Immune Response In Guinea Pigs Research Articles

Synthetic Particulate Subunit Vaccines for the Prevention of Q Fever.

Coxiella burnetti is an intracellular bacterium that causes Q fever, a disease of worldwide importance. Q-VAX® , the approved human Q fever vaccine, is a whole cell vaccine associated with safety concerns. Here a safe particulate subunit vaccine candidate is developed that is ambient-temperature stable and can be cost-effectively manufactured. Endotoxin-free Escherichia coli is bioengineered to efficiently self-assemble biopolymer particles (BPs) that are densely coated with either strings of 18 T-cell epitopes (COX-BP) or two full-length immunodominant antigens (YbgF-BP-Com1) all derived from C. burnetii. BP vaccine candidates are ambient-temperature stable. Safety and immunogenicity are confirmed in mice and guinea pig (GP) models. YbgF-BP-Com1 elicits specific and strong humoral immune responses in GPs with IgG titers that are at least 1000 times higher than those induced by Q-VAX® . BP vaccine candidates are not reactogenic. After challenge with C. burnetii, YbgF-BP-Com1 vaccine leads to reduced fever responses and pathogen burden in the liver and the induction of proinflammatory cytokines IL-12 and IFN-γ inducible protein (IP-10) when compared to negative control groups. These data suggest that YbgF-BP-Com1 induces functional immune responses reducing infection by C. burnetii. Collectively, these findings illustrate the potential of BPs as effective antigen carrier for Q fever vaccine development.

Dynamics of the immune response in laboratory animals to the introduction of a protective protein of the cause of respiratory syncytial infection in cattle

The authors in the article presented the results of assessing the immune response of guinea pigs using the protective protein of the bovine respiratory syncytial infection virus (RSI), obtained using recombinant E. coli BRSV-F1. The research was carried out in the branch laboratory of veterinary biotechnology and infectious animal diseases of the Vitebsk Order of the Badge of Honor of the State Academy of Veterinary Medicine. The average titer of specific antibodies of guinea pigs to the use of the protective protein of the bovine RSI virus was studied in 9 experimental groups and one control group. Blood sampling was carried out at the beginning of the experiment, before the second immunisation and 14 days after the second immunisation. The average titer of specific antibodies to the use of the protective protein of the bovine RSI virus was determined using an indirect hemagglutination reaction with an erythrocyte diagnosticum containing the bovine respiratory syncytial virus (RSV). With the introduction of samples of the prototype vaccine containing the gene encoding the production of the bovine RSI virus protein: group 1 - whole bacteria E. coli BRSV-F1 with an inducer of protein synthesis IPTG; 2nd, whole bacteria E. coli BRSV-F1 with protein synthesis inducer IPTG + 15% adjuvant IZA-15; 3rd, bacterial lysate E. coli BRSV-F1 with an inducer of protein synthesis IPTG; 4th – E. coli bacterial lysate BRSV-F1 with protein synthesis inducer IPTG + 15% adjuvant IZA-15; 5th - protective protein F1 + 15% adjuvant IZA-15; 6th - protective protein F1 + 2% cellulose, 7th - inactivated bovine RSV (JSC «Belvitunipharm») + 15% adjuvant IZA15; 8th - Hiprabovis 4 vaccines (HIPRA, Spain) and 9th - Bovi-shield Gold FP 5 L5 vaccines (Zoetis, USA) - in the experimental groups, an increase in antibody titer in paired blood sera was noted compared to the control group. From the data obtained, it was concluded that the studied samples of the vaccine prototype containing the gene encoding the production of the bovine RSV protein are not inferior in immunogenic activity to the vaccines Bovishield Gold FP 5 L5 and Hiprabovis 4. Therefore, the protective protein of the bovine RSI virus can be included in the composition of vaccines against this pathology to replace the cultural virus.

