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Comparative Genomics-Guided Epitope Prioritization and in Silico Design of a Multi-Epitope DNA Vaccine Candidate Against Megalocytivirus pagrus 1.

Megalocytivirus pagrus 1 infection is a World Organisation for Animal Health-listed aquatic animal disease caused by a virus species comprising the RSIV, ISKNV, and TRBIV genogroups. Here, we integrated comparative genomics and immunoinformatics to prioritize a multi-epitope protein construct, pMEV, and to design a DNA vaccine candidate encoding it, with emphasis on RSIV-type infection relevant to rock bream aquaculture. Analysis of 61 complete genomes identified 28 core gene clusters, from which myristoylated membrane protein (MMP) and major capsid protein (MCP) were prioritized as source antigens for epitope screening. Four cytotoxic T-cell, five helper T-cell, and five linear B-cell epitope candidates were selected based on sequence-based screening and exploratory peptide-MHC docking. The selected epitopes were assembled with rock bream beta-defensin-3, PADRE, and peptide linkers to generate the 283-aa pMEV construct. Sequence-based physicochemical analyses indicated properties relevant to subsequent structural and expression-based evaluation, while computationally refined structural modeling identified nine putative conformational B-cell epitope regions. TLR3 docking, normal mode analysis, and a 200-ns molecular dynamics simulation characterized the structural behavior of the selected computational complex without inferring receptor activation. C-ImmSim further generated model-dependent generic humoral and helper T-cell-associated response patterns within a mammalian-based simulation framework. Finally, the pMEV coding sequence was codon-optimized and incorporated into an in silico pcDNA3.1(+)-based DNA vaccine design. Collectively, this study provides a comparative genomics-guided framework for prioritizing an experimentally testable multi-epitope DNA vaccine candidate against M. pagrus 1, while construct expression, immunogenicity, and protective efficacy remain to be evaluated experimentally.

Animals

Synthetic long peptide and DNA personalized cancer vaccines induce robust neoantigen-specific T cell responses in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) is unresponsive to standard immunotherapies despite harboring cancer neoantigens capable of eliciting T cell responses. We completed two phase 1 clinical trials (NCT03956056 and NCT03122106) evaluating safety and immunogenicity of synthetic long peptide (SLP) and DNA personalized cancer vaccines (PCVs). PCVs were administered after resection and adjuvant chemotherapy. Tumor/normal whole-exome sequencing, RNA sequencing, and pVACtools were used to identify and prioritize candidate PCV neoantigens. PCVs were well tolerated without any grade ≥3 adverse events. Neoantigen-specific responses were demonstrated by interferon-γ enzyme-linked immunospot and intracellular cytokine staining. Expanded T cell receptor clonotypes were sequenced and transduced into autologous peripheral blood mononuclear cells to confirm neoantigen specificity. When compared with a contemporaneous institutional propensity-matched cohort, PCV patients demonstrated a trend toward prolonged median overall survival (4.4 versus 3.5 years, log-rank P = 0.23). Overall, PDAC PCVs are safe and feasible and elicit polyclonal T cell responses, linking prioritized cancer neoantigens to functional antitumor immunity.

Humans

[Analysis of cultures infected by vaccinal strains of viruses for detecting in them the integrated genome of these viruses (author's transl)].

White mice of 10-12 g were immunized with one of the three virus vaccines (vaccines against poliomyelitis, measles, smallpox) at various intervals. Poliovirus type II and measles virus, Edmonston strain, were labeled in tissue culture with 3H-uridine (30/uCi/ml). Smallpox virus (rabbit strain) was labeled with 3H-thymidine (30/uCi per ml) also in tissue culture. After purification and concentration of labeled poliomyelitis and measles viruses, viral RNA was isolated by double extraction with phenol, and precipitation with alcohol to which a yeast RNA-carrier was added. Isolation of 3H-thymidine-labeled viral RNA from smallpox virus was carried out by the same method with the addition of SDS to the final concentration of 1%. From the brain and lung cells of the vaccinated animals DNA was extracted by the kinetic reassociation method and hybridized with labeled viral nucleic acids. The formation of a hybrid with DNA-containing vaccine virus was controlled by chromatography in hydroxylapatite. No integration of viral and cell nucleic acids was demonstrated in our experiments, however, it cannot be ruled out completely, because this method does not detect homologous sequences if they occur in a small number of cells tested.

Animals

Development of a recombinant goose parvovirus VP2 neutralizing epitope-containing region vaccine adjuvanted with IL-2 and FliC for enhanced immune responses and protection against challenge.

