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Introduction: Hepatitis B is a disseminated liver inflammation from HBV, that causes diseases and a large number of deaths. Regarding the fact that some of the vaccinated people are non responder (NR), evaluation of immunity in vaccinated ones and identification of NR especially in high risk group is necessary.
Materials and Methods: In this descriptive study blood samples of all medical students of Borujerd Azad university at the age of 18-25 and vaccinated personnel of Borujerd Shariaty hospital were tested for Anti.HBS-Ab level by ELISA method with Radim kit (cat.KHB31). Results were analyzed according to the number of received vaccines, the duration of vaccination and demographic criteria using descriptive statistics.
Results: About 90% of samples had protective immunity and 10% were NR. 8% of immune group had more than 1000, 17.2 % between 500-1000 and 74.8 % between 10-500 miu/ml of Ab titer. About 75% of immune samples had received two vaccines. In NR group 53% had received three vaccines and 47% had two. 4% of samples were immune with the duration less than one month after vaccination which 85 % of them had two vaccines.
Conclusion: Herd immunity was 90% which is accordant to most studies. In some studies with different results the effective criteria were not differentiated. So regarding these differences, vaccinated people are recommended to evaluate their HBS.Ab level.

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Background: Brucella is a gram-negative intracellular bacterium. Since Brucella brings about health and socio-economic problems, its control is of primary importance. The common method of vaccination includes using live attenuated strains of this bacterium. This study was done to evaluate the immunogenicity of Brucella aburtus P39 gene in Balb/c mice. Materials and Methods: In this experimental study, P39 gene was amplified by polymerase chain reaction (PCR) method and after extraction, it was sub-cloned to eukaryotic expression vector pcDNA3. The intramuscular injection of the obtained plasmid to the Balb/c mice was done in three stages. After the last vaccination, immunologic tests, such as proliferative response in lymphocytes, IFN- assessment, IL-5, and determination of IgG2a and IgG1, were run. Results: The level of activation in splenic lymphocytes response was 3.6 and the measured IFN- was 3 ng/ml, whereas IL-5 was insignificant. IgG2a and IgG1 titers were 1.640 and 1.40, respectively. Conclusion: The findings of the immunological analysis show the appropriate immune response in Balb/c mice model after the injection of P39 gene containing plasmid. The immune system response was in Th1 form which decreased the number of bacteria in spleen. Therefore, P39 gene is of appropriate immunogenicity and DNA vaccination is efficient in the activation of cell immune response against this bacterium.

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Background: Noticing the sensitivity of measles virus to temperature and light, maintaining its stability is highly important in live vaccines. The aim of the study is to evaluate the stability of measles vaccine produced by AIK-C strain. Materials and Methods: In this experimental study, three lyophilized vaccine vials were incubated at 37˚C for one week and their stability was evaluated via accelerated test. In addition, reconstituted vaccines were incubated at 4˚C, 25˚C, and 37˚C for 0, 4, 8, 12, 16 hours after reconstitution and their remaining infectious virus titer was measured using CCID50 method. Half-life of the reconstituted measles vaccine was evaluated according to linear regression analysis. Results: When the reconstituted vaccine was incubated at 4˚C, 25˚C, and 37˚C, the titer loss per hour was equal to 0.05, 0.1 and 0.2 Log10 CCID50, respectively. Also, the half-life of this vaccine at these temperatures was 5.31, 2.26, and 1.36 hours, respectively. Conclusion: The loss of potency for measles vaccine produced by AIK-C strain is 0.33 Log after storage at 37°C for one week, while the reported amounts for commercial vaccines such as Mevilin-L, Attenuvax, Edmonston-Zagreb, and Rimevax are 0.7, 0.7, 1 and 0.78, respectively. Lyophilized and reconstituted vaccine containing AIK-C strain is more stable in comparison with Edmonston B, Schwartz, Biken-CAM, and Leningrad strains. The stability of the reconstituted AIK-C strain vaccine is similar to Moraten strain at 37˚C.