Aerosolized Zika virus infection in Guinea pigs

ABSTRACT Zika virus (ZIKV) is primarily transmitted through mosquito bites and sexual contact, and vertical transmission of ZIKV has also been observed in humans. In addition, ZIKV infection via unknown transmission routes has been frequently reported in clinical settings. However, whether ZIKV can be transmitted via aerosol routes remains unknown. In this study, we demonstrated that aerosolized ZIKV is fully infectious in vitro and in vivo. Remarkably, intratracheal (i.t.) inoculation with aerosolized ZIKV led to rapid viremia and viral secretion in saliva, as well as robust humoral and innate immune responses in guinea pigs. Transcriptome analysis further revealed that the expression of genes related to viral processes, biological regulation and the immune response was significantly changed. Together, our results confirm that aerosolized ZIKV can result in systemic infection and induce both innate and adaptive immune responses in guinea pigs, highlighting the possibility of ZIKV transmission via aerosols.

Porcine circovirus 2 capsid protein produced in N. benthamiana forms virus-like particles that elicit production of virus-neutralizing antibodies in guinea pigs

Porcine circovirus type 2 (PCV2) is a non-enveloped, icosahedral virus of the Circoviridae family, with a small, circular, single-stranded DNA genome. PCV2 infections cause substantial economic losses in the pig industry worldwide. Currently, commercially produced PCV2 vaccines are expensive, whereas plant-based expression systems can produce recombinant proteins at low cost for use as vaccines. In this study, recombinant PCV2 capsid protein (rCap) was transiently expressed in Nicotiana benthamiana and purified by metal affinity chromatography, with a yield of 102 mg from 1 kg plant leaves. Electron microscopy confirmed that purified rCap self-assembled into virus-like particles (VLPs) at neutral pH. It was shown to provoke a strong immune response in guinea pigs. The results indicate that plant systems can enable production of large amounts of proteins to serve as candidates for subunit vaccines.

Humoral and cell-mediated immune responses to plant-produced African horse sickness virus VP7 quasi-crystals

African horse sickness (AHS) is a devastating viral disease affecting equines and has resulted in many disastrous epizootics. To date, no successful therapeutic treatment exists for AHS, and commercially used live-attenuated vaccines have various undesirable side effects. Previous studies have shown that mice inoculated with insoluble African horse sickness virus (AHSV) VP7 crystals are protected from live challenge with a lethal dose of AHSV. This study investigates the humoral and cell-mediated immune responses in guinea-pigs to a safer monovalent vaccine alternative based on AHSV-5 VP7 quasi-crystals produced in plants. Guinea-pigs received prime- and boost-inoculations of between 10 and 50 μg of purified plant-produced AHSV VP7. Western immunoblot analysis of the humoral response showed stimulation of high titres of anti-VP7 antibodies 28 days after the boost-inoculation in sera from three of the five experimental animals. In addition, RNA-seq transcriptome profiling of guinea-pig spleen-derived RNA highlighted thirty significantly (q ≤ 0.05) differentially expressed genes involved in innate and adaptive immunity. Differential expression of genes involved in Th1, Th2 and Th17 cell differentiation suggest a cell-mediated immune response to AHSV-5 VP7. Upregulation of several important cytokines and cytokine receptors were noted, including TNFSF14, CX3CR1, IFNLR1 and IL17RA. Upregulation of IL17RA suggests a Th17 response which has been reported as a key component in AHSV immunity. While further investigation is needed to validate these findings, these results suggest that AHSV-5 VP7 quasi-crystals produced in N. benthamiana are immunogenic and induce both humoral and cell-mediated responses.

Repeat tick exposure elicits distinct immune responses in guinea pigs and mice

Ticks deposit salivary proteins into the skin during a bite to mediate acquisition of a blood meal. Acquired resistance to tick bites has been demonstrated to prevent Borrelia burgdorferi sensu lato (s.l.) transmission. However, the mechanism of resistance, as well as the protective antigens, have remained elusive. To address these unknowns, we utilized a guinea pig model of tick resistance and a mouse model of permissiveness. Guinea pigs developed immunity after multiple Ixodes scapularis tick infestations, characterized by rapid tick detachment and impaired feeding. In comparison, mice tolerated at least 6 infestations with no significant impact on feeding. We analyzed the bite sites by RNA-sequencing and histology, identifying several inflammatory pathways in tick immune animals, such as FcεRI signaling and complement activation, and activation of coagulation pathways that could impair local blood flow. Together, these results identify important pathways altered during tick rejection and potential tick proteins that could serve as vaccine candidates.

