Genetic Vaccination against Malaria Infection by Intradermal and Epidermal Injections of a Plasmid Containing the Gene Encoding the<i>Plasmodium berghei</i>Circumsporozoite Protein

Weiss, Richard; Leitner, Wolfgang W.; Scheiblhofer, Sandra; Chen, Defeng; Bernhaupt, Andrea; Mostböck, Sven; Thalhamer, Josef; Lyon, Jeffrey A.

doi:10.1128/iai.68.10.5914-5919.2000

Cited by 50 publications

(37 citation statements)

References 36 publications

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“…The dominant Th1 phenotype after i.m. gene immunisation and the preferential Th0 type of immune response induced by gene gun vaccination have been reported previously [28,29]. This difference in cytokine pattern has been attributed in part to the dose of DNA administered and the presence of immunostimulatory DNA sequences containing CpG motifs [30,31].…”

Section: Discussionmentioning

confidence: 95%

Induction of a Th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene

Leclerq¹,

Harms²,

Rosinha³

et al. 2002

Journal of Medical Microbiology

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The immunogenicity and protective ef®cacy of a DNA vaccine encoding the GroEL heatshock gene from Brucella abortus was tested in BALB/c mice immunised by intramuscular (i.m.) needle injection or epidermally by gene gun. The Brucella GroEL gene was ampli®ed by PCR and cloned into two different mammalian expression vectors pCMV-link and pCMV-tPA. The D17 cell line was transfected with both constructs and GroEL transcripts were detected by Northern blot. To determine the level of protein synthesised, transfected cell lysates were then submitted to Western blot. The nonsecreted form of the recombinant GroEL produced by the pCMV-link construct was detected in much greater amount than the secreted form of the protein produced by the pCMV-tPA construct. After immunisation, a strong anti-GroEL IgG response was detected in mice vaccinated by i.m. injection or gene gun only when the pCMV-link/ GroEL plasmid was used. Regarding the pattern of immune response induced, i.m. needle injection raised a predominantly Th1 response with mostly IgG2a-speci®c antiGroEL and high levels of IFN-ã produced by splenic T cells. Gene gun immunisation induced a Th0 type of immune response in mice characterised by a high IgG1/IgG2a ratio, and IL-4 and interferon (IFN)-ã production. Even though a distinct pattern of immune response was generated depending upon the immunisation route used, neither method engendered a signi®cant level of protection with the GroEL DNA vaccine.

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Section: Discussionmentioning

confidence: 95%

Induction of a Th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene

Leclerq¹,

Harms²,

Rosinha³

et al. 2002

Journal of Medical Microbiology

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“…DNA vaccine plasmids were prepared for gene gun injection as described in the Bio-Rad Gene Gun instruction manual and elsewhere (24,33). Briefly, 60 g of each plasmid was precipitated onto 0.5 to 1 mg of gold particles (0.5 to 2 m) (Deggussa, Washington, Pa.).…”

Section: Methodsmentioning

confidence: 99%

“…We believe that intramuscular and gene gun immunization may activate different arms of the immune system and that different antigens may protect by distinct immune effector mechanisms. Thus, it appears that for plasmid vaccination there is no single route which can produce the best protective response for all antigens (24,33).…”

Section: Vol 72 2004 Discovery Of Protective Malaria Parasite Antigmentioning

confidence: 99%

Novel Antigen Identification Method for Discovery of Protective Malaria Antigens by Rapid Testing of DNA Vaccines Encoding Exons from the Parasite Genome

