Specific IgG Antibody Responses Research Articles

Enhancement of immune responses by co-administration of bacterial ghosts-mediated Neisseria gonorrhoeae DNA vaccines.

Gonorrhoea remains a leading public health burden and the development of vaccine against gonorrhoea becomes more urgent. Here, a novel Neisseria gonorrhoeae DNA vaccine delivered by Salmonella enteritidis ghosts was developed and the immune responses of the vaccine candidate were evaluated. Neisseria gonorrhoeae nspA gene was cloned into the pVAX1 vector. The constructed recombinant plasmid pVAX1-nspA was loaded into the lyophilized SE ghosts to produce SE ghosts (pVAX1-nspA). Then, the immune responses induced by SE ghosts (pVAX1-nspA) alone and co-administrated with SE ghosts (pVAX1-porB) were evaluated in mouse model. Co-administered SE ghosts (pVAX1-nspA) and SE ghosts (pVAX1-porB) could elicited significantly higher levels of specific IgG antibody responses and lymphocyte proliferative responses than the control groups. Furthermore, the group co-administered SE ghosts (pVAX1-nspA) and SE ghosts (pVAX1-porB) had the highest bactericidal antibody titres. Co-administration of SE ghosts (pVAX1-nspA) and SE ghosts (pVAX1-porB) elicited significant specific humoral and cellular immune responses. This study demonstrates the potential of co-administration of SE ghosts (pVAX1-nspA) and SE ghosts (pVAX1-porB) as an attractive vaccination regimen for gonorrhoea.

Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2

SARS-CoV-2 has emerged as a previously unknown zoonotic coronavirus that spread worldwide causing a serious pandemic. While reliable nucleic acid–based diagnostic assays were rapidly available, only a limited number of validated serological assays were available in the early phase of the pandemic. Here, we evaluated a novel flow cytometric approach to assess spike-specific antibody responses.HEK 293T cells expressing SARS-CoV-2 spike protein in its natural confirmation on the surface were used to detect specific IgG and IgM antibody responses in patient sera by flow cytometry. A soluble angiotensin-converting-enzyme 2 (ACE-2) variant was developed as external standard to quantify spike-specific antibody responses on different assay platforms. Analyses of 201 pre-COVID-19 sera proved a high assay specificity in comparison to commercially available CLIA and ELISA systems, while also revealing the highest sensitivity in specimens from PCR-confirmed SARS-CoV-2-infected patients. The external standard allowed robust quantification of antibody responses among different assay platforms. In conclusion, our newly established flow cytometric assay allows sensitive and quantitative detection of SARS-CoV-2-specific antibodies, which can be easily adopted in different laboratories and does not rely on external supply of assay kits. The flow cytometric assay also provides a blueprint for rapid development of serological tests to other emerging viral infections

Parenterally Administered P24-VP8* Nanoparticle Vaccine Conferred Strong Protection against Rotavirus Diarrhea and Virus Shedding in Gnotobiotic Pigs.

Current live rotavirus vaccines are costly with increased risk of intussusception due to vaccine replication in the gut of vaccinated children. New vaccines with improved safety and cost-effectiveness are needed. In this study, we assessed the immunogenicity and protective efficacy of a novel P24-VP8* nanoparticle vaccine using the gnotobiotic (Gn) pig model of human rotavirus infection and disease. Three doses of P24-VP8* (200 μg/dose) intramuscular vaccine with Al(OH)3 adjuvant (600 μg) conferred significant protection against infection and diarrhea after challenge with virulent Wa strain rotavirus. This was indicated by the significant reduction in the mean duration of diarrhea, virus shedding in feces, and significantly lower fecal cumulative consistency scores in post-challenge day (PCD) 1–7 among vaccinated pigs compared to the mock immunized controls. The P24-VP8* vaccine was highly immunogenic in Gn pigs. It induced strong VP8*-specific serum IgG and Wa-specific virus-neutralizing antibody responses from post-inoculation day 21 to PCD 7, but did not induce serum or intestinal IgA antibody responses or a strong effector T cell response, which are consistent with the immunization route, the adjuvant used, and the nature of the non-replicating vaccine. The findings are highly translatable and thus will facilitate clinical trials of the P24-VP8* nanoparticle vaccine.

