1680:
immunostimulatory. CpG stimulatory (CpG-S) sequences occur twenty times more frequently in bacterially-derived DNA than in eukaryotes. This is because eukaryotes exhibit "CpG suppression" – i.e. CpG dinucleotide pairs occur much less frequently than expected. Additionally, CpG-S sequences are hypomethylated. This occurs frequently in bacterial DNA, while CpG motifs occurring in eukaryotes are methylated at the cytosine nucleotide. In contrast, nucleotide sequences that inhibit the activation of an immune response (termed CpG neutralising, or CpG-N) are over represented in eukaryotic genomes. The optimal immunostimulatory sequence is an unmethylated CpG dinucleotide flanked by two 5’
116:. DNA vaccines contain DNA that codes for specific antigens from a pathogen. The DNA is injected into the body and taken up by cells, whose normal metabolic processes synthesize proteins based on the genetic code in the plasmid that they have taken up. Because these proteins contain regions of amino acid sequences that are characteristic of bacteria or viruses, they are recognized as foreign and when they are processed by the host cells and displayed on their surface, the immune system is alerted, which then triggers immune responses. Alternatively, the DNA may be encapsulated in protein to facilitate cell entry. If this
1671:). However, the potential toxicity of prolonged cytokine expression is not established. In many commercially important animal species, cytokine genes have not been identified and isolated. In addition, various plasmid-encoded cytokines modulate the immune system differently according to the delivery time. For example, some cytokine plasmid DNAs are best delivered after immunogen pDNA, because pre- or co-delivery can decrease specific responses and increase non-specific responses.
28:
1011:
1795:. Unlike conventional DNA vaccinations alphavirus vectors kill transfected cells and are only transiently expressed. Alphavirus replicase genes are expressed in addition to the vaccine insert. It is not clear how alphavirus replicons raise an immune response, but it may be due to the high levels of protein expressed by this vector, replicon-induced cytokine responses, or replicon-induced apoptosis leading to enhanced antigen uptake by dendritic cells.
1742:(PkMSP1p42). They were then boosted with a recombinant canarypox virus encoding all four antigens (ALVAC-4). Immunized monkeys developed antibodies against sporozoites and infected erythrocytes, and IFN-γ-secreting T-cell responses against peptides from PkCSP. Partial protection against sporozoite challenge was achieved, and mean parasitemia was significantly reduced, compared to control monkeys. These models, while not ideal for extrapolation to
1517:
lymph node where they prime secondary B- and T-cell responses. In skeletal muscle, striated muscle cells are most frequently transfected, but seem to be unimportant in immune response. Instead, IM inoculated DNA "washes" into the draining lymph node within minutes, where distal dendritic cells are transfected and then initiate an immune response. Transfected myocytes seem to act as a "reservoir" of antigen for trafficking professional APCs.
651:
5700:
1477:
456:
1627:(PyCSP) enhanced protection against subsequent challenge (whereas plasmid-encoded PyCSP alone did not). It was proposed that GM-CSF caused dendritic cells to present antigen more efficiently and enhance IL-2 production and TH cell activation, thus driving the increased immune response. This can be further enhanced by first priming with a pPyCSP and pGM-CSF mixture, followed by boosting with a recombinant
1221:
1708:
been used as external adjuvants for both DNA and recombinant protein vaccination with variable success rates. Other organisms with hypomethylated CpG motifs have demonstrated the stimulation of polyclonal B-cell expansion. The mechanism behind this may be more complicated than simple methylation – hypomethylated murine DNA has not been found to mount an immune response.
1556:, which do not kill the cell, but limit viral infection by down-regulating the expression of viral components. DNA vaccinations can be used to curb viral infections by non-destructive IFN-mediated control. This was demonstrated for hepatitis B. IFN-γ is critically important in controlling malaria infections and is a consideration for anti-malarial DNA vaccines.
396:
inverted and tandem repeats, which are conspicuous in many commercially available cloning and expression vectors. Therefore, the reduction or complete elimination of extraneous noncoding backbone sequences would pointedly reduce the propensity for such events to take place and consequently the overall plasmid's recombinogenic potential.
1721:
envelope protein. Recombinant virus boosts have been shown to be very efficient at boosting DNA-primed CTL responses. Priming with DNA focuses the immune response on the required immunogen, while boosting with the recombinant virus provides a larger amount of expressed antigen, leading to a large increase in specific CTL responses.
1378:
profile of the immune response and thus the antibody isotype, which is not possible with either natural infection or recombinant protein immunization. Antibody responses generated by DNA are useful as a preparative tool. For example, polyclonal and monoclonal antibodies can be generated for use as reagents.
1539:(FDC), which are potent B-cell stimulators. T-cells can be stimulated by similar, germinal centre dendritic cells. FDC are able to generate an immune memory because antibodies production "overlaps" long-term expression of antigen, allowing antigen-antibody immunocomplexes to form and be displayed by FDC.
984:
The delivery method determines the dose required to raise an effective immune response. Saline injections require variable amounts of DNA, from 10 μg to 1 mg, whereas gene gun deliveries require 100 to 1000 times less. Generally, 0.2 μg – 20 μg are required, although quantities as
1728:
circumsporozoite surface protein (PyCSP), then boosted with a recombinant vaccinia virus expressing the same protein had significantly higher levels of antibody, CTL activity and IFN-γ, and hence higher levels of protection, than mice immunized and boosted with plasmid DNA alone. This can be further
1516:
IM and ID DNA delivery initiate immune responses differently. In the skin, keratinocytes, fibroblasts and
Langerhans cells take up and express antigens and are responsible for inducing a primary antibody response. Transfected Langerhans cells migrate out of the skin (within 12 hours) to the draining
1256:
Comparisons of antibody responses generated by natural (viral) infection, immunization with recombinant protein and immunization with pDNA are summarised in Table 4. DNA-raised antibody responses rise much more slowly than when natural infection or recombinant protein immunization occurs. As many as
1047:
The type of T-cell help raised is influenced by the delivery method and the type of immunogen expressed, as well as the targeting of different lymphoid compartments. Generally, saline needle injections (either IM or ID) tend to induce TH1 responses, while gene gun delivery raises TH2 responses. This
1720:
DNA-primed immune responses can be boosted by the administration of recombinant protein or recombinant poxviruses. "Prime-boost" strategies with recombinant protein have successfully increased both neutralising antibody titre, and antibody avidity and persistence, for weak immunogens, such as HIV-1
1377:
epitopes than recombinant protein-induced antibodies. In other words, DNA immunization induces a qualitatively superior response. Antibodies can be induced after one vaccination with DNA, whereas recombinant protein vaccinations generally require a boost. DNA immunization can be used to bias the TH
1074:
How these different methods operate, the forms of antigen expressed, and the different profiles of T-cell help is not understood. It was thought that the relatively large amounts of DNA used in IM injection were responsible for the induction of TH1 responses. However, evidence shows no dose-related
1711:
Most of the evidence for immunostimulatory CpG sequences comes from murine studies. Extrapolation of this data to other species requires caution – individual species may require different flanking sequences, as binding specificities of scavenger receptors vary across species. Additionally, species
1707:
Manipulation of CpG-S and CpG-N sequences in the plasmid backbone of DNA vaccines can ensure the success of the immune response to the encoded antigen and drive the immune response toward a TH1 phenotype. This is useful if a pathogen requires a TH response for protection. CpG-S sequences have also
1569:
An effective vaccine must induce an appropriate immune response for a given pathogen. DNA vaccines can polarise T-cell help towards TH1 or TH2 profiles and generate CTL and/or antibody when required. This can be accomplished by modifications to the form of antigen expressed (i.e. intracellular vs.
1507:
Besides direct transfection of dendritic cells or macrophages, cross priming occurs following IM, ID and gene gun DNA deliveries. Cross-priming occurs when a bone marrow-derived cell presents peptides from proteins synthesised in another cell in the context of MHC class 1. This can prime cytotoxic
1014:
Antigen presentation stimulates T cells to become either "cytotoxic" CD8+ cells or "helper" CD4+ cells. Cytotoxic cells directly attack other cells carrying certain foreign or abnormal molecules on their surfaces. Helper T cells, or Th cells, coordinate immune responses by communicating with other
404:
Once the plasmid inserts itself into the transfected cell nucleus, it codes for a peptide string of a foreign antigen. On its surface the cell displays the foreign antigen with both histocompatibility complex (MHC) classes I and class II molecules. The antigen-presenting cell then travels to the
1261:
DNA vaccine expressing HBV small and middle envelope protein was injected into adults with chronic hepatitis. The vaccine resulted in specific interferon gamma cell production. Also specific T-cells for middle envelop proteins antigens were developed. The immune response of the patients was not
395:
Structural instability phenomena are of particular concern for plasmid manufacture, DNA vaccination and gene therapy. Accessory regions pertaining to the plasmid backbone may engage in a wide range of structural instability phenomena. Well-known catalysts of genetic instability include direct,
1741:
were primed with a multicomponent, multistage DNA vaccine encoding two liver-stage antigens – the circumsporozoite surface protein (PkCSP) and sporozoite surface protein 2 (PkSSP2) – and two blood stage antigens – the apical merozoite surface protein 1 (PkAMA1) and merozoite surface protein 1
1679:
Plasmid DNA itself appears to have an adjuvant effect on the immune system. Bacterially derived DNA can trigger innate immune defence mechanisms, the activation of dendritic cells and the production of TH1 cytokines. This is due to recognition of certain CpG dinucleotide sequences that are
621:
immunization (ELI). Using this technique, potentially all the genes from a pathogen can be delivered at one time, which may be useful for pathogens that are difficult to attenuate or culture. ELI can be used to identify which genes induce a protective response. This has been tested with
77:
use. In some cases, protection from disease in animals has been obtained, in others not. Research is ongoing over the approach for viral, bacterial and parasitic diseases in humans, as well as for cancers. In August 2021, Indian authorities gave emergency approval to
175:
began tests of a different DNA vaccine against Zika in Miami. The NIH vaccine is injected into the upper arm under high pressure. Manufacturing the vaccines in volume remained unsolved as of August 2016. Clinical trials for DNA vaccines to prevent HIV are underway.
1604:, and GM-CSF) plus TH2-inducing cytokines increase antibody responses, whereas pro-inflammatory agents and TH1-inducing cytokines decrease humoral responses and increase cytotoxic responses (more important in viral protection). Co-stimulatory molecules such as
1699:
The innate system works with the adaptive immune system to mount a response against the DNA encoded protein. CpG-S sequences induce polyclonal B-cell activation and the upregulation of cytokine expression and secretion. Stimulated macrophages secrete IL-12,
1228:
Antibody responses elicited by DNA vaccinations are influenced by multiple variables, including antigen type; antigen location (i.e. intracellular vs. secreted); number, frequency and immunization dose; site and method of antigen delivery.
1767:) with DNA. Such DNA-coated microparticles can be as effective at raising CTL as recombinant viruses, especially when mixed with alum. Particles 300 nm in diameter appear to be most efficient for uptake by antigen presenting cells.
2716:
Böhm W, Kuhröber A, Paier T, Mertens T, Reimann J, Schirmbeck R (June 1996). "DNA vector constructs that prime hepatitis B surface antigen-specific cytotoxic T lymphocyte and antibody responses in mice after intramuscular injection".
195:
As of 2021 no DNA vaccines have been approved for human use in the United States. Few experimental trials have evoked a response strong enough to protect against disease and the technique's usefulness remains to be proven in humans.
3194:"Genetic immunization of BALB/c mice with a plasmid bearing the gene coding for a hybrid merozoite surface protein 1-hepatitis B virus surface protein fusion protects mice against lethal Plasmodium chabaudi chabaudi PC1 infection"
1051:
Generally the type of T-cell help raised is stable over time, and does not change when challenged or after subsequent immunizations that would normally have raised the opposite type of response in a naïve specimen. However, Mor
1448:
The 30kDa surface receptor binds specifically to 4500-bp DNA fragments (which are then internalised) and is found on professional APCs and T-cells. Macrophage scavenger receptors bind to a variety of macromolecules, including
4420:
Wolff JA, Dowty ME, Jiao S, Repetto G, Berg RK, Ludtke JJ, et al. (December 1992). "Expression of naked plasmids by cultured myotubes and entry of plasmids into T tubules and caveolae of mammalian skeletal muscle".
