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.
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16 resultados
Resultados da Pesquisa
Item LBSapSal-vaccinated dogs exhibit increased circulating T-lymphocyte subsets (CD4+ and CD8+) as well as a reduction of parasitism after challenge with Leishmania infantum plus salivary gland of Lutzomyia longipalpis.(2014) Soares, Rodrigo Dian de Oliveira Aguiar; Roatt, Bruno Mendes; Ker, Henrique Gama; Moreira, Nádia das Dores; Mathias, Fernando Augusto Siqueira; Cardoso, Jamille Mirelle de Oliveira; Gontijo, Nelder de Figueiredo; Romero, Oscar Bruna; Carvalho, Andréa Teixeira de; Martins Filho, Olindo Assis; Oliveira, Rodrigo Corrêa de; Giunchetti, Rodolfo Cordeiro; Reis, Alexandre BarbosaBackground: The development of a protective vaccine against canine visceral leishmaniasis (CVL) is an alternative approach for interrupting the domestic cycle of Leishmania infantum. Given the importance of sand fly salivary proteins as potent immunogens obligatorily co-deposited during transmission of Leishmania parasites, their inclusion in an anti-Leishmania vaccine has been investigated in the last few decades. In this context, we previously immunized dogs with a vaccine composed of L. braziliensis antigens plus saponin as the adjuvant and sand fly salivary gland extract (LBSapSal vaccine). This vaccine elicited an increase in both anti-saliva and anti-Leishmania IgG isotypes, higher counts of specific circulating CD8+ T cells, and high NO production. Methods: We investigated the immunogenicity and protective effect of LBSapSal vaccination after intradermal challenge with 1 × 107 late-log-phase L. infantum promastigotes in the presence of sand fly saliva of Lutzomyia longipalpis. The dogs were followed for up to 885 days after challenge. Results: The LBSapSal vaccine presents extensive antigenic diversity with persistent humoral and cellular immune responses, indicating resistance against CVL is triggered by high levels of total IgG and its subtypes (IgG1 and IgG2); expansion of circulating CD5+, CD4+, and CD8+ T lymphocytes and is Leishmania-specific; and reduction of splenic parasite load. Conclusions: These results encourage further study of vaccine strategies addressing Leishmania antigens in combination with proteins present in the saliva of the vector.Item Immunological profile of resistance and susceptibility innaturally infected dogs by Leishmania infantum.(2014) Leal, Gleisiane Gomes de Almeida; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Carneiro, Cláudia Martins; Giunchetti, Rodolfo Cordeiro; Carvalho, Andréa Teixeira de; Martins Filho, Olindo Assis; Francisco, Amanda Fortes; Cardoso, Jamille Mirelle de Oliveira; Mathias, Fernando Augusto Siqueira; Oliveira, Rodrigo Corrêa de; Carneiro, Mariângela; Vital, Wendel Coura; Reis, Alexandre BarbosaVisceral leishmaniasis has a great impact on public health, and dogs are considered the maindomestic reservoir of Leishmania infantum, the causal parasite. In this study, 159 animalsnaturally infected by L. infantum from an endemic area of Brazil were evaluated through ananalysis of cellular responses, using flow cytometry, and of the hematological parameters.The results confirmed that disease progression is associated with anemia and reductionsin eosinophils, monocytes and lymphocytes. The investigation of the immune response,based on the immunophenotypic profile of peripheral blood, showed declines in the abso-lute numbers of T lymphocytes CD5+and their subsets (CD4+and CD8+) and a drop of Blymphocytes in asymptomatic seropositive (AD-II) and symptomatic seropositive (SD) dogs.Neutrophils, when stimulated with soluble antigen of L. infantum, showed higher synthesisof interferon (IFN)- _+in AD-II and SD groups, with decreased production of interleukinItem Cytokine and nitric oxide patterns in dogs immunized with LBSap vaccine, before and after experimental challenge with Leishmania chagasi plus saliva of Lutzomyia longipalpis.