Comparative efficacy of virus like particle (VLP) vaccine of foot-and-mouth-disease virus (FMDV) type O adjuvanted with poly I:C or CpG in guinea pigs
Introduction
Foot-and-mouth disease (FMD) is a highly contagious viral disease of livestock that causes severe economic loss in susceptible cloven-hoofed animals, such as cattle, sheep, goats and pigs. FMD is a transboundary disease, caused by an RNA virus, Foot-and-Mouth Disease Virus (FMDV), a positive-sense single-stranded RNA virus belonging to genus Aphthovirus and family Picornaviridae [1]. The viral RNA is translated as a single open reading frame into a polyprotein, and cleaved predominantly by 3C viral protease to produce structural and non-structural proteins required for virus assembly and replication [2]. Among the seven known serotypes of FMDV, only three serotypes namely O, A and Asia1 are prevalent in India [3]. Use of inactivated FMD vaccine with oil adjuvant is the most effective way to control and eradicate FMD in endemic countries [4]. Although, the current FMD vaccine is chemically inactivated oil adjuvanted whole virus preparation that has been used to effectively control the disease in enzootic countries, its use may lead to new disease outbreaks due to a possible incomplete inactivation of the virus during manufacturing or inadvertent escape of virus [5]. To overcome these issues, new generation vaccines like virus-like particles (VLPs) are being developed as non-infectious, safe and marker vaccines. VLPs can mimic the epitopes present on the native virus, leading to stimulation of both B and T cells with increased cell mediated immunity (CMI) [6]. Baculovirus expression system (BVES) has been applied to developing veterinary vaccines based on VLPs produced in insect cells for blue tongue virus, bovine papillomavirus type1, chicken anemia virus, infectious bursal disease virus, porcine, mink and canine parvoviruses, and porcine encephalomyocarditis virus [6]. VLPs are also tested as vaccine candidates for FMDV type O [7] and type Asia1 viruses [8].
After translation of FMDV viral RNA, a 95 kDa polyprotein (P1) was produced inside host cell, which is cleaved by the viral 3C protease to yield VP0 (36 kDa), VP1 (32 kDa) and VP3 (27 kDa) to form the capsid. Auto-catalytic cleavage of VP0 into VP2 (28 kDa) and VP4 (8 kDa) occurs during encapsidation of the viral genome to produce the mature virus [9]. FMD virus capsids can be synthesized using recombinant techniques by expressing the P1 structural protein precursor and the 3C protease that cleaves it, since the capsid proteins spontaneously assemble to produce VLPs. The inherent problem is balancing the expression of P1 and 3C, the latter being toxic to insect cells, especially in the case of FMDV [9]. For FMD virus there are several report of VLPs produced for different serotypes [10], [11], including VLPs against FMDV Type O from our laboratory [7]. Earlier reports of generating VLPs of FMDV involved the use of wild type sequence of 3C [8], [10], [12]. However, considering the fact that 3C protease is toxic to cells [13], [14] and there is a need to reduce its toxicity, we introduced mutations in its critical amino acids. Amongst all serotypes, FMDV type O/IND/R2/75, the currently used Indian vaccine strain has huge potential as vaccine for use in Asian and Eurasian countries [15]. In this context, it is desirable to develop VLPs based vaccine for FMDV Type O/IND/R2/75.
Immuno-stimulating molecules such as polyriboinosinic:polyribocytidylic acid (Poly I:C) and CpG Oligo deoxy nucleotides (CpG ODN) are used to improve the potency and efficacy of vaccines [16], [17], [18]. The advantages of Poly I:C in vaccine formulations are increased humoral and/or cell mediated immune responses as they are potent inducers of IL-12 and type-I interferon secretion by the activation of innate immunity via the TLR3 receptor [19]. Cao et al. [20] reported that the Poly I:C mixed with multi-epitope peptide vaccine induced improved neutralizing antibody titre against FMDV in mice and suggested a dose of 100 μg of Poly I:C with antigen for mouse inoculation.
