Antibody repertoire development in fetal and neonatal piglets. XXIII: fetal piglets infected with a vaccine strain of PRRS Virus display the same immune dysregulation seen in isolator piglets

The Ig levels and antibody repertoire diversification in fetal piglets infected with an attenuated Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) were measured. Serum Ig levels were greatly elevated in PRRSV-infected fetuses; IgG was elevated >50-fold, IgM>5-15-fold and IgA>2-fold compared to control fetuses. Their IgM to IgG to IgA profile was the same as that in isolator piglets infected for the same period with wild-type PRRSV. Fetal animals showed less repertoire diversification than even isolator piglets that were maintained germfree (GF) while the repertoire diversification index (RDI) for PRRSV-infected isolator piglets was 10-fold higher and comparable to littermates infected with swine influenza (S-FLU). However, when expressed as the RDI:Ig ratio, infected fetuses appeared 10-fold less capable of repertoire diversification than uninfected littermates and GF isolator piglets. Compared to S-FLU isolator piglets that resolve the infection, the RDI:Ig of PRRSV-infected isolator piglets was 100-fold lower. Overall, infection of fetuses with an attenuated virus shows the same immune dysregulation seen postnatally in wild type infected isolator piglets, indicating that: (a) attenuation did not alter the ability of the virus to cause dysregulation and (b) the isolator infectious model reflects the fetal disease.     

Antibody responses following vaccination versus infection in a porcine circovirus-type 2 (PCV2) disease model show distinct differences in virus neutralization and epitope recognition

Porcine circovirus associated disease (PCVAD) encompasses a group of syndromes linked to infection with porcine circovirus type 2 (PCV2). Based on the hypothesis that the immune responses to vaccination versus infection are quantitatively and qualitatively different, the objective of this study was to evaluate immunity, virus replication and disease protection in pigs vaccinated with PCV2 capsid protein (CP) and during infection. The disease model included dual infection with PCV2 and porcine reproductive and respiratory syndrome virus (PRRSV), a virus known to enhance disease progression and severity. The principal effect of PRRSV infection was to increase peak PCV2 viremia by almost 40-fold; however, PCV2 failed to show a reciprocal effect on PRRSV. In vaccinated pigs, there was no evidence of disease or PCV2 replication following dual virus challenge. Immunity following vaccination favored PCV2 neutralizing activity; whereas, PCV2 infection and disease produced high levels of non-neutralizing antibody, primarily directed against a polypeptide in the C-terminal region of CP. These results support the notion that the magnitude of the total antibody response cannot be used as a measure of protective immunity. Furthermore, protection versus disease lies in the immunodominance of specific epitopes. Epitope specificity should be taken into consideration when designing PCV2 vaccines.     

Vaccination with porcine reproductive and respiratory syndrome killed virus vaccine and immunostimulatory oligodeoxynucleotides induces specific immunity in piglets

A large number of studies demonstrated the immunostimulatory effects of CpG oligonucleotides (ODN), particularly in mice. In present study, the objective was to investigate the immunoadjuvant effects of CpG ODN to porcine reproductive and respiratory syndrome (PRRS) killed virus vaccine (PRRSV) and its protective effects against PRRS virus in piglets. The PRRSV-specific antibodies titres and serum IgG1/IgG2 titres, the proliferation of lymphocytes, PRRSV-specific IFN-gamma, the expression of major histocompatibility complex class II (MHCII) of peripheral blood mononuclear cells (PBMCs) and post-challenge clinical protection were examined to identify the immune responses of the piglets. The results were found that the above-mentioned immune responses of the piglets inoculated with CpG ODN plus PRRSV were significantly stronger than those indued by PRRSV or PBS control groups. All these data suggested that CpG ODN could be employed as effective immunoadjuvant to raise the humoral and cellular responses of the piglets to PRRSV.     

