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The increasing significance of the aquaculture sector and commercially valuable species underscores the need to develop alternatives for controlling diseases such as Ichthyophthirius multifiliis‐induced ichthyophthiriasis. This ciliated protozoan parasite threatens nearly all freshwater fish species, causing substantial losses in the fishery industry. Despite this, effective large‐scale treatments are lacking, emphasizing the necessity of adopting preventive strategies. While the pathogenesis of ichthyophthiriasis and its immune stimulation allows for vaccination strategies, precise adjustments are crucial to ensure the production of an effective vaccine compound. Therefore, this study aimed to evaluate the impact of immunizing Astyanax lacustris with a genetic vaccine containing IAG52A from I. multifiliis and the molecular adjuvant IL‐8 from A. lacustris. Transcript analysis in immunized A. lacustris indicated mRNA production in fish muscles, demonstrating an expression of this mRNA. Fish were divided into five groups, receiving different vaccine formulations, and all groups received a booster dose 14 days after the initial immunization. Samples from vaccinated fish showed increased IL‐1β mRNA expression in the spleen within 6 h post the second dose and after 14 days. In the head kidney, IL‐1β mRNA expression showed no significant difference at 6 and 24 h but an increase was noted in fish injected with IAG and IAG + IL‐8 after 14 days. IL‐8 mRNA expression in the spleen and kidney did not significantly differ from the control group. Histological analysis revealed no variation in leukocyte concentration at 6 and 24 h post‐vaccination; however, after 14 days, the groups injected with IAG and IAG + IL‐8 exhibited a higher leukocyte density at the application sites than the control. The obtained data suggest that the used vaccine is transcribed, indicating its potential to stimulate innate immune response parameters through mRNA cytokine expression and leukocyte migration. [ABSTRACT FROM AUTHOR]

Purpose: The development of tumor vaccines has become a hot topic in immunotherapy for osteosarcoma (OS); however, more tumor antigens with stronger immunogenicity need to be identified. Methods: We downloaded six sets of gene expression profile data from online databases. The overexpressed genes were analyzed, intersected, and used to calculate the immune infiltration abundance in the TARGET OS dataset based on their expression matrix. Potential tumor antigen genes were identified based on whether they exhibited a high correlation with the antigen-presenting cells (APCs). A total of 1330 immune-related genes (IRGs) from the ImmPort website were retrieved based on their expression, and the Consensus Cluster method was used to obtain immune subtypes of the OS samples. Prognosis, immune microenvironment, and sensitivity to drugs were compared among the immune subtypes. Results: In total, 680 genes were overexpressed in at least two datasets, of which TREM2, TNFRSF12A, and THY1 were positively correlated with different APCs. Based on the expression matrix of 1330 IRGs in TARGET-OS, two immune subtypes, IS1 and IS2, were identified. The prognosis of the IS1 subtype was better than that of IS2, the expression of immune checkpoint (ICP)-related genes was higher in patients with the IS1 subtype, and immune cell infiltration and sensitivity to 16 drugs were generally higher in IS1 subtype patients. Conclusion: We identified three APC-correlated genes that can be considered to code for potential novel tumor antigens for OS vaccines. Two immune subtypes in patients with OS were identified to implement personalized treatments using mRNA vaccines.