Recently, several studies have shown that synthetic ODN containing unmethylated CpG dinucleotide (CpG ODN) is a potent adjuvant due to its capability of activating B cells, inhibiting apoptosis of B cells and inducing the maturation [21]. Further it was suggested that CpG ODN could boost the immune response elicited by vaccines by improving protein immunogenicity [22], inducing both humoral and cellular responses as well as shift in the isotype of IgG antibodies elicited by immunization, increase the magnitude of vaccine induced responses [23], accelerate the development of vaccine induced responses [24], [25], upregulate immunoglobulin gene expression [26] and inhibit Rous Sarcoma Virus induced allergy in guinea pigs [27]. Increased potency of vaccines with addition of CpG ODN as adjuvants targeting infectious diseases [29] are of BiothraxR anthrax vaccine [17], foot-and-mouth-disease inactivated virus vaccine [16], combination of CpG ODN and montanide ISA206 adjuvant combined with recombinant FMDV vaccine in cattle [24].
In this report, we tested the efficacy of ISA206, Poly I:C, and CpG adjuvanted FMDV VLP type O vaccine in two different vaccination regimens (primary and booster) in conferring protection against challenge in guinea pigs.
Section snippets
CpG ODN and poly(I:C)
CpG Oligodeoxynucleotides (or CpG ODN) are short single-stranded synthetic DNA molecules that contain a cytosine “C” followed by a guanine “G”. The “p” refers to the phosphodiester backbone of DNA, however some ODN have a modified phosphorothioate backbone. The CpG available from commercial source (CpG ODN 10109) were procured from M/S Integrated DNA Technology, Bangalore, India and the sequence of CpG used is 5′-TCGTcgTTTTAcgGCGCcgTGCCG-3′. The CpG was synthesized on phosphorothioate backbone
Screening for expression of capsid proteins of FMDV in insect cells by S-ELISA
The baculovirus expressed FMDV type O VLPs were tested in S-ELISA using anti-146s FMDV Type O polyclonal sera raised in rabbits and guinea pigs. The mean S-ELISA reactivity (A492 nm) of lysates and supernatants (n = 6) of six batches of VLPs produced in Sf9 cells were 2.288 for lysate and 0.591 for supernatant respectively, while the positive control (BHK21 cell grown inactivated 146S of FMDV Type O) showed A492 nm of 2.382 indicating the ability of the VLPs expressed in Sf9 could react with
Discussion
Foot- and- mouth disease (FMD) is the most devastating viral disease of cloven footed animals causing huge economic loss and disrupting the food security in developing countries. Though vaccination with the conventional vaccine remains the key factor in controlling spread of the virus, short duration of immunity, escape from production plant [5], requirement of booster dose weaken the immunogenic property of the current vaccine. Moreover the conventional vaccine preparation could contain
Conflict of interest statement
The authors do not have any conflict of interest to declare.
Acknowledgments
The authors thank Director, IVRI, Izatnagar and Joint Director, IVRI Campus, Hebbal, Bangalore for providing the necessary facilities to carry out the research work. Authors are also grateful to the assistance provided by laboratory staff of FMD Vaccine Production Laboratory and Isolation unit of Yelahanka, IVRI Campus, Bangalore, India. The financial assistance granted to the first author, M. Terhuja, in the form of Institute Fellowship for Master degree from IVRI is gratefully acknowledged.
References (32)
- et al.
Virus-like particles as a highly efficient vaccine platform: diversity of targets and production systems and advances in clinical development
Vaccine
(2012) - et al.
Novel immunogenic baculovirus expressed virus-like particles of foot-and-mouth disease (FMD) virus protect guinea pigs against challenge
Res Vet Sci
(2013) - et al.
Synthesis of empty capsid-like particles of Asia I foot-and-mouth disease virus in insect cells and their immunogenicity in guinea pigs
Vet Microbiol
(2009) - et al.
Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity
J Virol Methods
(2013) - et al.
Development of foot-and-mouth disease virus (FMDV) serotype O virus-like-particles (VLPs) vaccine and evaluation of its potency
Antivir Res
(2012) - et al.
Characterization of recombinant foot-and-mouth disease virus pentamer-like structures expressed by baculovirus and their use as diagnostic antigens in a blocking ELISA
Vaccine
(2007) - et al.
Increased potency of BioThraxR vaccine with the addition of the C-class CpG oligonucleotide adjuvant CPG 10109
Vaccine
(2007) - et al.
Toll-like receptors and RNA helicases: two parallel ways to trigger antiviral responses
Mol Cell
(2006) - et al.