Comparison of commercial and experimental porcine circovirus type 2 (PCV2) vaccines using a triple challenge with PCV2, porcine reproductive and respiratory syndrome virus (PRRSV), and porcine parvovirus (PPV)

The efficacies of commercial porcine circovirus type 2 (PCV2) vaccines and a live PCV1-2a chimeric vaccine were compared in conventional, PCV2-positive piglets using a PCV2–porcine reproductive and respiratory syndrome virus (PRRSV)–porcine parvovirus (PPV) coinfection challenge model. Seventy-three, 2-week-old pigs were randomized into seven groups including five vaccinated and two control groups. Pigs in the vaccinated groups were vaccinated at 3 weeks (one dose) or at 3 and 6 weeks (two dose) of age. All vaccine regimens tested were effective in reducing naturally occurring PCV2 viremia at 16 weeks of age and after PCV2 challenge, demonstrating the capability of the products to induce a lasting protective immunity despite the presence of PCV2 viremia at the time of vaccination.     

Children with otitis media mount a pneumococcal serotype specific serum IgG and IgA response comparable to healthy controls after pneumococcal conjugate vaccination

It has been suggested that otitis-prone children have an impaired antibody response. To investigate this in the context of pneumococcal vaccination, we used a multiplex bead-based assay to measure serum IgG and IgA levels against pneumococcal serotypes included in the 7-valent pneumococcal conjugate vaccine (PCV7; serotypes 4, 6B, 9V, 14, 18C, 19F and 23F) and 4 non-PCV7 serotypes (1, 5, 7F and 19A) in healthy (n=43) and otitis-prone children (n=75) before, 6 weeks after and 1 year after vaccination with one dose of PCV7. Pre-vaccination, otitis-prone children had significantly higher serum IgG levels against serotypes 4, 9V and 23F and against all non-PCV7 serotypes. One year following vaccination, there was no difference in IgG or IgA levels between healthy and otitis-prone children. The effect of the administration of one or two doses of PCV7 was investigated in otitis-prone children. After a second dose of PCV7, pneumococcal serotype specific IgG levels, but not IgA titres, were higher compared to the levels measured after the initial dose of PCV7. One year post PCV7 vaccination there was no difference in either IgG or IgA antibody levels to any of the PCV7 serotypes between children who received either one or two doses of PCV7. The finding that otitis-prone children do not have an impaired pneumococcal serotype-specific serum IgG or IgA response suggests that new pneumococcal conjugate vaccines may be immunogenic in otitis-prone children, however, further investigations are necessary to determine the clinical impact of such vaccines against the development of recurrent acute otitis media.     

A PCV2 vaccine based on genotype 2b is more effective than a 2a-based vaccine to protect against PCV2b or combined PCV2a/2b viremia in pigs with concurrent PCV2, PRRSV and PPV infection

The predominant genotype of porcine circovirus (PCV) in the pig population today is PCV2b yet PCV2a-based commercial vaccines are considered effective in protecting against porcine circovirus associated disease. The objective of this study was to compare the ability of PCV2a- and PCV2b-based vaccines to control PCV2b viremia in a challenge model that mimics the U.S. field situation. Sixty-three pigs were randomly assigned to one of eight groups. Sixteen pigs were vaccinated with an experimental live-attenuated chimeric PCV1-2a vaccine based on genotype 2a and another 16 pigs with a chimeric PCV1-2b vaccine based on genotype 2b. Challenge was done 28 days post vaccination (dpv) using PCV2b (or a combination of PCV2a and PCV2b), porcine reproductive and respiratory syndrome virus (PRRSV), and porcine parvovirus (PPV) to mimic what commonly occurs in the field. The experiment was terminated 21 days post challenge (dpc) or 49 dpv. Pigs vaccinated with the chimeric PCV1-2b vaccine had significantly higher levels of PCV1-2b viremia and shedding of the PCV1-2b vaccine virus in feces and nasal secretions but also a more robust humoral immune response as evidenced by significantly higher ELISA S/P ratios compared to the PCV1-2a vaccination. Regardless of challenge, the PCV1-2b vaccination significantly reduced the prevalence and amount of PCV2 viremia compared to the PCV1-2a vaccination. Interestingly, in the non-vaccinated pigs concurrent PCV2a infection resulted in clinical disease and increased macroscopic lung lesions compared to pigs challenged with PCV2b alone, further supporting the idea that concurrent PCV2a/PCV2b infection is necessary for optimal PCV2 replication.     