Effect of dietary zinc nanoparticle supplementation on mineral balance, tissue minerals status and immune response in Guinea pigs

The present study was designed to examine the effect of dietary supplementation of zinc nanoparticles on mineral balance, tissue minerals status and immune response in guinea pigs. Depending upon the source of Zn, 30 weaned male guinea pigs were randomly allocated to five different groups having 6 animals in each group in CRD. Animals under group T1 (control) were fed with basal diet with inorganic Zn (ZnSO4), whereas, animals were fed with organic Zn (Zn methionine) in group T2, commercial zinc nanoparticle in group T3 and two other zinc nanoparticles prepared in our laboratory in groups T4 and T5, respectively. They were supplemented with 20 mg Zn/kg feed on dry matter basis for 90 days. Results revealed that intake and balance of nitrogen and minerals (calcium, phosphorus, Zn and Cu) was not affected (P>0.05) by supplementation zinc nanoparticles. The level of Zn in liver and testes was significantly higher (P<0.05) in the zinc supplemented groups, when compared to that of control group, however, the mean values of Cu, Fe and Mn were comparable (P>0.05) among different groups, irrespective of the source of Zn. The mean antibody titre was significantly (P<0.05) higher in the commercial nanoparticle supplemented group as compared to other groups. As evidenced in our study spanning 90 days of trial, zinc nanoparticles can be safely supplemented up to 20 ppm level in the diet of guinea pigs.

Comparison of immune responses in guinea pigs by intranasal delivery with different nanoparticles-loaded FMDV DNA vaccine

To compare different nanoparticle-based nasal vaccines against foot-and-mouth disease (FMD), chitosan (CS)-coated poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) (CS/PLGA-NPs) and amino-functionalized mesoporous silica nanoparticles (Am/MSNs) loaded with FMDV recombinant plasmid (pP12A3C/IFN-CS/PLGA-NPs and pP12A3C/IFN-Am/MS-NPs) were used to induce mucosal and systemic immune responses in guinea pigs via intranasal delivery. Simultaneously, CpG oligodeoxy nucleotides (ODNs) as a vaccine adjuvant were encapsulated in chitosan-coated poly (lactic-co-glycolic acid) nanoparticles (CpG-CS/PLGA-NPs). The pP12A3C/IFN-CS/PLGA-NPs and CpG-CS/PLGA-NPs generated displayed good morphology, high stability, mean diameters of 500 and 400 nm and encapsulation efficiencies of 83.8% and 88.4%, respectively. Data from the in vitro release assay showed that plasmid and CpG were sustainably released from nanoparticles (up to 66.73% and 64%, respectively, of the total amount loaded). Guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs + CpG-CS/PLGA-NPs showed markedly higher mucosal, cellular and humoral immune responses than those administered pP12A3C/IFN-CS/PLGA-NPs or naked plasmid vaccine alone. FMDV-specific secretory immunoglobulin A (sIgA) antibodies in nasal washes were initially detected at 3 days post-vaccination with CS/PLGA-NPs loaded with plasmid. Guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs also displayed higher cellular and humoral immune responses than pP12A3C-CS/PLGA-NPs and naked plasmid vaccine alone. FMDV-specific immunoglobulin G (IgG) antibodies in serum were initially detected at 5 days post-vaccination (intramuscularly) with the naked plasmid. Finally, challenge experiments 42 days post-vaccine revealed 100% protection in guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs + CpG-CS/PLGA-NPs and pP12A3C/IFN-CS/PLGA-NPs. However, plasmid DNA was burst released from pP12A3C/IFN-Am/MS-NPs. Our attempts to use pP12A3C/IFN-Am/MS-NPs to immunize guinea pigs failed to induce immune responses. In conclusion, CpG and IFN-α adjuvant based FMD vaccines elicit protection in guinea pigs. Moreover, CS-coated PLGA NPs present an efficient and safe mucosal immune delivery system for FMDV DNA vaccine. Data from the current study provide a foundation for understanding and further evaluating protective immune responses in pigs.

