Jump to ContentJump to Main Navigation
Show Summary Details
More options …

European Journal of Nanomedicine

Editor-in-Chief: Hunziker, Patrick / Mollenhauer, Jan

Managing Editor: Löffler, Beat / Salieb-Beugelaar, Georgette

Editorial Board Member: Alexiou, Christoph / Balogh, Lajos / Barenholz, Yechezkel / Dawson, Kenneth / Fadeel, Bengt / Husseini, Ghaleb / Krol, Silke / Lee, Dong Soo / Lehr, Claus-Michael / Mangge, Harald / Müller, Bert / Peer, Dan / Scoles, Giacinto / Serruys, Patrick / Schwartz, Simo / Szebeni, Janos

4 Issues per year


CiteScore 2016: 1.06

SCImago Journal Rank (SJR) 2016: 0.411
Source Normalized Impact per Paper (SNIP) 2016: 0.360

Online
ISSN
1662-596X
See all formats and pricing
More options …

HIV-specific immunotherapy with DermaVir, the first pDNA/PEIm pathogen-like nanomedicine

Julianna Lisziewicz
  • Corresponding author
  • Genetic Immunity Berliniutca 47-49 Budapest 1045, Hungary and 8300 Greensboro Drive Mclean, VA 22102, USA
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Orsolya Lőrincz
  • Genetic Immunity Berliniutca 47-49 Budapest 1045, Hungary and 8300 Greensboro Drive Mclean, VA 22102, USA
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2012-12-19 | DOI: https://doi.org/10.1515/ejnm-2012-0011

Abstract

Eradication of HIV requires the clearance of latently infected cells that remained in the reservoirs after highly active antiretroviral therapy (HAART). DermaVir is the first nanomedicine that induces long-lasting cytotoxic T cells (CTL) capable to kill these HIV-infected cells. DermaVir is a synthetic “pathogen-like” nanomedicine mimicking the size, shape, surface properties, cellular entry, endosomal escape, and antigen expression features of pathogens (e.g., viruses). We can optimize the biological activity of DermaVir during the manufacturing processes by controlling the physico-chemical properties of the nanoparticles that influence its structure and intracellular mode-of-action. In the clinic, targeted delivery of DermaVir to epidermal Langerhans cells is achieved with the DermaPrep medical device. Three clinical trials consistently demonstrated long-lasting CTL induced by DermaVir in HIV-infected people and killing of HIV-infected cells compared to Placebo. Since HAART and DermaVir are complementary, we envision that their combination might be suitable to achieve the cure: HAART to potent viral load suppression and DermaVir to kill latently infected cells that get activated to produce HIV.

Keywords: eradication; HIV cure; intracellular trafficking; Langerhans cell-targeted delivery; lymph node; plasmid DNA; polyethylenimine (PEI); reservoir; vaccine

Nanomedicines in the space of HAART

HIV/AIDS is the leading infectious killer affecting more than 33 million people worldwide (1). HIV/AIDS is treated with the combination of three or more antiretroviral drugs (highly active antiretroviral therapy, HAART), because using any single drug has not been efficient in controlling infection due to the development of resistant strains of the virus. Currently, available HAART is potent in suppressing HIV replication and effective in decreasing HIV RNA level below the limit of detection (50 copies/mL) with only minimum side effects. Long-term HAART decreased morbidity and mortality associated with HIV infection (2). However, even optimal HAART, characterized by suppression of viral load to undetectable levels for years, has not provided a cure to the disease. Patients on optimal HAART have 12 years shorter life expectancy than HIV negative people (3, 4). In addition, increased AIDS-related and non-AIDS-related morbidity and mortality has been described in a significant proportion of individuals on optimal HAART due to the lack of normalization of their CD4+ T cell counts (5). Optimal HAART also failed to decrease the viral reservoirs, especially in the gut mucosa, where the residual low-level viral replication may be the cause of persistent immune activation that facilitates the progression to AIDS and death (6). One barrier of cure is the stable latent reservoirs of HIV-infected resting memory T cells that are able to produce HIV after cellular activation. HIV producing cells in the reservoirs that are not eliminated by antiretroviral drugs (ARVs) would be susceptible to immune clearance, but long-term optimal HAART diminishes HIV-specific T cell responses (7). Therefore, the immune system of successfully treated HIV-infected people is unprepared to kill infected cells, decrease viral reservoirs and control the virus replication.