Effect of high and low levels of maternally derived antibodies on porcine circovirus type 2 (PCV2) infection dynamics and production parameters in PCV2 vaccinated pigs under field conditions

The present study aimed to compare the efficacy of a porcine circovirus type 2 (PCV2) commercial vaccine in terms of average daily weight gain (ADWG) as well as infection dynamics in pigs with different maternally derived antibody (MDA) levels. A total of 337 animals from a PCV2 subclinically infected farm were distributed into two groups based on weight and PCV2 antibody levels (high [H] or low [L]) at 2 weeks of age. One week later, these animals were subdivided in four groups according to the treatment received. Vaccinated pigs (H-V and L-V) received 1 mL of a commercial vaccine and NV (H-NV and L-NV) received 1 mL of PBS. All piglets were subsequently bled at 7, 12, 18, 22 and 25 weeks of age and weighted at 12 and 25 weeks of age. V animals showed significantly lower PCV2 infection rates and viral load as well as higher ELISA S/P ratios and ADWG than NV ones. Compared with H-V piglets, L-V pigs showed numerically lower PCV2 infection rates, lower area under the curve of viral load, an earlier seroconversion and a numerically, but not significantly, higher ADWG. In this study, MDA did not seem to interfere with the effect of PCV2 vaccination on ADWG. However, only when a small subpopulation of pigs with the highest ELISA S/P ratios at vaccination was considered, an apparent interference of vaccine efficacy on ADWG was noticed. Therefore, the impact of the putative interference under field conditions is probably negligible for most farms.     

Role of IgA in the defense against respiratory infections IgA deficient mice exhibited increased susceptibility to intranasal infection with Mycobacterium bovis BCG

IgA is the predominant Ig isotype in mucosal tissue and is believed to be involved in defense against viral and bacterial infections at these sites. Here, we examined the role of IgA in the protection against intranasal (i.n.) infection with Mycobacterium bovis Bacillus Calmette-Guerin (BCG). IgA deficient (IgA-/-) mice and wild type non-targeted littermate (IgA+/+) mice were immunized by i.n. route with the mycobacterium surface antigen PstS-1 formulated with cholera toxin (CT). Our data showed that IgA-/- mice were more susceptible to BCG infection compared to IgA+/+ mice, as revealed by the higher bacterial loads in the lungs and bronchoalveolar lavage (BAL). Analysis of the Ig levels and the antibody responses to PstS-1 showed that IgA-/- mice had no detectable IgA either in the saliva or in the BAL. However, these mice displayed higher levels of total and specific IgM than IgA+/+ mice in both mucosal fluids. More importantly, analysis of the cytokine responses revealed a reduction in the IFN-gamma and TNF-alpha production in the lungs of IgA-/- compared to IgA+/+ mice. Altogether, our results suggest that IgA may play a role in protection against mycobacterial infections in the respiratory tract by blocking the pathogen entrance and/or by modulating the pro-inflammatory responses.     

Co-infection status of classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circoviruses (PCV2 and PCV3) in eight regions of China from 2016 to 2018

Classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circoviruses (PCV2 and PCV3) are economically important swine viruses that cause reproductive failure and/or respiratory symptoms in pigs. However, the co-infection status of these viruses in Chinese swine herds is not well clarified. In this study, we evaluated the co-infection of these four viruses in 159 pigs collected from 63 herds in eight regions of China from 2016 to 2018. CSFV, PRRSV, PCV2 and PCV3 were detected in 14, 56, 43 and 4 of the pigs, respectively. The percentage of singular infections was 32.71%, while the percentages of dual infections and multiple infections were 15.72% and 3.15%, respectively. The E2 of CSFV, ORF5 of PRRSV, ORF2s of PCV2 and PCV3 from all positive samples were determined and used for phylogenetic analyses. E2-based phylogenetic tree showed that all 14 CSFVs identified in this study belong to 2.1b subtype. ORF5-based phylogenetic tree showed that PRRSV2 is predominant in China while PRRSV1 can also be detected. In addition, 35, 16, 4 and 1 of our PRRSVs are clustered with highly pathogenic PRRSV2, NADC30-like PRRSV2, classical PRRSV2 and PRRSV1, respectively. ORF2-based phylogenetic trees showed that our PCVs are grouped with 2 PCV2 subtypes (PCV2d and PCV2b) and 3 PCV3 subtypes (PCV3a, PCV3b and PCV3c), respectively. Our results provide the latest co-infection status and the diversity of four important swine viruses in Chinese swine herds, which is beneficial for understanding the epidemiology of these viruses.     

Class-specific antibodies to bovine respiratory syncytial virus in experimentally infected lambs.

Enzyme-linked immunoabsorbent assay (ELISA) was used to titrate virus-specific IgG, IgM and IgA levels in nasal secretions, lung lavage fluids and serum samples sequentially obtained from lambs experimentally infected with bovine respiratory syncytial virus (RSV). Virus-specific IgG and IgM responses were measured by the indirect double antibody sandwich ELISA using anti-bovine RSV monoclonal antibody, as capture antibody, and peroxidase-conjugated anti-sheep IgG and anti-sheep IgM. Virus-specific IgA antibodies were measured by antibody capture assay using anti-sheep IgA (alpha-chain specific) and anti-bovine RSV monoclonal antibodies. Bovine RSV-specific IgM and IgA antibodies were detected in the serum samples within 6 days post-inoculation (p.i.). Virus-specific IgC antibodies appeared in serum samples 4 days later. In nasal secretions, IgA antibodies appeared 7 days p.i. but IgM antibodies were not detected until 12-16 days p.i. In serum samples, IgM titres were predominant for the first 2 weeks p.i. IgC titres becoming predominant thereafter. In nasal secretions and lung lavage fluids, IgA titres were significantly higher than IgM or IgG titres up to 21 days p.i. (0.01).     

Evaluation of novel recombinant porcine circovirus type 2d (PCV2d) vaccine in pigs naturally infected with PCV2d

IntroductionThe pathogenic porcine circovirus type 2 (PCV2) causes significant economic losses in pig production. Emergence of the PCV2d genotype has been linked with PCV2-associated disease (PCVAD) outbreaks. However, no study has been conducted efficacy of an experimental PCV2d-based subunit vaccine in pigs. Therefore, PCV2b- and PCV2d-based capsid (CP) proteins were generated using a baculovirus (Bac) expression system, and we evaluated the protective immune responses in a commercial pig farm where predominant PCV2d is circulating.MethodsEighteen 3-week-old pigs with maternal antibodies were randomly divided into four groups, and were immunized with purified Bac-2dCP, mixed 1:1 ratio with purified Bac-2bCP and Bac-2dCP (Bac-mCP), a commercial PCV2a-based subunit vaccine (VAC) or phosphate-buffered saline (PBS) as controls.ResultsThe Bac-2dCP and Bac-mCP groups had significantly higher PCV2b- or PCV2d- specific IgG and neutralizing antibody without interference by maternal antibody compared to control group in pigs naturally infected with PCV2d. Interestingly, not only serum IL-4 level was significantly increased in the Bac-2dCP group, but also PCV2d viremia level was significantly reduced than the control group.ConclusionsThe recombinant Bac-2dCP subunit vaccine is a good candidate for the effective reduction against PCV2d infection.     