Development of an anti-guinea pig CD4 monoclonal antibody for depletion of CD4+ T cells in vivo

The guinea pig serves as a useful animal model for a number of human diseases and has played an important role during development and testing of experimental vaccines and disease therapies. However, the availability of reagents to examine the immunological response in this species is very limited. Monoclonal antibodies (mAb) specific for cell surface proteins or products of immune cells have been useful tools for characterizing and quantifying immune responses in humans and in murine models of human disease, but very few similar reagents are available for characterizing and manipulating the immune response of guinea pigs. A rat IgG2a mAb specific for guinea pig CD4 has previously been described and was shown to inhibit T cell proliferation, but was inefficient at depleting CD4+ T cells in vivo. We hypothesized that the in vivo CD4+ T cell depletion function of this mAb could be improved by expression of the rat IgG2b heavy chain. We show that the purified mAb from an IgG2b class-switch variant, but not the parental IgG2a mAb, significantly depleted CD4+ T cells from secondary lymphoid tissue of guinea pigs. Further, treatment of guinea pigs with the IgG2b mAb at 2.0 mg/kg resulted in depletion of CD4+ T cells from peripheral blood and spleen. The use of this modified antibody to specifically alter the immune response of guinea pigs should prove useful in a number of guinea pig infectious disease models.

Immune response of a heat killed Brucella abortus vaccine in guinea pig

Background: Brucellosis is an endemic disease in Bangladesh which has economic impacts attributable to humans and animals. To control bovine brucellosis two types of vaccines are available- vaccine S19 and vaccine SRB51 but they have some adverse effects. On the other hand the heat killed vaccine produces less immunity but no adverse effect. Vaccination against brucellosis in Bangladesh has not yet been initiated and not recommended in subsistence management systems due to very low level of prevalence. But in commercial management systems the prevalence is reported to be higher and vaccination may be initiated. Before importing live vaccine which have some adverse effects locally prepared killed vaccine can be tested for its immune response. Hence this study was undertaken to evaluate the immune response of heat killed vaccine prepared from local isolate in guinea pig. Methods: Brucella abortus recently isolated from aborted fetal membranes (unpublished data) was used for vaccine production. Pour plate technique was used by tenfold serial dilution of the isolate to count cfu (colony forming unit)/ml of Brucella abortus for dose calculation of heat killed vaccine. Bacterial pellet was prepared by centrifugation of 200ml of the cultured broth at 10,000 rpm for 10 mins. The bacterial pellet was mixed with required amount of PBS (phosphate buffer saline) to obtain 40×1010 cfu organisms in 2ml dose for guinea pig inoculation. Then heat killed vaccine was prepared by heating the organism at 80˚C for 90 minutes and the prepared vaccine was inoculated subcutaneously 2ml (4×1010cfu) in each of the guinea pig. The sera of guinea pigs were collected at 1st, 2nd, 4th, 6th and 9th week after inoculation to determine the reciprocal antibody (Ab) titre by Rose Bengal test (RBT) and to examine the rise of antibody level by Enzyme-Linked Immunosorbent Assay (ELISA). Results: The antibody level started to rise significantly (p<0.01) from the 2nd week (OD value 0.2287, Reciprocal Ab titre 1:120) and reached a peak level at 4th week (OD value 0.2842, Reciprocal Ab titre 1:800) and then started to decline significantly (p<0.01) from 6th week (OD value 0.1832, Reciprocal Ab titre 1:35) to 9th week (OD value 0.1015, Reciprocal Ab titre 0). Conclusions: Heat killed vaccine without adjuvant induces immune response in guinea pigs which persists for a maximum period of 6 weeks. A further study to investigate the immune response of killed vaccine with adjuvant is recommended.

Novel avian paramyxovirus-based vaccine vectors expressing the Ebola virus glycoprotein elicit mucosal and humoral immune responses in guinea pigs

Paramyxovirus vaccine vectors based on human parainfluenza virus type 3 (HPIV-3) and Newcastle disease virus (NDV) have been previously evaluated against Ebola virus (EBOV) challenge. Although both the viral vectored vaccines efficiently induce protective immunity, some concerns remain to be solved. Since HPIV-3 is a common human pathogen, the human population has pre-existing immunity to HPIV-3, which may restrict the replication of the vaccine vector. For NDV, mesogenic (intermediate virulent) strain used in previous studies is currently classified as a Select Agent in the United States, thus making it unsuitable to be used as a vaccine vector. To overcome these concerns, we have developed a modified NDV vector based on a mesogenic NDV strain, in which the ectodomains of envelope glycoproteins were replaced with the corresponding ectodomains from avian paramyxovirus serotype 3 (APMV-3). The modified NDV vector was highly attenuated in chickens and was able to express the EBOV glycoprotein (GP) gene at high level. In addition, the recombinant APMV-3 was also evaluated as a vaccine vector to express the EBOV GP gene. Guinea pigs immunized with these two vector vaccines developed high levels of neutralizing GP-specific IgG and IgA antibodies.