Pathogenicity of porcine deltacoronavirus (PDCoV) strain NH and immunization of pregnant sows with an inactivated PDCoV vaccine protects 5‐day‐old neonatal piglets from virulent challenge

In this study, the pathogenicity of porcine deltacoronavirus (PDCoV) strain NH (passage 10, P10) was evaluated. We found that PDCoV strain NH is enteropathogenic in 5‐day‐old pigs. Pathogenicity experiments provided a challenge model for studying the protection efficiency of passive immunity. In order to investigate the protective efficacy of passive immunity in newborn piglets, pregnant sows were vaccinated with either a PDCoV‐inactivated vaccine at the Houhai acupoint (n = 5) or DMEM as a negative control (n = 2) using a prime/boost strategy 20 and 40 days before delivery. PDCoV spike (S)‐specific IgG and neutralizing antibody (NA) responses were detected in immunized sows and piglets born to immunized sows. PDCoV spike (S)‐specific sIgA was also detected in the colostrum and milk of immunized sows. Five days post‐farrowing, piglets were orally challenged with PDCoV strain NH (105 TCID50/piglet). Severe diarrhoea, high levels of viral RNA copies and substantial intestinal villus atrophy were detected in piglets born to unimmunized sows. Only 4 of 31 piglets (12.9%) born to immunized sows in the challenge group displayed mild to moderate diarrhoea, lower viral RNA copies and minor intestinal villi damage compared to piglets born to unimmunized sows post‐challenge. Mock piglets exhibited no typical clinical symptoms. The challenge experiment results indicated that the inactivated PDCoV vaccine exhibited 87.1% protective efficacy in the piglets. These findings suggest that the inactivated PDCoV vaccine has the potential to be an effective vaccine, providing protection against virulent PDCoV.

Immunization of cats to induce neutralizing antibodies against Fel d 1, the major feline allergen in human subjects

Cat allergy in human subjects is usually caused by the major cat allergen Fel d 1 and is found in approximately 10% of the Western population. Currently, there is no efficient and safe therapy for cat allergy available. Allergic patients usually try to avoid cats or treat their allergy symptoms. We developed a new strategy to treat Fel d 1-induced allergy in human subjects by immunizing cats against their own major allergen, Fel d 1. A conjugate vaccine consisting of recombinant Fel d 1 and a virus-like particle derived from the cucumber mosaic virus containing the tetanus toxin-derived universal T-cell epitope tt830-843 (CuMVTT) was used to immunize cats. Afirst tolerability and immunogenicity study, including a boost injection, was conducted by using the Fel-CuMVTT vaccine alone or in combination with an adjuvant. The vaccine was well tolerated and had no overt toxic effect. All cats induced a strong and sustained specific IgG antibody response. The induced anti-Fel d 1 antibodies were of high affinity and exhibited a strong neutralization ability tested both invitro and invivo. Areduction in the endogenous allergen level and a reduced allergenicity of tear samples, were observed. Vaccination of cats with Fel-CuMVTT induces neutralizing antibodies and might result in reduced symptoms of allergic cat owners. Both human subjects and animals could profit from this treatment because allergic cat owners would reduce their risk of developing chronic diseases, such as asthma, and become more tolerant of their cats, which therefore could stay in the households and not need to be relinquished to animal shelters.

CD40-targeted S1 subunit vaccine protects against MERS-CoV and S1-associated pulmonary immunopathology in transgenic human DPP4 mouse model

Abstract Middle East respiratory syndrome coronavirus (MERS-CoV) is a highly pathogenic respiratory virus that emerged in 2012. While infection control measures have played a major role in limiting human/camel-to-human transmissions, development of safe and effective human or camel vaccines is warranted. Here, we extended and optimized our previous rAd5-based vaccine platform characterized by in vivo amplified and CD40-mediated specific responses to generate MERS-CoV S1 subunit-based vaccine. We generated rAd5 constructs expressing CD40-targeted S1 fusion protein (rAd5-S1/F/CD40L), untargeted S1 (rAd5-S1), and GFP (rAd5-GFP), and evaluated their efficacy and safety in human DPP4 transgenic (hDPP4 Tg+) mice. Immunization of hDPP4 Tg+ mice showed that a single dose of rAd5-S1/F/CD40L elicited robust and significant specific IgG and neutralizing antibody responses as those induced in mice immunized with two-doses of rAd5-S1. After MERS-CoV challenge, both vaccines conferred complete protection against morbidity and mortality, as evidenced by significantly undetectable/reduced pulmonary viral loads as compared to control group. However, rAd5-S1 but not rAd5- S1/F/CD40L immunized mice exhibited marked pulmonary perivascular hemorrhage post MERS-CoV challenge despite the observed protection. Collectively, these data indicate that incorporating CD40L into this Ad5-based MERS-CoV S1 subunit vaccine as targeting molecule and molecular adjuvant can not only enhance immunogenicity and efficacy but also prevents induction of inadvertent pulmonary pathology in immunized and challenged mice, thereby offering a promising strategy to enhance the safety and potency of antigen-specific immune responses.