One dose of a porcine circovirus 2 (PCV2) sub-unit vaccine administered to 3-week-old conventional piglets elicits cell-mediated immunity and significantly reduces PCV2 viremia in an experimental model

The immunogenicity and efficacy generated by one dose of a PCV2 sub-unit vaccine (Porcilis PCV^®) were evaluated in 3-week-old conventional piglets. Vaccination induced both humoral and cell-mediated responses against PCV2, which were increased after the challenge with a PCV2 genotype “b” isolate. High levels of maternally derived antibodies (IPMA ≥ 10 log₂) at the time of vaccination were found to interfere with the active seroconversion, whereas titres below 8 log₂ allowed the development of a proper antibody response. Nevertheless, the immunity induced by one dose of the product was partly protective against PCV2 infection, since viremia, shedding and viral load in tissues were significantly reduced in vaccinated pigs compared to controls.     

Poly (d,l-lactide-co-glycolide) nanoparticle-entrapped vaccine induces a protective immune response against porcine epidemic diarrhea virus infection in piglets

Porcine epidemic diarrhea (PED) causes 80–100% mortality in neonatal piglets, and its causative agent, the porcine epidemic diarrhea virus (PEDV), poses an important threat to the swine industry worldwide. In this study, we prepared biodegradable poly (d,l-lactide-co-glycolide) (PLGA) nanoparticle–entrapped PEDV killed vaccine antigens (KAg) (PLGA-KAg). Late-term pregnant sows were intranasally inoculated with PLGA-KAg, and the mortality resulting from challenge with highly virulent PEDV was investigated in their passively immunized suckling piglets. PEDV-specific IgG and IgA antibody titers were enhanced in pregnant sows immunized with PLGA-KAg relative to the titers in sows inoculated with KAg. Similar results were seen in the passively immunized suckling piglets of these sows. Improved lymphocyte proliferation responses and IFN-γ levels were induced in pregnant sows immunized with PLGA-KAg compared with those vaccinated with KAg or with Montanide™ ISA 201 VG emulsified killed PEDV vaccine (201-KAg). Importantly, there was less piglet mortality in the group vaccinated with PLGA-KAg than in the groups vaccinated with KAg or 201-KAg. These results demonstrate that PLGA-KAg is a promising candidate vaccine that can provide protective immunity against PEDV infection in suckling piglets.     

Formalin-inactivated bovine RSV vaccine influences antibody levels in bronchoalveolar lavage fluid and disease outcome in experimentally infected calves

Respiratory syncytial virus (RSV) causes severe respiratory disease in calves and human infants. In response to outbreaks, formalin inactivated (FI)-RSV vaccines were developed and found to exacerbate disease following a live RSV infection. We have reproduced vaccination induced disease enhancement in calves and screened various antibody isotypes in bronchoalveolar lavage fluid (BALF) from two studies: one with disease enhancement and another where moderate protection resulted from FI-bovine RSV (BRSV) vaccination. Semi-protected vaccinated calves produced BRSV-specific BALF IgG1, but not IgA and IgG2 prior to infection; whereas, calves with enhanced disease failed to develop BRSV-specific IgG1 in BALF. Ultimately, the formulation and delivery of RSV vaccines influences protective antibody levels in respiratory secretions.     

In vivo oral administration effects of various oligodeoxynucleotides containing synthetic immunostimulatory motifs in the immune response to pseudorabies attenuated virus vaccine in newborn piglets