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Background: In recent years Influenza viruses have caused widely spread moderate to severe infection in all around the world and there is no Influenza vaccine which can protect people only with one dose injection till now. Therefore , producing a universal vaccine based on virus like particle (VLP) could be ideal. In this study one of the molecular structures was considered for VLP based Influenza vaccine. Materials and Methods: In this experimental study, the human influenza virus (A /New Caledonia 20/1999/ (H1N1)) was propagated in MDCK cell culture. Viral RNA was extracted using RNX-plus solution. Complementary DNA synthesis was carried out using uni-12 primer and random hexamer as specific and general primers, respectively. Neuraminidase open reading frame (1413-bp) was amplified by PCR and cloned into pBlue-script SK. Neuraminidase coding frame sub cloned into pFastBac11 plasmid through SalI/XhoI sites. After verification of cloned Neuraminidase by restriction analysis, it was subjected to automated sequencing bi-directionally. The recombinant pFastBac Neuraminidase vector was transformed to E.coli DH10Bac cells which harbor bacmid DNA and helper plasmid to create Neuraminidase recombinant bacmid. Results: Neuraminidase recombinant bacmid was created by homologous recombination between pFastBacNA and bacmid and was verified by PCR using Neuraminidase specific and M13 universal primers. Conclusion: Recombinant baculovirus expressing Neuraminidase gene can be also used with other individual recombinant baculoviruses expressing HA and M1 genes in production of influenza VLPs or proteins resulting from this structure could be purified in specific insects for vaccine research studies.

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Background: Today, the use of nano-materials is one of the most common methods of making modern medications these materials are very useful in increasing the accessibility of drugs to target. The aim of this study is to obtain immunogenic nano-vaccine against meningitis caused by Haemophilus influenza.

Materials and Methods: In this experimental study, the PRP (polyribosylribitol phosphate) antigen of Haemophilus influenza was conjugated to keyhole limpet hemocyanin (KLH), a powerful immunogen molecule, and a nanoparticle with high adsorption called poly lactic co-glycolic acid (PLGA). The two-part and three-part conjugated antigens were injected into male SW1 race mice. The animals were randomly divided into five groups, which received PRP, PRP+KLH, PRP-KLH, PRP-KLH-PLGA, and PRP-TT intramuscularly together with complete Freund’s adjuvant, respectively. Twenty eight days after injection, blood samples were obtained and increases in serum antibody titer were determined with ELISA technique. For evaluation of the amount of the produced antigen cell entrance into immune cells, immune cells uptake assay and flow cytometry technique were used.

Results: The results showed increases in serum IgG antibody titers of animals immunized with conjugate vaccines. The findings also suggest the higher phagocytosis of conjugated triplex-containing nanoparticle by host immune cells.

Conclusion: The findings of this study indicate that antigen-containing PLGA has considerably higher absorption and immunogenicity and can be more powerful vaccines against Haemophilus meningitis.

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Background: Delivery of antigens directly to dendritic cells enhances the immune responses to the antigen and is an attractive approach for eliciting cellular immune responses against mutagenic pathogens like HIV virus. So the aim of this study is evaluation of immune responses elicited by delivered multi-epitopic HIV-1 tat/pol/gag/env recombinant protein to dendritic cells in sito using &alphaDEC-205 mAb.

Materials and Methods: In this study, recombinant protein expressed by pET23a-HIVtop4 plasmid including HIVtop4 sequence (Gag158-186, Pol150-190, ENV296-323, ENV577-610, Tat1-20 and Tat44-61) in BL21 E. coli cells was used as vaccine model. To exploiting dendritic cells, properties for immunization purposes, we conjugated this recombinant protein chemically to anti body against DEC-205 receptor on these cells. Balb/c mice immunized subcutaneously (s.c.) with conjugated multi-epitopic protein or un-conjugated one (as control) simultaneously with Poly I: C as dendritic cell maturation factor. Lymphocyte proliferation was measured by bromo di uridine assay, Cytotoxicity by Grenzyme B production activity, IL-4, IL-17, IFN- cytokines production and total antibody by direct and indirect ELISA methods in order.

Results: Immunization by anti DEC-205 conjugated peptide led to a significant increase in the proliferative responses of lymphocytes, production of Gr-B, IFN-&gamma, IL-4 and IL-17 cytokines and total antibody titer in comparison with the none targeted groups.

Conclusion: It is concluded that targeting of protein antigens to DEC-205+dendritic cells significantly enhances immune responses in compare to non-targeting strategies.