Immunotherapy improves the reactivity of the immune system to guard the body from internal and external intruders as infections (e.g., HIV), allergens and malignancies. The innate arm, composed of the phagocytic cells, circulating macrophages and the complement system, is responsible for the immediate defense. Later, the adaptive arm responds in a highly specific manner against molecular determinants of the intruders. Nanoparticles (NPs) made from different types of materials, having various sizes, shapes and surface charges activate the innate and adaptive immune system (8–10). Therefore, nanotechnology is suitable for improving efficacy and increasing specificity of products indicated as vaccine and immunotherapy. The immunotherapeutic efficacy of NPs is achieved by specific activation (e.g., cancer, infectious diseases) or suppression (e.g., allergy, autoimmune diseases) of the immune system. Efficacy of immune responses is improved by targeted delivery of antigens to professional immune cells specialized for either stimulation or suppression of immune reactivity. This is achieved by using NPs as antigen (virus-like particles; VLPs), NP formulation of soluble antigens (DNA, peptides, proteins) and new administration routes (transdermal, nasal, intratracheal). Increasing the specificity of immunotherapy is achieved by personalization of the antigens in the NP taking into account the diversity of the disease (e.g., tumor cells) and genomic background of the individual (e.g., HLA type).

More than 30 ARVs and drug combinations are currently available to achieve long-term suppression of HIV RNA to <50 copies/mL and control HIV disease. The failure to develop a vaccine for HIV has recently changed the treatment strategy from the long-term suppression of viral load to the cure of the disease. Approaches for HAART intensification with additional potent drugs also failed to provide a cure or any additional treatment benefits (4, 11–15). Consequently, it became evident that alternatives to vaccines and to HAART are required to eradicate HIV disease. The first HIV cure approach is eradication of the virus. Eradication was demonstrated in one HIV+ patient after bone marrow transplantation with donor cells resistant to HIV infection (16). Alternative approach for HIV cure is the induction of a long-term remission, similarly to the cure of Hepatitis C infection. We have previously described the remission of an HIV+ patient (Berlin patient, 1999) whose immune system was activated to kill infected cells by short interruptions of HAART (17). This work has led to the identification of “elite controllers” representing a model for remission. Elite controllers have large numbers of cells containing replication competent HIV and their viral load is suppressed by the cellular arm of the immune system (18, 19). Recently, Siliciano and his team have demonstrated that boosting of HIV-specific T cell responses (CTL) prior to reactivating latent HIV will be essential for eradication of the virus (20, 21). These results suggest that HIV-specific immunotherapy is essential for both remission and eradication of HIV, consequently for the cure of HIV.

Immunotherapeutic nanomedicines

Immunotherapeutic nanomedicines are new, complex, multi-modular, targeted products that provide superior therapeutic effects compared to all previous vaccine approaches. Their physical size is usually over 50 nm, which is the approximate threshold of immune recognition (22). Soluble antigens, <50 nm in size, are generally not recognized by the immune system consequently poorly immunogenic. In fact, the size range of immunotherapeutic nanomedicines corresponds to the size range of viruses. Nature developed an effective and specific immune surveillance against viruses. Accordingly, triggering the immune system with nanomedicines provides exceptional immunogenicity since our body considers NPs as harmful viruses that need to be eliminated (23). The best examples for the superior immune recognition of nanomedicines are the human papilloma virus (HPV) vaccines, Gardasil and Cervarix. These vaccines are composed from one surface protein (L1) of the HPV that self-assemble to VLPs. These VLPs, morphologically similar to the wild type HPV, induce potent immune responses in the absence of adjuvants. In contrast, the L1 protein purified from bacteria remains a soluble protein, does not assemble to VLPs, and does not induce immune responses (24). These VLP vaccines are safe and protect young uninfected people from cancer. However, none of these vaccines prevent the development of cancer in HPV-infected people, since they unsuccessfully induce therapeutically beneficial T cell responses (25).

Creating “particulate vaccines” has recently been recognized in the HIV field to improve the immunogenicity of small soluble antigens. This approach involves an increase in the physical size of the antigen to the size of pathogens. There are so-called “natural” particulate vaccines, based on VLPs that induce both humoral and cellular immune responses against HIV (26, 27). HIV VLPs are essentially non-infective viruses consisting of self-assembled viral envelope proteins without the accompanying genetic material. A different approach is to use an adjuvant that increases the size of the antigen. One of the several proposed mechanism of aluminum salts, the adjuvant approved in the US and EU, is attributed to their particulate nature, however recently concerns have been raised regarding their safety (27).

A new approach in vaccine development is the use of a plasmid DNA (pDNA) that can express one or more protein antigens in the body. pDNA is attractive for immunotherapeutic nanomedicine development because (i) it has excellent safety profile, (ii) intracellularly expressed antigens are processed and presented on the host MHC molecules, and (iii) recently improved large-scale manufacturing capabilities provide cost effective production. Unfortunately, promising animal studies demonstrating the induction of immune responses with naked DNA injected intramuscularly or intradermally could not be reproduced in human subjects. Possible reasons of the weak immunogenicity is that the naked DNA poorly enters into the cells, does not reach the nucleus, and the expressed soluble protein antigens are not recognized by the immune system, similarly to the previously described soluble L1 protein of the HPV.