1779:
have been used to improve DNA vaccination efficiency. The gene encoding the antigen of interest is inserted into the alphavirus replicon, replacing structural genes but leaving non-structural replicase genes intact. The
1547:
Both helper and cytotoxic T-cells can control viral infections by secreting interferons. Cytotoxic T cells usually kill virally infected cells. However, they can also be stimulated to secrete antiviral cytokines such as
1372:
Additionally, the titres of specific antibodies raised by DNA vaccination are lower than those obtained after vaccination with a recombinant protein. However, DNA immunization-induced antibodies show greater affinity to
2167:
White LO, Gibb E, Newham HC, Richardson MD, Warren RC (July 1979). "Comparison of the growth of virulent and attenuated strains of
Candida albicans in the kidneys of normal and cortison-treated mice by chitin assay".
2213:"Construction of live vaccines using genetically engineered poxviruses: biological activity of vaccinia virus recombinants expressing the hepatitis B virus surface antigen and the herpes simplex virus glycoprotein D"
507:. Immune responses to this method can be affected by factors including needle type, needle alignment, speed of injection, volume of injection, muscle type, and age, sex and physiological condition of the recipient.
1257:
12 weeks may be required to reach peak titres in mice, although boosting can decrease the interval. This response is probably due to the low levels of antigen expressed over several weeks, which supports both
1468:, and other delivery methods bypass this entry method, but understanding it may be useful in reducing costs (e.g. by reducing the requirement for cytofectins), which could be important in animal husbandry.
337: – is the "vehicle" from which the immunogen is expressed, optimising vector design for maximal protein expression is essential. One way of enhancing protein expression is by optimising the
1115:. In autoimmune diseases, the goal is to shift the self-destructive TH1 response (with its associated cytotoxic T cell activity) to a non-destructive TH2 response. This has been successfully applied in
159:
demonstrated that direct injection of mice with plasmid DNA encoding a flu antigen protected the animals against subsequent experimental infection with influenza virus. In 2016 a DNA vaccine for the
3146:
Huebener N, Fest S, Strandsby A, Michalsky E, Preissner R, Zeng Y, et al. (July 2008). "A rationally designed tyrosine hydroxylase DNA vaccine induces specific antineuroblastoma immunity".
2669:"Immune responses induced by intramuscular or gene gun injection of protective deoxyribonucleic acid vaccines that express the circumsporozoite protein from Plasmodium berghei malaria parasites"
1027:
profiles. A major advantage of DNA vaccines is the ease with which they can be manipulated to bias the type of T-cell help towards a TH1 or TH2 response. Each type has distinctive patterns of
73:. DNA vaccines have theoretical advantages over conventional vaccines, including the "ability to induce a wider range of types of immune response". Several DNA vaccines have been tested for
989:. Saline injections require more DNA because the DNA is delivered to the extracellular spaces of the target tissue (normally muscle), where it has to overcome physical barriers (such as the
3099:"Targeting of HIV-1 antigens for rapid intracellular degradation enhances cytotoxic T lymphocyte (CTL) recognition and the induction of de novo CTL responses in vivo after immunization"
1184:
proteins, but the principle should also be applicable to DNA vaccines. Targeting antigens for intracellular degradation (and thus entry into the MHC class I pathway) by the addition of
2279:
Ulmer JB, Donnelly JJ, Parker SE, Rhodes GH, Felgner PL, Dwarki VJ, et al. (March 1993). "Heterologous protection against influenza by injection of DNA encoding a viral protein".
443:
of the protein can also affect antibody responses. "Ordered" structures (such as viral particles) are more effective than unordered structures. Strings of minigenes (or MHC class I
3864:"Characterization of humoral and CD4+ cellular responses after genetic immunization with retroviral vectors expressing different forms of the hepatitis B virus core and e antigens"
3278:
1570:
secreted), the method and route of delivery or the dose. It can also be accomplished by the co-administration of plasmid DNA encoding immune regulatory molecules, i.e. cytokines,
5110:"Improving protective immunity induced by DNA-based immunization: priming with antigen and GM-CSF-encoding plasmid DNA and boosting with antigen-expressing recombinant poxvirus"
5058:"Genetic vaccination against malaria infection by intradermal and epidermal injections of a plasmid containing the gene encoding the Plasmodium berghei circumsporozoite protein"
1504:
to present antigens. After IM and ID injections, dendritic cells present antigen in the draining lymph node and transfected macrophages have been found in the peripheral blood.
388:
tripartite leader (TPL) sequences and modifications to the polyadenylation and transcriptional termination sequences. An example of DNA vaccine plasmid is pVAC, which uses SV40
1015:
cells. In most cases, T cells only recognize an antigen if it is carried on the surface of a cell by one of the body's own MHC, or major histocompatibility complex, molecules.
202:
DNA immunization is also being investigated as a means of developing antivenom sera. DNA immunization can be used as a technology platform for monoclonal antibody induction.
1729:
enhanced by priming with a mixture of plasmids encoding PyCSP and murine GM-CSF, before boosting with recombinant vaccinia virus. An effective prime-boost strategy for the
3970:"Activation of cutaneous dendritic cells by CpG-containing oligodeoxynucleotides: a role for dendritic cells in the augmentation of Th1 responses by immunostimulatory DNA"
1623:. Plasmid encoding B7-1 (a ligand on APCs) successfully enhanced the immune response in tumour models. Mixing plasmids encoding GM-CSF and the circumsporozoite protein of
1048:
is true for intracellular and plasma membrane-bound antigens, but not for secreted antigens, which seem to generate TH2 responses, regardless of the method of delivery.
1640:
1 (C-terminus)-hepatitis B virus surface protein (PcMSP1-HBs) abolished protection against challenge, compared to protection acquired by delivery of pPcMSP1-HBs alone.
1212:. Co-inoculation with plasmids encoding co-stimulatory molecules IL-12 and TCA3 were shown to increase CTL activity against HIV-1 and influenza nucleoprotein antigens.
5005:
Sato Y, Roman M, Tighe H, Lee D, Corr M, Nguyen MD, et al. (July 1996). "Immunostimulatory DNA sequences necessary for effective intradermal gene immunization".
1990:
4084:"Protective cellular immunity: cytotoxic T-lymphocyte responses against dominant and recessive epitopes of influenza virus nucleoprotein induced by DNA immunization"
330:
vectors (with multiple genes of interest) are sometimes constructed to express more than one immunogen, or to express an immunogen and an immunostimulatory protein.
199:
A veterinary DNA vaccine to protect horses from West Nile virus has been approved. Another West Nile virus vaccine has been tested successfully on
American robins.
3242:
1237:
Humoral responses after a single DNA injection can be much longer-lived than after a single injection with a recombinant protein. Antibody responses against
372:(RSV) promoter had much higher expression rates. More recently, expression and immunogenicity have been further increased in model systems by the use of the
5201:
Krieg AM, Yi AK, Matson S, Waldschmidt TJ, Bishop GA, Teasdale R, et al. (April 1995). "CpG motifs in bacterial DNA trigger direct B-cell activation".
4382:
Mancini-Bourgine M, Fontaine H, Bréchot C, Pol S, Michel ML (May 2006). "Immunogenicity of a hepatitis B DNA vaccine administered to chronic HBV carriers".
1724:
Prime-boost strategies have been successful in inducing protection against malarial challenge in a number of studies. Primed mice with plasmid DNA encoding
2400:"A West Nile virus DNA vaccine utilizing a modified promoter induces neutralizing antibody in younger and older healthy adults in a phase I clinical trial"
1241:
virus (HBV) envelope protein (HBsAg) have been sustained for up to 74 weeks without boost, while lifelong maintenance of protective response to influenza
3047:"DNA immunization: ubiquitination of a viral protein enhances cytotoxic T-lymphocyte induction and antiviral protection but abrogates antibody induction"
2926:
Oliveira PH, Prather KJ, Prazeres DM, Monteiro GA (September 2009). "Structural instability of plasmid biopharmaceuticals: challenges and implications".
5768:
6823:
955:
Intranasally delivered liposome-DNA complexes can result in expression in distal mucosa as well as nasal muscosa and the generation of IgA antibodies
4504:"Antigen presentation by dendritic cells after immunization with DNA encoding a major histocompatibility complex class II-restricted viral epitope"
4912:"IL-12 and NK cells are required for antigen-specific adaptive immunity against malaria initiated by CD8+ T cells in the Plasmodium yoelii model"
1759:
The efficiency of DNA immunization can be improved by stabilising DNA against degradation, and increasing the efficiency of delivery of DNA into
4703:"Specific immune induction following DNA-based immunization through in vivo transfection and activation of macrophages/antigen-presenting cells"
1526:
5966:
4291:"Enhancement of cell-mediated immunity against HIV-1 induced by coinnoculation of plasmid-encoded HIV-1 antigen with plasmid expressing IL-12"
3815:"Role of different lymphoid tissues in the initiation and maintenance of DNA-raised antibody responses to the influenza virus H1 glycoprotein"
3397:"Induction of CD8+ T cell responses to dominant and subdominant epitopes and protective immunity to Sendai virus infection by DNA vaccination"
5855:
1168:
molecules. These peptides are derived from cytosolic proteins that are degraded and delivered to the nascent MHC class I molecule within the
463:
DNA vaccines have been introduced into animal tissues by multiple methods. In 1999, the two most popular approaches were injection of DNA in
2323:
6006:
6001:
3444:
Lode HN, Huebener N, Zeng Y, Fest S, Weixler S, Gaedicke G (December 2004). "DNA minigene vaccination for adjuvant neuroblastoma therapy".
4961:"Immunization with plasmid DNA encoding for the measles virus hemagglutinin and nucleoprotein leads to humoral and cell-mediated immunity"
2620:"Complexity of the cytokine and antibody response elicited by immunizing mice with Plasmodium yoelii circumsporozoite protein plasmid DNA"
5648:"Multistage multiantigen heterologous prime boost vaccine for Plasmodium knowlesi malaria provides partial protection in rhesus macaques"
2568:"Intranasal administration of plasmid DNA nanoparticles yields successful transfection and expression of a reporter protein in rat brain"
4456:
Anderson RG, Kamen BA, Rothberg KG, Lacey SW (January 1992). "Potocytosis: sequestration and transport of small molecules by caveolae".
601:
inner core and poly(beta-amino ester) outer coat function synergistically to increase efficiency by addressing barriers associated with
2887:"Novel engineered HIV-1 East African Clade-A gp160 plasmid construct induces strong humoral and cell-mediated immune responses in vivo"
985:
low as 16 ng have been reported. These quantities vary by species. Mice for example, require approximately 10 times less DNA than
997:) before it is taken up by the cells, while gene gun deliveries drive/force DNA directly into the cells, resulting in less "wastage".
831:
IM; IV (to transfect tissues systemically); intraperitoneal; oral immunization to the intestinal mucosa; nasal/lung mucosal membranes
104:
from a pathogen, or attenuated viruses which stimulate an immune response in the vaccinated organism. DNA vaccines are members of the
2961:
Oliveira PH, Mairhofer J (September 2013). "Marker-free plasmids for biotechnological applications - implications and perspectives".
4242:"A plasmid encoding murine granulocyte-macrophage colony-stimulating factor increases protection conferred by a malaria DNA vaccine"
1488:
mice have shown that antigen is presented by bone-marrow derived cells, which include dendritic cells, macrophages and specialised
557:, and topical administration of pDNA to the eye and vaginal mucosa. Mucosal surface delivery has also been achieved using cationic
1763:. This has been demonstrated by coating biodegradable cationic microparticles (such as poly(lactide-co-glycolide) formulated with
6403:
249:
expression ensures protein more closely resembles normal eukaryotic structure, with accompanying post-translational modifications
6187:
2042:
Alarcon JB, Waine GW, McManus DP (1999). "DNA Vaccines: Technology and
Application as Anti-parasite and Anti-microbial Agents".
1530:
6016:
4022:"Preferential induction of a Th1 immune response and inhibition of specific IgE antibody formation by plasmid DNA immunization"
265:
Potential when using nasal spray administration of plasmid DNA nanoparticles to transfect non-target cells, such as brain cells
5365:"CpG motifs present in bacteria DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12, and interferon gamma"
405:
lymph nodes and presents the antigen peptide and costimulatory molecule signalling to T-cell, initiating the immune response.
6874:
6802:
2792:
1266:
Table 4. Comparison of T-dependent antibody responses raise by DNA immunisations, protein inoculations and viral infections
447:) from different pathogens raise cytotoxic T-cell responses to some pathogens, especially if a TH epitope is also included.
6408:
413:
Immunogens can be targeted to various cellular compartments to improve antibody or cytotoxic T-cell responses. Secreted or
6440:
6192:
6118:
5971:
120:
protein is included in the DNA, the resulting vaccine can combine the potency of a live vaccine without reversion risks.