BACKGROUND: Due to low price, safety and effectiveness, anti caries DNA vaccine is expected to become an important means of human anti caries. Scholars are looking for an effective immune pathway to induce antibody production or a sustained-release system and adjuvant to protect vaccine titer, which has become a research focus in recent years. OBJECTIVE: To observe the immune effect of the sustained release hydrogel poly(lactic/glycolic acid)-polyethylene glycol-poly(lactic/glycolic acid) loaded with anti-caries gene vaccine pVAX1-SpaP/P. METHODS: The recombinant plasmid pVAX1-SpaP/P and the empty plasmid pVAX1 were loaded with poly(lactic/glycolic acid)-polyethylene glycol-poly(lactic/ glycolic acid) hydrogel, respectively. Twenty-eight New Zealand white rabbits were randomly divided into seven groups. Quadriceps femoris injection, submandibular gland subcutaneous injection, oral administration, and nasal drip injection of hydrogel loaded with recombinant plasmid pVAX1-SpaP/P were performed in four groups. The remaining groups were the control groups. In the negative control group 1, quadriceps muscle was injected with hydrogel loaded with empty plasmid pVAX1. In the negative control group 2, hydrogel loaded with empty plasmid pVAX1 was orally taken. The quadriceps femoris of the blank control group was injected with saline. The immunization was enhanced once a week after the initial immunization. The second immunization was conducted one week apart, and immunized for the third time after an interval of two weeks. ELISA was used to detect anti-SpaP IgA antibody in saliva and anti-SpaP IgG antibody in serum before and after immunization. At 3 days after the end of the three immunizations, the expression of pVAX1-SpaP/P protein was observed by immunohistochemistry technique. RESULTS AND CONCLUSION: (1) Two weeks after immunization with hydrogels loaded with recombinant plasmid pVAX1-SpaP/P via different pathways, the levels of anti-SpaP IgA antibody in saliva and anti-SpaP IgG antibody in serum were significantly increased in rabbits. Anti-SpaP IgG antibody level reached a peak at 10 weeks; anti-SpaP IgA antibody level reached a peak at 6 weeks. Anti-SpaP IgG antibody levels in the oral group were higher than those in the other six groups at 6-12 weeks (P < 0.05). Anti-SpaP IgA antibody levels were higher in the submandibular gland group than those in the other six groups at 4-8 weeks (P < 0.05). (2) One week after immunization with hydrogels loaded with recombinant plasmid pVAX1-SpaP/P via different pathways, anti-SpaP IgA antibody levels increased and peaked at 8 weeks. The antibody levels were higher in the submandibular gland group than those in the other six groups at 1-6 weeks (P < 0.05). The antibody levels were higher in the oral group than those in the other six groups at 8-12 weeks (P < 0.05). Anti-SpaP IgG antibody level in rabbit saliva began to increase at 2 weeks after immunization, and reached a peak at 10 weeks. The antibody levels of the oral group were higher than those in the other six groups at 2, 3, and 6 weeks (P < 0.05). The antibody levels of the quadriceps femoris group at 10 weeks were higher than those of the other six groups (P < 0.05). (3) After immunization with hydrogels loaded with recombinant plasmid pVAX1-SpaP/P via different pathways, immunohistochemical staining showed that positive pVAX1-SpaP/P protein expression in the injection site tissues. (4) The results showed that the poly(lactic/glycolic acid)-polyethylene glycolpoly( lactic/glycolic acid) hydrogel loaded pVAX1-SpaP/P anti caries vaccine could induce specific antibody production in the four ways to induce the immune effect and maintain long time. [ABSTRACT FROM AUTHOR]

Purpose: The development of tumor vaccines has become a hot topic in immunotherapy for osteosarcoma (OS); however, more tumor antigens with stronger immunogenicity need to be identified., Methods: We downloaded six sets of gene expression profile data from online databases. The overexpressed genes were analyzed, intersected, and used to calculate the immune infiltration abundance in the TARGET OS dataset based on their expression matrix. Potential tumor antigen genes were identified based on whether they exhibited a high correlation with the antigen-presenting cells (APCs). A total of 1330 immune-related genes (IRGs) from the ImmPort website were retrieved based on their expression, and the Consensus Cluster method was used to obtain immune subtypes of the OS samples. Prognosis, immune microenvironment, and sensitivity to drugs were compared among the immune subtypes., Results: In total, 680 genes were overexpressed in at least two datasets, of which TREM2, TNFRSF12A, and THY1 were positively correlated with different APCs. Based on the expression matrix of 1330 IRGs in TARGET-OS, two immune subtypes, IS1 and IS2, were identified. The prognosis of the IS1 subtype was better than that of IS2, the expression of immune checkpoint (ICP)-related genes was higher in patients with the IS1 subtype, and immune cell infiltration and sensitivity to 16 drugs were generally higher in IS1 subtype patients., Conclusion: We identified three APC-correlated genes that can be considered to code for potential novel tumor antigens for OS vaccines. Two immune subtypes in patients with OS were identified to implement personalized treatments using mRNA vaccines., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