Oral immunization of BALB/c mice with Helicobacter pylori lysate combined with dmLT adjuvant for the induction of mucosal and system immune responses

Objective To observe the protective effects of oral immunization with Helicobacter pylori (Hp) lysates in combination with mucosal adjuvant dmLT (double mutant heat-labile toxin) against Hp infection in a BALB/c mouse model and to analyze the features of induced immune responses. Methods BALB/c mice were orally immunized with Hp lysate (Sydney strain 1, SS1 strain) and dmLT adjuvant, and then innoculated with live Hp strains through oral gavage. A control group was set up by oral administration of normal saline (200 μl/mouse). The colonization of Hp strains in the stomachs of mice was measured six weeks after bacterial inoculation. Samples of serum, spleen, mesenteric lymph node (MLN), small intestine, cecum and feces were collected from mice to analyze the features of induced immune responses. Results The colonization of Hp strains in the stomachs of the immunized mice was significantly decreased as compared with that of the control group. Increased specific IgG antibody responses which were predominantly of IgG1 subtype were detected in the serum samples of the immunized mice and the IgG1/IgG2a ratio was significantly higher than that of the control group. Elevated secretory IgA (sIgA) was detected in the samples of small intestine, cecum and feces in the immunization group, especially in the small intestine samples, while no significant change in sIgA secretion was observed in the control group. The percentages of IL-17+ CD4+ T cells in spleen and mesenteric lymph nodes of the immunization group were significantly higher than those of the control group. Conclusions Oral immunization with Hp lysates in combination with adjuvant dmLT induced mucosal and systemic immune responses and enhanced the resistance to Hp colonization in BALB/c mice, which was associated with the significantly increased Th17 immune responses and Th2 polarization. This study provided reference for further evaluation of dmLT as a mucosal adjuvant in the development of recombinant protein vaccines against Hp infection. Key words: Sydney strain 1 lysate; Double mutant heat-labile toxin (dmLT); BALB/c mouse; Immune response; Helicobacter pylori vaccine

Extracellular overproduction of E7 oncoprotein of Iranian human papillomavirus type 16 by genetically engineered Lactococcus lactis

BackgroundWe aimed at constructing Lactococcus lactis strains expressing HPV-16 recombinant E7 (rE7) oncoprotein and examining its overproduction ability followed by optimizing batch and fed-batch fermentations. Thereafter, in order to assess the immunogenicity of recombinant L. lactis cells, C57BL/6 mice were immunized by oral gavage.ResultsThe results suggested that recombinant strains harboring optiE7 and E7 genes produced a maximum of 4.84 and 1.91 μg/mL of rE7 in static flask experiments, while the corresponding strains gave a maximum yield of 35.49 and 14.24 μg/mL in batch experiments, respectively. Fed-batch study indicated that the concentration of rE7 protein significantly increased after feeding yeast extract plus GM17 medium. The rE7 production of the best performing strains was 2.09- and 1.48-fold higher than that of the strains during the batch fermentation. Furthermore, biomass levels were 1.98- and 1.92-fold higher than those in batch cultivation. Oral immunization of C57BL/6 mice with recombinant L. lactis produced significant specific IgG and IgA antibody responses in serum and vaginal fluids, respectively. Our outcomes suggest that vaccination with L. lactis expressing rE7 can generate significant protective effects against E7-expressing cell line. Also, our study provides evidence that the presence of large amounts of E7-specific CD4+ T helper and CD8+ T cell precursors was stimulated. Significantly higher frequencies of HPV-16 E7 specific IL-2- and IFN-γ-secreting T cells were detected in antigen-stimulated splenocytes and intestinal mucosal lymphocytes, when compared to the control groups.ConclusionsWe conclude that optimization of culture conditions along with recombinant protein expression can highly stimulate both specific humoral and cell-mediated immune responses in mice after oral immunization. These promising results represent a step towards fast-tracking a vaccine against HPV-16-associated cervical cancer.

Oral Immunization against PEDV with Recombinant Lactobacillus casei Expressing Dendritic Cell-Targeting Peptide Fusing COE Protein of PEDV in Piglets.

