EFAR - Escola de Farmácia

URI permanente desta comunidadehttp://www.hml.repositorio.ufop.br/handle/123456789/451

Notícias

O curso de Farmácia em Ouro Preto foi criado em 1839, sendo a mais antiga Escola de Farmácia da América Latina.

Navegar

Resultados da Pesquisa

Agora exibindo 1 - 10 de 50

Vaccination with formulation of nanoparticles loaded with Leishmania amazonensis antigens confers protection against experimental visceral leishmaniasis in hamster.
(2023) Cabrera González, Marco Antonio; Gonçalves, Ana Alice Maia; Ottino, Jennifer; Leite, Jaqueline Costa; Resende, Lucilene Aparecida; Melo Júnior, Otoni Alves de Oliveira; Silveira, Patricia; Cardoso, Mariana Santos; Fujiwara, Ricardo Toshio; Bueno, Lilian Lacerda; Santos, Renato Lima; Carvalho, Tatiane Furtado de; Garcia, Giani Martins; Paes, Paulo Ricardo de Oliveira; Galdino, Alexsandro Sobreira; Chávez Fumagalli, Miguel Angel; Melo, Marilia Martins; Lemos, Denise da Silveira; Martins Filho, Olindo Assis; Dutra, Walderez Ornelas; Mosqueira, Vanessa Carla Furtado; Giunchetti, Rodolfo Cordeiro
Visceral leishmaniasis (VL) is a fatal disease caused by the protozoa Leishmania infantum for which dogs are the main reservoirs. A vaccine against canine visceral leishmaniasis (CVL) could be an important tool in the control of human and CVL by reducing the infection pressure of L. infantum. Despite the CVL vaccine available on the market, the Brazilian Ministry of Health did not implement the use of it in their control programs. In this sense, there is an urgent need to develop more efficient vaccines. In this study, the association between two polymeric nanoformulations, (poly (D, L-lactic) acid (PLA) polymer) loading Leishmania amazonensis antigens, was evaluated as a potential immunobiological agent against VL using golden hamsters as an experimental model. The results indicated that no significant adverse reactions were observed in animals vaccinated with LAPSmP. LAPSmP presented similar levels of total anti-Leishmania IgG as compared to LAPSmG. The LAPSmP and LAPSmG groups showed an intense reduction in liver and spleen parasitic load by qPCR. The LAPSmP and LAPSmG vaccines showed exceptional results, indicating that they may be promising candidates as a VL vaccine.
Nanoformulations with Leishmania braziliensis antigens triggered controlled parasite burden in vaccinated golden hamster (Mesocricetus auratus) against visceral leishmaniasis.
(2022) Ottino, Jennifer; Leite, Jaqueline Costa; Melo Júnior, Otoni Alves de Oliveira; Cabrera González, Marco Antonio; Carvalho, Tatiane Furtado de; Garcia, Giani Martins; Batista, Maurício Azevedo; Silveira, Patrícia; Cardoso, Mariana Santos; Bueno, Lilian Lacerda; Fujiwara, Ricardo Toshio; Santos, Renato Lima; Paes, Paulo Ricardo de Oliveira; Lemos, Denise da Silveira; Martins Filho, Olindo Assis; Galdino, Alexsandro Sobreira; Chávez Fumagalli, Miguel Angel; Dutra, Walderez Ornelas; Mosqueira, Vanessa Carla Furtado; Giunchetti, Rodolfo Cordeiro
Leishmaniasis is a widespread vector-borne disease in Brazil, with Leishmania (Leishmania) infantum as the primary etiological agent of visceral leishmaniasis (VL). Dogs are considered the main reservoir of this parasite, whose treatment in Brazil is restricted to the use of veterinary medicines, which do not promote a parasitological cure. Therefore, efficient vaccine development is the best approach to Canine Visceral Leishmaniasis (CVL) control. With this in mind, this study used hamsters (Mesocricetus auratus) as an experimental model in an anti-Leishmania preclinical vaccine trial to evaluate the safety, antigenicity, humoral response, and effects on tissue parasite load. Two novel formulations of nanoparticles made from poly(D, L-lactic) acid (PLA) polymer loading Leishmania braziliensis crude antigen (LB) exhibiting two different particle sizes were utilized: LBPSmG (570 nm) and LBPSmP (388 nm). The results showed that the nanoparticles were safe and