Proteins

Composition and Surface Charge of DNA-Loaded Microparticles Determine Maturation and Cytokine Secretion in Human Dendritic Cells

Induction of Potent Immune Responses by Cationic Microparticles with Adsorbed Human Immunodeficiency Virus DNA Vaccines

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