Porcine epidemic diarrhea (PED) is a highly contagious disease in newborn piglets. In our previous study, a genetically engineered Lactobacillus casei oral vaccine (pPG-COE-DCpep/L393) expressing a dendritic cell (DC)-targeting peptide fused with porcine epidemic diarrhea virus (PEDV) COE antigen was developed. This vaccine induced significant levels of anti-PEDV specific IgG and IgA antibody responses in mice, indicating a potential strategy against PEDV infection. In this study, pPG-COE-DCpep/L393 was used for oral vaccination of newborn piglets against PEDV. We then assessed the immune responses and protection efficacy of pPG-COE-DCpep/L393. An indirect enzyme-linked immunosorbent assay (ELISA) showed that the recombinant Lactobacillus vaccine elicits a specific systemic and mucosal immune response. The T-helper cells mediated by pPG-COE-DCpep/L393 and PEDV infection display a Th1 phenotype. The histopathological results showed that pPG-COE-DCpep/L393 promotes lymphocyte proliferation and effectively protects piglets against PEDV infection. The transforming growth factor-β level indicated that the recombinant Lactobacillus vaccine plays a role in anti-inflammatory responses in mesenteric lymph nodes during PEDV infection. These results show that pPG-COE-DCpep/L393 is a potential vaccine against PEDV infection.

Transfer of Anti-Rotavirus Antibodies during Pregnancy and in Milk Following Maternal Vaccination with a Herpes Simplex Virus Type-1 Amplicon Vector.

Rotaviruses (RVs) are important enteric pathogens of newborn humans and animals, causing diarrhea and in rare cases death, especially in very young individuals. Rotavirus vaccines presently used are modified live vaccines that lack complete biological safety. Previous work from our laboratory suggested that vaccines based on in situ produced, non-infectious rotavirus-like particles (RVLPs) are efficient while being entirely safe. However, using either vaccine, active mucosal immunization cannot induce protective immunity in newborns due to their immature immune system. We therefore hypothesized that offspring from vaccinated dams are passively immunized either by transfer of maternal antibodies during pregnancy or by taking up antibodies from milk. Using a codon optimized polycistronic gene expression cassette packaged into herpesvirus particles, the simultaneous expression of the RV capsid genes led to the intracellular formation of RVLPs in various cell lines. Vaccinated dams developed a strong RV specific IgG antibody response determined in sera and milk of both mother and pups. Moreover, sera of naïve pups nursed by vaccinated dams also had RV specific antibodies suggesting a lactogenic transfer of antibodies. Although full protection of pups was not achieved in this mouse model, our observations are important for the development of improved vaccines against RV in humans as well as in various animal species.

Evaluation of Classical Swine Fever Virus (CSFV)-Specific IgA, IgG, and IgM Antibody response in Swine Vaccinated with Alphavirus Replicon Particles-Expressed Antigens

Classical swine fever virus (CSFV) E2 glycoprotein was expressed in an alphavirus based replicon particles (RP) expression system. A fluorescent microsphere immunoassay (FMIA) has been developed for the detection of CSFV E2-specific antibody in swine vaccinated with alphavirus RP. CSFV full length E2 (aa 1-376) was fragmented into several pieces and recombinant proteins were expressed in Escherichia coli. Purified proteins were conjugated to microsphere beads, the target antigens were assembled into a single multiplex, and tested against sera vaccinated with alphavirus-expressed antigens. The results reported as mean fluorescence intensity (MFI) obtained from the median value for at least 100 microspheres and the MFI values converted to positive per sample (S/P) ratio. Of the eight recombinant E2 proteins evaluated in this study, the highest MFI values were obtained for E2 (aa 1-181). CSFV E2 glycoprotein was expressed in alphavirus based replicon expression system. The results show that vaccinated animals had CSFV-specific IgA, IgG and IgM in serum and oral fluids. The MFI values for the negative serum sample showed 20-70-fold reduction compared to positive serum sample. Antibody response to CSFV antigens were IgG>IgM>IgA. The results demonstrated that the simultaneous detection of IgG, IgM and IgA antibodies could provide an improved diagnostic tool.

Resistance to Chronic Toxoplasma gondii Infection Induced by a DNA Vaccine Expressing GRA16.