harmless to hamsters and were antigenic with the induction in LBSap, LBPSmG, and LBPSmG groups of total anti-Leishmania IgG antibodies 30 days after challenge, which persists 200 days in LBSap and LBPSmP. At the same time, a less pronounced hepatosplenomegaly in LBSap, LBPSmG, and LBPSmP was found when compared to control groups, as well as a less pronounced inflammatory infiltrate and granuloma formation in the spleen. Furthermore, significant reductions of 84%, 81%, and 90% were observed in spleen parasite burden accessed by qPCR in the LBSap, LBPSmG, and LBPSmP groups, respectively. In this way, LBSap, LBPSmG, and LBPSmP formulations showed better results in vaccinated and L. infantum-challenged animals in further reducing parasitic load in the spleen and attenuating lesions in liver and splenic tissues. This results in safe, harmless nanoformulation vaccines with significant immunogenic and infection control potential. In addition, animals vaccinated with LBPSmP had an overall reduction in parasite burden in the spleen, indicating that a smaller nanoparticle could be more efficient in targeting antigen-presenting cells.
LBMPL vaccine therapy induces progressive organization of the spleen microarchitecture, improved Th1 adaptative immune response and control of parasitism in Leishmania infantum naturally infected dogs.
(2022) Roatt, Bruno Mendes; Cardoso, Jamille Mirelle de Oliveira; Reis, Levi Eduardo Soares; Moreira, Gabriel José Lucas; Gonçalves, Letícia Captein; Marques, Flávia de Souza; Moreira, Nádia das Dores; Vieira, Paula Melo de Abreu; Soares, Rodrigo Dian de Oliveira Aguiar; Giunchetti, Rodolfo Cordeiro; Reis, Alexandre Barbosa
The spleen plays a central role in human and canine visceral leishmaniasis, where the activation of the immune response occurs in one of the tissues where Leishmania infantum reproduces. Therefore, this organ is both a target to understand the mechanisms involved in the parasite control and a parameter for assessing the therapeutic response. In this sense, this study aimed to evaluate the main histological, immunological and parasitological aspects in the spleen of symptomatic dogs naturally infected by L. infantum treated with the therapeutic vaccine LBMPL. For this, dogs were divided into four groups: dogs uninfected and untreated (NI group); L. infantum-infected dogs that were not treated (INT group); L. infantum-infected dogs that received treatment only with monophosphoryl lipid A adjuvant (MPL group); and L. infantum-infected dogs that received treatment with the vaccine composed by L. braziliensis promastigote proteins associated with MPL adjuvant (LBMPL group). Ninety days after the therapeutics protocol, the dogs were euthanized and the spleen was collected for the proposed evaluations. Our results demonstrated a reduction of hyperplasia of red pulp and follicular area of white pulp, increased mRNA expression of IFN-γ, TNF-α, IL-12 and iNOS, and decreased IL-10 and TGF-β1, and intense reduction of splenic parasitism in dogs treated with the LBMPL vaccine. These results possibly suggest that the pro-inflammatory environment promoted the progressive organization of the splenic architecture favoring the cellular activation, with consequent parasite control. Along with previously obtained data, our results propose the LBMPL vaccine as a possible treatment strategy for canine visceral leishmaniasis (CVL).
Down regulation of IL-10 and TGF-β1 mRNA expression associated with reduced inflammatory process correlates with control of parasitism in the liver after treating L. infantum infected dogs with the LBMPL vaccine therapy.
(2022) Roatt, Bruno Mendes; Cardoso, Jamille Mirelle de Oliveira; Brito, Rory Cristiane Fortes de; Reis, Levi Eduardo Soares; Moreira, Gabriel José Lucas; Vieira, Paula Melo de Abreu; Souza, Flávia Marques de; Lima, Wanderson Geraldo de; Soares, Rodrigo Dian de Oliveira Aguiar; Giunchetti, Rodolfo Cordeiro; Reis, Alexandre Barbosa