Background: Arg-gingipain A ( rgpA ) and Arg-gingipain B ( rgpB ) are crucial virulence factors associated with Porphyromonas gingivalis ( P. gingivalis ) and have been recognized as promising targets for antibacterial vaccines. Although vaccines containing rgpA have shown efficacy, the incorporation of rgpB , which lacks the haemagglutinin adhesin ( HA ) domain, diminishes the vaccine's effectiveness. This study aims to assess the immunogenicity of the functional HA domain of rgpA in mouse periodontitis models., Methods: A total of 24 mice were randomly divided into four groups, each receiving different immune injections: group A received phosphate-buffered saline (PBS) as an empty control; group B received pVAX1 as a negative control (NC); group C received pVAX1- HA ; and group D received pVAX1- rgpA . The mice were subjected to intramuscular injections every two weeks for a total of three administrations. Prior to each immunization, blood samples were collected for antibody detection under isoflurane anesthesia. Following the final immunization, periodontitis was induced two weeks later by using sutures soaked in a P. gingivalis solution. The mice were euthanized after an additional two-week period. To assess the safety of the procedure, major organs were examined through hematoxylin-eosin (HE) staining. Subsequently, the levels of IgG, IgG1, and IgG2a in the serum were quantified via enzyme-linked immunosorbent assay (ELISA). Additionally, the expression of inflammatory factors in the gingiva, including interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor alpha (TNF-α), was determined using quantitative real-time reverse transcript PCR (qRT-PCR). The extent of bone loss in periodontal tissues was evaluated using micro-computed tomography (micro-CT) and HE staining., Results: HE staining of the organs confirmed the absence of vaccine-induced toxicity in vivo . After the second immunization, both the rgpA and HA groups displayed significantly higher specific IgG titers in comparison to the NC and PBS groups ( p < 0.05). Furthermore, the rgpA and HA groups exhibited a noteworthy predominance of IgG1 antibodies after three immunization doses, while there was a noticeable reduction in IgG2a levels observed following ligation with P. gingivalis sutures, as opposed to the NC and PBS groups ( p < 0.05). Additionally, both the HA and rgpA groups showed a significant decrease in the expression of inflammatory factors such as IL-6, IL-1β, and TNF-α, as well as a reduction in bone loss around periodontitis-affected teeth, when compared to the NC and PBS groups ( p < 0.05)., Conclusions: The results of this study demonstrate that the rgpA -engineered/functionalized HA gene vaccine is capable of eliciting a potent prophylactic immune response against P. gingivalis -induced periodontitis, effectively serving as an immunogenic and protective agent in vivo .

Objective The aim of this study was to investigate the reversibility and safety of KISS1 metastasis suppressor (KISS1) gene vaccine in immunocastration. Methods Six eight-week old ram lambs were randomly divided into vaccinated and control groups. The vaccine (1 mg/ram lamb) was injected at weeks 0, 3, and 6 of the study. Blood samples were collected from the jugular vein before primary immunization and at weeks 2, 4, 6, 10, 14, 22, and 30 after primary immunization. All ram lambs were slaughtered at 38 weeks of age, and samples were collected. Results The specific anti-KISS1 antibody titers in vaccinated animals were significantly higher and the serum testosterone level was significantly lower than those in the control groups from week 4 to 14 after primary immunization (p

[Display omitted] The concept of using mRNA to produce its own medicine in situ in the body makes it an ideal drug candidate, holding great potential to revolutionize the way we approach medicine. The unique characteristics of mRNA, as well as its customizable biomedical functions, call for the rational design of delivery systems to protect and transport mRNA molecules. In this review, a nanoparticle toolkit is presented for the development of mRNA-based therapeutics from a drug delivery perspective. Nano-delivery systems derived from either natural systems or chemical synthesis, in the nature of organic or inorganic materials, are summarised. Delivery strategies in controlling the tissue targeting and mRNA release, as well as the role of nanoparticles in building and boosting the activity of mRNA drugs, have also been introduced. In the end, our insights into the clinical and translational development of mRNA nano-drugs are presented. [ABSTRACT FROM AUTHOR]

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OBJECTIVE To observe enhancement of anti-tumor immunity by gene vaccine using nucleofection technology.METHODS The technique of nucleofection was used to transfereffectively plasmid DNA into immature dendritic cells (iDCs); we studied immune responses regulated by DNA vaccine using real-time quantitative polymerase chain reaction (PCR) and Western-blotting to optimize the follow-up lymphocyte activation. The anti-tumor capacity of lymphocytes primed by DCs was analyzed using lactate dehydrogenase with a non-radioactive cytotoxicity assay.RESULTS Human monocyte-derived dendritic cells (hMoDCs) were induced by interleukin (IL)-4 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in vitro from human monocytes for 5 or 6 days. DNA vaccine was transfected to iDCs with high transfection (35.73%) using nucleofection. Compared with the iDC group, the expression of Th1 cell cytokine IL-12, IL-18 and Th2 cell cytokine IL-4 increased after stimulation. CD86 and CD83 were upregulated compared with non-nucleofected groups 48 hours after nucleofection with DC-pVAX-PRA. The result of the cytotoxicity assay showed that DCs-pVAX-PRA primed non-adherent peripheral blood mononuclear cells (PBMCs) exhibit their highest cytotoxicity against target cells.CONCLUSION The results show that DNA vaccine was transfected to iDC with high transfection efficiency using nucleofection, priming autologous lymphocytes for anti-tumor immunity by upregulated expression of co-stimulatory molecules, adhesion molecules and cytokines. These results provided a basis to explore the molecular mechanism of DNA vaccine in vivo.