Toxoplasma gondii can infect all warm-blooded animals including human beings. T. gondii dense granule protein 16 (TgGRA16) as a crucial virulence factor could modulate the host gene expression. Here, a DNA vaccine expressing TgGRA16 was constructed to explore the protective efficacy against T. gondii infection in Kunming mice. The immune responses induced by pVAX-GRA16 were also evaluated. Mice immunized with pVAX-GRA16 could elicit higher levels of specific IgG antibody and strong cellular response compared to those in controls. The DNA vaccination significantly increased the levels of cytokines (IFN-γ, IL-2, IL-4, and IL-10) and the percentages of CD4+ and CD8+ T cells in mice. After lethal challenge, mice immunized with pVAX-GRA16 (8.4 ± 0.78 days) did not show a significant longer survival time than that in controls (7.1 ± 0.30 days) (p > 0.05). However, in chronic toxoplasmosis model (administration of 10 brain cysts of PRU strain orally), numbers of tissue cysts in mice immunized with pVAX-GRA16 were significantly reduced compared to those in controls (p < 0.05) and the rate of reduction could reach 43.89%. The results indicated that the TgGRA16 would be a promising vaccine candidate for further development of effective epitope-based vaccines against chronic T. gondii infection in mice.

Commentary on 'Dissecting the hemagglutinin head and stalk specific IgG antibody response in healthcare workers following pandemic H1N1 vaccination'.

Commentary on ‘Dissecting the hemagglutinin head and stalk specific IgG antibody response in healthcare workers following pandemic H1N1 vaccination’

Arabidopsis thaliana plants expressing Rift Valley fever virus antigens: Mice exhibit systemic immune responses as the result of oral administration of the transgenic plants

The zoonotic Rift Valley fever virus affects livestock and humans in Africa and on the Arabian Peninsula. The economic impact of this pathogen due to livestock losses, as well as its relevance to public health, underscores the importance of developing effective and easily distributed vaccines. Vaccines that can be delivered orally are of particular interest.Here, we report the expression in transformed plants (Arabidopsis thaliana) of Rift Valley fever virus antigens. The antigens used in this study were the N protein and a deletion mutant of the Gn glycoprotein. Transformed lines were analysed for specific mRNA and protein content by RT-PCR and Western blotting, respectively. Furthermore, the plant-expressed antigens were evaluated for their immunogenicity in mice fed the transgenic plants. After oral intake of fresh transgenic plant material, a proportion of the mice elicited specific IgG antibody responses, as compared to the control animals that were fed wild-type plants and of which none sero-converted.Thus, we show that transgenic plants can be readily used to express and produce Rift Valley Fever virus proteins, and that the plants are immunogenic when given orally to mice. These are promising findings and provide a basis for further studies on edible plant vaccines against the Rift Valley fever virus.

DNA prime-protein boost based vaccination with a conserved region of leptospiral immunoglobulin-like A and B proteins enhances protection against leptospirosis.

Leptospirosis is a zoonotic disease caused by pathogenic spirochetes of theLeptospira genus. Vaccination with bacterins has severe limitations. Here, we evaluated the N-terminal region of the leptospiral immunoglobulin-like B protein (LigBrep) as a vaccine candidate against leptospirosis using immunisation strategies based on DNA prime-protein boost, DNA vaccine, and subunit vaccine. Upon challenge with a virulent strain ofLeptospira interrogans, the prime-boost and DNA vaccine approaches induced significant protection in hamsters, as well as a specific IgG antibody response and sterilising immunity. Although vaccination with recombinant fragment of LigBrep also produced a strong antibody response, it was not immunoprotective. These results highlight the potential of LigBrep as a candidate antigen for an effective vaccine against leptospirosis and emphasise the use of the DNA prime-protein boost as an important strategy for vaccine development.

Poor association of allergen-specific antibody, T- and B-cell responses revealed with recombinant allergens and a CFSE dilution-based assay.

BackgroundThe adaptive immunity underlying allergy comprises two components, the allergen‐specific antibody (i.e. IgE, IgG) and the T‐cell response. These two components are responsible for different disease manifestations and can be targeted by different therapeutic approaches. Here, we investigated the association of allergen‐specific antibody and T‐ as well as B‐cell responses in pollen‐allergic patients using recombinant (r) major birch pollen allergen rBet v 1 and major timothy grass pollen allergen rPhl p 5 as defined antigens.MethodsAllergen‐specific IgE and IgG antibody responses were determined by ELISA, and allergen‐specific T‐ and B‐cell responses were measured in peripheral blood mononuclear cells using a carboxyfluorescein‐diacetate‐succinimidylester (CFSE) dilution assay.ResultsCFSE staining in combination with T‐cell‐ and B‐cell‐specific gating allowed discriminating between allergen‐specific T‐cell and B‐cell responses. Interestingly, we identified patients where mainly T cells and others where mainly B cells proliferated in response to allergen stimulation. No association between the level of allergen‐specific Ig responses and B‐ or T‐cell proliferation was observed.ConclusionPurified recombinant allergens in conjunction with CFSE staining allow the dissection of allergen‐specific B‐ and T‐cell responses. The dissociation of allergen‐specific antibody, and B‐ and T‐cell responses may explain the occurrence of selective IgE‐ and T‐cell‐mediated manifestations of allergic inflammation and may be important for the development of diagnostic and therapeutic strategies selectively targeting B cells and T cells.