Various biodegradable and non-biodegradable polymeric and liposomal delivery systems have been explored for transforming HIV-antigens to synthetic NPs in order to increase their immunogenicity and to protect them against extra- and intracellular degradation (28, 29). Targeting dendritic cells (DCs) that are essential for initiating immune responses, can be achieved by different nanomedicine size; >100 nm nanomedicines target the peripheral immature DCs, and the smaller size ∼50 nm nanomedicines drain to the lymph node resident DCs (30). Modification of the surface of the nanomedicine with DC-specific receptor ligands has been shown to increase the targeting specificity (31). However, several challenges including crossing physical barriers like the cell and nuclear membranes or adhesion to non-target tissues still need to be overcome during the development of a synthetic delivery system.

Pathogen-like features of DermaVir nanomedicine

DermaVir is the first synthetic pathogen-like nanomedicine that has been developed for HIV-specific immunotherapy. The active pharmaceutical ingredient (API) is a 12.5 kBp pDNA encoding 15 antigens of the HIV (32). The high number of antigens encoded in the DNAsupport the broadest epitope selection, and the assembly of replication-, transcription- and integration defective ‘complex virus-like particles’ (VLP+s).

DermaVir nanomedicine composed from the pDNA core that is covered by a synthetic polymer, a mannobyosilated polyethylenimine-mannobiose (PEIm) (33, 34). DermaVir NP not only looks like as a virus but also functions as a virus: it enters cells via receptor-mediated endocytosis, escapes from endosome and delivers the pDNA to the nucleus. The pDNA expresses fifteen HIV proteins that self-assemble to a complex VLP+.

The pathogen-like character of DermaVir nanomedicine is based on the structure and function of the NP (Figure 1). It ensures the targeting of antigen presenting cells (APCs) of the immune system and effective expression of antigens. The PEIm polymer that contains covalently linked mannobiose residues outside of the NPscreatesa pathogen-like surface resembling viruses and other pathogens because they usually have sugar-like residues (e.g., glycoproteins) on their surface (35–39). Beside the pathogen-like surface elements, the cationic nature of the PEIm ensures the positive surface charge likewise specific for viruses. PEIm when mixed with the pDNA in solution spontaneously forms 70–300 nm spherical NPs, which resembles the size range of viruses (40, 41). The mannobiose residues not only responsible for the pathogen-like surface but also the pathogen-like entry via receptor-mediated endocytosis. Once entering the cell, the NPs translocate to the endosomes, where the cells degrade ‘invaders’ by lowering the pH with the use of proton pumps. In the edosomes the polymer envelope protects the pDNA core from the hydrolytic degradation since the PEIm is able to buffer the low pH with its proton sponge effect (34, 42). This way the loosened NPsare released into the cytoplasm and the free pDNA can reach the nucleus (43, 44). After the pDNA enters the nucleus, the encoded antigens can be transcribed and the expressed viral antigens can self-assemble to replication-, reverse transcription- and integration deficient VLP+. These VLP+s and also non-VLP+-associated antigens then stimulate naïve T cells that will further proliferate to HIV-specific CTLs.

Pathogen-like features of DermaVir nanomedicine compared to a DNA virus.
Figure 1

Pathogen-like features of DermaVir nanomedicine compared to a DNA virus.

Controlling the biological activity of DermaVir during the manufacturing

During the development of DermaVir nanomedicine we explored the structure-activity relationship of the pathogen-like NPs and found that their fine structure determines the biological activity. As shown by atomic force microscopy the different DermaVir formulations have different fine structures; one has smooth, unified surface as envelop, the other has no coherent surface and the pDNA protrudes from the envelop (34). When we investigated the biological activity of these formulations we found that the coherent structured NPs have ∼30% higher potency compared to the other one. The difference between the formulations was only the ionic strength and the pH of the nanomedicine solution thus these physico-chemical properties are responsible for the structure of the evolving NPs (34). At low pH and/or high ionic strength loose structure NPs form, because under these circumstances the pDNA and PEIm components have limited interactions. In this case, the biological activity will decrease since the loose NP cannot survive the endosome and the pDNA degrades in the lysosome (Figure 2A). At higher pH the pDNA and PEIm components can form compact NP with smooth surface, where the pDNA is protected from the extra- and intracellular degradation, the NP survives the endosome, and the pDNA is delivered to the nucleus (Figure 2B) (34).

The fine structure of pDNA/PEIm NPs defines the intracellular trafficking and biological activity. (A) Loose structure NPs cannot survive the endosome. (B) Compact NPs survive the endosome and deliver the pDNA to the nucleus.
Figure 2

The fine structure of pDNA/PEIm NPs defines the intracellular trafficking and biological activity. (A) Loose structure NPs cannot survive the endosome. (B) Compact NPs survive the endosome and deliver the pDNA to the nucleus.

Taking into account that both pDNA and PEIm are polyions explains why the ionic conditions influence their interactions; the dissociation and relative charge density of the components is determined by their ionic environment this way the compactness of the forming NPs also depends on these physico-chemical parameters. By fine-tuning the ionic properties of the formulation we can control the intracellular trafficking and biological activity of the NP: setting the optimal pH and ionic strength of the nanomedicine solution during the manufacturing processes makes us able to control the biological activity of pDNA/PEIm nanomedicines (33, 34).