et al. 2004

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We describe a novel approach for identifying target antigens for preerythrocytic malaria vaccines. Our strategy is to rapidly test hundreds of DNA vaccines encoding exons from the Plasmodium yoelii yoelii genomic sequence. In this antigen identification method, we measure reduction in parasite burden in the liver after sporozoite challenge in mice. Orthologs of protective P. y. yoelii genes can then be identified in the genomic databases of Plasmodium falciparum and Plasmodium vivax and investigated as candidate antigens for a human vaccine. A pilot study to develop the antigen identification method approach used 192 P. y. yoelii exons from genes expressed during the sporozoite stage of the life cycle. A total of 182 (94%) exons were successfully cloned into a DNA immunization vector with the Gateway cloning technology. To assess immunization strategies, mice were vaccinated with 19 of the new DNA plasmids in addition to the well-characterized protective plasmid encoding P. y. yoelii circumsporozoite protein. Single plasmid immunization by gene gun identified a novel vaccine target antigen which decreased liver parasite burden by 95% and which has orthologs in P. vivax and P. knowlesi but not P. falciparum. Intramuscular injection of DNA plasmids produced a different pattern of protective responses from those seen with gene gun immunization. Intramuscular immunization with plasmid pools could reduce liver parasite burden in mice despite the fact that none of the plasmids was protective when given individually. We conclude that high-throughput cloning of exons into DNA vaccines and their screening is feasible and can rapidly identify new malaria vaccine candidate antigens.The year 2002 saw the publication of the genomic sequences of the human malaria parasite Plasmodium falciparum (12) and the rodent parasite Plasmodium yoelii yoelii (5). The hope is that this information will bring insights into parasite biology and lead to the development of new vaccines and drugs. However, novel research approaches are required to efficiently study the thousands of genes. This paper describes the development of a high-throughput technique for the identification of vaccine target antigens among newly annotated malaria genes. Our method rapidly produces large numbers of DNA vaccines carrying P. y. yoelii exons and measures their ability to reduce parasite load in mice. We call this screening technique the antigen identification method.The novelty and efficiency of the antigen identification method come from a combination of rapid production of DNA vaccines and sensitive measurement of parasite killing. With the annotated P. y. yoelii genomic sequence, we identify P. y. yoelii genes expressed during the sporozoite stage by comparison with expressed sequence tags (ESTs) generated from a cDNA library of P. yoelii sporozoites (20). PCR primers for these sporozoite P. y. yoelii genes are synthesized to be compatible with the Gateway cloning system, which allows rapid production of DNA vaccine plasmids. Mice are immunized with the DNA...

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“…The latter has been suggested to induce a Th1 milieu with 100-1000 times less amount of DNA than intramuscular injection [29]. However, gene gun bombardment was questioned for allergen immunotherapy by several studies [22,[30][31][32][33].…”

Section: Discussionmentioning

confidence: 99%

A mimotope gene encoding the major IgE epitope of allergen Phl p 5 for epitope-specific immunization

et al. 2009

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A gene vaccine based on a mammalian expression vector containing the sequence of a peptide mimotope of Phl p 5 was constructed. To test whether mimotope gene vaccines can induce allergen-specific antibody responses via molecular mimicry, BALB/c mice were immunized using the mimotope construct with or without a tetanus toxin T-helper epitope. Moreover, intradermal injection was compared to epidermal application via gene gun immunization.Immunization with both mimotope gene constructs elicited allergen-specific antibody responses. As expected, gene gun bombardment induced a Th2-biased immune response, typically associated with IgG1 and IgE antibody production. In contrast, intradermal injection of the vaccine triggered IgG2a antibody expression without any detectable IgE levels, thus biasing the immune response towards Th1. In an RBL assay, mimotope-specific IgG antibodies were able to prevent crosslinking of allergen-specific IgE by Phl p 5. A construct coding for the complete Phl p 5 induced Tcell activation, IFN-γ and IL-4 production. In contrast, the mimotope-DNA construct being devoid of allergen-specific T-cell epitopes had no capacity to activate allergen-specific T cells.Taken together, our data show that it is feasible to induce blocking IgG antibodies with a mimotope-DNA construct when applied intradermally. Thus the mimotope-DNA strategy has two advantages: (1) the avoidance of IgE induction and (2) the avoidance of triggering allergenspecific T-lymphocytes. We therefore suggest that mimotope gene vaccines are potential candidates for epitope-specific immunotherapy of type I allergy.

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Genetic Vaccination against Malaria Infection by Intradermal and Epidermal Injections of a Plasmid Containing the Gene Encoding thePlasmodium bergheiCircumsporozoite Protein

Cited by 50 publications

References 36 publications

Induction of a Th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene

Induction of a Th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene

Novel Antigen Identification Method for Discovery of Protective Malaria Antigens by Rapid Testing of DNA Vaccines Encoding Exons from the Parasite Genome

A mimotope gene encoding the major IgE epitope of allergen Phl p 5 for epitope-specific immunization

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