5798:
4556:"DNA binding to human leukocytes. Evidence for a receptor-mediated association, internalization, and degradation of DNA"
6850:
6157:
5790:
6262:
2143:
2059:
698:
609:
escape and intracellular cargo concentration. Tested in mice, the hybrid vector was found to induce immune response.
69:(s) against which an immune response is sought, so the cells directly produce the antigen, thus causing a protective
4343:"Long-term maintenance of B cell immunity to influenza virus hemagglutinin in mice following DNA-based immunization"
680:
6943:
6891:
6829:
5799:
1643:
The advantages of genetic adjuvants are their low cost and simple administration, as well as avoidance of unstable
426:
240:
Obviates need for peptide synthesis, expression and purification of recombinant proteins and use of toxic adjuvants
2838:"Increased immune response elicited by DNA vaccination with a synthetic gp120 sequence with optimized codon usage"
6450:
6418:
6413:
6242:
5424:"IFN-gamma promotes IL-6 and IgM secretion in response to CpG motifs in bacterial DNA and oligodeoxynucleotides"
5151:"Augmentation and suppression of immune responses to an HIV-1 DNA vaccine by plasmid cytokine/Ig administration"
4853:"DNA-mediated immunization in a transgenic mouse model of the hepatitis B surface antigen chronic carrier state"
1631:
expressing PyCSP. However, co-injection of plasmids encoding GM-CSF (or IFN-γ, or IL-2) and a fusion protein of
1099:(IL-12) production, while the gene gun bombards the DNA directly into the cell, thus bypassing TH1 stimulation.
6928:
6508:
6113:
676:
1712:
such as ruminants may be insensitive to immunostimulatory sequences due to their large gastrointestinal load.
380:. Additional modifications to improve expression rates include the insertion of enhancer sequences, synthetic
6306:
6011:
5530:"Potent, protective anti-HIV immune responses generated by bimodal HIV envelope DNA plus protein vaccination"
164:
128:
5589:"Boosting with recombinant vaccinia increases immunogenicity and protective efficacy of malaria DNA vaccine"
3345:"Suppression of ongoing ocular inflammatory disease by topical administration of plasmid DNA encoding IL-10"
1172:(ER). Targeting gene products directly to the ER (by the addition of an ER insertion signal sequence at the
4605:"The production and characterization of murine monoclonal antibodies to a DNA receptor on human leukocytes"
672:
259:
Limited to protein immunogens (not useful for non-protein based antigens such as bacterial polysaccharides)
4192:"Enhanced CTL responses mediated by plasmid DNA immunogens encoding costimulatory molecules and cytokines"
2774:
425:
responses can be improved by targeting antigens for cytoplasmic degradation and subsequent entry into the
1553:
6428:
5865:
5848:
3759:"Different T helper cell types and antibody isotypes generated by saline and gene gun DNA immunization"
1536:
4804:"Pathogenetic effector function of CD4-positive T helper 1 cells in hepatitis B virus transgenic mice"
1535:
DNA vaccination generates an effective immune memory via the display of antigen-antibody complexes on
6478:
1153:. This may prove to be a useful tool in assessing CTL epitopes and their role in providing immunity.
1146:
1142:
756:
4754:
2822:
1399:
cells, these cells were thought to be unique because of their extensive network of T-tubules. Using
1075:
differences in TH type. The type of T-cell help raised is determined by the differentiated state of
6433:
6398:
6393:
6388:
1493:
1141:(CTL) without the inherent risk associated with live vaccines. CTL responses can be raised against
1076:
661:
291:
5056:
Weiss R, Leitner WW, Scheiblhofer S, Chen D, Bernhaupt A, Mostböck S, et al. (October 2000).
3573:
Jones CH, Ravikrishnan A, Chen M, Reddinger R, Kamal Ahmadi M, Rane S, et al. (August 2014).
108:, because they contain a genetic information (DNA or RNA) that codes for the cellular production (
6879:
6812:
6793:
6589:
6561:
6523:
6518:
6498:
6021:
5528:
Letvin NL, Montefiori DC, Yasutomi Y, Perry HC, Davies ME, Lekutis C, et al. (August 1997).
1760:
1664:
1637:
1056:. (1995) immunized and boosted mice with pDNA encoding the circumsporozoite protein of the mouse
740:
665:
602:
5725:
Hooper JW, Thompson E, Wilhelmsen C, Zimmerman M, Ichou MA, Steffen SE, et al. (May 2004).
4755:"Differential dependence on target site tissue for gene gun and intramuscular DNA immunizations"
4240:
Weiss WR, Ishii KJ, Hedstrom RC, Sedegah M, Ichino M, Barnhart K, et al. (September 1998).
1407:(or, non-clathrin coated pits). However, subsequent research revealed that other cells (such as
1245:
was demonstrated in mice after gene gun delivery. Antibody-secreting cells (ASC) migrate to the
1191:, or mutation of other signal sequences, was shown to be effective at increasing CTL responses.
6933:
6860:
6789:
5783:
2501:"Protection against malaria by immunization with plasmid DNA encoding circumsporozoite protein"
1088:
1036:
377:
168:
70:
59:
5709:
5149:
Barouch DH, Santra S, Steenbeke TD, Zheng XX, Perry HC, Davies ME, et al. (August 1998).
2885:
Muthumani K, Zhang D, Dayes NS, Hwang DS, Calarota SA, Choo AY, et al. (September 2003).
282:
DNA vaccines elicit the best immune response when high-expression vectors are used. These are
6918:
6855:
6689:
6684:
6135:
6081:
5991:
1601:
1453:
and are thus candidates for DNA uptake. Receptor-mediated DNA uptake could be facilitated by
1169:
744:
583:
vectors for oral administration to the intestinal mucosa and recombinant adenovirus vectors.
333:
Because the plasmid – carrying relatively small genetic code up to about 200 K
295:
5473:"B cell activation and plasma cell differentiation are inhibited by de novo DNA methylation"
4136:
Restifo NP, Bacík I, Irvine KR, Yewdell JW, McCabe BJ, Anderson RW, et al. (May 1995).
2667:
Leitner WW, Seguin MC, Ballou WR, Seitz JP, Schultz AM, Sheehy MJ, Lyon JA (December 1997).
2269:
US Patent 4722848 - Method for immunizing animals with synthetically modified vaccinia virus
1066:(PyCSP) and found that the initial TH2 response changed, after boosting, to a TH1 response.
6738:
6679:
6470:
6108:
5986:
5937:
5904:
5838:
5600:
5541:
5484:
5471:
Barwick BG, Scharer CD, Martinez RJ, Price MJ, Wein AN, Haines RR, et al. (May 2018).
5376:
5314:
5210:
5014:
4864:
4702:
4603:
Bennet RM, Hefeneider SH, Bakke A, Merritt M, Smith CA, Mourich D, Heinrich MC (May 1988).
4465:
4191:
4033:
3923:
3702:
3645:
3586:
3508:
3453:
3254:
2668:
2618:
Mor G, Klinman DM, Shapiro S, Hagiwara E, Sedegah M, Norman JA, et al. (August 1995).
2512:
2288:
2224:
1785:
1776:
1644:
1119:
440:
144:
109:
5253:
5150:
5108:
Sedegah M, Weiss W, Sacci JB, Charoenvit Y, Hedstrom R, Gowda K, et al. (June 2000).
4803:
4604:
4290:
4241:
4137:
3969:
3758:
3691:"DNA vaccines: protective immunizations by parenteral, mucosal, and gene-gun inoculations"
3396:
3344:
2619:
1176:) should thus enhance CTL responses. This was successfully demonstrated using recombinant
167:. The study was planned to involve up to 120 subjects aged between 18 and 35. Separately,
8:
6661:
6627:
6513:
6336:
6279:
6274:
6220:
6051:
5919:
5891:
5843:
5646:
Rogers WO, Baird JK, Kumar A, Tine JA, Weiss W, Aguiar JC, et al. (September 2001).
5423:
4959:
Cardoso AI, Blixenkrone-Moller M, Fayolle J, Liu M, Buckland R, Wild TF (November 1996).
4911:
1921:
1792:
1764:
1633:
1400:
1112:
752:
748:
287:
74:
6331:
6071:
5705:
5604:
5545:
5488:
5380:
5318:
5301:
Krieg AM, Wu T, Weeratna R, Efler SM, Love-Homan L, Yang L, et al. (October 1998).
5214:
5018:
4868:
4469:
4037:
3927:
3706:
3649:
3590:
3551:
3512:
3457:
3299:
Widera G, Austin M, Rabussay D, Goldbeck C, Barnett SW, Chen M, et al. (May 2000).
3258:
2516:
2292:
2228:
268:
Cross-contamination when manufacturing different types of live vaccines in same facility
6694:
6528:
6269:
6230:
6215:
6091:
6076:
6061:
5981:
5899:
5505:
5472:
5453:
5283:
5234:
5180:
5038:
4941:
4833:
4784:
4732:
4678:
4653:
4634:
4528:
4503:
4320:
4271:
4219:
4162:
3999:
3947:
3911:
3788:
3671:
3609:
3574:
3532:
3477:
3426:
3374:
3171:
3123:
3098:
3022:
2997:
2810:
2698:
2649:
2597:
2530:
2473:
2448:
2424:
2399:
2372:
2345:
2193:
1964:
1939:
1893:
1868:
1704:, TNF-α, IFN-α, IFN-β and IFN-γ, while stimulated B-cells secrete IL-6 and some IL-12.
1485:
1465:
618:
389:
369:
361:
315:
303:
5751:
5726:
5082:
5057:
4580:
4555:
4108:
4083:
3888:
3863:
3839:
3814:
3266:
3218:
3193:
3071:
3046:
2903:
2886:
2862:
2837:
2784:
2566:
Harmon BT, Aly AE, Padegimas L, Sesenoglu-Laird O, Cooper MJ, Waszczak BL (May 2014).
2247:
2212:
2135:
2051:
1194:
CTL responses can be enhanced by co-inoculation with co-stimulatory molecules such as
6938:
6817:
6784:
6617:
6458:
6348:
6237:
6098:
5923:
5886:
5776:
5756:
5677:
5672:
5647:
5628:
5623:
5588:
5569:
5564:
5529:
5510:
5445:
5404:
5399:
5364:
5342:
5337:
5303:"Sequence motifs in adenoviral DNA block immune activation by stimulatory CpG motifs"
5302:
5275:
5226:
5172:
5131:
5087:
5073:
5030:
4982:
4933:
4892:
4887:
4852:
4825:
4776:
4724:
4683:
4626:
4585:
4533:
4481:
4438:
4399:
4364:
4312:
4289:
Tsuji T, Hamajima K, Fukushima J, Xin KQ, Ishii N, Aoki I, et al. (April 1997).
4263:
4211:
4167:
4113:
4061:
4056:
4021:
3991:
3939:
3893:
3862:
Sällberg M, Townsend K, Chen M, O'Dea J, Banks T, Jolly DJ, et al. (July 1997).
3844:
3830:
3780:
3730:
3725:
3690:
3689:
Fynan EF, Webster RG, Fuller DH, Haynes JR, Santoro JC, Robinson HL (December 1993).
3663:
3614:
3524:
3469:
3418:
3366:
3322:
3270:
3223:
3209:
3163:
3128:
3076:
3062:
3027:
2978:
2943:
2908:
2867:
2798:
2788:
2734:
2730:
2690:
2641:
2589:
2548:
2543:
2500:
2478:
2429:
2377:
2304:
2252:
2185:
2149:
2139:
2065:
2055:
1969:
1898:
1842:
1652:
1582:
mixture of 2 plasmids, one encoding the immunogen and the other encoding the cytokine
1208:
1062:
994:
637:. Even partial expression libraries can induce protection from subsequent challenge.
464:
299:
180:
83:
6763:
5742:
5663:
5587:
Sedegah M, Jones TR, Kaur M, Hedstrom R, Hobart P, Tine JA, Hoffman SL (June 1998).
5457:
5287:
5184:
4945:
4837:
4788:
4736:
4638:
4324:
4275:
4223:
4099:
4003:
3879:
3792:
3536:
3481:
3378:
3175:
2853:
2702:
2653:
2197:
952:
Intravenously delivered liposome-DNA complexes can potentially transfect all tissues
841:
Table 3. Advantages and disadvantages of commonly used DNA vaccine delivery methods
179:
In August 2021, Indian authorities gave emergency approval to ZyCoV-D. Developed by
55:
sequence into the cells of an organism as a mechanism to induce an immune response.