The liver plays an important role in human and canine visceral leishmaniasis, then it is considered as target to understand the mechanisms involved in the parasite control and a parameter to assess therapeutic responses. In this sense, our study focuses on evaluating the major alterations in the liver by histological (morphometric parenchyma inflammation/semi-quantitative portal inflammation), immunohistochemical assays (parasitism), and qPCR (parasitism and cytokine gene expression) in Leishmania infantum naturally infected dogs and treated with LBMPL vaccine. Animals were divided in four groups: NI group (n = 5): uninfected and untreated dogs; INT group (n = 7): L. infantum-infected dogs and not treated; MPL group (n = 6): L. infantum-infected dogs that received only monophosphoryl lipid A adjuvant, and LBMPL group (n = 10): L. infantum-infected dogs that received treatment with the vaccine composed by L. braziliensis disrupted promastigotes associated with MPL adjuvant. Ninety days after the end of treatments, the dogs were euthanized, and the liver was collected for the proposed evaluations. Significantly lower portal inflammatory reactions, and lower parenchyma inflammation were observed in the LBMPL group compared to INT and MPL groups. iNOS mRNA expression was higher in LBMPL group and in contrast, IL-10 and TGF-β1 mRNA expression was lower in this group when compared to INT group. Immunohistochemical and qPCR analysis showed significant parasite load reduction in LBMPL group compared to INT and MPL animals. Our data suggest that in naturally Leishmania-infected dogs, LBMPL vaccine reduces the damage in the hepatic tissue, being able to attenuate the type 2 immune response. It could be associated with a marked reduction in the parasitism decreasing liver inflammation in treated dogs. Along with previously obtained data, our results suggest that LBMPL vaccine can significantly contribute to the therapy strategy for L. infantum infected dogs.
In vitro infectivity of strains isolated from dogs naturally infected with Leishmania infantum present a distinct pathogenic profile in hamsters.
(2020) Resende, Lucilene Aparecida; Soares, Rodrigo Dian de Oliveira Aguiar; Moreira, Nádia das Dores; Ferreira, Sidney de Almeida; Lanna, Mariana Ferreira; Cardoso, Jamille Mirelle de Oliveira; Mathias, Fernando Augusto Siqueira; Vital, Wendel Coura; Mariano, Reysla Maria da Silveira; Leite, Jaqueline Costa; Silveira, Patricia; Carvalho, Tatiane Furtado de; Santos, Renato Lima; Lemos, Denise da Silveira; Martins Filho, Olindo Assis; Dutra, Walderez Ornelas; Reis, Alexandre Barbosa; Giunchetti, Rodolfo Cordeiro
Visceral leishmaniasis (VL) is a severe disease caused by Leishmania infantum. Dogs are the parasite’s main reservoir, favoring its transmission in the urban environment. The analysis of L. infantum from infected dogs contributes to the identification of more virulent parasites, thereby supporting basic and applied studies such as vaccinal and therapeutic strategies. We proposed the in vitro and in vivo characterization of L. infantum strains from naturally infected dogs from a VL endemic area based on an infectivity and pathogenicity analysis. DH82 canine macrophages were infected in vitro with different strains for infectivity analysis, showing distinct infectivity profiles. The strains that showed greater and lesser infectivity using in vitro analyses (616 and 614, respectively) were used to infect hamsters for pathogenicity analysis. The group infected with strain 616 showed 100% survival while the group infected with strain 614 showed 50% after seven months of follow up. Furthermore, the 614 strain induced more noticeable clinicopathological changes and biochemical abnormalities in liver function, along with high inflammation and parasite load in the liver and spleen. We confirmed high variability of infectivity and pathogenicity in L. infantum strains from infected dogs. The results support the belief that screening for L. infantum infectivity using in vitro experiments is inadequate when it comes to selecting the most pathogenic strain.
