Gene gun-mediate DNA vaccination against foot-and-mouth disease virus
Introduction
Foot-and-mouth disease (FMD) is one of the most dangerous disease of cloven-hoofed animals. The disease is enzootic in Asia (including the Middle-East), Africa, Eastern Europe and some parts of South America. Due to successful vaccination campaigns, Western-Europe and parts of South America are free of the disease. In addition to North America, New Zealand and Australia. Morbidity is extremely high, but mortality is low except in young calves. FMDV, the causative agent of the disease belongs to the aphthoviruses genus of the picornaviridae and like other viruses of that family, it is a nonenveloped virus. The virion is composed of 60 capsomers made of four viral proteins, VP1-4. The genome is a 8.5 kb long ssRNA in the positive orientation. The genome carries one open reading frame resulting in translation of one large polypeptide which undergoes autoproteolytic cleavage to form mature viral proteins [1], [2], [3].
Vaccination campaigns have reduced the number of disease outbreaks throughout the world resulting in the eradication of the disease in several areas, as mentioned above. On the other hand, there are large number of disease outbreaks in the rest of the world which calls for a better campaign for controlling the disease in terms of vaccine efficacy, reducing production cost and vaccine administration.
In recent years, DNA vaccination has emerged as one of the most promising venues for recombinant vaccine [4], [5]. The concept is based on inoculation of plasmids encoding a gene or genes under a strong eukaryotic promoter, e.g. the early CMV promoter/enhancer or alphavirus [6]. Uptake of the DNA and expression of the recombinant proteins initiates both a cellular and humoral immune response. Following vaccination and DNA entry into the cell, expression of the gene occurs. This permits the entry of the newly expressed protein into the class I major histocompatibility complex (MHC) antigen presentation pathway through the post-translational process of cleaving the endogenous protein into 9–11 aa long peptides. Presentation of the processed proteins elicits a CD8+ cytotoxic T lymphocyte (CTL) response [7], [8]. The induction and involvement of antibodies upon DNA vaccination has also been demonstrated in mice, bovine and nonhuman primates [9], [10], [11]. Thus, the immune response to DNA vaccines seems to be analogous to that induced by conventional live attenuated virus (LAV) vaccination. In both cases, viral antigens are expressed in the cells (LAVs through normal viral replication, and DNA vaccines as a result of expressing the recombinant antigen) and presented to the immune system in such a way that it induces both humoral as well as cell-mediated immunity.
In this manuscript, we describe the building of FMDV expression constructs which were used for DNA vaccination of pigs. We could show that a construct carrying viral P1, 2A and 3CD sequences can partially protect pigs from FMDV challenge. On the other hand, a construct carrying the O1 and Asia1 VP1 genes together with 3C protease from the O1 Geshur (G) isolate could not protect vaccinated animals against virus challenge.
Section snippets
Animals
Large White combined with Landrace and Pietrin (Belgium) pigs, males and females, 3 weeks old weighing between 8.9 and 13.3 kg were used vaccination and challenge experiments.
Viruses
FMDV, O1 Geshure and Asia 1 Shamir were the source of RNA for RT PCR and cloning. Israeli EMCV isolate obtained from the Virology Department in Shiba Hospital. O1 Geshure was used in challenge experiments.
RNA extraction
Total FMDV RNA was isolated from FMDV infected primary pig kidney cells (PKCs) by using Tri ReagentTM (Molecular
Expression of viral proteins
To test viral proteins expression, BHK-21 cells were transfected with the two plasmids (pKVI326 and 335). Forty eight hour post-transfection cells were analyzed by reaction with the a propitiate antibodies and visualize after reacting with fluorescent conjugated antibody. As a transfection efficiency control, pEGFP (Clonetec, Palto Alto, CA) transfected cells were monitored for GFP expression and usually 70–80% transfection efficiency was achieved.
Cells transfected with pKVI326 were stained by
Discussion
In this communication, we describe the construction of a plasmid carrying FMDV sequences that can confirm partial protection against FMDV challenge. The advantage of such a construct is its ability to express and process all structural proteins to form empty viral capsid which will be a superior immunogenic entity as it can initiate the immune system better than separate viral proteins like VP1 itself.
To do so the L, the entire P1 region and together 2A as one fragment and 3CD fragment were
Acknowledgements
This work was supported by a grant from BARD no. US-2417-94.
References (21)
- et al.