6743:
6728:
6584:
6445:
6358:
5746:
5738:
5667:
5659:
5618:
5608:
5559:
5549:
5500:
5492:
5435:
5394:
5384:
5332:
5322:
5265:
5238:
5218:
5162:
5121:
5077:
5069:
5042:
5022:
4972:
4923:
4882:
4872:
4851:
Mancini M, Hadchouel M, Davis HL, Whalen RG, Tiollais P, Michel ML (October 1996).
4815:
4766:
4714:
4673:
4665:
4616:
4575:
4567:
4523:
4515:
4473:
4430:
4391:
4354:
4302:
4253:
4203:
4157:
4149:
4103:
4095:
4051:
4041:
3981:
3951:
3931:
3883:
3875:
3834:
3826:
3770:
3720:
3710:
3675:
3653:
3604:
3594:
3575:"Hybrid biosynthetic gene therapy vector development and dual engineering capacity"
3516:
3461:
3430:
3408:
3356:
3312:
3262:
3213:
3205:
3155:
3118:
3110:
3066:
3058:
3017:
3009:
2970:
2935:
2898:
2857:
2849:
2780:
2726:
2680:
2631:
2601:
2579:
2538:
2520:
2468:
2460:
2419:
2411:
2395:
2367:
2357:
2296:
2242:
2232:
2177:
2131:
2047:
1959:
1951:
1888:
1880:
1549:
1419:
1138:
422:
136:
6284:
4395:
3159:
2974:
2939:
518:
delivery ballistically accelerates plasmid DNA (pDNA) that has been absorbed onto
471:
delivery. Several other techniques have been documented in the intervening years.
368:
promoter was conventionally used until research showed that vectors driven by the
139:
into vaccines that may be able to prevent other diseases. They altered the DNA of
6748:
6546:
6353:
6311:
6197:
6177:
6147:
6130:
6056:
6046:
5976:
5915:
5911:
5860:
5126:
5109:
5026:
4771:
4719:
4207:
3520:
2685:
2449:"DNA vaccination of American robins (Turdus migratorius) against West Nile virus"
1497:
1450:
1438:
1080:
1032:
488:
480:
414:
373:
311:
132:
105:
5270:
5167:
4820:
4621:
4307:
4258:
4153:
3986:
3775:
3413:
3361:
3317:
3300:
2636:
1253:
for long-term antibody production, and generally localise there after one year.
1129:
and is somewhat successful in shifting the response for an established disease.
487:(ID), delivering DNA to extracellular spaces. This can be assisted either 1) by
310:
stability and hence increase protein expression. Plasmids also include a strong
6885:
6612:
6503:
6463:
6343:
6164:
6103:
5593:
Proceedings of the
National Academy of Sciences of the United States of America
5534:
Proceedings of the
National Academy of Sciences of the United States of America
5496:
5440:
5369:
Proceedings of the
National Academy of Sciences of the United States of America
5307:
Proceedings of the
National Academy of Sciences of the United States of America
4928:
4857:
Proceedings of the
National Academy of Sciences of the United States of America
4026:
Proceedings of the
National Academy of Sciences of the United States of America
3695:
Proceedings of the National Academy of Sciences of the United States of America
3634:"Protection against mycoplasma infection using expression-library immunization"
3579:
Proceedings of the National Academy of Sciences of the United States of America
2505:
Proceedings of the National Academy of Sciences of the United States of America
2217:
Proceedings of the National Academy of Sciences of the United States of America
1738:
1620:
1589:
1508:
T-cell responses and seems to be important for a full primary immune response.
1242:
1188:
1126:
319:
156:
3301:"Increased DNA vaccine delivery and immunogenicity by electroporation in vivo"
2398:, Pierson TC, Hubka SA, Desai N, Rucker S, Gordon IJ, et al. (May 2011).
6912:
6768:
6753:
6723:
6718:
6573:
6326:
6321:
6225:
5870:
5327:
4877:
3715:
3497:"Attenuated Shigella as a DNA delivery vehicle for DNA-mediated immunization"
1955:
1869:"DNA immunization as a technology platform for monoclonal antibody induction"
1781:
1408:
776:(abdominal skin); vaginal mucosa; surgically exposed muscle and other organs
562:
350:
323:
124:
6125:
5613:
5554:
4477:
4046:
4020:
Raz E, Tighe H, Sato Y, Corr M, Dudler JA, Roman M, et al. (May 1996).
3599:
3465:
2525:
2415:
2362:
2300:
2211:
Paoletti E, Lipinskas BR, Samsonoff C, Mercer S, Panicali D (January 1984).
1023:
responses, including lymphoproliferation and the generation of a variety of
6923:
6845:
6488:
6383:
6316:
6257:
6252:
5932:
5823:
5803:
5760:
5681:
5514:
5389:
5135:
5091:
4977:
4960:
4937:
4669:
4519:
4434:
4403:
4359:
4342:
3914:, Steinman RM (March 1998). "Dendritic cells and the control of immunity".
3618:
3473:
3326:
3274:
3227:
3167:
3031:
2982:
2947:
2912:
2802:
2593:
2482:
2433:
2381:
2153:
2069:
1973:
1902:
1819:
1814:
1597:
1430:
1374:
1095:
into the dendritic cell, which is then stimulated to differentiate for TH1
990:
606:
354:
219:
44:
22:
5727:"Smallpox DNA vaccine protects nonhuman primates against lethal monkeypox"
5632:
5573:
5449:
5408:
5346:
5279:
5230:
5176:
5034:
4986:
4896:
4829:
4780:
4728:
4687:
4630:
4589:
4537:
4485:
4442:
4368:
4316:
4267:
4215:
4171:
4117:
4065:
3995:
3943:
3897:
3848:
3784:
3734:
3667:
3528:
3422:
3370:
3132:
3114:
3080:
2871:
2836:
André S, Seed B, Eberle J, Schraut W, Bültmann A, Haas J (February 1998).
2738:
2694:
2645:
2552:
2464:
2308:
2256:
2237:
1596:
In general, co-administration of pro-inflammatory agents (such as various
1527:
Follicular dendritic cells § Antigen capturing, memory B-cell support
27:
6632:
6622:
6483:
6289:
6172:
6140:
6066:
5819:
5811:
4138:"Antigen processing in vivo and the elicitation of primary CTL responses"
2189:
1809:
1434:
1412:
1246:
1238:
1165:
1161:
1123:
1092:
1084:
496:
215:
148:
95:
4802:
Franco A, Guidotti LG, Hobbs MV, Pasquetto V, Chisari FV (August 1997).
4190:
Iwasaki A, Stiernholm BJ, Chan AK, Berinstein NL, Barber BH (May 1997).
2324:"The U.S. government has begun testing its first Zika vaccine in humans"
1884:
1422:) could also internalize DNA. The mechanism of DNA uptake is not known.
6758:
6733:
6423:
6373:
4502:
Casares S, Inaba K, Brumeanu TD, Steinman RM, Bona CA (November 1997).
2584:
2567:
2534:
2181:
1991:"India gives emergency approval for world's first COVID-19 DNA vaccine"
1804:
1789:
1685:
1628:
1571:
1501:
1443:
1416:
1173:
1116:
1028:
1010:
624:
567:
551:
547:
500:
430:
429:(MHC) class I pathway. This is usually accomplished by the addition of
417:-bound antigens are more effective at inducing antibody responses than
385:
346:
342:
160:
4571:
3343:
Daheshia M, Kuklin N, Kanangat S, Manickan E, Rouse BT (August 1997).
1137:
One of the advantages of DNA vaccines is that they are able to induce
6807:
6378:
6246:
5815:
5222:
3658:
3633:
1575:
1437:, or through specific receptors. These might include a 30kDa surface
1185:
1181:
1150:
820:(cationic); microspheres; recombinant adenovirus vectors; attenuated
817:
492:
433:
418:
357:
more commonly used in the target species may improve its expression.
334:
152:
3496:
3013:
1619:
This concept was applied in topical administration of pDNA encoding
1471:
1381:
650:
262:
Potential for atypical processing of bacterial and parasite proteins
6649:
6642:
6637:
6607:
6363:
6299:
6294:
4958:
4654:"Gene vaccination with naked plasmid DNA: mechanism of CTL priming"
1843:"Developing Snake Antivenom Sera by Genetic Immunization: A Review"
1689:
1668:
1461:
1404:
1177:
1157:
1108:
1096:
1024:
986:
821:
592:
579:
573:
558:
523:
515:
468:
444:
283:
184:
101:
5363:
Klinman DM, Yi AK, Beaucage SL, Conover J, Krieg AM (April 1996).
5254:"Contribution of CpG motifs to the immunogenicity of DNA vaccines"
4381:
3968:
Jakob T, Walker PS, Krieg AM, Udey MC, Vogel JC (September 1998).
3935:
479:
Injection in saline is normally conducted intramuscularly (IM) in
6368:
5807:
5724:
2565:
2128:
DNA vaccines for viral infections: basic studies and applications
1840:
1693:
1656:
1489:
1476:
1456:
1391:
When DNA uptake and subsequent expression was first demonstrated
1333:
1057:
605:
gene delivery which include cellular uptake and internalization,
597:
543:
539:
504:
381:
327:
113:
79:
66:
62:
48:
40:
2210:
1688:. Additionally, flanking regions outside this immunostimulatory
6671:
6656:
5942:
4189:
4082:
Fu TM, Friedman A, Ulmer JB, Liu MA, Donnelly JJ (April 1997).
2925:
1730:
1701:
1681:
1585:
single bi- or polycistronic vector, separated by spacer regions
1396:
1250:
1203:
634:
630:
554:
527:
484:
455:
140:
117:
4701:
Chattergoon MA, Robinson TM, Boyer JD, Weiner DB (June 1998).
4700:
3757:
Feltquate DM, Heaney S, Webster RG, Robinson HL (March 1997).
3756:
3572:
3342:
3145:
1429:
uptake of DNA occurs non-specifically, in a method similar to
875:
Inefficient site for uptake due to morphology of muscle tissue
6666:
6551:
6152:
6086:
5527:
5251:
3861:
1613:
825:
338:
5055:
4801:
4602:
4501:
3813:
Boyle CM, Morin M, Webster RG, Robinson HL (December 1996).
3298:
2499:
Sedegah M, Hedstrom R, Hobart P, Hoffman SL (October 1994).
2498:
172:
135:
vaccines by using genetic engineering to transform ordinary
5961:
5470:
5148:
5107:
4850:
4752:
3191:
2446:
2394:
2130:. Advances in Virus Research. Vol. 55. pp. 1–74.
1660:
1648:
1609:
1605:
1496:(APC). After gene gun inoculation to the skin, transfected
1220:
1199:
1195:
1091:(which supports TH2 responses). pDNA injected by needle is
1069:
519:
365:
307:
4455:
2447:
Kilpatrick AM, Dupuis AP, Chang GJ, Kramer LD (May 2010).
1531:
Follicular dendritic cells § Interaction with B-cells
1102:
4753:
Torres CA, Iwasaki A, Barber BH, Robinson HL (May 1997).
4288:
4239:
4135:
3812:
2617:
2278:
2166:
1647:
and potentially toxic, "conventional" adjuvants (such as
1336:
949:
Can increase transfection of intravenously delivered pDNA
928:
Significant shearing of DNA after high-pressure expulsion
921:
DNA can be delivered to cells mm to cm below skin surface
591:
A hybrid vehicle composed of bacteria cell and synthetic
52:
5586:
5362:
5200:
3688:
3192:
Wunderlich G, Moura IC, del Portillo HA (October 2000).
2884:
2715:
2666:
5252:
Klinman DM, Yamshchikov G, Ishigatsubo Y (April 1997).
3494:
526:
microparticles into the target cells, using compressed
467:: by using a standard hypodermic needle, or by using a
5422:
Yi AK, Chace JH, Cowdery JS, Krieg AM (January 1996).
2835:
1696:-rich to ensure binding and uptake into target cells.
4419:
3495:
Sizemore DR, Branstrom AA, Sadoff JC (October 1995).
3044:
1746:
in humans, will be important in pre-clinical trials.
1403:, it was proposed that DNA uptake was facilitated by
1202:
for DNA vaccines against influenza nucleoprotein, or
1107:
Polarisation in T-cell help is useful in influencing
143:
virus by inserting a gene from other viruses (namely
5645:
5300:
3967:
3443:
459:
DNA vaccine and Gene therapy techniques are similar.
5421:
4652:Corr M, Lee DJ, Carson DA, Tighe H (October 1996).
4553:
3394:
2041:
931:
10-fold lower expression, and lower immune response
86:, it is the first DNA vaccine approved for humans.
4651:
4554:Bennett RM, Gabor GT, Merritt MM (December 1985).
4081:
3910:
3045:Rodriguez F, Zhang J, Whitton JL (November 1997).