Kinetics of phenotypic and functional changes in mouse models of sponge implants : rational selection to optimize protocols for specific biomolecules screening purposes.
(2020) Lanna, Mariana Ferreira; Resende, Lucilene Aparecida; Soares, Rodrigo Dian de Oliveira Aguiar; Miranda, Marina Barcelos de; Mendonça, Ludmila Zanandreis de; Melo Júnior, Otoni Alves de Oliveira; Mariano, Reysla Maria da Silveira; Leite, Jaqueline Costa; Silveira, Patricia; Oliveira, Rodrigo Corrêa de; Dutra, Walderez Ornelas; Reis, Alexandre Barbosa; Martins Filho, Olindo Assis; Moura, Sandra Aparecida Lima de; Lemos, Denise da Silveira; Giunchetti, Rodolfo Cordeiro
The sponge implant has been applied as an important in vivo model for the study of inflammatory processes as it induces the migration, proliferation, and accumulation of inflammatory cells, angiogenesis, and extracellular matrix deposition in its trabeculae. The characterization of immune events in sponge implants would be useful in identifying the immunological events that could support the selection of an appropriate experimental model (mouse strain) and time post-implant analysis in optimized protocols for novel applications of this model such as in biomolecules screening. Here, the changes in histological/morphometric, immunophenotypic and functional features of infiltrating leukocytes (LEU) were assessed in sponge implants for Swiss, BALB/c, and C57BL/6 mice. A gradual increase of fibrovascular stroma and a progressive decrease in LEU infiltration, mainly composed of polymorphonuclear cells with progressive shift toward mononuclear cells at late time-points were observed over time. Usually, Swiss mice presented a more prominent immune response with late mixed pattern (pro-inflammatory/anti-inflammatory: IL-2/IFN-γ/IL-4/IL-10/IL-17) of cytokine production. While BALB/c mice showed an early activation of the innate response with a controlled cytokine profile (low inflammatory potential), C57BL/6 mice presented a typical early pro-inflammatory (IL-6/TNF/IFN-γ) response with persistent neutrophilic involvement. A rational selection of the ideal time-point/mouse-lineage would avoid bias or tendentious results. Criteria such as low number of increased biomarkers, no recruitment of cytotoxic response, minor cytokine production, and lower biomarker connectivity (described as biomarker signature analysis and network analysis) guided the choice of the best time-point for each model (Day5/Swiss; Day7/BALB/c; Day6/C57BL/6) with wide application for screening purposes, such as identification of therapeutic biomolecules, selection of antigens/adjuvants, and follow-up of innate and adaptive immune response to vaccines candidates.
Phase I and II clinical trial comparing the LBSap, Leishmune®, and Leish-Tec® vaccines against canine visceral leishmaniasis.
(2020) Soares, Rodrigo Dian de Oliveira Aguiar; Roatt, Bruno Mendes; Mathias, Fernando Augusto Siqueira; Reis, Levi Eduardo Soares; Cardoso, Jamille Mirelle de Oliveira; Brito, Rory Cristiane Fortes de; Ker, Henrique Gama; Oliveira, Rodrigo Corrêa de; Giunchetti, Rodolfo Cordeiro; Reis, Alexandre Barbosa
In this study, we performed a phase I and II clinical trial in dogs to evaluate the toxicity and immunogenicity of LBSap-vaccine prototype, in comparison to Leishmune® and Leish-Tec® vaccines. Twenty-eight dogs were classified in four groups: (i) control group received 1 mL of sterile 0.9% saline solution; (ii) LBSap group received 600 µg of Leishmania braziliensis promastigotes protein and 1 mg of saponin adjuvant; (iii) Leishmune®; and (iv) Leish-Tec®. The safety and toxicity of the vaccines were measured before and after three immunizations by clinical, biochemical, and hematological parameters. The clinical examinations revealed that some dogs of LBSap and