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Background: The outbreaks of new antigenic variants of influenza viruses in human populations have increased necessity the improvement of controlling programs. Influenza vaccines are formulated with adjuvant to enhance and direct the host immune responses. Currently, much effort is devoted to designing molecular adjutants. Hemokinin-1 (HK-1) activates T and B cells for proliferation, survival, differentiation into plasma cells, and antibody production. In this study, the effect of HK-1 as a molecular adjuvant for inducing humoral immune response against influenza virus was investigated.

Materials and Methods: The HK-1 coding sequence was cloned into pcDNA3.1 vector and used as adjuvant. Groups of mice were immunized with an inactivated influenza vaccine formulated with HK-1. The sera of vaccinated mice were collected prior to priming and boosting injections and at defined weeks, and analyzed with serological assays.

Results: The results showed that HK-1 was able to increase antibody titer against virus vaccine. The mice immunized with the adjuvanted vaccine produced higher antibody titers against influenza comparing to vaccine alone immunized group. Number of boosting had no effect on the enhancing of antibody titer.

Conclusion: These data revealed that HK-1 as a molecular adjuvant induces stronger humoral and memory responses against influenza immunization.

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Background: Hypervariability of hepatitis C virus (HCV) proteins is an important obstacle to design an efficient vaccine for the infection. To construct a protective vaccine against HCV, a DNA vaccine containing conserved epitopes of the virus was designed. To enhance the induced immune responses, adjuvant activity of N-terminal domain of gp96 (NT(gp96)) was used.

Materials and Methods: A multi-epitope (PT) DNA vaccine encoding four HCV immunodominant cytotoxic T lymphocyte epitopes (HLA-A2 and H2-D^d) from Core, E2, NS3 and NS5B antigens in addition to a T-helper CD4+ epitope from NS3 protein and a B-cell epitope from E2 protein was designed and constructed. Then, NT(gp96) was fused to the PT DNA (PT-NT(gp96)). The stimulated cellular and humoral immune responses of PT and PT-NT(gp96) were evaluated in mice model.

Results: According to multicolor flow cytometry assay, the frequency of CD8+ T-cells producing IFNγ and TNFα in the splenocytes of immunized mice with PT-NT(gp96) (6.8%, 4%) was significantly higher than those of immunized with PT (0.9% , 0.8%), respectively. The same results have obtained in hepatic lymphocytes of the vaccinated mice. The level of IgG, IgG1 and IgG2a in the mice vaccinated with PT-NT (gp96) was significantly higher than the value obtained from the mice immunized with PT.

Conclusion: The results showed that PT DNA vaccine induces immune responses in mice model. Fusion of NT (gp96) to PT DNA vaccine causes to enhance cellular and humoral immune responses against HCV compared to sole PT vaccine.

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Background: Nef is one of the HIV-1 critical proteins, because it is essential for viral replication and AIDS disease progression and induction of immune response against it can partially inhibit viral infection. Moreover, a domain of the HIV-1 Trans-Activator of Transcription (Tat, 48-60 aa) could act as a cell penetrating peptide (CPP). In current study, cloning and expression of Tat-Nef fusion protein was performed in E. coli for the first time. The protein expression was confirmed by western blot analysis and was purified using reverse staining method.

Materials and Methods: In this experimental study, primarily, cloning of Tat-Nef fusion gene was done in pGEX6p2 expression vector. Then, the expression of Tat-Nef recombinat protein in E.coli BL21 (DE3) strain was performed by using IPTG inducer. The protein expression was confirmed by SDS-PAGE and western blotting using anti-Nef monoclonal antibody. Then, the recombinant fusion protein was purified from gel using reverse staining method.

Results: The results of PCR analysis and enzyme digestion showed a clear band of ~ 726 bp in agarose gel indicating the correct Tat-Nef fusion cloning in pGEX6p2 prokaryotic expression vector. In addition, a 54 kDa band of Tat-Nef on SDS-PAGE revealed Tat-Nef protein expression that western blot analysis using anti-Nef monoclonal antibody confirmed it.

Conclusion: The purified Tat-Nef recombinant fusion protein will be used as an antigen for protein vaccine design against HIV infection.

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