1259:primary and secondary phases of antibody response.
1206:for DNA vaccines against the murine malaria model
1035:, patterns of lymphocyte trafficking and types of
228:Immune response focused on the antigen of interest
6824:Cedillo v. Secretary of Health and Human Services
5004:
3808:
3806:
3804:
3802:
3187:
3185:
2960:
2779:. Vol. 54. Academic Press. pp. 129–88.
1520:
1472:Antigen presentation by bone marrow-derived cells
1382:Mechanistic basis for DNA-raised immune responses
934:Requires large amounts of DNA (up to 300 μg)
712:Table 2. Summary of plasmid DNA delivery methods
376:(CMV) immediate early promoter, and a retroviral
6910:
4340:
4336:
4334:
4019:
3752:
3750:
3748:
3746:
3744:
3631:
1232:
491:; 2) by temporarily damaging muscle fibres with
225:Polarise T-cell response toward type 1 or type 2
5358:
5356:
4694:
3338:
3336:
3096:
2125:
2121:
2119:
2117:
2115:
2113:
2111:
2109:
2107:
2105:
2103:
2101:
2099:
1674:
946:High levels of immune response can be generated
595:has been employed for DNA vaccine delivery. An
5103:
5101:
4549:
4547:
4497:
4495:
4185:
4183:
4181:
4131:
4129:
4127:
3904:
3799:
3632:Barry MA, Lai WC, Johnston SA (October 1995).
3395:Chen Y, Webster RG, Woodland DL (March 1998).
3182:
2494:
2492:
2346:"DNA Immunization for HIV Vaccine Development"
2097:
2095:
2093:
2091:
2089:
2087:
2085:
2083:
2081:
2079:
2037:
2035:
2033:
2031:
1847:Clinical Toxinology in Asia Pacific and Africa
1340:
1215:
762:Large amounts (approximately 100-200 μg)
100:Conventional vaccines contain either specific
5784:
4909:
4748:
4746:
4331:
3741:
3554:. The State University of New York at Buffalo
3240:
3092:
3090:
3038:
2995:
2878:
2768:
2029:
2027:
2025:
2023:
2021:
2019:
2017:
2015:
2013:
2011:
1749:
1659:toxin, cationic and mannan-coated liposomes,
1559:
1224:Schematic diagram of an antibody and antigens
1145:and immunorecessive CTL epitopes, as well as
1132:
1042:
903:Th2 response may not be the response required
881:Th1 response may not be the response required
640:
5353:
5196:
5194:
4449:
4415:
4413:
4235:
4233:
3963:
3961:
3855:
3549:
3488:
3333:
2766:
2764:
2762:
2760:
2758:
2756:
2754:
2752:
2750:
2748:
1574:or co-stimulatory molecules. These "genetic
1149:, in a manner that appears to mimic natural
1031:and chemokine expression, specific types of
155:). In 1993, Jeffrey Ulmer and co-workers at
5580:
5245:
5142:
5098:
5000:
4998:
4996:
4952:
4844:
4544:
4492:
4178:
4124:
3390:
3388:
2772:
2660:
2489:
2076:
1916:
1914:
1912:
679:. Unsourced material may be challenged and
533:
5791:
5777:
5639:
5521:
4903:
4795:
4743:
4596:
4015:
4013:
3446:Annals of the New York Academy of Sciences
3292:
3234:
3087:
2613:
2611:
2343:
2008:
1005:
5750:
5671:
5622:
5612:
5563:
5553:
5504:
5439:
5398:
5388:
5336:
5326:
5294:
5269:
5191:
5166:
5125:
5081:
5049:
4976:
4927:
4886:
4876:
4819:
4770:
4718:
4677:
4620:
4579:
4527:
4410:
4358:
4341:Justewicz DM, Webster RG (October 1996).
4306:
4282:
4257:
4230:
4161:
4107:
4077:
4075:
4055:
4045:
3985:
3958:
3887:
3838:
3774:
3724:
3714:
3682:
3657:
3625:
3608:
3598:
3412:
3360:
3316:
3217:
3122:
3070:
3021:
2902:
2861:
2829:
2745:
2684:
2635:
2583:
2542:
2524:
2472:
2423:
2371:
2361:
2246:
2236:
1963:
1892:
1866:
1087:(which supports TH1 cell development) or
699:Learn how and when to remove this message
349:than the target species, so altering the
65:containing the DNA sequence encoding the
4993:
4645:
3385:
2709:
2559:
2321:
1909:
1475:
1219:
1070:Basis for different types of T-cell help
1009:
868:pDNA spreads rapidly throughout the body
779:Small amounts (as little as 16 ng)
454:
408:
399:
26:
4010:
2608:
1933:
1931:
1841:Henrique Roman Ramos and Paulo Lee Ho.
1836:
1834:
1386:
1262:robust enough to control HBV infection
1103:Practical uses of polarised T-cell help
857:Intramuscular or Intradermal injection
617:Another approach to DNA vaccination is
360:Another consideration is the choice of
286:that usually consist of a strong viral
6911:
5415:
4072:
3097:Tobery TW, Siliciano RF (March 1997).
2996:Kutzler MA, Weiner DB (October 2008).
2344:Chen Y, Wang S, Lu S (February 2014).
1940:"DNA vaccines: roles against diseases"
1564:
1332:Number of inoculations to obtain high
865:Permanent or semi-permanent expression
633:lung pathogen with a relatively small
345:cells. Pathogens often have different
183:, it is the first DNA vaccine against
5772:
4560:The Journal of Clinical Investigation
2998:"DNA vaccines: ready for prime time?"
2315:
1862:
1860:
1770:
1715:
1019:DNA immunization can raise multiple T
306:may sometimes be included to improve
4658:The Journal of Experimental Medicine
4508:The Journal of Experimental Medicine
3552:"A hybrid vehicle that delivers DNA"
3103:The Journal of Experimental Medicine
1985:
1983:
1937:
1928:
1831:
1788:have been used to build recombinant
1122:for the desired type of response in
878:Relatively large amounts of DNA used
677:adding citations to reliable sources
644:
4910:Doolan DL, Hoffman SL (July 1999).
3241:Weiner DB, Kennedy RC (July 1999).
1511:
906:Requires inert particles as carrier
795:Very high (as much as 300 μg)
612:
474:
13:
6851:Eradication of infectious diseases
6608:Androvax (androstenedione albumin)
5691:
2453:Vector Borne and Zoonotic Diseases
2404:The Journal of Infectious Diseases
2046:. Vol. 42. pp. 343–410.
2044:Advances in Parasitology Volume 42
1873:Emerging Microbes & Infections
1867:Liu S, Wang S, Lu S (April 2016).
1857:
1156:Cytotoxic T-cells recognise small
1000:
809:Small amounts (up to 100 μg)
586:
378:cis-acting transcriptional element
272:
243:Long-term persistence of immunogen
234:Stability for storage and shipping
231:Ease of development and production
14:
6955:
6441:Respiratory syncytial virus (RSV)
3550:Nealon, Cory (25 November 2014).
3267:10.1038/scientificamerican0799-50
1980:
1304:Duration of antigen presentation
893:DNA bombarded directly into cells
790:
773:
6830:Alternative vaccination schedule
5800:Artificial induction of immunity
5698:
5074:10.1128/IAI.68.10.5914-5919.2000
3831:10.1128/JVI.70.12.9074-9078.1996
3210:10.1128/IAI.68.10.5839-5845.2000
3063:10.1128/JVI.71.11.8497-8503.1997
2719:Journal of Immunological Methods
2326:. MIT Technology Review Magazine
2126:Robinson HL, Pertmer TM (2000).
649:
427:major histocompatibility complex
353:of the immunogen to reflect the
277:
253:
5743:10.1128/JVI.78.9.4433-4443.2004
5664:10.1128/IAI.69.9.5565-5572.2001
5464:
4375:
4100:10.1128/JVI.71.4.2715-2721.1997
3880:10.1128/JVI.71.7.5295-5303.1997
3566:
3543:
3437:
3139:
2989:
2954:
2919:
2854:10.1128/JVI.72.2.1497-1503.1998
2440:
2388:
2337:
2272:
2263:
1360:C’-dependent or C’-independent
190:
163:began testing in humans at the
58:DNA vaccines work by injecting
6662:Ovandrotone albumin (Fecundin)
2204:
2160:
1765:cetyltrimethylammonium bromide
1542:
1521:Maintenance of immune response
1318:Kinetics of antibody response
734:Injection (hypodermic needle)
341:usage of pathogenic mRNAs for
131:devised a strategy to produce
1:
6193:Group B streptococcal disease
6012:Vaccines for Children Program
4396:10.1016/j.vaccine.2005.08.013
3160:10.1158/1535-7163.MCT-08-0109
3148:Molecular Cancer Therapeutics
2975:10.1016/j.tibtech.2013.06.001
2940:10.1016/j.tibtech.2009.06.004
2904:10.1016/S0042-6822(03)00459-8
2785:10.1016/S0065-3527(08)60367-X
2136:10.1016/S0065-3527(00)55001-5
2052:10.1016/S0065-308X(08)60152-9
1825:
1775:Recombinant alphavirus-based
1425:Two theories dominate – that
1233:Kinetics of antibody response
1083:can differentiate to secrete
862:No special delivery mechanism
824:vector; aerosolised cationic
542:instillation of naked DNA on
214:Antigen presentation by both
205:
165:National Institutes of Health
129:New York Department of Health
5127:10.4049/jimmunol.164.11.5905
5027:10.1126/science.273.5273.352
4772:10.4049/jimmunol.158.10.4529
4720:10.4049/jimmunol.160.12.5707
4208:10.4049/jimmunol.158.10.4591
3521:10.1126/science.270.5234.299
2773:Lewis PJ, Babiuk LA (1999).
2731:10.1016/0022-1759(96)00035-X
2686:10.4049/jimmunol.159.12.6112
2322:Regalado A (2 August 2016).
1924:. World Health Organization.
1737:has also been demonstrated.
1675:Immunostimulatory CpG motifs
1578:" can be administered as a:
23:messenger RNA (mRNA) vaccine
7:
5271:10.4049/jimmunol.158.8.3635
5168:10.4049/jimmunol.161.4.1875
4821:10.4049/jimmunol.159.4.2001
4622:10.4049/jimmunol.140.9.2937
4308:10.4049/jimmunol.158.8.4008
4259:10.4049/jimmunol.161.5.2325
4154:10.4049/jimmunol.154.9.4414
3987:10.4049/jimmunol.161.6.3042
3776:10.4049/jimmunol.158.5.2278
3414:10.4049/jimmunol.160.5.2425
3362:10.4049/jimmunol.159.4.1945
3318:10.4049/jimmunol.164.9.4635
2637:10.4049/jimmunol.155.4.2039
1798:
1466:cationic liposome packaging
1357:Ab isotype (murine models)
1290:Amount of inducing antigen
1216:Humoral (antibody) response
941:Liposome-mediated delivery
737:Aqueous solution in saline
510:
450:
326:polyadenylation sequences.
157:Merck Research Laboratories
127:and Dennis Panicali at the
31:The making of a DNA vaccine
10:
6960:
5497:10.1038/s41467-018-04234-4
5441:10.4049/jimmunol.156.2.558
4929:10.4049/jimmunol.163.2.884
1750:Enhancing immune responses
1655:, monophosphoryl lipid A,
1560:Immune response modulation
1537:follicular dendritic cells
1524:
1133:Cytotoxic T-cell responses
1043:Other types of T-cell help
784:Pneumatic (jet) injection
641:Helpful tabular comparison
93:
89:
20:
6869:
6838:
6777:
6709:
6600:
6572:
6539:
6208:
6039:
6032:
5951:
5879:
5831:
1638:merozoite surface protein
1273:
1270:
979:
851:
848:
845:
813:
799:
730:
716:
565:microspheres, attenuated
6188:Clostridioides difficile
5328:10.1073/pnas.95.21.12631
4878:10.1073/pnas.93.22.12496
3716:10.1073/pnas.90.24.11478
3002:Nature Reviews. Genetics
1956:10.11599/germs.2013.1034
1761:antigen-presenting cells
1500:migrate to the draining
1494:antigen presenting cells
1147:subdominant CTL epitopes
1077:antigen presenting cells
965:Ineffectiveness in serum
534:Mucosal surface delivery
6944:21st-century inventions
6451:Tick-borne encephalitis
5614:10.1073/pnas.95.13.7648
5555:10.1073/pnas.94.17.9378
4478:10.1126/science.1310359
4423:Journal of Cell Science
4047:10.1073/pnas.93.10.5141
3600:10.1073/pnas.1411355111
3466:10.1196/annals.1322.012
2963:Trends in Biotechnology
2928:Trends in Biotechnology
2526:10.1073/pnas.91.21.9866
2363:10.3390/vaccines2010138
2301:10.1126/science.8456302
1665:carboxymethyl cellulose
1457:polyguanylate sequences
1274:Method of Immunization
1139:cytotoxic T lymphocytes
1037:innate immune responses
1006:Helper T cell responses
759:with variable success)
603:antigen-presenting cell
503:solutions of saline or
6861:List of vaccine topics
5856:Mathematical modelling
5652:Infection and Immunity
5390:10.1073/pnas.93.7.2879
5062:Infection and Immunity
4978:10.1006/viro.1996.0603
4670:10.1084/jem.184.4.1555
4520:10.1084/jem.186.9.1481
4435:10.1242/jcs.103.4.1249
4360:10.1006/viro.1996.0501
3198:Infection and Immunity
2776:DNA vaccines: a review
1938:Khan KH (March 2013).