OBJECTIVE To observe anti-tumor effects of PVAX-PSMA gene vaccine. METHODS The PSMA gene was inserted into a mammalian expression vector, PVAX-1, to construct the DNA vaccine candidate, and was then used to vaccinate C57BL/6 mice. Animals vaccinated with PVAX-1 and NaCl were used as controls. Anti-PSMA antibody was detected in sera of the animals. The proliferation and cytotoxicity of the spleen cells were observed. The immunized mice were inoculated with RM-1 cells. The mice were inoculated with RM-1 cells, and then the mice were immunized. The anti-tumor efficacy of the gene vaccine was evaluated by the ratio of tumor formation, tumor volume, tumor mass before and a er gene vaccination and evaluated by survival rate of the immunized mice.RESULTS High level of anti-PSMA antibody was induced in the PVAX-PSMA group. The splenocytes from PVAX-PSMA group were stimulated to produce strong proliferation responses and significant cytotoxic T-cells (CTL) activity. After the mice were immunized with PVAX-PSMA gene, tumor occurrence was decreased, and the growth velocity of tumor was markedly reduced, resulting in prolonged tumor-free time (P < 0.05).CONCLUSION PVAX-PSMA gene vaccine has significant anti- tumor effects and provides an experimental basis for primary prevention and immunotherapy of prostate cancer.

Here we described nano-polyplexes (NPs) made of oleoyl-carboxymethy-chitosan (OCMCS)/hyaluronic acid (HA) as novel potential carriers for oral gene vaccines delivery. Aerolysin gene (aerA) of Aeromonas hydrophila as microbial antigen was efficiently loaded to form OCMCS-HA/aerA (OHA) NPs. OHA NPs performed the optimal parameters, i.e. smallest (154.5 ± 9.4 nm), positive charged (+7.9 ± 0.5 mV) and monodispersed system with the N/P ratio of 5 and OCMCS/HA weight ratio of 4. Upon the introduction of HA, OHA NPs was beneficial for the DNA release in intestinal environments in comparison to OA NPs. The mean fluorescence intensity detected in Caco-2 cells incubated with OHA NPs was about 2.5-fold higher than that of OA NPs; however, it decreased significantly in the presence of excess free HA. The OHA NPs and OA NPs decreased the transepithelial electric resistance (TEER) of Caco-2 monolayers obviously and induced increasing the apparent permeability coefficient ( P app ) of DNA by 5.45–6.09 folds compared with free DNA. Significantly higher (P < 0.05) antigen-specific antibodies were detected in serum after orally immunized with OHA NPs than that immunized with OA NPs and DNA alone in carps. These results enable the OHA NPs might resolve challenges arising from gastrointestinal damage to gene antigens, and offer an approach applicable for oral vaccination. [ABSTRACT FROM AUTHOR]

Nanoparticles are becoming promising carriers for gene delivery because of their high capacity in gene loading and low cell cytotoxicity. In this study, a chitosan-based nanoparticle encapsulated within a recombinant pcDNA3.1-dsNKG2D-IL-15 plasmid was generated. The fused dsNKG2D-IL-15 gene fragment consisted of double extracellular domains of NKG2D with IL-15 gene at downstream. The average diameter of the gene nanoparticles ranged from 200 nm to 400 nm, with mean zeta potential value of 53.8 ± 6.56 mV. The nanoparticles which were loaded with the dsNKG2D-IL-15 gene were uptaken by tumor cells with low cytotoxicity. Tumor cells pre-transfected by gene nanopartilces stimulated NK and T cells in vitro. Intramuscular injection of gene nanoparticles suppressed tumor growth and prolonged survival of tumor-bearing mice through activation of NK and CD8 + T cells. Thus, chitosan-based nanoparticle delivery of dsNKG2D-IL-15 gene vaccine can be potentially used for tumor therapy. [ABSTRACT FROM AUTHOR]

Objective MTDH/AEG-1 could act as an oncogene by regulating cellular transformation, proliferation, invasion, metastasis, and angiogenesis. This study aims to explore the mechanism by which MTDH/AEG-1 inhibits cancer growth and metastasis and enhances chemosensitivity. Methods Mouse model was established using orally immunized mice exposed to attenuated Salmonella containing vectors carrying full length MTDH/AEG-1 gene, and we were able to enhance the immune response and inhibit the growth and metastasis of prostate cancer through activation of cellular and humoral immunities and induction of CD8+ T cells. Immunohistochemistry and TUNEL assay, CD4+ and CD8+ T cell analysis by flow cytometry, HE staining, RT-PCR analysis, Western-blot analysis and quantitative polymerase chain reaction were performed. Results The MTDH/AEG-1 gene vaccine induced the anti-tumor function of cytotoxic T lymphocytes and CD8+ T cells and inhibited tumor growth and metastasis of prostate cancer. In the therapy model, the MTDH/AEG-1 gene vaccine significantly enhanced chemosensitivity to paclitaxel, inhibited tumor growth, promoted tumor cell apoptosis, and prolonged the survival time of tumor-bearing mice without any apparent side effects. Conclusions Our results demonstrated that MTDH/AEG-1-based DNA vaccines could used for the treatment of prostate cancer in terms of the inhibition of tumor growth, the lifespan of tumor-bearing animals. Combined with chemotherapy, MTDH/AEG-1-based DNA vaccines may produce highly favorable outcomes in the prevention and treatment of prostate cancer, suggesting the immune efficacy of MTDH/AEG-1-based DNA should be further analyzed in other cancers. [ABSTRACT FROM AUTHOR]