Gosling plague (GP), caused by goose parvovirus (GPV), is a highly contagious and fatal viral disease. Vaccination is essential for disease prevention; however, conventional attenuated and inactivated vaccines have several limitations. Genetically engineered vaccines based on defined antigenic regions represent a promising alternative strategy. This study aimed to identify neutralizing epitope-containing regions within the GPV VP2 protein and develop effective recombinant vaccines. The GPV VP2 protein was divided into 11 overlapping fragments, and the anchored periplasmic expression (APEx) bacterial display system combined with flow cytometry (FCM) was used for antigenic region screening. GPV VP2-specific single-domain antibodies (VHHs) were further applied to identify neutralizing epitope-containing regions. Six neutralizing epitope-containing regions were identified and linked together to construct the VP2M recombinant antigen. The VP, VP2M, interleukin-2 (IL-2), and flagellin (FliC) genes were inserted into prokaryotic and eukaryotic expression vectors to generate protein and DNA vaccines. Three-day-old goslings were randomly assigned into 15 experimental groups for immunization. Immune responses were evaluated by measuring anti-GPV antibody levels, IgG, IgM, and IgA production, IFN-γ levels, immune-related gene expression, splenocyte proliferation, neutralizing activity, and protective efficacy against GPV challenge. The results showed that vaccines containing neutralizing epitope-containing regions induced stronger immune responses than control vaccines. Vaccinated groups exhibited increased anti-GPV antibody levels, IgG, IgM, IgA production, IFN-γ levels, immune-related gene expression, and splenocyte proliferation. Following GPV challenge, VP2M-based vaccines significantly reduced viral genome copies in the bursa of Fabricius, spleen, thymus, and intestinal tissues, accompanied by decreased histopathological lesions based on semi-quantitative scoring. Furthermore, the protective efficacy exceeded 50% in vaccines without adjuvants and reached 90% in groups containing combined IL-2 and FliC adjuvants. In conclusion, this study identifies novel neutralizing epitope-containing regions within GPV VP2 and provides a potential strategy for developing safe and effective recombinant vaccines against GP infection.

GP

Antibody and cell-mediated immunity to a DNA free herpes simplex subunit vaccine.

The immunogenicity of a DNA free herpes simplex subunit vaccine was evaluated in chimpanzees and rabbits. The results clearly demonstrate that 1 injection of 3 micrograms/kg elicited antibodies as well as cell-mediated immunity in all the animals studied. These antibodies persisted for at least 6 months. Furthermore the vaccine also protected 50% of the animals against an experimental infection and reduced the rate of latent infection in nervous sensory ganglia.

Animals

In vitro DNA synthesis in lymphocytes from turkeys vaccinated with LaSota, TC, and inactivated Newcastle disease vaccines.

In vitro cultures of peripheral blood lymphocytes from turkeys vaccinated and revaccinated with Newcastle disease (ND) vaccines were stimulated to transformation when exposed to the homologous and heterologous strains of ND virus. The mitogenesis was measured by the uptake of 3H-thymidine into newly synthesized DNA. There was considerable difference in DNA synthesis by lymphocytes drawn 0, 2, 5, and 10 days after vaccination and revaccination with the three vaccines. Stimulation of DNA synthesis, evident as early as the 2nd day, was highest in lymphocytes from turkeys vaccinated or revaccinated with TCND intramuscularly. Stimulation was least in lymphocytes from turkeys vaccinated and revaccinated with LaSota vaccine by aersol. Stimulation was intermediate from an inactivated vaccine given subcutaneously. DNA synthesis was greater with the homologous than with the heterologous strains of NDV. Synthesis was even greater when the same strain was used as a viral suspension in allantoic or cell-culture fluid than the commercial vaccine. The bovine paramyxovirus (PI3) resulted in a minimum DNA synthesis or completely inhibited it. A many-fold (order of magnitude) stimulatory effect was observed when PHA was used as an antigen. The stimulation of DNA synthesis did not parallel the HI antibody response.

Animals

The occurrence of antibodies against single-stranded DNA in the sera of patients with acute and chronic leukaemia.

One hundred and seventy-five sera from thirty-three patients with acute myeloid leukaemia, forty-two patients with chronic myeloid leukaemia and twelve patients with acute lymphatic leukaemia were examined by a radioimmunological technique for the presence of antibodies against single-stranded and double-stranded DNA. The levels of single-stranded DNA binding activity was significantly higher in all three types of leukaemia compared to those of healthy controls. In contrast, none of these sera exhibited a positive reaction with double-stranded DNA. In some cases the level of serum anti-DNA antibodies increased after the decrease of the leucocyte count. The presence of anti-DNA antibodies in leukaemic patients may have some biological significance.