1754:
1481:
1225:
1016:
770:DNA-coated gold beads
546:surfaces, such as the
538:Alternatives included
460:
211:No risk for infections
169:Inovio Pharmaceuticals
96:Vaccine § History
71:immunological response
60:genetically engineered
32:
16:Vaccine containing DNA
6929:Nucleic acid vaccines
6856:Vaccinate Your Family
6307:Japanese encephalitis
5477:Nature Communications
5428:Journal of Immunology
5258:Journal of Immunology
5155:Journal of Immunology
5114:Journal of Immunology
4916:Journal of Immunology
4808:Journal of Immunology
4759:Journal of Immunology
4707:Journal of Immunology
4609:Journal of Immunology
4295:Journal of Immunology
4246:Journal of Immunology
4196:Journal of Immunology
4142:Journal of Immunology
3974:Journal of Immunology
3763:Journal of Immunology
3401:Journal of Immunology
3349:Journal of Immunology
3305:Journal of Immunology
3115:10.1084/jem.185.5.909
2673:Journal of Immunology
2624:Journal of Immunology
2465:10.1089/vbz.2009.0029
2416:10.1093/infdis/jir054
2238:10.1073/pnas.81.1.193
1645:recombinant cytokines
1602:tumor necrosis factor
1479:
1223:
1170:endoplasmic reticulum
1013:
993:and large amounts of
918:No particles required
806:Ocular; intravaginal
458:
409:Vaccine insert design
400:Mechanism of plasmids
290:to drive the in vivo
30:
6739:John Franklin Enders
1786:Semliki Forest virus
1616:are sometimes used.
1492:called professional
1446:scavenger receptors.
1387:DNA uptake mechanism
1282:Recombinant protein
814:Cytofectin-mediated
800:Topical application
673:improve this section
173:GeneOne Life Science
145:Herpes simplex virus
110:protein biosynthesis
5920:Virus-like particle
5844:Vaccine ingredients
5731:Journal of Virology
5605:1998PNAS...95.7648S
5546:1997PNAS...94.9378L
5489:2018NatCo...9.1900B
5381:1996PNAS...93.2879K
5319:1998PNAS...9512631K
5313:(21): 12631–12636.
5215:1995Natur.374..546K
5019:1996Sci...273..352S
4869:1996PNAS...9312496M
4863:(22): 12496–12501.
4470:1992Sci...255..410A
4088:Journal of Virology
4038:1996PNAS...93.5141R
3928:1998Natur.392..245B
3868:Journal of Virology
3819:Journal of Virology
3707:1993PNAS...9011478F
3701:(24): 11478–11482.
3650:1995Natur.377..632B
3591:2014PNAS..11112360J
3585:(34): 12360–12365.
3513:1995Sci...270..299S
3458:2004NYASA1028..113L
3259:1999SciAm.281a..50W
3247:Scientific American
3051:Journal of Virology
2842:Journal of Virology
2517:1994PNAS...91.9866S
2293:1993Sci...259.1745U
2287:(5102): 1745–1749.
2229:1984PNAS...81..193P
1885:10.1038/emi.2016.27
1592:, or fusion protein
1565:Cytokine modulation
1401:electron microscopy
1267:
1180:viruses expressing
1113:autoimmune diseases
968:Risk of disease or
846:Method of delivery
842:
713:
561:-DNA preparations,
6896:Never to phase III
6695:Hexavalent vaccine
6509:Epstein–Barr virus
6404:Oxford–AstraZeneca
6092:NmVac4-A/C/Y/W-135
5708:has a profile for
3243:"Genetic vaccines"
2585:10.1038/gt.2014.28
2182:10.1007/bf00470753
1771:Alphavirus vectors
1716:Alternative boosts
1482:
1464:delivery systems,
1285:Natural infection
1265:
1226:
1017:
840:
720:Formulation of DNA
717:Method of delivery
711:
619:expression library
530:as an accelerant.
461:
370:Rous Sarcoma Virus
316:termination signal
237:Cost-effectiveness
33:
6906:
6905:
6818:Vaccines and SIDS
6705:
6704:
6479:Hepatitis A and B
6455:Varicella zoster
5912:Subunit/component
5714:
5599:(13): 7648–7653.
5540:(17): 9378–9383.
5209:(6522): 546–549.
5120:(11): 5905–5912.
5068:(10): 5914–5919.
5013:(5273): 352–354.
4765:(10): 4529–4532.
4713:(12): 5707–5718.
4572:10.1172/JCI112226
4464:(5043): 410–411.
4390:(21): 4482–4489.
4202:(10): 4591–4601.
4032:(10): 5141–5145.
3922:(6673): 245–252.
3825:(12): 9074–9078.
3644:(6550): 632–635.
3507:(5234): 299–302.
3204:(10): 5839–5845.
3057:(11): 8497–8503.
2794:978-0-12-039854-6
2679:(12): 6112–6119.
2511:(21): 9866–9870.
2410:(10): 1396–1404.
1726:Plasmodium yoelii
1653:calcium phosphate
1370:
1369:
1339:and migration of
1063:Plasmodium yoelii
995:connective tissue
977:
976:
896:Small amounts DNA
838:
837:
803:Aqueous solution
787:Aqueous solution
709:
708:
701:
499:; or 3) by using
318:, such as bovine
314:/transcriptional
300:complementary DNA
181:Cadila Healthcare
84:Cadila Healthcare
6951:
6797:MMR autism fraud
6744:Maurice Hilleman
6729:Hilary Koprowski
6037:
6036:
5793:
5786:
5779:
5770:
5769:
5764:
5754:
5737:(9): 4433–4443.
5712:
5702:
5701:
5686:
5685:
5675:
5658:(9): 5565–5572.
5643:
5637:
5636:
5626:
5616:
5584:
5578:
5577:
5567:
5557:
5525:
5519:
5518:
5508:
5468:
5462:
5461:
5443:
5419:
5413:
5412:
5402:
5392:
5375:(7): 2879–2883.
5360:
5351:
5350:
5340:
5330:
5298:
5292:
5291:
5273:
5264:(8): 3635–3639.
5249:
5243:
5242:
5223:10.1038/374546a0
5198:
5189:
5188:
5170:
5161:(4): 1875–1882.
5146:
5140:
5139:
5129:
5105:
5096:
5095:
5085:
5053:
5047:
5046:
5002:
4991:
4990:
4980:
4956:
4950:
4949:
4931:
4907:
4901:
4900:
4890:
4880:
4848:
4842:
4841:
4823:
4814:(4): 2001–2008.
4799:
4793:
4792:
4774:
4750:
4741:
4740:
4722:
4698:
4692:
4691:
4681:
4664:(4): 1555–1560.
4649:
4643:
4642:
4624:
4615:(9): 2937–2942.
4600:
4594:
4593:
4583:
4566:(6): 2182–2190.
4551:
4542:
4541:
4531:
4514:(9): 1481–1486.
4499:
4490:
4489:
4453:
4447:
4446:
4429:(4): 1249–1259.
4417:
4408:
4407:
4379:
4373:
4372:
4362:
4338:
4329:
4328:
4310:
4301:(8): 4008–4013.
4286:
4280:
4279:
4261:
4252:(5): 2325–2332.
4237:
4228:
4227:
4187:
4176:
4175:
4165:
4148:(9): 4414–4422.
4133:
4122:
4121:
4111:
4094:(4): 2715–2721.
4079:
4070:
4069:
4059:
4049:
4017:
4008:
4007:
3989:
3980:(6): 3042–3049.
3965:
3956:
3955:
3908:
3902:
3901:
3891:
3874:(7): 5295–5303.
3859:
3853:
3852:
3842:
3810:
3797:
3796:
3778:
3769:(5): 2278–2284.
3754:
3739:
3738:
3728:
3718:
3686:
3680:
3679:
3661:
3659:10.1038/377632a0
3629:
3623:
3622:
3612:
3602:
3570:
3564:
3563:
3561:
3559:
3547:
3541:
3540:
3492:
3486:
3485:
3441:
3435:
3434:
3416:
3407:(5): 2425–2432.
3392:
3383:
3382:
3364:
3355:(4): 1945–1952.
3340:
3331:
3330:
3320:
3311:(9): 4635–4640.
3296:
3290:
3289:
3287:
3286:
3277:. Archived from
3238:
3232:
3231:
3221:
3189:
3180:
3179:
3154:(7): 2241–2251.
3143:
3137:
3136:
3126:
3094:
3085:
3084:
3074:
3042:
3036:
3035:
3025:
2993:
2987:
2986:
2958:
2952:
2951:
2923:
2917:
2916:
2906:
2882:
2876:
2875:
2865:
2848:(2): 1497–1503.
2833:
2827:
2826:
2820:
2816:
2814:
2806:
2770:
2743:
2742:
2713:
2707:
2706:
2688:
2664:
2658:
2657:
2639:
2630:(4): 2039–2046.
2615:
2606:
2605:
2587:
2563:
2557:
2556:
2546:
2528:
2496:
2487:
2486:
2476:
2444:
2438:
2437:
2427:
2392:
2386:
2385:
2375:
2365:
2341:
2335:
2334:
2332:
2331:
2319:
2313:
2312:
2276:
2270:
2267:
2261:
2260:
2250:
2240:
2208:
2202:
2201:
2164:
2158:
2157:
2123:
2074:
2073:
2039:
2006:
2005:
2003:
2002:
1987:
1978:
1977:
1967:
1935:
1926:
1925:
1918:
1907:
1906:
1896:
1864:
1855:
1854:
1838:
1588:plasmid-encoded
1512:Target site role
1498:Langerhans cells
1480:A dendritic cell
1460:
1455:the presence of
1447:
1420:Langerhans cells
1342:
1268:
1264:
1260:
1189:signal sequences
971:
970:immune reactions
843:
839:
792:
775:
714:
710:
704:
697:
693:
690:
684:
653:
645:
613:ELI immunization
475:Saline injection
423:cytotoxic T-cell
421:antigens, while
298:of the gene (or
137:smallpox vaccine
106:genetic vaccines
6959:
6958:
6954:
6953:
6952:
6950:
6949:
6948:
6909:
6908:
6907:
6902:
6901:
6886:Clinical trials
6865:
6834:
6773:
6749:Stanley Plotkin
6711:
6701:
6613:Cancer vaccines
6596:
6590:Schistosomiasis
6568:
6562:Trypanosomiasis
6535:
6499:Cytomegalovirus
6409:Pfizer–BioNTech
6204:
6028:
5977:Vaccine wastage
5947:
5875:
5827:
5797:
5767:
5720:
5719:
5718:
5703:
5699:
5694:
5692:Further reading
5689:
5644:
5640:
5585:
5581:
5526:
5522:
5469:
5465:
5420:
5416:
5361:
5354:
5299:
5295:
5250:
5246:
5199:
5192:
5147:
5143:
5106:
5099:
5054:
5050:
5003:
4994:
4957:
4953:
4908:
4904:
4849:
4845:
4800:
4796:
4751:
4744:
4699:
4695:
4650:
4646:
4601:
4597:
4552:
4545:
4500:
4493:
4454:
4450:
4418:
4411:
4380:
4376:
4339:
4332:
4287:
4283:
4238:
4231:
4188:
4179:
4134:
4125:
4080:
4073:
4018:
4011:
3966:
3959:
3909:
3905:
3860:
3856:
3811:
3800:
3755:
3742:
3687:
3683:
3630:
3626:
3571:
3567:
3557:
3555:
3548:
3544:
3493:
3489:
3442:
3438:
3393:
3386:
3341:
3334:
3297:
3293:
3284:
3282:
3239:
3235:
3190:
3183:
3144:
3140:
3095:
3088:
3043:
3039:
3014:10.1038/nrg2432
3008:(10): 776–788.