Improved neutralising antibody response against foot-and-mouth-disease virus in mice inoculated with a multi-epitope peptide vaccine using polyinosinic and poly-cytidylic acid as an adjuvant
J Virol Methods
(2012) - et al.
CpG oligodeoxynucleotide and montanide ISA 206 adjuvant combination augments the immune responses of a recombinant FMDV vaccine in cattle
Vaccine
(2011)
Immunostimulatory CpG oligonucleotides: effect on gene expression and utility as vaccine adjuvants
Vaccine
CpG oligonucleotides as adjuvants for vaccines targeting infectious diseases
Adv drug Deliv Rev
Recombinant protein vaccines produced in insect cells
Vaccine
Picornaviruses and their replication
Foot-and-mouth disease
Clin Microb Rev
Annual report 2009–2010 project directorate on foot and mouth disease
Cited by (20)
B cell memory responses induced by foot-and-mouth disease virus-like particles in BALB/c mice
2022, Veterinary Immunology and ImmunopathologyCitation Excerpt :Control and eradication of the disease from regions of endemicity involves the use of inactivated vaccines (Pacheco et al., 2010). Nevertheless, FMDV inactivated vaccines have their immunological limitation providing relatively short-term humoral immune protection, so the titer of protective antibody can only be maintained through routine repeated vaccine (de Los Santos et al., 2018; Grant et al., 2016; Terhuja et al., 2015). Long-term serological protection, providing protection for months to years, is dependent on long-lived plasma cells (LLPCs) and memory B cells (Ndungu et al., 2009; Nguyen et al., 2019).
SARS-CoV-2 vaccine research and development: Conventional vaccines and biomimetic nanotechnology strategies
2021, Asian Journal of Pharmaceutical SciencesCitation Excerpt :Consequently, both T cell response and antibody production are conferred to provide comprehensive protection of immunized individuals from virus infection [124–129]. The CpG adjuvanted VLP vaccines were shown to enhance cellular immunity in preclinical models of infections for the foot-and-mouth-disease virus and influenza virus [130,131]. The combination of a CpG adjuvant is an important strategy to enhance the efficacy, potency, and safety of VLP vaccines.
Comparison of immune responses in guinea pigs by intranasal delivery with different nanoparticles-loaded FMDV DNA vaccine
2020, Microbial PathogenesisCitation Excerpt :In addition, CpG oligodeoxy-nucleotides (ODNs) that activate plasmacytoid dendritic cells and stimulate IFN-a production could be advantageous as a vaccine adjuvant. Stimulation with CpG ODN has been shown to protect against several viral infections to date, including FMDV [41–43], birnavirus [44], influenza virus [45]and orthopox virus [46]. FMDV consists of the RNA genome surrounded by a protein shell or capsid.
Generation of acid resistant virus like particles of vaccine strains of foot-and-mouth disease virus (FMDV)
2019, BiologicalsCitation Excerpt :Sf-21 insect cells (Spodoptera frugiperda IPLB-Sf21-AE clone; Invitrogen, USA), Sf-9 (clone of Sf21, Invitrogen) and Tn5 insect cells (Trichoplusia ni, High-Five™ BTI-TN-5B1-4; Invitrogen) were maintained in Sf-900 II SFM (Invitrogen) serum free medium at 27 °C, pH 6.2 were used for production of VLPs. P1-2A-3C genes of FMDV serotype O/IND/R2/75 was cloned as reported elsewhere [16]. Briefly, OP1-2A and O3C genes were amplified from the plasmid DNA by PCR amplification and mutations were introduced at position 38 and 48 (m3CG38SF48S).
Adjuvant formulations for virus-like particle (VLP) based vaccines
2017, Clinical ImmunologyCitation Excerpt :Cytidine monophosphate guanosine oligodeoxynucleotides (CpG or CpG ODN) are a class of DNA analogs composed of unmethylated cytosine and guanine motifs that mimic bacterial DNA and stimulate the innate immune system by binding to the TLR9 and triggering B cell proliferation, dendritic APC maturation, and Th1-mediated immune activation [86]. A foot-and-mouth-disease virus (FMDV) VLP-based vaccine was shown to enhance cell mediated immunity in a guinea pig model [87]. In particular, CpG adjuvant markedly tilted the response toward a Th1 rather than Th2 type of immunity.