Langerhans cell-targeting delivery

Optimizing the biophysical properties of a nanomedicine is only one important issue when a product candidate is developed. As DermaVir nanomedicine was designed to target antigen-presenting cells (APCs) and mimic a natural ‘invasion of pathogens’ it was obvious to target the first line defense organ of the body, the skin. Both intradermal injection and skin scarification would target the epidermal Langerhans cells (LCs) but we designed an alternative route of administration to enhance the number of targeted APCs. This CE marked medical device, DermaPrep combines the empty patch technology with a skin preparation method to activate and successfully target nanomedicines into the LCs of the epidermal layer of the skin. Using DermaPrep device various liquid nanomedicine formulation may be administered. The skin preparation method, applied before DermaPrep patch application, is essential (a) to create the “danger signal” for the LCs required to pick up the nanoparticles and migrate to the lymph nodes, and (b) to interrupt the stratum corneumenhancing nanoparticle penetration (45, 46). The area of a patch is 80 cm2, this way the nanomedicine targets ∼8 million LCs (47). Under each patch ∼8×1012 DermaVir nanoparticles (correspond to 0.1 mg pDNA) are administered in liquid formulation, which means that about 1 million nanoparticles could be picked up by every LC. Activated LCs in the epidermis are looking for pathogens and pick up any “suspicious” nanoparticles such as pathogen-like nanoparticles like DermaVir (48). After taking them up they migrate to the local lymph nodes (49). Here, they express pDNA-encoded antigens and present most HIV epitopes to the passing naïve T cells (50, 51). HIV-specific CTLs primed in the lymph nodes seek to kill the virus-infected target cells.

The most unique property of DermaPrep device is the ability to target millions of LCs by modernizing the method of skin scarification, which is known be used for vaccination against smallpox, one of the very few diseases which were eradicated (52). The nowadays most studied vaccine administration systems such as electroporation and microneedles are able to target only a few mm2 of the skin which allows them to deliver material to 3 orders of magnitude less APCs compared to DermaPrep (53–56).

DermaVir for the Cure of HIV/AIDS

DermaVir has successfully completed Phase II clinical development, therefore it is the most advanced nanomedicine developed for HIV-specific immunotherapy. A single DermaVir immunization in HIV-infected subjects on fully suppressive HAART demonstrated dose-dependent expansion of long lasting HIV-specific CTL with high proliferation capacity (57). However, frequency of DermaVir-boosted CTL decreased within a year suggesting that repeated DermaVir immunizations are required for durable CTL activity. Consequently, all subsequent clinical trials investigated repeated DermaVir doses. On HIV-infected adults receiving fully suppressive HAART DermaVir was as safe as placebo and potent CTL were induced in the 0.4 mg DNA dose group (58, 59). These results were confirmed in an independent placebo controlled randomized trial demonstrating the excellent safety and superior immunogenicity of the 0.4 mg DNA dose on HIV-infected, treatment naïve adults (60). In this dose group the medium HIV RNA significantly decreased by 70% compared to Placebo suggesting the killing of HIV infected cells. Viral load suppression occurred slowly, as predicted by DermaVir mechanism of action, similarly to cancer vaccines (61).

Based on the presently available preclinical and clinical results we hypothesize that DermaVir immunotherapy will overcome the following limitations of present HAART (Table 1): (i) reconstitution of HIV-specific immune responses that decreased during optimal HAART (ii) depletion of HIV-infected cells in contrast to current HAART that inhibit only one step in HIV life cycle. Compared to HAART the antiviral activity of DermaVir is delayed, slow and less potent, because recognizing and killing of millions of infected cells with CTL in the presence of millions of new infections take more time to decrease viral load than blocking HIV replication with drugs. The effectiveness of killing infected cells is revealed by their capacity to manage the infection for ca. 15 years in the absence of any treatment. Therefore, 0.5 log reduction of HIV RNA in 24 weeks, demonstrated with DermaVir, should be sufficient to decrease the amount of HIV-infected cells that are not eliminated by HAART.

Table 1

Comparison of the features of DermaVir and HAART.

Cure of HIV from infected patient is defined by the absence of HIV rebound after HAART interruption. The different, complementary mechanism of action of HAART and DermaVir is suitable to achieve cure. The rapid and potent viral load reduction with HAART is essential to block HIV replication and reach undetectable viral load. After that, DermaVir immunotherapy could address what HAART intensification could not achieve; kill HIV-infected cells that remained in reservoirs and boost HIV-specific T cells to reconstitute immune responses. Since the immune system is slow to kill infected cells it will take time to substantially decrease the infected cells from the reservoirs and fully reconstitute HIV-specific immune responses. We envision that repeated DermaVir immune intensification could eliminate significant amount of infected cells from the reservoirs. Consequently, patients could decrease drug exposure and their immune system could maintain a low or undetectable HIV RNA level. Undetectable HIV RNA for 6 months after interruption of HAART could demonstrate cure of HIV similarly to the cure of HCV infection.