Background and Objective: The M2 gene expressing the conserved protein in influenza virus can be used to make a single-dose vaccine with permanent immunity. Material and Methods: The mice were allocated to one case group immunized with pcDNA3-M2 and two control groups with pcDNA and PBS, in three dozes with interval of two weeks. Two weeks after the last injection, Cellular immunity was analyzed by MTT lymphocyte proliferation, interferon gamma (IFN-gamma) and interleukin 4 (IL-4) ratio assays. The remaining animals were challenged with PR8 virus. Results: The production rate of IFN8 and IL4 in pcDNA - M2 group was higher than that of control groups (P >0.0001). Given the results of lymphocyte proliferation, Stimulation index (SI) in vaccinated mice was significantly higher than that of control groups (P<0.05). In comparison with mortality rate of 100% in control groups , the animals Challenged with PR8 vaccine had a 50% fatal rate implying a high protection level for this vaccine. Conclusion: The pcDNA3-M2 Vaccine can be considered as a promising vaccine against influenza infections. [ABSTRACT FROM AUTHOR]

PURPOSE: To investigate the efficiency of pcDNA3-PAc on Japanese long-eared white rabbits by intranasal immunization, and observe the appreciate gene vaccine dose in rabbit immunization. METHODS: Thirty Japanese long-eared white rabbits were randomly divided into 5 groups (6 in each group) as follows: 200, 400, 600 µg pcDNA3-PAc plasmid group; 400 µg pcDNA3 group and inactivated whole-cell vaccine positive control group. The rabbits were immunized twice, and plasmid groups and whole-cell group were coupled Freund's adjuvant with 1:1 ratio. The specific IgG and S-IgA antibodies in serum and saliva were detected with indirect ELISA method. The data was analyzed with SPSS 17.0 software package. RESULTS: ➀ The peak time of the antibodies appeared between 8-10 weeks after the first immunization; ➁ The specific antibodies IgG and S-IgA could be detected 2 weeks after immunization; ➂ The level of salivary specific S-IgA and serum specific IgG of pcDNA3-PAc were significantly higher than negative groups (P<0.05). The titer of the 200 µg was significantly lower than those of 400 µg and 600 µg group (P<0.05). CONCLUSIONS: ➀ The recombinant plasmid pcDNA3-PAc has immunogenicity, which can induce specific immune responses for 14 weeks in rabbits. ➁ The results of the present study show that 200 µg, 400 µg and 600 µg are effective immunizational dosage to 1.5 kg Japanese long-eared white rabbits. ➂ 400 µg and 600 µg pcDNA3-PAc can be considered as the optimal dosage than 200 µg at present experimental condition. [ABSTRACT FROM AUTHOR]

Induction of local (pulmonary) immunity plays a critical role in preventing dissemination of Mycobacterium tuberculosis (M. tb) during the early infection stage. To induce specific mucosal immunity, chitosan, a natural cationic polysaccharide, was employed as a mucosal gene carrier and complexed with pHSP65pep, our previously constructed multi-epitope gene vaccine, which induces splenic gamma-interferon (IFN-γ)+ T helper cell 1 responses. The resultant chitosan-pHSP65pep was administered intranasally to BALB/c mice with four doses of 50 μg DNA followed by mycobacterial challenge 4 weeks after the final immunization. It was found that the chitosan formulation significantly induced production of secretory immunoglobulin A ( P < 0.05) as determined by measuring its concentrations in lung lavage fluid and enhanced pulmonary CD4+ and CD8+IFN-γ+ T cell responses ( P < 0.001) compared with naked gene vaccine. Improved protection against Mycobacterium bovis bacillus Calmette-Guérin (BCG) challenge was consistently achieved by the chitosan-DNA formulation both as the vaccine alone or in a BCG prime-vaccine boost immunization scenario. Our study shows that mucosal delivery of gene vaccine in a chitosan formulation remarkably enhances specific SIgA concentrations and mucosal IFN-γ+ T cell response, which correlated positively with immunological protection. [ABSTRACT FROM AUTHOR]

