Programa de Pós-graduação em Biotecnologia e Biodiversidade Rede Pró-Centro-Oeste
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Navegando Programa de Pós-graduação em Biotecnologia e Biodiversidade Rede Pró-Centro-Oeste por Por Orientador "Kipnis, Ana Paula Junqueira"
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Item Uso de nanopartículas metálicas na vacinologia: implicações para o desenvolvimento de vacinas contra doenças infecciosas(Universidade Federal de Goiás, 2018-10-09) Marques Neto, Lázaro Moreira; Kipnis, André; http://lattes.cnpq.br/4434965360286741; Kipnis, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987; Kipnis, Ana Paula Junqueira; Guilo, Lídia Andreu; Campos, Helioswilton Sales de; Fonseca, Simone Gonçalves da; Silva, Roosevelt Alves daThe search for new adjuvants is the main goal in vaccinology. Along with this, understanding the impact of using nanoparticles as a delivery system and immunomodulator in vaccine systems directly impacts the development of new vaccines. In this work, we seek to study and elucidate the adjuvanticity of magnetic nanoparticles, as well as its immunogenicity and protection of the vaccine systems. Initially, a literature review was made seeking scientific bases that demonstrated the possibility of using metallic nanoparticles (MeNPs) as innate immune system stimulators. It was also sought to find elements in which metallic nanoparticles could aid in the generation Th1, Th17 and T CD8 type cellular response. From this review, it was verified that the magnetic nanoparticles, or with metallic ions, were able to stimulate the activation of costimulatory molecules (CD80, CD40 and CD86), to induce secretion of cytokines (IL-1, IL-6, IFN-γ and TNF-α) as well as the humoral immune response, but no work demonstrated whether these nanoparticles were able to induce cellular response. Consequently, in the second part of the study, tuberculosis was used as model to verify if a vaccine formulation with a magnetic nanoparticle of manganese ferrite combined with recombinant fusion protein would have the ability to induce a protective cellular immune response, without adding other adjuvants. The nanoparticle was coated with recombinant CMX fusion protein and BALB/c mice were vaccinated with this formulation, in protocol with three vaccinations with 21-day intervals. Subsequently, the vaccinated animals were infected with Mycobacterium tuberculosis (H37Rv) to evaluate the protection conferred by the vaccine. The results showed that the nanoparticle was able to generate cellular immune responses of Th1, Th17 and T CD8 types, depending on the route of inoculation (subcutaneous, intranasal and mixed). The most preeminent response was Tc1 which was recalled after infection was able to protect against the challenge with Mtb. In addition, there was no appearance of side effects or damage to organs of infected animals, demonstrating that the formulation is safe. Finally, the vaccine formulations with MeNPs, more specifically with manganese ferrite, demonstrate potential application in vaccinology, and may be applied in vaccine formulations to generate cellular immune response, but the route must be considered and in case of use other adjuvants it should consider the possible interaction of NP with the molecule and their ligand.Item Estudo da formação de biofilmes bacterianos em endopróteses (“stent”) e inibição da formação de biofilme por peptídeos antimicrobianos de venenos de artrópodes(Universidade Federal de Goiás, 2019-11-04) Neves, Rogério Coutinho das; Kipnis, André; http://lattes.cnpq.br/4434965360286741; Kipnis, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987; Faria, Fabrícia Paula de; Carollo, Carlos Alexandre; Bataus, Luiz Artur Mendes; Fonseca, Simone Gonçalves da; Silva Junior, Nelson Jorge daThe use of synthetic materials as temporary or permanent insertion in the body can result in infections associated with the colonization of these materials. The colonization of these materials can result in bacterial biofilms formation. Acinetobacter baumannii infections are difficult to treat due to biofilm formation and resistance to multiple drugs. Staphylococcus strains are able to form biofilm and to resist against antibiotics, in addition of being associated with endocarditis. Thus, in this thesis we hypothesize that hospital acquired bacteria were able to form biofilm in vascular stent. In this study, the biofilm formation in polystyrene plates and on coronary stents of three isolates of multidrug-resistant A. baumannii (MDR) AB 02, AB 53 and AB 72 were evaluated. The biofilm formation in coronary stents were analyzed by scanning electron microscopy. The antimicrobial peptides from wasps (Agelaia-MPI, Polydia-MPII, Polydim-I) and scorpion (Con10 and NDBP 5.8) were used to determine the minimum inhibitory concentration (MIC) and biofilm eradication (MBEC) of A. baumannii on polystyrene plates. Additionally, methicillin resistant Staphylococcus spp was also used to evaluate the ability of peptides to inhibit biofilm formation. A. baumannii MDR in contact with the vascular stent adhered to the biomaterial and initiated the formation of bacterial biofilm. The MIC of the wasp peptides against strains of A. baumannii MDR ranged from 3.12 to 6.25 μM and those of scorpion from 6.25 to 25 μM. The MBEC of Agelaia-MPI and Polybia-MPII was 6.25 μM. Con10 presented a 6.25 μM MBEC for the AB 72 strain and 12.5 μM for the AB 02 and AB 53 strains and the NBDP 5.8 inhibited at the 25 μM concentration. Agelaia-MPI inhibited the biofilm dispersion of AB 02 and AB 53 at 6.25 μM concentration and of AB 72 at 12.5 μM. Polybia-MPII inhibited at the concentration 6.25 μM. Agelaia-MPI and Polybia-MPII inhibited mature biofilms at 6.25 μM. Because Agelaia and Polybia peptides presented the best inhibitory performance they were tested against Staphylococcus ssp. Polybia-MPII and Agelaia-MPI showed MIC and MBEC of 12.5 μM. We conclude that A. baumannii forms biofilm in both plates and on vascular stents. The AMPs from wasp venoms (Agelaia-MPI and Polydia-MPII) prevent the biofilm formation of both A. baumannii and Staphylococcus epidermidis. These peptides were also able to reduce the bacterial load of biofilm-containing biomaterials. Therefore, we suggest that the Agelaia-MPI and Polybia-MPII antimicrobial peptides may be modified to coat biomaterials and prevent biofilm formation as well as for the treatment of individuals afflicted with contaminated biomaterials.