Potential advantages of DermaVir immunotherapy compared to HAART include its excellent safety, higher specificity, the longevity of an immune response and likely cost-savings as well as, at least theoretically, the chance to achieve a cure. However, despite decades of research, no therapeutic vaccine has reached the market. Challenges include (i) repeated failures of prophylactic vaccines, (ii) immune escape from T cell recognition based on the high genetic diversity of the virus and the HLA diversity of the host; (iii) the shortage of funding compared to vaccine and drug development. Any treatment that can eradicate HIV from infected patients or cure the disease by remission would have a huge commercial opportunity. Nanotechnology offers opportunities to develop new treatment approaches to the cure of HIV/AIDS. We envision that HIV-specific immunotherapy with synthetic pathogen-like nanomedicines might have the safety, efficacy and cost features to contribute to the cure of HIV/AIDS and significantly improve public health.

Disclosures and conflict of interest

Lisziewicz and Lőrincz hold shares in Genetic Immunity (PWRV). This work was supported by grants: HIKC05, FIBERSCN and DVCLIN01 of the National Office for Research and Technology (NKTH) in Hungary.

References

  • 1.

    Global epidemic report on HIV infection: http://www.unaids.org/globalreport/default.htm (Accessed 29 November, 2012). 

  • 2.

    Palella FJ, Delaney KM, Moorman AC, Loveless MO, Fuhrer J, Satten GA, et al. Declining morbidity and mortality among patients with advanced Human Immunodeficiency Virus infection. New Engl J Med 1998;338:853–60. Google Scholar

  • 3.

    Holtgrave DR. Causes of the decline in AIDS deaths, United States, 1995–2002: prevention, treatment or both? Int J STD AIDS 2005;16:777–81.Google Scholar

  • 4.

    Losina E, Schackman BR, Sadownik SN, Gebo KA, Walensky RP, Chiosi JJ, et al. Racial and sex disparities in life expectancy losses among HIV-infected persons in the United States: impact of risk behavior, late initiation, and early discontinuation of antiretroviral therapy. CID 2009;49:1570–8.CrossrefGoogle Scholar

  • 5.

    Baker JV, Peng G, Rapkin J, Abrams DI, Silverberg MJ, MacArthur RD, et al. CD4+ count and risk of non-AIDS diseases following initial treatment for HIV infection. AIDS 2008;22:841–8. CrossrefGoogle Scholar

  • 6.

    Mavigner M, Delobel P, Cazabat M, Dubois M, L’faqihi-Olive FE, Raymond S, et al. HIV-1 residual viremia correlates with persisitent T-cell activation in poor immunological responders to combination antiretroviral therapy. PLoS ONE 2009;4:e7658. CrossrefGoogle Scholar

  • 7.

    Casazza JP, Betts MR, Picker LJ, Koup RA. Decay kinetics of Human Immunodeficiency Virus-specific CD8+ T cells in peripheral blood after initiation of highly active antiretroviral therapy. J Virol 2001;75:6508–16. CrossrefGoogle Scholar

  • 8.

    Goldsmith M, Mizrahy S, Peer D. Grand challenges in modulating the immune response with RNAi nanomedicines. Nanomedicine 2011;6:1771–85.CrossrefGoogle Scholar

  • 9.

    Dobrovolskaia MA, McNeil SE. Immunological properties of engineered Nanomaterials. Nat Nanotech 2007;2:469–78.CrossrefGoogle Scholar

  • 10.

    Zolnik BS, González-Fernández A, Sadrieh N, Dobrovolskaia MA. Nanoparticles and the immune system. Endocrinology 2010;151:458–65.Google Scholar

  • 11.

    Haase AT. Perils at mucosal front lines for HIV and SIV and their hosts. Nat Rev Immunol 2005;5:783–92.CrossrefPubMedGoogle Scholar

  • 12.

    Shen L, Siliciano RF. Viral reservoirs, residual viremia, and the potential of highly active antiretroviral therapy to eradicate HIV infection. J Allergy ClinImmunol 2008;122:22–8.Google Scholar

  • 13.

    Dinosoa JB, Kima SY, Wiegandc AM, Palmerc SE, Ganged SJ, Cranmer L, et al. Treatment intensification does not reduce residual HIV-1 viremia in patients on highly active antiretroviral therapy. PNAS 2009;106:9403–8.Google Scholar

  • 14.

    Gandhi RT, Zheng L, Bosch RZ, Chan ES, Margolis DM, Read S, et al. The effect of Raltegravir intensification on low-level residual viremia in HIV-infected patients on antiretroviral therapy: a randomized controlled trial. PLoS Med 2011;7:1–11.Google Scholar

  • 15.