We developed cationic liposomes containing DNA through a conventional process involving steps of (i) preformation of liposomes, (ii) extrusion, (iii) drying and rehydration and (iv) DNA complexation. Owing to its high prophylactic potentiality against tuberculosis, which had already been demonstrated in preclinical assays, we introduced modifications into the conventional process towards getting a simpler and more economical process for further scale-up. Elimination of the extrusion step, increasing the lipid concentration (from 16 to 64 mM) of the preformed liposomes and using good manufacturing practice bulk lipids (96-98% purity) instead of analytical grade purity lipids (99.9-100%) were the modifications studied. The differences in the physico-chemical properties, such as average diameter, zeta potential, melting point and morphology of the liposomes prepared through the modified process, were not as significant for the biological properties, such as DNA loading on the cationic liposomes, and effective immune response in mice after immunisation as the control liposomes prepared through the conventional process. Beneficially, the modified process increased productivity by 22% and reduced the cost of raw material by 75%. [ABSTRACT FROM AUTHOR]

PURPOSE: An anticaries DNA vaccine pEGFP-N1-Srv+ was used to immunize rats by different immune pathways. The expression of recombinant plasmid in different tissues in vivo and the specific immune response and protection effects against dental caries were observed. METHODS: 20 SD rats were divided into 4 groups, immunized with the recombinant plasmid pEGFP-N1-SrV + by submandibular gland-target injection (TSG), and intramuscular injection, respectively; then the expression of recombinant plasmid in different tissues were detected by immunohistochemistry technique. 24 SD rats were divided into 4 groups, immunized with recombinant plasmid pEGFP-N1-SrV+ of 100 µg, then boosted once after two weeks, through the same routes as above;then indirect ELISA technique was used to detect the specific antibodies. Keyes caries scores were used to evaluate the anticaries effects. The data was analyzed by using SPSS17.0 software package. RESULTS: ➀ Recombinant plasmid was positively expressed in the muscle fibers and submandibular glands. ➁ The specific salivary anti-SR IgA and serum IgG were detected, and the peak time of the antibodies level appeared 4 weeks after initial . At the 4th week, the levels of specific anti-SR antibodies were higher in the experimental group than that in the negative control group. The levels of salivary specific anti-SR IgA were significantly higher in TSG immunization group than that in the intramuscular injection group. ➂ Keyes caries scores were not significantly different between the experimental groups and negative control groups. CONCLUSIONS: The recombinant plasmid pEGFP-N1-Srv+ expressed in vivo and effectively increased specific salivary anti-SR IgA and serum IgG, and TSG immunization route significantly increased the specific salivary anti-SR IgA compared with the intramuscular immunization route; however, the recombinant plasmid pECFP-N1-Srv+ alone can not protect the rats from dental caries. [ABSTRACT FROM AUTHOR]

Active immunization holds great promise for the treatment of Alzheimer's disease but the infiltration of T-lymphocytes and associated meningoencephalitis observed in clinical trials needs to be overcome. To avoid this toxicity, previous studies have used synthetic truncated derivatives of Aβ to promote humoral immunity. In this study, we developed a novel vaccine [p(Aβ3-10)10-MT] that expresses ten repeats of Aβ3-10 with melatonin (MT) as an adjuvant, and administered it intramuscularly in three-month-old Tg-APPswe/PSEN1dE9 (Tg) mice by in vivo electroporation. The p(Aβ3-10)10-MT vaccine induced high titers of anti-Aβ antibodies, which in turn reduced Aβ deposits in the mouse brains and decreased cognitive impairment. Immunoglobulin isotyping revealed a predominantly IgG1 response, indicating a Th2 anti-inflammatory response. Ex vivo cultured splenocytes exhibited a low IFN-γ and high IL-4 response. Immunohistochemical analysis revealed that glial cell activation was also attenuated. These results indicate that p(Aβ3-10)10-MT may potentially be an effective vaccine to reduce accumulated Aβ and attenuate cognitive deficits. [ABSTRACT FROM AUTHOR]

Abstract: We report the effects of a synthetic peptide designed to act as a nuclear localization signal on the treatment of tuberculosis. The peptide contains 21 amino acid residues with the following specific domains: nuclear localization signal from SV 40T, cationic shuttle sequence, and cysteamide group at the C-terminus. The peptide was complexed with the plasmid DNAhsp65 and incorporated into cationic liposomes, forming a pseudo-ternary complex. The same cationic liposomes, composed of egg chicken l-α-phosphatidylcholine, 1,2-dioleoyl-3-trimethylammonium-propane, and 1,2-dioleoyl-3-trimethylammonium-propane (2:1:1M), were previously evaluated as a gene carrier for tuberculosis immunization protocols with DNAhsp65. The pseudo-ternary complex presented a controlled size (250nm), spherical-like shape, and various lamellae in liposomes as evaluated by transmission electron microscopy. An assay of fluorescence probe accessibility confirmed insertion of the peptide/DNA into the liposome structure. Peptide addition conferred no cytotoxicity in vitro, and similar therapeutic effects against tuberculosis were seen with four times less DNA compared with naked DNA treatment. Taken together, the results indicate that the pseudo-ternary complex is a promising gene vaccine for tuberculosis treatment. This work contributes to the development of multifunctional nanostructures in the search for strategies for in vivo DNA delivery. [Copyright &y& Elsevier]

