Safety and Immunogenicity of an In Vivo Muscle Electroporation Delivery System for DNA-hsp65 Tuberculosis Vaccine in Cynomolgus Monkeys.

A Bacille Calmette-Guérin (BCG) is still the only licensed vaccine for the prevention of tuberculosis, providing limited protection against Mycobacterium tuberculosis infection in adulthood. New advances in the delivery of DNA vaccines by electroporation have been made in the past decade. We evaluated the safety and immunogenicity of the DNA-hsp65 vaccine administered by intramuscular electroporation (EP) in cynomolgus macaques. Animals received three doses of DNA-hsp65 at 30-day intervals. We demonstrated that intramuscular electroporated DNA-hsp65 vaccine immunization of cynomolgus macaques was safe, and there were no vaccine-related effects on hematological, renal, or hepatic profiles, compared to the pre-vaccination parameters. No tuberculin skin test conversion nor lung X-ray alteration was identified. Further, low and transient peripheral cellular immune response and cytokine expression were observed, primarily after the third dose of the DNA-hsp65 vaccine. Electroporated DNA-hsp65 vaccination is safe but provides limited enhancement of peripheral cellular immune responses. Preclinical vaccine trials with DNA-hsp65 delivered via EP may include a combination of plasmid cytokine adjuvant and/or protein prime-boost regimen, to help the induction of a stronger cellular immune response.

Animal Welfare in Radiation Research: The Importance of Animal Monitoring System.

Long-term research into radiation exposure significantly expanded following World War II, driven by the increasing number of individuals falling ill after the detonation of two atomic bombs in Japan. Consequently, researchers intensified their efforts to investigate radiation's effects using animal models and to study disease models that emerged post-catastrophe. As a result, several parameters have been established as essential in these models, encompassing radiation doses, regimens involving single or multiple irradiations, the injection site for transplantation, and the quantity of cells to be injected. Nonetheless, researchers have observed numerous side effects in irradiated animals, prompting the development of scoring systems to monitor these animals' well-being. The aim of this review is to delve into the historical context of using animals in radiation research and explore the ethical considerations related to animal welfare, which has become an increasingly relevant topic in recent years. These concerns have prompted research groups to adopt measures aimed at reducing animal suffering. Consequently, for animal welfare, the implementation of a scoring system for clinical and behavioral monitoring is essential. This represents one of the primary challenges and hurdles in radiation studies. It is concluded that implementing standardized criteria across all institutions is aimed at ensuring result reproducibility and fostering collaboration within the scientific community.

Dietary supplementation with hybrid palm oil alters liver function in the common Marmoset.

Hybrid palm oil, which contains higher levels of oleic acid and lower saturated fatty acids in comparison with African palm oil, has been proposed to be somehow equivalent to extra virgin olive oil. However, the biological effects of its consumption are poorly described. Here we have explored the effects of its overconsumption on lipid metabolism in a non-human primate model, the common marmoset. Dietary supplementation of marmoset with hyperlipidic diet containing hybrid palm oil for 3 months did not modify plasma lipids levels, but increased glucose levels as compared to the supplementation with African palm oil. Liver volume was unexpectedly found to be more increased in marmosets consuming hybrid palm oil than in those consuming African palm oil. Hepatic total lipid content and circulating transaminases were dramatically increased in animals consuming hybrid palm oil, as well as an increased degree of fibrosis. Analysis of liver miRNAs showed a selective modulation of certain miRNAs by hybrid palm oil, some of which were predicted to target genes involved in cell adhesion molecules and peroxisomal pathways. Our data suggest that consumption of hybrid palm oil should be monitored carefully, as its overconsumption compared to that of African palm oil could involve important alterations to hepatic metabolism.