    McMahon D, Jones J, Wiegand A, Gange SJ, Kearney M, Palmer S, et al. Short-course Raltegravir intensification does not reduce persistent low-level viremia in patients with HIV-1 suppression during receipt of combination antiretroviral therapy. CID 2010;50:912–9.Google Scholar

  • 16.

    Allers K, Hütter G, Hofmann J, Loddenkemper C, Rieger K, Thiel E, et al. Evidence for the cure of HIV infection by CCR5D32/D32 stem cell transplantation. Blood 2011;117:2791–9. Google Scholar

  • 17.

    Lisziewicz J, Rosenberg E, Lieberman J, Jessen H, Lopalco L, Siliciano R, et al. Control of HIV despite the discontinuation of antiretroviral therapy. New Engl J Med 1999;340:1683–4. Google Scholar

  • 18.

    Deeks SG, Walker BD. Human immunodeficiency virus controllers: mechanisms of durable virus control in the absence of antiretroviral therapy. Immunity 2007;27:406–16.PubMedCrossrefGoogle Scholar

  • 19.

    Ndhlovu ZM, Proudfoot J, Cesa K, Alvino DM, McMullen A, Vine S, et al. Elite controllers with low to absent effector CD8+ T cell responses maintain highly functional, broadly directed central memory responses. J Virol 2012;86:6959–69.CrossrefGoogle Scholar

  • 20.

    Shan L, Dend K, Shroff NS, Durand CM, Rabi SA, Yang H-C, et al. Stimulation of HIV-1-specific cytolytic T lymphocytes facilitates elimination of latent viral reservoir after virus reactivation. Immunity 2012;36:1–11.Google Scholar

  • 21.

    Vaccari M, Trindade CJ, Venzon D, Zanetti M, Franchini G. Vaccine-induced CD8+ central memory T cells in protection from simian AIDS. J Immunol 2005;175:3502–7.Google Scholar

  • 22.

    Xiang SD, Scholzen A, Minigo A, David C, Apostolopoulos V, Mottram PL, et al. Pathogen recognition and development of particulate vaccines: does size matter? Methods 2006;40:1–9. CrossrefPubMedGoogle Scholar

  • 23.

    Lisziewicz J, Szebeni J. The janus face of immune stimulation by nanomedicines: examples for the good and the bad. Eur J Nanomed 2010;3:13–8. Google Scholar

  • 24.

    Christensen ND, Höpfl R, DiAngelo SL, Cladel NM, Patrick SD, Welsh PA, et al. Assembled baculovirus-expressed human papillomavirus type 11 L1 capsid protein virus-like particles are recognized by neutralizing monoclonal antibodies and induce high titres of neutralizing antibodies. J Gen Virol 1994;75:2271–6. CrossrefGoogle Scholar

  • 25.

    Hildesheim A, Herrero R, Wacholder S, Rodriguez AC, Solomon D, Bratti MC, et al. Effect of human papillomavirus 16/18 L1 viruslike particle vaccine among young women with preexisting infection – A randomized trial. J Am Med Assoc 2007;298:743–53. Google Scholar

  • 26.

    Martin SJ, Vyakarnam A, Cheingsong-Popov R, Callow D, Jones KL, Senior JM, et al. Immunization of human HIV-seronegative volunteers with recombinant p17/p24:Ty virus-like particles elicits HIV-1 p24-specific cellular and humoral immune responses. AIDS 1993;7:1315–23.Google Scholar

  • 27.

    Peek LJ, Middaugh CR, Berkland C. Nanotechnology in vaccine delivery. Adv Drug Deliv Rev 2008;60:915–28. PubMedCrossrefGoogle Scholar

  • 28.

    Lamalle-Bernard D, Munier S, Compagnon C, Charles MH, Kalyanaraman VS, Delair T, et al. Coadsorption of HIV-1 p24 and gp120 proteins to surfactant-free anionic PLA nanoparticles preserves antigenicity and immunogenicity. J Control Rel 2006;115:57–67.Google Scholar

  • 29.

    Fairman J, Moore J, Lemieux M, Van Rompay K, Geng Y, Warner J, et al. Enhanced in vivo immunogenicity of SIV vaccine candidates with cationic liposome-DNA complexes in a rhesus macaque pilot study. Hum Vaccin 2009;5:141–50.CrossrefGoogle Scholar

  • 30.

    Reddy ST, Rehor A, Schmoekel HG, Hubbell JA, Swartz MA. In vivo targeting of dendritic cells in lymph nodes with poly(propylene sulfide) nanoparticles. J Control Rel 2006;112:26–34. Google Scholar

  • 31.

    Diebold SS, Kursa M, Wagner E, Cotton M, Zenke M. Mannose Polyethylenimine conjugates for targeted DNA delivery into dendritic cell. J Biol Chem 1999;274:19087–94. Google Scholar

  • 32.