Hollow mesoporous silica nanoparticles as delivery vehicle of foot-and-mouth disease virus-like particles induce persistent immune responses in guinea pigs.

Foot‐and‐mouth disease (FMD) is an acute and febrile infectious disease, which can cause great economic losses. Virus‐like particles (VLPs) as an advantageous antigen can induce significant specific immune response. To improve immunity of VLPs, especially, make it induce persistent immune response, the hollow mesoporous silica nanoparticles (HMSNs) as a potential nano‐adjuvant were synthesized and loaded the FMD virus (FMDV) VLPs. They were injected into guinea pigs and the specific immune response was detected. The results confirmed that HMSNs/VLPs can induce persistent humoral immunity with high‐level antibody titer for more than three months. HMSNs also improve the T‐lymphocyte proliferation and IFN‐γ induced by FMDV VLPs, and provides the ideal protection against FMDV challenge. These consequences indicated that HMSNs were good protein delivery vehicle and potential nano‐adjuvant of vaccines.

Adjuvantation of inactivated Foot and Mouth Disease Virus vaccine with IL-15 expressing plasmid improves the immune response in Guinea Pigs

Foot and mouth disease is a highly contagious disease affecting cloven footed animals. Vaccination using inactivated virus is followed to control the disease. As the immune response conferred by the inactivated vaccine is short lived, there is a need for an alternate vaccine with increased duration of immunity. Inclusion of adjuvant which enhances B and T cell responses is one of the strategies to increase the duration of immune responses of the vaccine. Interleukin 15 is one such a cytokine which improves the cell mediated immune response and also involved in the maintenance of memory T and B cells. In the present communication, we evaluated the role of bovine IL-15 as an adjuvant to inactivated FMD vaccine in guinea pig model. Animals injected with FMD inactivated vaccine and IL-15 plasmid showed improved levels of neutralizing antibodies which were maintained up to 6 months (as the level of neutralizing antibodies is more >1.5 which is considered to give protection). Increased Th1 and Th2 responses (by measuring the level of IL-4 and IFN- gamma responses) were seen in IL-15 adjuvanted guinea pigs compared to animals injected with inactivated vaccine alone.

Intranasal delivery of a bivalent norovirus vaccine formulated in an in situ gelling dry powder.

The global health community is beginning to understand the burden of norovirus-associated disease, which has a significant impact in both developed and developing countries. Norovirus virus like particle (VLP)-based vaccines are currently under development and have been shown to elicit systemic and mucosal immune responses when delivered intranasally. In the present study, we describe the use of a dry powder formulation (GelVac™) with an in situ gelling polysaccharide (GelSite™) extracted from Aloe vera for nasal delivery of a bivalent vaccine formulation containing both GI and GII.4 norovirus VLPs. Dose-ranging studies were performed to identify the optimal antigen dosages based on systemic and mucosal immune responses in guinea pigs and determine any antigenic interference. A dose-dependent increase in systemic and mucosal immunogenicity against each of the VLPs were observed as well as a boosting effect for each VLP after the second dosing. A total antigen dose of ≥50 μg of each GI and GII.4 VLPs was determined to be the maximally immunogenic dose in guinea pigs. The immunogenicity results of this bivalent formulation, taken together with previous work on monovalent GelVac™ norovirus vaccine formulation, provides a basis for future development of this norovirus VLP vaccine.