Numerous studies have demonstrated that oligonucleotides containing CpG motifs (CpG ODN) are efficient immunoadjuvants to various antigens administered by parenteral routes to mice. Recently, it has been found that CpG ODNs also is a promising mucosal adjuvant in mice. To date, there have been no studies to screen the optimal CpG sequence and modified ODN backbone to piglets in vivo, when delivered by oral route. We have previously demonstrated that human-specific CpG ODN is a potent adjuvant to pseudorabies live attenuated virus (PRV) vaccine when administered subcutaneously (SC) or ocularly in piglets. In this study, we screened and evaluated the optimal CpG sequences (porcine-specific, human-specific, mouse-specific ODN) and optimal backbone (SOS-backbone consisting of a nuclease-resistant phosphorothioate guanosines at the 5' and the 3'-end and with a phosphodiester (O) in the center and phosphorothioate (S) backbone (S-backbone)) to PRV vaccine delivered orally in piglets. The proliferation of peripheral blood mononuclear cells (PBMCs), IFN-gamma and IL-4 in serum, and the titre of IgG, IgG2/IgG1 isotype in serum and IgA in intestinal washings and feces to PRV vaccine were tested at different time-points. The results suggested that, CpG ODNs augmented systemic (IgG in serum, T-cell proliferation) and mucosal (IgA in intestinal washings and feces) immune responses against antigen. CpG ODNs stimulated both T-helper type1 (Th1) (IgG2) and Th2 (IgA) responses when delivered orally. With the same backbone, the porcine-specific ODN-induced responses were comparable with human-specific ODNs, but stronger than mouse-specific CpG ODNs. SOS-backbone induced a stronger IFN-gamma and proliferative responses than S-backbone, while antibody responses induced by SOS-backbones were slightly less or similar with S-backbone. The in vivo data demonstrate for the first time that porcine-specific and human-specific ODNs both are optimal sequences for mucosal system in piglets.     

Antibody isotype responses to Schistosoma japonicum antigens in subjects from a schistosomiasis area with repeated praziquantel chemotherapy compared with a new endemic zone in Hunan Province,P.R. China

To demonstrate the dynamics of specific antibody isotypes against schistosome adult worm (AWA) and soluble egg (SEA) antigens, we evaluated (in 1999-2000) 112 subjects infected with Schistosoma japonicum from 2 regions of Hunan Province, China. Fifty-eight subjects were from Area A, a well-known endemic area with repeated chemotherapy. Area B (n = 54) is a new endemic focus in another part of the same province. Serum samples were collected prior to praziquantel (PZQ) chemotherapy, and at 2 and 12 months post-treatment. IgM, IgA, IgG, IgG2, IgG4 and IgE antibodies to AWA and SEA were measured by quantitative enzyme-linked immunosorbent assay (ELISA). Pre-treatment antibody isotype levels from Area A, except IgA against AWA and SEA, were significantly higher than those from Area B. In response to chemotherapy, most antibody isotype levels fell or remained stable. However, in Area A there was a significant increase in the IgA, IgE and IgG4 responses to AWA 2 months after PZQ--which fell to approach pre-treatment levels by 12 months. A similar response was seen in Area B with IgE and IgG4 to AWA. Levels of all AWA-specific IgE and IgG4 were significantly higher in subjects from Area A compared with Area B at all time-points. AWA-IgE levels demonstrated significant linear correlations with age and number of previous PZQ treatments in Area A only. All SEA-specific isotypes in both areas fell significantly in response to treatment--except IgE, which remained stable in both area. All SEA-specific isotype levels (except IgA) were significantly higher from Area A at baseline. This significant difference was maintained through 12-months follow-up for IgE, IgG2 and IgG4 only. This study suggests that multiple episodes of schistosome infection may be required to generate antibody isotype levels that have been associated with resistance to re-infection in other studies. Further, a surrogate marker of successful chemotherapy (AWA-IgG4) performed less effectively in patients with previous treatment courses.     

Serum immunoglobulin levels and humoral immune competence in coalworkers

Serum immunoglobulin (Ig) levels were measured in 788 coalworkers and 121 nonmining controls for comparison with the radiological category of pneumoconiosis after taking into account age and smoking habit. In addition a simple assessment of humoral immune competence was made by estimating the titre of serum antibody against the common gut commensal Escherichia coli. Smoking was found to depress serum IgA and IgM whilst levels of IgG and IgA increased slightly with age. Men with radiological signs of coalworkers pneumoconiosis (CWP) had significantly raised levels of IgA and IgG with increasing pneumoconiosis category. Even coalworkers with less than category 1 simple pneumoconiosis had raised levels of IgA, suggesting that increased production of this immunoglobulin occurs before radiologically identifiable pathological changes have occurred in the lung tissue. No association between reduced humoral immune competence and radiological category of pneumoconiosis was found. Whether high Ig levels in men exposed to coal dust are merely a passive response to dusted lung tissue or whether they indicate that an immunological process is important in the development of pneumoconiotic lesions remains uncertain.     

