Vaccines-2013 - OMICS Group

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D. N. Rao, J Vaccines Vaccin 2013, 4:5http://dx.doi.org/10.4172/2157-7560.S1.0183 rd International Conference onVaccines & VaccinationJuly 29-31, 2013 Embassy Suites Las Vegas, NV, USAMultiple peptide antigen (MAP) approach for V antigen of Y. pestis as a vaccine candidateD. N. RaoAIIMS, New DelhiYersin pestis is the causative agent of the most deadly disease plague. F1 & V antigens are the vaccine candidates. We were thefirst to map the B and T cell epitopes on F1 and V antigen and when we linked B-T and studied humoral, cellular and mucosalimmune responses and in vivo protective study during challenge experiments, few B-T constructs showed complete protection ascompared to native antigens. In this study, we followed multiple antigen peptide (MAP) approach towards development of subunitvaccine in which protective epitopes were assembled on a lysine backbone. MAP was synthesized containing seven peptidesof length varying from 15-25 aa on the lysine backbone. The authenticity of MAP was verified by amino acid analysis, SDS-PAGEand immunoblot. Palmitate was coupled at the end of amino terminus. MAP was encapsulated in microspheres (polylactide/glycolide beads) and immunized via intranasal with two immunoadjuvants murabutide (MB) and CpG ODN 1826 (CpG), in twostrains of mice while humoral and mucosal immune responses were studied till day 120 and memory response was checked byimmunizing with native V antigen.Epitope specific serum, mucosal washes IgG, IgA, IgG subclasses and specific activity were measured by indirect ELISA andsandwich ELISA respectively. MAP in saline and MAP in microspheres showed similar titres with maximum on day 60 witha titre of 1,02,400 which maintained till day 90 and declined to 51,200 on day 120. On day 135 the titres increased to 1,02,400showing good memory response after challenge with native V antigen. Out of, MAP +MB and MAP +CpG, and MAP +MB+CpGformulations MAP in CpG showed maximum peak titres 4,05,600 on day 60. The MAP +MB and MAP +CpG showed similarpeak titres with maximum 2,04,800 on day 60. The specific activity levels, which indicate local antibody production, correlatedwell with the antibody levels. When IgG subclasses were estimated, the MAP in saline and MAP +MB showed higher levels ofIgG1 indicating Th1 directing immune response whereas MAP in microspheres, and MAP+CpG+MB showed a mixed IgG1 andIgG2a/2b indicating a mixed Th1/Th2 directing immune response.In the mucosal washes, MAP+CpG showed maximum IgG peak titres of 6,400 and 3,200 while MAP +MB and MAP+MB+CpG showed 3,200 and 1,600 on day 60 in intestinal washes and lung washes respectively and the IgA peak titres forMAP+CpG were maximum with 3,200 and 1,600 while MAP +MB and MAP +MB+CpG showed 1,600 and 800 in intestinalwashes and lung washes respectively. Secretory component in mucosal washes was also detected showing the presence of SIgA.This study revealed that MAP is highly immunogenic with high and long lasting antibody titres in serum as well as in mucosalwashes with good recall response and this approach can be used for sub unit vaccine development for plague. Currently, we areperforming in vivo protective study.dnrao311@rediffmail.comJ Vaccines Vaccin 2013ISSN: 2157-7560, JVV an open access journalVaccines-2013July 29-31, 2013Volume 4 Issue 5Page 132
Ghaffarifar F et al., J Vaccines Vaccin 2013, 4:5http://dx.doi.org/10.4172/2157-7560.S1.0183 rd International Conference onVaccines & VaccinationJuly 29-31, 2013 Embassy Suites Las Vegas, NV, USAEvaluating DNA vaccine encoding GRA5 antigen of Toxoplasma gondii in BALB/c miceGhaffarifar F 1 , Naserifar R 1 , Dalimi A 1 and Sharifi Z 21Tarbiat Modares University, Iran2High Institute for Research and Education in Transfusion Medicine, IranBackground: Severe or lethal damages of toxoplasmosis clearly indicate the need for the development of a more effective vaccine.Immunization with recombinant plasmid encoding protective proteins is a promising vaccination technique. Therefore, thisstudy aimed to evaluate the immunization with plasmid encoding GRA5 antigen of Toxoplasma gondii in BALB/c mice.Materials and Methods: In this experimental study, three groups of BALB/c mice (n=10 in each group) were selected usingsimple random sampling. GRA5 gene was cloned into pcDNA3 plasmid and purified by plasmid purification kits and thenthe product was injected (IM). To determine the status of cellular and humoral immunity, the Il-4, IFN-y and IgG, IgG2a, IgGsubtypes were evaluated respectively using the ELISA-based assay.Results: The group immunized with pcGRA5 indicated a significant augmented response in humoral and cellular immunity(P≤0.05) which was confirmed by MTT test. The mean survival time for the experimental and control groups were 9 and 6 days,respectively.Conclusion: The immunized mice by pcGRA5 produce the higher titers of IFNγ indicated a Th1 response which is confirmedby the high level of IgG2a. Findings of this study demonstrate that GRA5 gene of T. gondii can be a potential vaccine candidateagainst the toxoplasmosis.Keywords: Toxoplasma gondii, DNA vaccine, GRA5 antigen, humoral immunity, cellular immunity.ghafarif@modares.ac.irJ Vaccines Vaccin 2013ISSN: 2157-7560, JVV an open access journalVaccines-2013July 29-31, 2013Volume 4 Issue 5Page 133
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