Leishmune® groups presented changes at the site of vaccination inoculum, such as nodules, mild edema, and local pain, which were transient and disappeared seventy-two hours after vaccination, but these results indicate that adverse changes caused by the immunizations are tolerable. The immunogenicity results demonstrate an increase of B lymphocytes CD21+ regarding the Leishmune® group and monocytes CD14+ concerning LBSap and Leishmune® groups. In the in vitro analyses, an increase in lymphoproliferative activity in LBSap and Leishmune® groups was observed, with an increase of antigen-specific CD4+ and CD8+ T lymphocytes in the LBSap group. A second approach of in vitro assays aimed at evaluating the percentage of antigen-specific CD4+ and CD8+ T lymphocytes producers of IFN-γ and IL-4, where an increase in both IFN-γ producing subpopulations in the LBSap group was observed, also showed an increase in IFN-γ producers in CD8+ lymphocytes in the Leish-Tec® group. Our data regarding immunogenicity indicate that the vaccination process, especially with the LBSap vaccine, generated a protective immune response compatible with L. infantum parasite control. Based on the foregoing, the LBSap vaccine would be suitable for further studies of phase III clinical trial in endemic areas with high prevalence and incidence of canine visceral leishmaniasis (VL) cases.
Effect on cellular recruitment and the innate immune response by combining saponin, monophosphoryl lipid-A and incomplete freund’s adjuvant with Leishmania (Viannia) braziliensis antigens for a vaccine formulation.
(2019) Souza, Juliana Vitoriano de; Mathias, Fernando Augusto Siqueira; Moreira, Nádia das Dores; Soares, Rodrigo Dian de Oliveira Aguiar; Vieira, Paula Melo de Abreu; Carvalho, Andréa Teixeira de; Carneiro, Cláudia Martins; Giunchetti, Rodolfo Cordeiro; Brito, Rory Cristiane Fortes de; Fujiwara, Ricardo Toshio; Roatt, Bruno Mendes; Melo, Maria Norma; Reis, Alexandre Barbosa
The poor immunogenicity displayed by some antigens has encouraged the development of strategies to improve the immune response and safety of vaccine candidates, resulting in an intense search for substances that potentiate vaccine response. Adjuvants have these properties helping vaccine candidates to induce a strong, durable, and fast immune response. In this study, we evaluated the specific immune response of adjuvants alone, Saponin (SAP), Incomplete Freund’s Adjuvant (IFA) and Monophosphoryl lipid-A SE (MPL-SE ) and in combination with total antigen of L. braziliensis (LB): LBSAP, LBIFA and LBMPL. The specific immune response induced by these compositions demonstrated that they were powerfully immunogenic, increasing cellular infiltration in the skin. Draining lymph nodes cultures showed that LBIFA and LBMPL have higher ability to increase the capacity of APCs to present antigens, with increased frequency of CD11c+ CD86+ cells. SAP, MPL, LBSAP, LBIFA and LBMPL could activate lymphocytes increasing expression of CD69 and CD25. LBSAP group was an excellent inducer of pro-inflammatory cytokines at 24 h. At 48 h, higher cytokines production was observed in IFA, LBIFA, MPL and LBMPL groups. Our data demonstrate that LBSAP and LBMPL are potential formulations to be tested in other experimental models. Also, the data obtained could expand the knowledge about immune response after sensitization and also contribute to the development of safe, immunogenic and effective vaccines.
Canine visceral leishmaniasis biomarkers and their employment in vaccines.
(2019) Giunchetti, Rodolfo Cordeiro; Silveira, Patricia; Resende, Lucilene Aparecida; Leite, Jaqueline Costa; Melo Júnior, Otoni Alves de Oliveira; Alves, Marina Luiza Rodrigues; Costa, Laís Moreira; Lair, Daniel Ferreira; Chaves, Vinícius Rossi; Soares, Ingrid dos Santos; Mendonça, Ludmila Zanandreis de; Lanna, Mariana Ferreira; Ribeiro, Helen Silva; Gonçalves, Ana Alice Maia; Santos, Thaiza Aline Pereira; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Souza, Juliana Vitoriano de; Moreira, Nádia das Dores; Siqueira, Fernando Augusto Mathias; Cardoso, Jamille Mirelle de Oliveira; Vital, Wendel Coura; Galdino, Alexsandro Sobreira; Viana, Kelvinson Fernandes; Martins Filho, Olindo Assis; Lemos, Denise da Silveira; Dutra, Walderez Ornelas; Reis, Alexandre Barbosa