The article focuses on the study regarding adenovirus vaccine construction containing CpG DNA gene adjuvant and 10 x Aβ3-10. It employed methods such as nasal immunization, anti-Aβ42 IgG1 induction to transgenic Alzheimer's disease (AD) mice. Results show that 10 x Aβ3-10 immunized mice showed improved memories than control mice.

Abstract: The licensed smallpox vaccine, comprised of infectious vaccinia, is no longer popular as it is associated with a variety of adverse events. Safer vaccines have been explored such as further attenuated viruses and component designs. However, these alternatives typically provide compromised breadth and strength of protection. We conducted a genome-level screening of cowpox, the ancestral poxvirus, in the broadly immune-presenting C57BL/6 mouse as an approach to discovering novel components with protective capacities. Cowpox coding sequences were synthetically built and directly assayed by genetic immunization for open-reading frames that protect against lethal pulmonary infection. Membrane and non-membrane antigens were identified that partially protect C57BL/6 mice against cowpox and vaccinia challenges without adjuvant or regimen optimization, whereas the 4-pox vaccine did not. New vaccines might be developed from productive combinations of these new and existing antigens to confer potent, broadly efficacious protection and be contraindicated for none. [Copyright &y& Elsevier]

Abstract: We present a comparative study of the physico-chemical properties, in vitro cytotoxicity and in vivo antibody production of surface-complexed DNA in EPC/DOTAP/DOPE (50/25/25% molar) liposomes and DOTAP/DOPE (50/50% molar) lipoplexes. The study aims to correlate the biological behavior and structural properties of the lipid carriers. We used DNA-hsp65, whose naked action as a gene vaccine against tuberculosis has already been demonstrated. Additionally, surface-complexed DNA-hsp65 in EPC/DOTAP/DOPE (50/25/25% molar) liposomes was effective as a single-dose tuberculosis vaccine. The results obtained showed that the EPC inclusion stabilized the DOTAP/DOPE structure, producing higher melting temperature and lower zeta potential despite a close mean hydrodynamic diameter. Resemblances in morphologies were identified in both structures, although a higher fraction of loaded DNA was not electrostatically bound in EPC/DOTAP/DOPE. EPC also induced a striking reduction in cytotoxicity, similar to naked DNA-hsp65. The proper immune response lead to a polarized antibody production of the IgG2a isotype, even for the cytotoxic DOTAP/DOPE. However, the antibody production was detected at 15 and 30 days for DOTAP/DOPE and EPC/DOTAP/DOPE, respectively. Therefore, the in vivo antibody production neither correlates with the in vitro cytotoxicity, nor with the structural stability alone. The synergistic effect of the structural stability and DNA electrostatic binding upon the surface of structures account for the immunological effects. By adjusting the composition to generate proper packing and cationic lipid/DNA interaction, we allow for the optimization of liposome formulations for required immunization or gene therapy. In a specific manner, our results contribute to studies on the tuberculosis therapy and vaccination. [Copyright &y& Elsevier]

The article presents a study which discusses the construction of DNa vaccine which are used to immunize mice in order to assess their immunogenicity with IL-15 gene.

Despite its young life, genetic immunization (GI) has seen both fame and infamy. Initially under the limelight because of its appeal as a simple method of delivering vaccines, some experiments displayed disappointing immune potency and, consequently, excitement dimmed. Newer focus on the flexibility of GI, afforded by its foundation in molecular biology, has recently rekindled activity in the field. Approached as a recombinant DNA technology, deficiencies become addressable. Unlike any other subunit vaccine modality, such as protein or carbohydrate, DNA is chemically simple, stable, consistent, easily amplified and the base material of a vast array of bioconstruction and biocontrol techniques. GI provides scientists with a simple platform for merging other disciplines, such as molecular biology, biochemistry, genetics, chemistry, informatics and microbiology, into the development of superior vaccine products. [ABSTRACT FROM AUTHOR]