Recombinant Pvs48/45 antigen expressed in E. coli generates antibodies that block malaria transmission in Anopheles albimanus mosquitoes.

Transmission of malaria parasites from humans to Anopheles mosquitoes can be inhibited by specific antibodies elicited during malaria infection, which target surface Plasmodium gametocyte/gamete proteins. Some of these proteins may have potential for vaccine development. Pvs48/45 is a P. vivax gametocyte surface antigen orthologous to Pfs48/45, which may play a role during parasite fertilization and thus has potential for transmission blocking (TB) activity. Here we describe the expression of a recombinant Pvs48/45 protein expressed in Escherichia coli as a ∼60kDa construct which we tested for antigenicity using human sera and for its immunogenicity and transmission blocking activity of specific anti-mouse and anti-monkey Pvs48/45 antibodies. The protein reacted with sera of individuals from malaria-endemic areas and in addition induced specific IgG antibody responses in BALB/c mice and Aotus l. griseimembra monkeys. Sera from both immunized animal species recognized native P. vivax protein in Western blot (WB) and immunofluorescence assays. Moreover, sera from immunized mice and monkeys produced significant inhibition of parasite transmission to An. Albimanus mosquitoes as shown by membrane feeding assays. Results indicate the presence of reactive epitopes in the Pvs48/45 recombinant product that induce antibodies with TB activity. Further testing of this protein is ongoing to determine its vaccine potential.

Peak TMA Specific IgG Responses May Predict the Likelihood of TMA Exposed Workers Developing TMA Specific IgE Responses

Workplace exposure to Trimellitic Anhydride (TMA) can elicit specific IgG and IgE antibody responses leading to occupational asthma (OA). An immuno-surveillance program to identify and remove workers with TMA-specific serum IgE, who are at risk for OA, has been previously developed. This purpose of this study was to determine whether the kinetics of specific antibody responses can differentiate workers who only develop TMA specific IgG from those that progress to specific IgE.

Analysis of humoral immune response and cytokines in chickens vaccinated with Eimeria brunetti apical membrane antigen-1 (EbAMA1) DNA vaccine

This study aimed to determine the changes of cytokines, specific serum IgG and several parameters in chickens vaccinated with DNA vaccine encoding Eimeria brunetti apical membrane antigen-1 (EbAMA1) antigen. Two-week-old chickens were divided into five groups (four groups for experiment) randomly. Experimental groups of chickens were immunized with DNA vaccine while control group of chickens were injected with pVAX1 plasmid alone or TE buffer solution. All immunizations were boosted 2weeks later. The EbAMA1 specific IgG antibody responses were measured at weeks 1–6 post-second immunizations and several parameters were also identified. The result showed that the antibody titers in chickens vaccinated with DNA vaccines were significantly different from those of the control groups 1week after the second immunization and reached the maximum values 3weeks post-second immunization. IFN-γ concentration was increased the highest level against EbAMA1 of all chickens vaccinated with vaccines up to 56-fold, follow by the specific IgG antibody levels were increased 10–17-fold compared with those of TE solution and plasmid (pVAX1) control chickens 1–6weeks post-second immunization. In case of the levels of IL-10 and IL-17 was increased in experimental chickens with 4–5-fold. Even though it was statistically significant, TGF-β and IL-4 levels were higher in vaccinated than unvaccinated chickens. The results suggested that DNA vaccines encoding E. brunetti apical membrane antigen-1 (EbAMA1) could increase serum specific IgG antibody and cytokines concentration and could give protection against E. brunetti infection.

Microarrayed dog, cat, and horse allergens show weak correlation between allergen-specific IgE and IgG responses⋆

Microarrayed dog, cat, and horse allergens show weak correlation between allergen-specific IgE and IgG responses⋆