Avaliação da segurança e imunogenicidade da vacina candidata ao controle da tuberculose pVAXhsp65 administrada por eletroporação em macacos cynomolgus (Macaca fascicularis) / Evaluation of safety and immunogenicity of the vaccine candidate for tuberculosis control pVAX-hsp65 administered by electroporation in cynomolgus monkeys (macaca fascicularis)

A tuberculose é uma doença infecciosa crônica causada pelo Mycobacterium tuberculosis e foi declarada emergência em saúde pública mundial pela Organização Mundial de Saúde. A tuberculose persiste como um problema de saúde mundial, em parte, porque os indivíduos infectados, muitas vezes, não aderem ao longo tratamento de forma devida. A ampla vacinação com a BCG reduziu a ocorrência das formas mais graves de tuberculose em crianças, porém, a forma adulta pulmonar é responsável pela principal causa de morte no mundo. A validação de novas vacinas para utilização na clínica humana, passa pela necessidade de se testá-las, ainda em fase pré-clínica, em modelo animal que desenvolva e reproduza de forma semelhante a doença humana. Avaliamos, neste estudo, os aspectos referentes à segurança do método de eletroporação e a imunogenicidade da vacina pVAX-hsp65, administrada em 3 doses com intervalo de 1 mês cada, em macacos cynomolgusForam realizadas análises clínicas, hemograma, teste de função renal, hepática, além da avaliação das subpopulações celulares (TCD4, TCD8, NK, linfócitos B, células dendríticas mielóide e plasmacitóide), marcador de ativação celular (HLA-DR e CD69), grânulos citotóxicos (granzima B/perforina), citocinas (IFN-gama, TNF-alpha, IL-12, IL-10, IL-2, IL-4, IL-5 e IL-6), marcadores de proliferação (Ki-67) e ligados a apoptose (BcL-2). A vacina se mostrou segura, sem causar efeitos adversos relacionados ao local da inoculação, não induziu disfunção hepática ou renal nem alterações hematológicas. A vacinação não induziu conversão ao teste tuberculínico. Observamos um aumento de células T CD4+ de memória central, o que caracteriza ativação celular...

Tuberculosis is a chronic infectious disease caused by Mycobacteriumtuberculosis and was declared a public health emergency by World HealthOrganization. Tuberculosis remains a worldwide health problem, partly becauseinfected individuals often refuse the long- treatment. Widespread vaccinationwith BCG reduced the occurrence of severe forms of tuberculosis in children;however, pulmonary tuberculosis in adult is the main cause of deathworldwide. To validate new vaccines for clinical use in human, preclinical testsin animal model to reproduce human disease is necessary. Our mean goal wasto evaluated, the safety and immunogenicity of a new vaccine pVAX-hsp65,administrated by electroporation in cynomolgus monkeys in three doses withone month apart. Clinical analyzes were performed: Red and white blood cellscount, renal and liver functional test, evaluation of lymphocyte subsets (CD4,CD8, NK, B lymphocytes, and myeloid and plasmacytoide dendritic cells),markers for cell activation (HLA-DR and CD69), activation of cytotoxic granules(granzyme B / perforin), cytokines (IFN-gamma, TNF-alpha, IL-12, IL-10, IL-2, Il-4, IL-5and IL-6), proliferation (Ki-67) and anti-apoptosis (BCL-2) markers. The vaccineproved to be safe, with no adverse effects related to the inoculation site and didnot induce liver or kidney dysfunction or hematological changes. Thevaccination did not convert the tuberculin skin test. We observed anenhancement of central memory TCD4 lymphocytes which indicates cellactivation...

Avaliação do fenótipo e estágio de ativação celularem primatas não-humanos (Macaca fascicularise Macaca mulatta) mantidos em cativeiro como base para estudo pré-clínico de vacinas / Evaluation of phenotype and activation stage celularem nonhuman primates (Macaca fascicularise Macaca mulatta) maintained in captivity as a basis for preclinical study of vaccines

Os primatas não-humanos vêm sendo utilizados na pesquisa biomédica há muitos anos, e tem provado ser um modelo relevante, devidoao alto nível de homologia com os humanos, principalmente em mecanismos fisiológicos, hematológicos e imunológicos. Nas últimas décadas, os macacos cynomolgus (Macaca fascicularis) e macacos rhesus (Macaca mulatta), estão entre os primatas não-humanos mais utilizados como modelo experimental, para o desenvolvimento de diversas doenças, incluindo o estudo da patogênese na tuberculose, contribuindo para numerosos avanços terapêuticos e testes vacinais. Apesar da existência de fármacos eficazes para o combate à maioria dos casos de TB, a vacinação representa uma alternativa importante para o seu controle. A avaliação da imunogenicidade de uma nova vacina para a TB, em estudo pré-clínico com primatas não-humanos, será o primeiro passo para se averiguar o posterior uso em humanosPara esta finalidade, é essencial, uma seleção apurada dos animais empregados, levandoem consideração o status clínico e imunológico...