Abstract: Background: HCV is a major cause of chronic liver disease, cirrhosis, hepatocellular carcinoma and death of end-stage liver disease worldwide. Therapy regimen based on IFN-α combined with ribavirin can induce a sustained virological and biochemical response in 20–60% of treated patients depending on the HCV genotype, the virus load and the age of the patients. So the development of a preventive or a therapeutic vaccine is very desirable. Methods: Thirty female BALB/c mice of 6–8 weeks old were randomly divided into five groups of six each to receive injection with experimental vaccine (pHCV-NS3, pHCV-NS3-Th1) and experimental controls (saline, pCI-neo, pCI-neo-Ii), respectively. After the fifth immunization, humoral and cellular immune responses were estimated. The therapeutic efficacy was also evaluated with BALB/c mice carried tumor cells expressing HCV-NS3 protein. Results: Specific antibodies to HCV-NS3 could be detected only in pHCV-NS3 immunized group. The antibody titers reach up to 1/1024. For CD4+ Th cell proliferation assay, only the pHCV-NS3 and pHCV-NS3-Th1 treated groups were positive according to absorbance assayed at 450nm. The absorbance of the pHCV-NS3-Th1 treated group was significant higher than that of pHCV-NS3 treated group (P <0.01, 0.002). Only the pHCV-NS3-Th1 immunized group produced detectable IFN-γ, the concentration was 33.65pg/ml. For IL-4 detection, only pHCV-NS3 immunized group produced tiny IL-4 cytokine, the concentration was 4.55pg/ml. pHCV-NS3 and pHCV-NS3-Th1 immunized mice showed significantly lower tumorigenesis rate and higher survival rate compared to experimental controls, but no significant differences were observed in our experiment between the two vaccine immunized groups. Conclusions: Minigene vaccine based on invariant chain CLIP genetic substitution might be a potential candidate for HCV therapeutic vaccine development. The results might also have some inspiring significance for the therapeutic vaccine development against other chronic infectious diseases and tumor. [Copyright &y& Elsevier]

Objective: The aim of this study was to investigate the reversibility and safety of KISS1 metastasis suppressor ( KISS1 ) gene vaccine in immunocastration., Methods: Six eight-week old ram lambs were randomly divided into vaccinated and control groups. The vaccine (1 mg/ram lamb) was injected at weeks 0, 3, and 6 of the study. Blood samples were collected from the jugular vein before primary immunization and at weeks 2, 4, 6, 10, 14, 22, and 30 after primary immunization. All ram lambs were slaughtered at 38 weeks of age, and samples were collected., Results: The specific anti- KISS1 antibody titers in vaccinated animals were significantly higher and the serum testosterone level was significantly lower than those in the control groups from week 4 to 14 after primary immunization (p<0.05). No significant difference was observed at weeks 22 and 30 after the primary immunization. Similar results were also found for scrotal circumference, testicular weight, length, breadth, and spermatogenesis in seminiferous tubules in week 30 after primary immunization. KS ( KISS1 -hepatitis B surface antigen S ) fusion fragment of KISS1 gene vaccine was not detected in host cell genomic DNA of 9 tissues of the vaccinated ram lambs by polymerase chain reaction., Conclusion: The effects of KISS1 gene vaccine in immunocastration were reversible and no integration events were recorded.

Objective: This study aims to use Arginine-gingipain A gene vaccine (pVAX1-rgpA) to immunize adult Beagle dogs and to evaluate its effect during peri-implantitis progression and development., Methods: Plasmid pVAX1-rgpA was constructed. The second and third bilateral mandible premolars of 15 adult Beagle dogs were extracted, and the implants were placed immediately. After 3 months, the animals were randomly divided into groups A, B, and C. Afterward, the animals were immunized thrice with plasmid pVAX1-rgpA, with heat-killed Porphyromonas gingivalis, or pVAX1, respectively. IgG in the serum and secretory IgA (sIgA) in saliva were quantitatively analyzed by enzyme-linked immunosorbent assay before and after 2 weeks of immunization. Peri-implantitis was induced with cotton ligatures fixed around the neck of implants. Probing depth (PD) and bleeding on probing were recorded. All animals were sacrificed after ligaturation for 6 weeks. Decalcified sections with thickness of 50 μm were prepared and dyed with methylene blue to observe the bone phenotype around implants., Results: Levels of serum IgG and sIgA in saliva were higher in groups A and B after immunization than before the process (P<0.05) and higher than those in group C (P<0.05). However, no difference was observed between groups A and B (P>0.05). At 4 and 6 weeks after ligaturation, PD of the ligatured side in group C was higher than that in groups A and B (P<0.05). On the other hand, no difference was identified between groups A and B (P>0.05). Bone loss in group A was significantly lower than that of the other groups (P<0.05). Abundant inflammatory cells and bacteria were present in the bone loss area around the implants in the three groups, as identified through hard tissue section observation. However, group C presented the most number of inflammatory cells and bacteria in the bone loss area around the implants., Conclusions: IgG and sIgA can be generated by immunity with rgpA DNA vaccine, which can significantly slow down bone loss during experimental peri-implantitis in dogs.