    Somogyi E, Xu J, Gudics A, Tóth J, Kovács AL, Lori F, et al. A plasmid DNA immunogen expressing fifteen protein antigens and complex virus-like particles (VLP+) mimicking naturally occurring HIV. Vaccine 2011;29:744–53. CrossrefPubMedGoogle Scholar

  • 33.

    Toke ER, Lőrincz O, Somogyi E, Lisziewicz J. Rational development of a stable liquid formulation for nanomedicine products. Int J Pharm 2010;392:261–7.Google Scholar

  • 34.

    Lőrincz O, Toke ER, Somogyi E, Horkay F, Chandran PL, Douglas JF, et al. Structure and biological activity of pathogen-like synthetic nanomedicines. Nanomedicine: NBM 2012;8:497–506. PubMedGoogle Scholar

  • 35.

    Shim MS, Kwon YJ. Controlled cytoplasmic and nuclear localization of plasmid DNA and siRNA by differentially tailored polyethylenimine. J Control Rel 2009;133:206–13.Google Scholar

  • 36.

    Ogay ID, Lihoradova OA, Azimova SS, Abdukarimov AA, Slack JM, Lynn DE. Transfection of insect cell lines using polyethylenimine. Cytotechnology 2006;51:89–98.CrossrefPubMedGoogle Scholar

  • 37.

    Salehi N, Peng CA. Gene transfection of Toxoplasma gondii using PEI/DNA polyplexes. J Microbiol Methods 2012;91:133–7.CrossrefPubMedGoogle Scholar

  • 38.

    Han X, Fang Q, Yao F, Wang X, Wang J. The heterogeneous nature of polyethylenimine-DNA complex formation affects transient gene expression. Cytotechnology 2009;60:63–75.CrossrefGoogle Scholar

  • 39.

    Reddy ST, Swartz MA, Hubbell JA. Targeting dendritic cells with biomaterials: developing the next generation of vaccines. Trends Immunol 2006;27:573–9.PubMedCrossrefGoogle Scholar

  • 40.

    Bruewer M, Utech M, Ivanov AI, Hopkins AM, Parkos CA. Nusrat A. Interferon-gamma induces internalization of epithelial tight junction proteins via a macropinocytosis-like process. FASEB J 2005;19:923–33.CrossrefGoogle Scholar

  • 41.

    Racoosin EL, Swanson JA. M-CSF-induced macropinocytosis increases solute endocytosis but not receptor-mediated endocytosis in mouse macrophages. J Cell Sci 1992;102:867–80.PubMedGoogle Scholar

  • 42.

    Boussif O, Lezoualc’h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. PNAS 1995;92:7297–301.CrossrefGoogle Scholar

  • 43.

    Schaffer DV, Fidelman NA, Dan N, Lauffenburger DA. Vector unpacking as a potential barrier for receptor-mediated polyplex gene delivery. BiotechnolBioen 2000;67:598–606.Google Scholar

  • 44.

    Suh J, Wirtz D, Hanes J. Efficient active transport of gene nanocarriers to the nucleus. PNAS 2003;100:3878–82.CrossrefGoogle Scholar

  • 45.

    Matzinger P. The danger model: a renewed sense of self. Science 2002;296:301–5.Google Scholar

  • 46.

    Nicolas JF, Guy B. Intradermal, epidermal and transcutaneous vaccination: from immunology to clinical practice. Expert Rev Vaccines 2008;7:1201–14.PubMedCrossrefGoogle Scholar

  • 47.

    Bauer J, Bahmer FA, Worl J, Neuhuber W, Schuler G, Fartasch M. A strikingly constant ratio exists between Langerhans cells and other epidermal cells in human skin. A stereologic study using the optical disector method and the confocal laser scanning microscope. J Invest Dermat 2001;116:313–8. CrossrefGoogle Scholar

  • 48.

    Lisziewicz J, Tőke ER. Nanomedicine applications towards the cure of HIV. Nanomedicine: NBM 2012; in press: doi:10.1016/j.nano.2012.05.012. CrossrefGoogle Scholar

  • 49.

    Lisziewicz J, Gabrilovich DI, Varga G, Xu J, Greenberg PD, Arya SK, et al. Induction of potent human immunodeficiency virus type 1-specific T-cell-restricted immunity by genetically modified dendritic cells. J Virol 2001;75:7621–8. CrossrefGoogle Scholar

  • 50.

    Lisziewicz J, Trocio J, Xu J, Whitman L, Ryder A, Bakare N, et al. Control of viral rebound through therapeutic immunization with DermaVir. AIDS 2005;19:35–43.PubMedCrossrefGoogle Scholar

  • 51.

    Lisziewicz J, Bakare N, Calarota SA, Bánhegyi D, Szlávik J, Újhelyi E, et al. Single DermaVir Immunization: dose-dependent expansion of precursor/memory T cells against all HIV antigens in HIV-1 infected individuals. PLoS ONE 2012;7:e35416.CrossrefGoogle Scholar

  • 52.