2994:
2990:
2959:
2955:
2924:
2920:
2883:
2879:
2834:
2830:
2818:
2817:
2808:
2807:
2795:
2771:
2746:
2714:
2710:
2665:
2661:
2616:
2609:
2564:
2560:
2497:
2490:
2445:
2441:
2393:
2389:
2342:
2338:
2329:
2327:
2320:
2316:
2277:
2273:
2268:
2264:
2209:
2205:
2165:
2161:
2146:
2124:
2077:
2062:
2040:
2009:
2000:
1998:
1989:
1988:
1981:
1936:
1929:
1920:
1919:
1910:
1865:
1858:
1839:
1832:
1828:
1801:
1773:
1757:
1752:
1733:malarial model
1718:
1677:
1567:
1562:
1545:
1533:
1523:
1514:
1474:
1454:
1451:ribonucleotides
1442:
1389:
1384:
1366:C’-independent
1343:to bone marrow
1258:
1235:
1218:
1164:) complexed to
1135:
1105:
1081:Dendritic cells
1072:
1045:
1033:immunoglobulins
1022:
1008:
1003:
1001:Immune response
982:
969:
757:intraperitoneal
705:
694:
688:
685:
670:
654:
643:
615:
589:
587:Polymer vehicle
536:
513:
489:electroporation
481:skeletal muscle
477:
453:
415:plasma membrane
411:
402:
374:cytomegalovirus
312:polyadenylation
302:) of interest.
280:
275:
273:Plasmid vectors
256:
208:
193:
133:recombinant DNA
98:
92:
82:. Developed by
25:
17:
12:
11:
5:
6957:
6947:
6946:
6941:
6936:
6931:
6926:
6921:
6904:
6903:
6900:
6899:
6898:
6897:
6894:
6883:
6877:
6871:
6870:
6867:
6866:
6864:
6863:
6858:
6853:
6848:
6842:
6840:
6836:
6835:
6833:
6832:
6827:
6820:
6815:
6810:
6805:
6800:
6787:
6781:
6779:
6775:
6774:
6772:
6771:
6766:
6764:Katalin Karikó
6761:
6756:
6751:
6746:
6741:
6736:
6731:
6726:
6721:
6715:
6713:
6707:
6706:
6703:
6702:
6700:
6699:
6698:
6697:
6692:
6687:
6682:
6674:
6669:
6664:
6659:
6654:
6653:
6652:
6647:
6646:
6645:
6640:
6630:
6625:
6620:
6610:
6604:
6602:
6598:
6597:
6595:
6594:
6593:
6592:
6587:
6578:
6576:
6570:
6569:
6567:
6566:
6565:
6564:
6556:
6555:
6554:
6543:
6541:
6537:
6536:
6534:
6533:
6532:
6531:
6526:
6521:
6519:Herpes simplex
6516:
6511:
6506:
6501:
6493:
6492:
6491:
6486:
6481:
6473:
6468:
6467:
6466:
6461:
6453:
6448:
6443:
6438:
6437:
6436:
6431:
6426:
6421:
6419:Sinopharm BIBP
6416:
6411:
6406:
6401:
6396:
6391:
6386:
6381:
6376:
6371:
6369:Bharat Biotech
6366:
6356:
6351:
6346:
6341:
6340:
6339:
6334:
6324:
6319:
6314:
6309:
6304:
6303:
6302:
6297:
6287:
6282:
6277:
6272:
6267:
6266:
6265:
6260:
6255:
6250:
6235:
6234:
6233:
6223:
6218:
6212:
6210:
6206:
6205:
6203:
6202:
6201:
6200:
6195:
6190:
6182:
6181:
6180:
6175:
6167:
6162:
6161:
6160:
6155:
6145:
6144:
6143:
6133:
6128:
6123:
6122:
6121:
6116:
6106:
6101:
6096:
6095:
6094:
6089:
6079:
6074:
6069:
6064:
6059:
6054:
6049:
6043:
6041:
6034:
6030:
6029:
6027:
6026:
6025:
6024:
6019:
6014:
6009:
6004:
5996:
5995:
5994:
5992:Vaccine injury
5989:
5984:
5979:
5974:
5969:
5964:
5955:
5953:
5952:Administration
5949:
5948:
5946:
5945:
5940:
5935:
5926:
5909:
5908:
5907:
5902:
5894:
5889:
5883:
5881:
5877:
5876:
5874:
5873:
5868:
5863:
5858:
5853:
5852:
5851:
5841:
5835:
5833:
5829:
5828:
5796:
5795:
5788:
5781:
5773:
5766:
5765:
5721:
5704:
5697:
5696:
5695:
5693:
5690:
5688:
5687:
5638:
5579:
5520:
5463:
5434:(2): 558–564.
5414:
5352:
5293:
5244:
5190:
5141:
5097:
5048:
4992:
4971:(2): 293–299.
4951:
4922:(2): 884–892.
4902:
4843:
4794:
4742:
4693:
4644:
4595:
4543:
4491:
4448:
4409:
4374:
4330:
4281:
4229:
4177:
4123:
4071:
4009:
3957:
3903:
3854:
3798:
3740:
3681:
3624:
3565:
3542:
3487:
3452:(1): 113–121.
3436:
3384:
3332:
3291:
3233:
3181:
3138:
3109:(5): 909–920.
3086:
3037:
2988:
2969:(9): 539–547.
2953:
2934:(9): 503–511.
2918:
2897:(1): 134–146.
2877:
2828:
2819:|journal=
2793:
2744:
2708:
2659:
2607:
2578:(5): 514–521.
2558:
2488:
2459:(4): 377–380.
2439:
2387:
2356:(1): 138–159.
2336:
2314:
2271:
2262:
2223:(1): 193–197.
2203:
2176:(3): 173–177.
2170:Mycopathologia
2159:
2144:
2075:
2060:
2007:
1979:
1927:
1922:"DNA vaccines"
1908:
1856:
1829:
1827:
1824:
1823:
1822:
1817:
1812:
1807:
1800:
1797:
1772:
1769:
1756:
1753:
1751:
1748:
1739:Rhesus monkeys
1717:
1714:
1676:
1673:
1594:
1593:
1586:
1583:
1566:
1563:
1561:
1558:
1544:
1541:
1522:
1519:
1513:
1510:
1484:Studies using
1473:
1470:
1388:
1385:
1383:
1380:
1368:
1367:
1364:
1361:
1358:
1354:
1353:
1350:
1347:
1344:
1329:
1328:
1325:
1322:
1319:
1315:
1314:
1313:several weeks
1311:
1308:
1307:several weeks
1305:
1301:
1300:
1297:
1294:
1291:
1287:
1286:
1283:
1280:
1276:
1275:
1272:
1243:haemagglutinin
1234:
1231:
1217:
1214:
1143:immunodominant
1134:
1131:
1111:responses and
1104:
1101:
1071:
1068:
1044:
1041:
1020:
1007:
1004:
1002:
999:
981:
978:
975:
974:
973:
972:
966:
963:
958:
957:
956:
953:
950:
947:
942:
938:
937:
936:
935:
932:
929:
924:
923:
922:
919:
914:
913:Jet injection
910:
909:
908:
907:
904:
899:
898:
897:
894:
889:
885:
884:
883:
882:
879:
876:
871:
870:
869:
866:
863:
858:
854:
853:
850:
847:
836:
835:
832:
829:
815:
811:
810:
807:
804:
801:
797:
796:
793:
788:
785:
781:
780:
777:
771:
768:
764:
763:
760:
738:
735:
732:
728:
727:
726:Amount of DNA
724:
721:
718:
707:
706:
657:
655:
648:
642:
639:
614:
611:
588:
585:
535:
532:
512:
509:
476:
473:
452:
449:
410:
407:
401:
398:
320:growth hormone
279:
276:
274:
271:
270:
269:
266:
263:
260:
255:
252:
251:
250:
244:
241:
238:
235:
232:
229:
226:
223:
212:
207:
204:
192:
189:
91:
88:
15:
9:
6:
4:
3:
2:
6956:
6945:
6942:
6940:
6937:
6935:
6934:Gene delivery
6932:
6930:
6927:
6925:
6922:
6920:
6917:
6916:
6914:
6895:
6893:
6890:
6889:
6887:
6884:
6881:
6878:
6876:
6873:
6872:
6868:
6862:
6859:
6857:
6854:
6852:
6849:
6847:
6844:
6843:
6841:
6837:
6831:
6828:
6826:
6825:
6821:
6819:
6816:
6814:
6811:
6809:
6806:
6804:
6801:
6798:
6796:
6791:
6788:
6786:
6783:
6782:
6780:
6776:
6770:
6769:Drew Weissman
6767:
6765:
6762:
6760:
6757:
6755:
6754:H. Fred Clark
6752:
6750:
6747:
6745:
6742:
6740:
6737:
6735:
6732:
6730:
6727:
6725:
6724:Louis Pasteur
6722:
6720:
6719:Edward Jenner
6717:
6716:
6714:
6708:
6696:
6693:
6691:
6690:DTwP-HepB-Hib
6688:
6686:
6685:DTaP-IPV-HepB
6683:
6681:
6678:
6677:
6676:combination:
6675:
6673:
6670:
6668:
6665:
6663:
6660:
6658:
6655:
6651:
6648:
6644:
6641:
6639:
6636:
6635:
6634:
6631:
6629:
6626:
6624:
6621:
6619:
6616:
6615:
6614:
6611:
6609:
6606:
6605:
6603:
6599:
6591:
6588:
6586:
6583:
6582:
6580:
6579:
6577:
6575:
6574:Helminthiasis
6571:
6563:
6560:
6559:
6557:
6553:
6550:
6549:
6548:
6545:
6544:
6542:
6538:
6530:
6527:
6525:
6522:
6520:
6517:
6515:
6512:
6510:
6507:
6505:
6502:
6500:
6497:
6496:
6494:
6490:
6487:
6485:
6482:
6480:
6477:
6476:
6475:combination:
6474:
6472:
6469:
6465:
6462:
6460:
6457:
6456:
6454:
6452:
6449:
6447:
6444:
6442:
6439:
6435:
6432:
6430:
6427:
6425:
6422:
6420:
6417:
6415:
6412:
6410:
6407:
6405:
6402:
6400:
6397:
6395:
6392:
6390:
6387:
6385:
6382:
6380:
6377:
6375:
6372:
6370:
6367:
6365:
6362:
6361:
6360:
6357:
6355:
6352:
6350:
6347:
6345:
6342:
6338:
6335:
6333:
6330:
6329:
6328:
6325:
6323:
6320:
6318:
6315:
6313:
6310:
6308:
6305:
6301:
6298:
6296:
6293:
6292:
6291:
6288:
6286:
6283:
6281:
6278:
6276:
6273:
6271:
6268:
6264:
6261:
6259:
6256:
6254:
6251:
6248:
6244:
6241:
6240:
6239:
6236:
6232:
6229:
6228:
6227:
6224:
6222:
6219:
6217:
6214:
6213:
6211:
6207:
6199:
6196:
6194:
6191:
6189:
6186:
6185:
6183:
6179:
6176:
6174:
6173:DPT/DTwP/DTaP
6171:
6170:
6169:combination:
6168:
6166:
6163:
6159:
6156:
6154:
6151:
6150:
6149:
6146:
6142:
6139:
6138:
6137:
6134:
6132:
6129:
6127:
6124:
6120:
6117:
6115:
6112:
6111:
6110:
6107:
6105:
6102:
6100:
6097:
6093:
6090:
6088:
6085:
6084:
6083:
6082:Meningococcus
6080:
6078:
6075:
6073:
6072:Leptospirosis
6070:
6068:
6065:
6063:
6060:
6058:
6055:
6053:
6050:
6048:
6045:
6044:
6042:
6038:
6035:
6031:
6023:
6020:
6018:
6015:
6013:
6010:
6008:
6007:Vaccine court
6005:
6003:
6000:
5999:
5997:
5993:
5990:
5988:
5985:
5983:
5980:
5978:
5975:
5973:
5970:
5968:
5965:
5963:
5962:GAVI Alliance
5960:
5959:
5957:
5956:
5954:
5950:
5944:
5941:
5939:
5936:
5934:
5930:
5927:
5925:
5921:
5917:
5913:
5910:
5906:
5903:
5901:
5898:
5897:
5895:
5893:
5890:
5888:
5885:
5884:
5882:
5878:
5872:
5869:
5867:
5864:
5862:
5859:
5857:
5854:
5850:
5847:
5846:
5845:
5842:
5840:
5837:
5836:
5834:
5830:
5825:
5821:
5817:
5813:
5809:
5805:
5801:
5794:
5789:
5787:
5782:
5780:
5775:
5774:
5771:
5762:
5758:
5753:
5748:
5744:
5740:
5736:
5732:
5728:
5723:
5722:
5716:
5715:
5707:
5683:
5679:
5674:
5669:
5665:
5661:
5657:
5653:
5649:
5642:
5634:
5630:
5625:
5620:
5615:
5610:
5606:
5602:
5598:
5594:
5590:
5583:
5575:
5571:
5566:
5561:
5556:
5551:
5547:
5543:
5539:
5535:
5531:
5524:
5516:
5512:
5507:
5502:
5498:
5494:
5490:
5486:
5482:
5478:
5474:
5467:
5459:
5455:
5451:
5447:
5442:
5437:
5433:
5429:
5425:
5418:
5410:
5406:
5401:
5396:
5391:
5386:
5382:
5378:
5374:
5370:
5366:
5359:
5357:
5348:
5344:
5339:
5334:
5329:
5324:
5320:
5316:
5312:
5308:
5304:
5297:
5289:
5285:
5281:
5277:
5272:
5267:
5263:
5259:
5255:
5248:
5240:
5236:
5232:
5228:
5224:
5220:
5216:
5212:
5208:
5204:
5197:
5195:
5186:
5182:
5178:
5174:
5169:
5164:
5160:
5156:
5152:
5145:
5137:
5133:
5128:
5123:
5119:
5115:
5111:
5104:
5102:
5093:
5089:
5084:
5079:
5075:
5071:
5067:
5063:
5059:
5052:
5044:
5040:
5036:
5032:
5028:
5024:
5020:
5016:
5012:
5008:
5001:
4999:
4997:
4988:
4984:
4979:
4974:
4970:
4966:
4962:
4955:
4947:
4943:
4939:
4935:
4930:
4925:
4921:
4917:
4913:
4906:
4898:
4894:
4889:
4884:
4879:
4874:
4870:
4866:
4862:
4858:
4854:
4847:
4839:
4835:
4831:
4827:
4822:
4817:
4813:
4809:
4805:
4798:
4790:
4786:
4782:
4778:
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3281:on 2009-03-25
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2396:Ledgerwood JE
2391:
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2171:
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2145:9780120398553
2141:
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2114:
2112:
2110:
2108:
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2100:
2098:
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2071:
2067:
2063:
2061:9780120317424
2057:
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2038:
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2034:
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2030:
2028:
2026:
2024:
2022:
2020:
2018:
2016:
2014:
2012:
1996:
1992:
1986:
1984:
1975:
1971:
1966:
1961:
1957:
1953:
1949:
1945:
1941:
1934:
1932:
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1813:
1811:
1808:
1806:
1803:
1802:
1796:
1794:
1791:
1787:
1783:
1782:Sindbis virus
1778:
1768:
1766:
1762:
1747:
1745:
1744:P. falciparum
1740:
1736:
1732:
1727:
1722:
1713:
1709:
1705:
1703:
1697:
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1458:
1452:
1445:
1440:
1436:
1432:
1428:
1423:
1421:
1418:
1414:
1410:
1409:keratinocytes
1406:
1402:
1398:
1394:
1379:
1376:
1365:
1363:C’-dependent
1362:
1359:
1356:
1355:
1351:
1348:
1345:
1338:
1335:
1331:
1330:
1326:
1323:
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1197:
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1187:
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1171:
1167:
1163:
1159:
1154:
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1144:
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1128:
1125:
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1118:
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1082:
1078:
1067:
1065:
1064:
1059:
1055:
1049:
1040:
1038:
1034:
1030:
1026:
1012:
998:
996:
992:
988:
967:
964:
961:
960:
959:
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948:
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939:
933:
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927:
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920:
917:
916:
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911:
905:
902:
901:
900:
895:
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890:
887:
886:
880:
877:
874:
873:
872:
867:
864:
861:
860:
859:
856:
855:
852:Disadvantage
844:
833:
830:
828:formulations
827:
823:
819:
816:
812:
808:
805:
802:
798:
794:
789:
786:
783:
782:
778:
772:
769:
766:
765:
761:
758:
754:
750:
746:
742:
739:
736:
733:
729:
725:
723:Target tissue
722:
719:
715:
703:
700:
692:
682:
678:
674:
668:
667:
663:
658:This section
656:
652:
647:
646:
638:
636:
632:
628:
626:
620:
610:
608:
604:
600:
599:
594:
584:
582:
581:
576:
575:
570:
569:
564:
563:biodegradable
560:
556:
553:
549:
545:
541:
531:
529:
525:
521:
517:
508:
506:
502:
498:
494:
490:
486:
485:intradermally
482:
472:
470:
466:
457:
448:
446:
442:
437:
435:
432:
428:
424:
420:
416:
406:
397:
393:
391:
387:
383:
379:
375:
371:
367:
363:
358:
356:
352:
351:gene sequence
348:
344:
340:
336:
331:
329:
328:Polycistronic
325:
324:beta-globulin
321:
317:
313:
309:
305:
301:
297:
293:
292:transcription
289:
285:
278:Vector design
267:
264:
261:
258:
257:
254:Disadvantages
248:
245:
242:
239:
236:
233:
230:
227:
224:
221:
217:
213:
210:
209:
203:
200:
197:
188:
186:
182:
177:
174:
170:
166:
162:
158:
154:
150:
146:
142:
138:
134:
130:
126:
125:Enzo Paoletti
121:
119:
115:
111:
107:
103:
97:
87:
85:
81:
76:
72:
68:
64:
61:
56:
54:
50:
46:
42:
39:is a type of
38:
29:
24:
19:
6919:DNA vaccines
6846:Epidemiology
6822:
6794:
6680:DTaP-IPV/Hib
6471:Yellow fever
6384:EpiVacCorona
6136:Tuberculosis
6109:Pneumococcal
6077:Lyme disease
5928:
5905:Heterologous
5804:Immunization
5734:
5730:
5711:DNA vaccine
5710:
5655:
5651:
5641:
5596:
5592:
5582:
5537:
5533:
5523:
5480:
5476:
5466:
5431:
5427:
5417:
5372:
5368:
5310:
5306:
5296:
5261:
5257:
5247:
5206:
5202:
5158:
5154:
5144:
5117:
5113:
5065:
5061:
5051:
5010:
5006:
4968:
4964:
4954:
4919:
4915:
4905:
4860:
4856:
4846:
4811:
4807:
4797:
4762:
4758:
4710:
4706:
4696:
4661:
4657:
4647:
4612:
4608:
4598:
4563:
4559:
4511:
4507:
4461:
4457:
4451:
4426:
4422:
4387:
4383:
4377:
4353:(1): 10–17.
4350:
4346:
4298:
4294:
4284:
4249:
4245:
4199:
4195:
4145:
4141:
4091:
4087:
4029:
4025:
3977:
3973:
3919:
3915:
3912:Banchereau J
3906:
3871:
3867:
3857:
3822:
3818:
3766:
3762:
3698:
3694:
3684:
3641:
3637:
3627:
3582:
3578:
3568:
3556:. Retrieved
3545:
3504:
3500:
3490:
3449:
3445:
3439:
3404:
3400:
3352:
3348:
3308:
3304:
3294:
3283:. Retrieved
3279:the original
3253:(1): 50–57.
3250:
3246:
3236:
3201:
3197:
3151:
3147:
3141:
3106:
3102:
3054:
3050:
3040:
3005:
3001:
2991:
2966:
2962:
2956:
2931:
2927:
2921:
2894:
2890:
2880:
2845:
2841:
2831:
2775:
2725:(1): 29–40.
2722:
2718:
2711:
2676:
2672:
2662:
2627:
2623:
2575:
2572:Gene Therapy
2571:
2561:
2508:
2504:
2456:
2452:
2442:
2407:
2403:
2390:
2353:
2349:
2339:
2328:. Retrieved
2317:
2284:
2280:
2274:
2265:
2220:
2216:
2206:
2173:
2169:
2162:
2127:
2043:
1999:. Retrieved
1997:. 2021-08-20
1994:
1950:(1): 26–35.
1947:
1943:
1876:
1872:
1850:
1846:
1820:mRNA vaccine
1815:Gene therapy
1774:
1758:
1743:
1734:
1725:
1723:
1719:
1710:
1706:
1698:
1678:
1642:
1632:
1624:
1618:
1598:interleukins
1595:
1568:
1546:
1534:
1515:
1506:
1483:
1426:
1424:
1392:
1390:
1371:
1310:< 1 week
1279:DNA vaccine
1255:
1236:
1227:
1207:
1193:
1155:
1136:
1106:
1073:
1061:
1053:
1050:
1046:
1018:
991:basal lamina
983:
753:subcutaneous
743:(skeletal);
695:
689:October 2021
686:
671:Please help
659:
623:
616:
596:
590:
578:
572:
566:
537:
514:
478:
462:
441:conformation
438:
412:
403:
394:
359:
332:
281:
246:
201:
198:
194:
191:Applications
178:
122:
99:
57:
36:
34:
18:
6882:from market
6778:Controversy
6712:researchers
6628:Hepatitis B
6514:Hepatitis C
6459:Chicken pox
6285:Hepatitis E
6280:Hepatitis B
6275:Hepatitis A
6221:Chikungunya
6198:Shigellosis
6052:Brucellosis
5938:Therapeutic
5892:Inactivated
5832:Development
5820:Inoculation
5812:Vaccination
5483:(1): 1900.
3558:16 December
1810:HIV vaccine
1735:P. knowlesi
1686:pyrimidines
1684:and two 3’
1634:P. chabaudi
1572:lymphokines
1543:Interferons
1435:pinocytosis
1413:fibroblasts
1327:rapid rise
1324:rapid rise
1247:bone marrow
1239:hepatitis B
1166:MHC class I
1162:amino acids
1124:preclinical
1093:endocytosed
731:Parenteral
497:bupivacaine
347:AT-contents
296:translation
216:MHC class I
149:hepatitis B
47:a specific
37:DNA vaccine
6913:Categories
6813:Thiomersal
6759:Paul Offit
6734:Jonas Salk
6710:Inventors/
6581:research:
6558:research:
6495:research:
6424:Skycovione
6414:Sanofi–GSK
6359:SARS-CoV-2
6270:Hantavirus
6231:rVSV-ZEBOV
6216:Adenovirus
6184:research:
6062:Diphtheria
5900:Attenuated
3285:2007-11-21
2330:2016-08-06
2001:2021-08-22
1879:(4): e33.
1853:: 401–414.
1826:References
1805:Vector DNA
1790:alphavirus
1525:See also:
1502:lymph node
1444:macrophage
1417:epithelial
1321:slow rise
1299:? (ng-μg)
1174:N-terminus
1117:predisease
1029:lymphokine
849:Advantage
625:Mycoplasma
607:phagosomal
568:Salmonalla
501:hypertonic
431:N-terminal
386:adenovirus
343:eukaryotic
322:or rabbit
206:Advantages
161:Zika virus
94:See also:
75:veterinary
45:transfects
21:See also:
6892:Phase III
6880:Withdrawn
6808:Pox party
6618:ALVAC-CEA
6540:Protozoan
6429:Sputnik V
6379:CoronaVac
6349:Rotavirus
6247:Pandemrix
6099:Pertussis
6040:Bacterial
5924:Synthetic
5887:Conjugate
5839:Adjuvants
5816:Infection
5713:(Q578537)
2821:ignored (
2811:cite book
1793:replicons
1625:P. yoelii
1576:adjuvants
1209:P. yoelii
1186:ubiquitin
1182:influenza
1151:infection
1060:parasite
888:Gene gun
834:variable
818:Liposomes
767:Gene gun
660:does not
493:myotoxins
436:signals.
434:ubiquitin
419:cytosolic
222:molecules
153:influenza
123:In 1983,
6939:Virology
6650:Prostvac
6643:Gardasil
6638:Cervarix
6585:Hookworm
6464:Shingles
6446:Smallpox
6364:Corbevax
6300:Gardasil
6295:Cervarix
6033:Vaccines
5987:Schedule
5958:Global:
5866:Timeline
5808:Vaccines
5761:15078924
5682:11500430
5515:29765016
5458:42145608
5288:41861994
5185:36488254
5136:10820272
5092:10992502
4965:Virology
4946:41651105
4938:10395683
4838:20528634
4789:45069087
4737:33499198
4639:22923379
4404:16310901
4347:Virology
4325:38099483
4276:21038927
4224:41779568
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