Generation and immunogenicity of porcine circovirus type 2 chimeric virus-like particles displaying porcine reproductive and respiratory syndrome virus GP5 epitope B

Virus-like particles (VLPs) can be used as transfer vehicles carrying foreign proteins or antigen epitopes to produce chimeric VLPs for bivalent or multivalent vaccines. Based on the crystal structure of porcine circovirus type 2 (PCV2) capsid protein (Cap), in addition to alignment of the Cap sequences collected from various isolates of PCV2 and PCV1, we predicted that Loop CD of the PCV2 Cap should tolerate insertion of foreign epitopes, and furthermore that such an insertion could be presented on the surface of PCV2 VLPs. To validate this, the GP5 epitope B of porcine reproductive and respiratory syndrome virus (PRRSV) was inserted into Loop CD of the PCV2 Cap. The 3D structure of the recombinant PCV2 Cap (rCap) was simulated by homology modeling; it appeared that the GP5 epitope B was folded as a relatively independent unit, separated from the PCV2 Cap backbone. Furthermore, based on transmission electron microscopy, the purified PCV2 rCap self-assembled into chimeric VLPs which entered PK-15 cells. In addition, PCV2 chimeric VLPs induced strong humoral (neutralizing antibodies against PCV2 and PRRSV) and cellular immune responses in mice. We concluded that the identified insertion site in the PCV2 Cap had great potential to develop PCV2 VLPs-based bivalent or multivalent vaccines; furthermore, it would also facilitate development of a nano-device to present a functional peptide on the surface of the VLPs that could be used for therapeutic purposes.     

Recombination in Vaccine and Circulating Strains of Porcine Reproductive and Respiratory Syndrome Viruses

Em2007, a porcine reproductive and respiratory syndrome virus (PRRSV) variant with a unique 68 aa deletion in Nsp2, was recently isolated in China. Phylogenetic and molecular evolutionary analyses indicated that Em2007 is a natural recombinant between a vaccine strain of PRRSV and circulating virus. We also tested its pathogenicity in piglets.     

Synthetic B- and T-cell epitope peptides of porcine reproductive and respiratory syndrome virus with Gp96 as adjuvant induced humoral and cell-mediated immunity

Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) has recently caused huge economic losses in the pig industry worldwide. Commercial vaccines, including inactivated vaccines and attenuated live vaccines, are available but fail to provide sustainable protection, especially against genetically heterologous strains. Thus several approaches have been used to develop more effective PRRSV vaccines and/or immune modulators to accelerate and magnify immune responses to PRRSV vaccines. Heat shock protein Gp96 is one such modulator that enhances both the innate and adaptive immune responses. In the present study, two B-cell epitopes and seven T-cell epitopes from PRRSV and a Pan DR T-helper cell epitope were synthesized and mixed with the N-terminal 22–355 aa of Gp96 (Gp96N) as an adjuvant, and immune responses were evaluated. Our results show that Gp96N activated PRRSV-specific humoral immune responses elicited by BCE-peptides and promoted the PRRSV-specific cellular immunity induced by TCE-peptides. Moreover, higher levels of IL-12 and TNF-α and lower levels of IL-4 and IL-10 were observed in the serum of Gp96N-vaccinated piglets compared to piglets immunized with no Gp96N, displaying a predominant Th1 type of immune response induced by Gp96N. Following challenge with the virulent HP-PRRSV isolate JXwn06, piglets vaccinated with the mixture of peptides and Gp96N presented with milder clinical symptoms, lower viremia, and less pathological lesions in their lungs, however, this vaccine could not provide lasting and effective protection against HP-PRRSV infection. These data provide important bases for the development of PRRSV epitope-based synthetic peptide vaccines combined with Gp96N as attractive immunomodulators in swine.