DNA immunization can stimulate florid local inflammation, and the anti viral immunity induced varies depending on injection site
Vaccine
(1997) - et al.
DNA vaccination of mice with a plasmid encoding Puumala hantavirus nucleocapsid protein mimics the B-cell response induced by virus infection
J. Biotechnol.
(2000) - et al.
Effective DNA vaccination of cattle with the mycobacterial antigens MPB83 and MPB70 does not compromise the specificity of the comparative intradermal tuberculin skin test
Vaccine
(2000) - et al.
DNA vaccination of Macaques with several different nef sequences induces multispecific T cell responses
Virology
(2001) - et al.
Expression in Escherichia coli and purification of biologically active L proteinase of foot-and-mouth disease virus
Virus. Res.
(1995) - et al.
Plasmid encoding foot-and-mouth disease virus VP1 epitoes elicited immune response in mice and swine and protect swine against viral infection
Virology
(2000) - et al.
Development of DNA vaccines for foot-and-mouth disease, evaluation of vaccines encoding replicating and non-replicating nucleic acids in swine
J. Biotechnol.
(1999) - et al.
Recombinant viruses expressing the foot-and-mouth disease virus capsid precursor polypeptide (P1) induce cellular but not humoral antiviral immunity and partial protection in pigs
Virology
(1999) - Bachrach HL. Foot and mouth disease virus: properties, molecular biology, and immunogenicity. Beltsville symposia in...
- et al.
The complete nucleotide sequence of the RNA for the primary translation product of foot and mouth disease virus
Nucl. Acids. Res.
(1984)
Cited by (53)
A DNA vaccination regime including protein boost and electroporation protects cattle against foot-and-mouth disease
2012, Antiviral ResearchCitation Excerpt :These publications were shortly followed by the successful generation of the first DNA vaccine against FMDV (Wong et al., 2000). However, despite this early success, to date, there has only been four documented cases which have demonstrated full protective efficacy against FMDV in the naturally susceptible species sheep and pigs (Niborski et al., 2006; Li et al., 2008; Wong et al., 2000, 2002), along with another four where partial protection was observed (Cedillo-Barron et al., 2001; Ward et al., 1997; Benvenisti et al., 2001; Mingxiao et al., 2007; Borrego et al., 2011). Here we have demonstrated for the first time that clinical protection against FMDV can also be achieved in cattle following the use of a DNA prime-protein boost strategy and that this protection was best achieved by use of an electroporation delivery device and the inclusion of the immune modulator bovine GM-CSF.
Foot-and-Mouth Disease
2009, Vaccines for Biodefense and Emerging and Neglected DiseasesDramatic improvement in FMD DNA vaccine efficacy and cross-serotype antibody induction in pigs following a protein boost
2008, VaccineCitation Excerpt :FMD O type virus was detected in both blood and nasal swab samples from these control pigs from 2-day post-challenge until termination (supplemental table). A number of FMD DNA vaccines have been developed and protection against challenge has been reported [19,32–36]. However, the development of neutralising antibodies in these animals was slower and at a lower level than that of a conventional vaccine [19,32,33].
A recombinant plasmid co-expressing swine ubiquitin and the GP5 encoding-gene of porcine reproductive and respiratory syndrome virus induces protective immunity in piglets
2008, VaccineCitation Excerpt :Over the past decade, DNA vaccination has emerged as a promising alternative vaccination method because it has the capacity to induce both humoral and cellular immune responses [9]. Used alone, however, DNA vaccines are often inferior to protein immunogens, especially for the induction of antibodies [10] and cellular immune responses in large animals [11]. Numerous studies have explored methods for improving immune responses induced by DNA immunization by altering routes of vaccine administration, enhancing antigen expression, and modulating processing and immune activation steps [9,12,13].
Optimization strategy for plasmid DNAs containing multiple-epitopes of foot-and-mouth disease virus by cis-expression with IL-2
2008, VaccineCitation Excerpt :These two methods, tandem repeat and cis-expression of epitopes of FMDV may be considered at the same time in the future designation. Researches on immune style, such as pre-injection of muscles with a relatively large volume of hypertonic sucrose [34] or injecting with gene gun [11], made DNA vaccine easier to effect. But those methods were not adaptive in extensive clinical usage.
Immune responses of swine inoculated with a recombinant fowlpox virus co-expressing P12A and 3C of FMDV and swine IL-18
2008, Veterinary Immunology and Immunopathology