The natural history of canine visceral leishmaniasis (CVL) has been well described, particularly with respect to the parasite load in different tissues and immunopathological changes according to the progression of clinical forms. The biomarkers evaluated in these studies provide support for the improvement of the tools used in developing vaccines against CVL. Thus, we describe the major studies using the dog model that supplies the rationale for including different biomarkers (tissue parasitism, histopathology, hematological changes, leucocytes immunophenotyping, cytokines patterns, and in vitro co-culture systems using purified T-cells subsets and macrophages infected with L. infantum) for immunogenicity and protection evaluations in phases I and II applied to pre-clinical and clinical vaccine trials against CVL. The search for biomarkers related to resistance or susceptibility has revealed a mixed cytokine profile with a prominent proinflammatory immune response as relevant for Leishmania replication at low levels as observed in asymptomatic dogs (highlighted by high levels of IFN-γ and TNF-α and decreased levels in IL-4, TGF-β and IL-10). Furthermore, increased levels in CD4+ and CD8+ T-cell subsets, presenting intracytoplasmic proinflammatory cytokine balance, have been associated with a resistance profile against CVL. In contrast, a polyclonal B-cell expansion towards plasma cell differentiation contributes to high antibody production, which is the hallmark of symptomatic dogs associated with high susceptibility in CVL. Finally, the different studies used to analyze biomarkers have been incorporated into vaccine immunogenicity and protection evaluations. Those biomarkers identified as resistance or susceptibility markers in CVL have been used to evaluate the vaccine performance against L. infantum in a kennel trial conducted before the field trial in an area known to be endemic for visceral leishmaniasis. This rationale has been a guiding force in the testing and selection of the best vaccine candidates against CVL and provides a way for the veterinary industry to register commercial immunobiological products.
Neutrophil properties in healthy and Leishmania infantum-naturally infected dogs.
(2019) Wardini, Amanda Brito; Silva, Lucia Helena Pinto da; Nadaes, Natalia Rocha; Nascimento, Michelle Tanny Cunha do; Roatt, Bruno Mendes; Reis, Alexandre Barbosa; Viana, Kelvinson Fernandes; Giunchetti, Rodolfo Cordeiro; Saraiva, Elvira Maria
Visceral leishmaniasis is a chronic disease that afects humans and dogs as well. Dogs, the domestic reservoir of Leishmania, play a central role in the transmission of visceral leishmaniasis, the most severe form of this disease. Neutrophils are the most abundant leukocytes in blood and interact with the parasite after infection. Here, we evaluate the efector properties of neutrophils from healthy and naturally Leishmania infantum-infected dogs. Our results showed that the parasite induced neutrophil extracellular trap (NET) release from neutrophils in both groups. Additionally, phagocytosis and NETs contributed diferently to parasite killing by neutrophils from healthy and infected animals, and IFN-γ, IL-8, IL-4 and TNF-α production by neutrophils from both groups were diferentially modulated by the parasite. Our results contribute to a better understanding of the complex role played by neutrophils in canine visceral leishmaniasis, which may favor the development of more efective therapies.

EFAR - Escola de Farmácia

Notícias

Navegar

Filtros

Configurações

Ordenar por

Resultados por página

Resultados da Pesquisa