Active immunization with Brucella abortus S19 phage lysate elicits serum IgG that protects guinea pigs against virulent B. abortus and protects mice by passive immunization

Brucellosis is an economically important zoonosis of worldwide significance. Earlier (Jain et al., 2015) we reported methodology for generation of phage lysate preparations against Brucella abortus S19 using brucellaphage ‘ϕLd’. In this study, using a fixed dose (Two mouse PD100) of lysates, the prophylactic efficacies of both plain and alum gel adjuvanted lysates were evaluated in guinea pig by direct virulent challenge and passive mouse protection test (PMPT). Strong humoral and cell mediated immune responses in guinea pigs and protection comparable to S19 vaccine was observed with low dose (1.0 μg protein and 120 μg carbohydrate adsorbed on 0.1% aluminium gel). Passive transfer of antibodies to mice using d 90 post immunization sera of guinea pig protected the animals against challenge. The study suggested the significance of humoral immunity in murine brucellosis. Further, the methodology can be explored to produce a new class of immunotherapeutic agents against bovine brucellosis.

Detection of herpes simplex virus type 2 (HSV-2) -specific cell-mediated immune responses in guinea pigs during latent HSV-2 genital infection

Genital infections with herpes simplex virus type 2 (HSV-2) are a source of considerable morbidity and are a health concern for newborns exposed to virus during vaginal delivery. Additionally, HSV-2 infection diminishes the integrity of the vaginal epithelium resulting in increased susceptibility of individuals to infection with other sexually transmitted pathogens. Understanding immune protection against HSV-2 primary infection and immune modulation of virus shedding events following reactivation of the virus from latency is important for the development of effective prophylactic and therapeutic vaccines. Although the murine model of HSV-2 infection is useful for understanding immunity following immunization, it is limited by the lack of spontaneous reactivation of HSV-2 from latency. Genital infection of guinea pigs with HSV-2 accurately models the disease of humans including the spontaneous reactivation of HSV-2 from latency and provides a unique opportunity to examine virus-host interactions during latency. Although the guinea pig represents an accurate model of many human infections, relatively few reagents are available to study the immunological response to infection. To analyze the cell-mediated immune response of guinea pigs at extended periods of time after establishment of HSV-2 latency, we have modified flow-cytometry based proliferation assays and IFN-γ ELISPOT assays to detect and quantify HSV-specific cell-mediated responses during latent infection of guinea pigs. Here we demonstrate that a combination of proliferation and ELISPOT assays can be used to quantify and characterize effecter function of virus-specific immune memory responses during HSV-latency.

Ebolavirus Glycoprotein Fc Fusion Protein Protects Guinea Pigs against Lethal Challenge.

Ebola virus (EBOV), a member of the Filoviridae that can cause severe hemorrhagic fever in humans and nonhuman primates, poses a significant threat to the public health. Currently, there are no licensed vaccines or therapeutics to prevent and treat EBOV infection. Several vaccines based on the EBOV glycoprotein (GP) are under development, including vectored, virus-like particles, and protein-based subunit vaccines. We previously demonstrated that a subunit vaccine containing the extracellular domain of the Ebola ebolavirus (EBOV) GP fused to the Fc fragment of human IgG1 (EBOVgp-Fc) protected mice against EBOV lethal challenge. Here, we show that the EBOVgp-Fc vaccine formulated with QS-21, alum, or polyinosinic-polycytidylic acid-poly-L-lysine carboxymethylcellulose (poly-ICLC) adjuvants induced strong humoral immune responses in guinea pigs. The vaccinated animals developed anti-GP total antibody titers of approximately 105−106 and neutralizing antibody titers of approximately 103 as assessed by a BSL-2 neutralization assay based on vesicular stomatitis virus (VSV) pseudotypes. The poly-ICLC formulated EBOVgp-Fc vaccine protected all the guinea pigs against EBOV lethal challenge performed under BSL-4 conditions whereas the same vaccine formulated with QS-21 or alum only induced partial protection. Vaccination with a mucin-deleted EBOVgp-Fc construct formulated with QS-21 adjuvant did not have a significant effect in anti-GP antibody levels and protection against EBOV lethal challenge compared to the full-length GP construct. The bulk of the humoral response induced by the EBOVgp-Fc vaccine was directed against epitopes outside the EBOV mucin region. Our findings indicate that different adjuvants can eliciting varying levels of protection against lethal EBOV challenge in guinea pigs vaccinated with EBOVgp-Fc, and suggest that levels of total anti-GP antibodies elicit by protein-based GP subunit vaccines do not correlate with protection. Our data further support the development of Fc fusions of GP as a candidate vaccine for human use.