    Liu L, Zhong Q, Tian T, Dubin K, Athale SK, Kupper TS. Epidermal injury and infection during poxvirus immunization is crucial for the generation of highly protective T cell-mediated immunity. Nat Med 2010;16:224–7. PubMedCrossrefGoogle Scholar

  • 53.

    Elisabeth TT, Dietmar R, Christian O, Iacob M, Rune K. Taking electroporation-based delivery of DNA vaccination into humans: a generic clinical protocol. Methods Mol Biol 2008;423:497–507.Google Scholar

  • 54.

    Lee JW, Park JH, Prausnitz MR. Dissolving microneedles for transdermal drug delivery. Biomaterials 2008;29:2113–24.PubMedCrossrefGoogle Scholar

  • 55.

    Albrecht MT, Livingston BD, Pesce JT, Bell MG, Hannaman D, Keane-Myers AM. Electroporation of a multivalent DNA vaccine cocktail elicits a protective immune response against anthrax and plague. Vaccine 2012;30:4872–83.PubMedCrossrefGoogle Scholar

  • 56.

    Kumar A, Wonganan P, Sandoval MA, Li X, Zhu S, Cui Z. Microneedle-mediated transcutaneous immunization with plasmid DNA coated on cationic PLGA nanoparticles. J Control Rel 2012;163:230–9.Google Scholar

  • 57.

    Lisziewicz J, Calarota S, Banhegyi D, Lisziewicz Zs, Ujhelyi E, Lori F. Single DermaVir Patch treatment of HIV+ individuals induces long-lasting, high-magnitude, and broad HIV-specific T cell responses. Paper presented at the 15th Conference on Retroviruses and Opportunistic Infections, Boston, MA, USA. 2008; Poster #715. Google Scholar

  • 58.

    Rodriguez B, Asmuth D, Matining R, Spritzler J, Li X-D, Jacobson J, et al. Repeated-dose transdermal administration of DermaVir, a candidate plasmid DNA-based therapeutic HIV vaccine, is safe and well-tolerated: a 61-week analysis of ACTG Study 5176, paper presented at XVIIIth International AIDS Conference, Vienna, Austria. 2010; Abstract # A-240-0111-10145.Google Scholar

  • 59.

    Clinical trials of DermaVir nanomedicine: http://www.clinicaltrials.gov/ct2/results?term=dermavir&Search=Search (Accessed 29 November, 2012).

  • 60.

    vanLunzen J. et al. DermaVir for initial treatment of HIV-infected subjects demonstrates preliminary safety, immunogenicity and HIV-RNA reduction versus placebo immunization. Paper presented at the XVIIIth International AIDS Conference, Vienna, Austria. 2010; Abstract #A-240-0111-12561. Google Scholar

  • 61.

    Kantoff PW, Higano CS, Shore ND, Berger ER, Small EJ, Penson DF, et al. Sipuleucel-T immunotherapy for castration-resistant prostate cancer. N Engl J Med 2010;363:411–22. Google Scholar

About the article

Julianna Lisziewicz

Julianna Lisziewicz, PhD, President, Chief Executive Officer, Genetic Immunity. Dr. Julianna Lisziewicz co-founded Genetic Immunity in 1998 where she serves as the President and Chief Executive Officer and directs the translational research program on therapeutic vaccines from discovery to clinical trials. In 1994, Dr. Lisziewicz co-founded the non-profit Research Institute for Genetic and Human Therapy and directed its research and business affairs in the USA. RIGHT was focusing on the treatment of HIV/AIDS from multiple perspectives: virology, molecular biology, immunology and medicine. From 1990 to 1995, she was Head of the Antiviral Unit in the Laboratory of Tumor Cell Biology at the NCI, NIH in Bethesda, Maryland, USA. While at NIH, she discovered and developed antisense oligonucleotide therapy and gene therapy for HIV/AIDS. In 2005, she was awarded by the EU the Marie Curie Chair. She received her PhD in molecular biology from the Max-Planck Institute, Goettingen.

Orsolya Lőrincz

Orsolya Lőrincz, MSc, Head of Quality Control Laboratory, Genetic Immunity. Orsolya Lőrincz graduated from university in 2007 as a chemist and since then she has been working at Genetic Immunity’s product development department. She takes part in formulation and also the analytical development. From 2010 she is the head of the quality control laboratory.


Corresponding author: Julianna Lisziewicz, Genetic Immunity Berliniutca 47-49 Budapest 1045, Hungary and 8300 Greensboro Drive Mclean, VA 22102, USA


Published Online: 2012-12-19

Published in Print: 2012-12-01


Citation Information: European Journal of Nanomedicine, ISSN (Online) 1662-596x, ISSN (Print) 1662-5986, DOI: https://doi.org/10.1515/ejnm-2012-0011.

Export Citation

©2012 by Walter de Gruyter Berlin Boston. Copyright Clearance Center

Comments (0)

Please log in or register to comment.
Log in