Antibodies, Neoplasm

Intestinal immunity and vaccine development: a WHO memorandum.

As part of the research component of the WHO Diarrhoeal Diseases Control Programme, a Scientific Working Group met in August 1978 to review recent advances in knowledge of intestinal immunity, the application of genetic techniques in enteric vaccine development, the status of currently available immunizing agents against cholera, typhoid fever, and Shigella dysentery, and the prospects for the development of new or improved vaccines against the well known and newly recognized agents such as rotavirus and enterotoxigenic Escherichia coli. In each of these areas, the Group made specific recommendations for further research that should be considered for support under the WHO research programme in the field of diarrhoeal diseases.

Animals

Study of DNA metabolism of lymph-node cells by direct lymphatic administration of tritiated thymidine.

A method for studying DNA metabolism in lymph-node cells by injecting tritiated thymidine intralymphatically is described. The administration of [3H]thymidine through a lymph vessel enabled a high concentration to be attained with only a small quantity of the precursor in close proximity to the cells. The significance of the method is that it may also be used in studies of metabolic processes in human lymph-nodes.

Animals

Tumor immunology.

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Antigens, Neoplasm

Comparative studies of different strains of BCG vaccine in mice: T-cell dependent immune responses.

Three tests have been selected to demonstrate the close relationship between the innate immunogenicity and immunopotentiating capacity of different strains of BCG vaccines grown under the same conditions. The five strains of BCG which have been most extensively studied were PASTEUR, PHIPPS, TICE, CONNAUGHT and GLAXO. The first four are alike, they have similar growth characteristics and closely resemble each other, in immunological properties-lymphoproliferative response in vivo and T-cell potentiating effect-PASTEUR BCG being the strongest. GLAXO strain is different in these respects and is shown to have atypical properties.

Animals

Effect of BCG on cytostatic activity of peritoneal macrophages from normal and tumor-bearing rats.

In tumor-bearing rats inoculated intramuscularly with 5 x 10(4) SLC cells, the cytostatic activity of peritoneal macrophages was elevated in the early stage (7 days after transplantation) and decreased in the advanced stage (21 days after transplantation). When BCG was intraperitoneally administered into normal and tumor-bearing rats, peritoneal macrophages showed higher cytostatic activity than the untreated macrophages. This elevated cytostatic activity of the marcophages obtained from BCG-treated tumor-bearing rats was maintained even at an advanced stage.

Animals

Therapeutic melanoma vaccines: Platforms, neoantigen strategies, and emerging combination immunotherapies.

Melanoma has emerged as a major focus of cancer immunotherapy research because of its highly immunogenic nature and responsiveness to immune-based treatments. Therapeutic melanoma vaccines are designed to stimulate tumor-specific immune responses through the delivery of Tumor-Associated Antigens (TAAs), Tumor-Specific Antigens (TSAs), and personalized neoantigens. This narrative review provides an overview of current melanoma vaccine strategies, including peptide-based vaccines, dendritic cell vaccines, nucleic acid-based platforms such as mRNA, DNA, and viral vector vaccines. Recent advances in vaccine engineering and tumor genomics have accelerated the development of personalized neoantigen vaccines capable of targeting mutations unique to individual tumors. In parallel, Artificial Intelligence (AI) and Machine Learning (ML) are increasingly being incorporated into neoantigen identification pipelines to improve epitope prediction and optimize vaccine design. Combination strategies involving Immune Checkpoint Inhibitors (ICIs), particularly anti-PD-1 and anti-CTLA-4 therapies, have further enhanced interest in melanoma vaccines by helping overcome tumor-induced immune suppression and augment T-cell activation. In addition to reviewing vaccine mechanisms and emerging technologies, this manuscript examines the evolving clinical trial landscape through analysis of melanoma vaccine studies registered on ClinicalTrials.gov. Although many studies have reported encouraging safety and immunogenicity findings, challenges related to tumor heterogeneity, immune evasion, biomarker selection, and manufacturing complexity continue to limit widespread clinical implementation. Ongoing advances in computational immunology, biomaterial engineering, and precision oncology are expected to further refine melanoma vaccine development and improve therapeutic efficacy. Collectively, these innovations may help establish melanoma vaccines as an increasingly important component of future personalized cancer immunotherapy strategies.

DNA vaccines