Studying of S- and L- form Brucella’s Thermo-Extracts Immunogenic Characteristics at Cavies Guinea Pigs

Relevance. In the Russian Federation is noted the negative dynamics of epizootic process of brucellosis among epidemiologically important species of farm animals (cattle and small ruminants), which represents a threat to the population. Used in Russia live vaccine based on a strain of Brucella abortus 19 the BA has reduced virulence, but capable at high doses (108 -2 109 m.c.) cause a generalized infection in guinea pigs and humans, and in violation of the rules cause post-vaccination complications. Goal. Assess possibility of the thermo-extract derived from the S- and L-forms of Brucella, get an immune response in guinea pigs, and reduce the risk of infection with virulent brucella. Materials and methods. Two series of experiments were carried out on guinea pigs. Immunized guinea pigs thermo-extracts (TE) from a strain of B. abortus I-206 in the S- and L-forms, and live brucellosis vaccine (Scientific and Production Association for Immunological Preparations «Microgen», Russia). To infect guinea pigs using virulent B. abortus 544 (Reference) and B. melitensis I-203 from the museum of living cultures of the Irkutsk Scientific Research Anti-Plague Institute. Results. In the first and second experiment after immunization L TE in dose 5 mg and 10 mg after infection with B. abortus and B. melitensis 5441-203 were approximately similar results. Immunization of Brucella in TE S-form or complex of L + S TE either of two doses (5 mg or 10 mg) cultures after infection B. abortus and B. melitensis 5441-203 gave the same result as a vaccine B. abortus 19VA. Conclusions. The results indicate the prospects of further study of experimental steps for using immunizing agents TE S, L TE and TE S + L on the laboratory animals.

Preclinical dose-ranging studies of a novel dry powder norovirus vaccine formulation

Norovirus is the primary cause of viral gastroenteritis in humans with multiple genotypes currently circulating worldwide. The development of a successful norovirus vaccine is contingent on its ability to induce both systemic and mucosal antibody responses against a wide range of norovirus genotypes. Norovirus virus-like particles (VLPs) are known to elicit systemic and mucosal immune responses when delivered intranasally. Incorporation of these VLPs into an intranasal powder vaccine offers the advantage of simplicity and induction of neutralizing systemic and mucosal antibodies. Nasal immunization, which provides the advantage of ease of administration and a mucosal delivery mechanism, faces the real issue of limited nasal residence time due to mucociliary clearance. Herein, we describe a novel dry powder (GelVac™) formulation of GI or GII.4 norovirus VLPs, two dominant circulating genotypes, to identify the optimal antigen dosages based on systemic and mucosal immune responses in guinea pigs. Systemic and mucosal immunogenicity of each of the VLPs was observed in a dose-dependent manner. In addition, a boosting effect was observed after the second dosing of each VLP antigen. With the GelVac™ formulation, a total antigen dose of ≥15μg was determined to be the maximally immunogenic dose for both GI and GII.4 norovirus VLPs based on evaluation for 56 days. Taken together, these results indicate that norovirus VLPs could be used as potential vaccine candidates without using an immunostimulatory adjuvant and provide a basis for the development of a GelVac™ bivalent GI/GII.4 norovirus VLP vaccine.

Protection against Foot-and-Mouth Disease Virus in Guinea Pigs via Oral Administration of Recombinant Lactobacillus plantarum Expressing VP1.

Mucosal vaccination is an effective strategy for generating antigen-specific immune responses against mucosal infections of foot-and-mouth disease virus (FMDV). In this study, Lactobacillus plantarum strains NC8 and WCFS1 were used as oral delivery vehicles containing a pSIP411-VP1 recombinant plasmid to initiate mucosal and systemic immune responses in guinea pigs. Guinea pigs were orally vaccinated (three doses) with NC8-pSIP411, NC8-pSIP411-VP1, WCFS1-pSIP411, WCFS1-pSIP411-VP1 or milk. Animals immunized with NC8-pSIP411-VP1 and WCFS1-pSIP411-VP1 developed high levels of antigen-specific serum IgG, IgA, IgM, mucosal secretory IgA (sIgA) and neutralizing antibodies, and revealed stronger cell-mediated immune responses and enhanced protection against FMDV challenge compared with control groups. The recombinant pSIP411-VP1 effectively improved immunoprotection against FMDV in guinea pigs.