(2013) Resende, Lucilene Aparecida; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Viana, Kelvinson Fernandes; Mendonça, Ludmila Zanandreis de; Lanna, Mariana Ferreira; Lemos, Denise da Silveira; Oliveira, Rodrigo Corrêa de; Martins Filho, Olindo Assis; Fujiwara, Ricardo Toshio; Carneiro, Cláudia Martins; Reis, Alexandre Barbosa; Giunchetti, Rodolfo CordeiroIn the studies presented here, dogs were vaccinated against Leishmania (Leishmania) cha-gasi challenge infection using a preparation of Leishmania braziliensis promastigote proteinsand saponin as adjuvant (LBSap). Vaccination with LBSap induced a prominent type 1immune response that was characterized by increased levels of interleukin (IL-) 12 andinterferon gamma (IFN- _) production by peripheral blood mononuclear cells (PBMC) uponstimulation with soluble vaccine antigen. Importantly, results showed that this type ofresponsiveness was sustained after challenge infection; at day 90 and 885 after L. chagasichallenge infection, PBMCs from LBSap vaccinated dogs produced more IL-12, IFN- _ andconcomitant nitric oxide (NO) when stimulated with Leishmania antigens as comparedto PBMCs from respective control groups (saponin, LB- treated, or non-treated controldogs). Moreover, transforming growth factor (TGF)- _ decreased in the supernatant of SLcA-stimulated PBMCs in the LBSap group at 90 days. Bone marrow parasitological analysisrevealed decreased frequency of parasitism in the presence of vaccine antigen. It is con-cluded that vaccination of dogs with LBSap vaccine induced a long-lasting type 1 immuneresponse against L. chagasi challenge infection.Item Systemic and compartmentalized immune response in canine visceral leishmaniasis.(2009) Reis, Alexandre Barbosa; Martins Filho, Olindo Assis; Carvalho, Andréa Teixeira de; Giunchetti, Rodolfo Cordeiro; Carneiro, Cláudia Martins; Mayrink, Wilson; Tafuri, Washington Luiz; Oliveira, Rodrigo Corrêa deHuman visceral leishmaniasis (VL) and canine visceral leishmaniasis (CVL) are the most important emerging diseases with high prevalence in Latin American countries and are mainly caused by Leishmania (L.) chagasi (Syn = L. infantum). CVL has a great impact on Brazilian public health because domestic dogs are the most important VL peri-domicile reservoirs in both urban and peri-urban areas. Our findings highlight the complexity of cellular immunological events related to the natural infection from dogs by L. chagasi, additionally correlating major peripheral blood phenotypic markers with clinical status and tissues parasite density. Our main results demonstrated that lower frequency ofcirculating B cells and monocytes are important markers of severe CVL, whereas increased levels of CD8+ lymphocytes appear to be the major phenotypic feature of asymptomatic disease. Determination of the isotypes patterns during CVL demonstrated thatasymptomatic dogs and those with low parasitism are associated with an increase of IgG1, while the symptomatic dogs and those with high parasitism are associated with an increase of IgG, IgG2, IgM, IgA and IgE immunoglobulins. Pioneer findings obtained by our group showed a correlation between clinical status of CVL with degree of tissue parasite density. This data demonstrated that asymptomatic dogs presented low parasitism while symptomatic dogs are associated with high parasite load in various tissues such as skin, bone marrow and spleen. We have also investigated the association between tissue parasitism and CVL clinical forms. Regardless of clinical status, skin and spleen are the major sites of high parasite density during ongoing CVL. Furthermore, we demonstrated that bone marrow and spleen parasite density are the most reliable parasitological markers to decode the clinical status of CVL. In this article, we have reviewed some aspectsof the histopathological and immunological events occurring in natural and experimentalL. chagasi/L. infantum infection, pointing out the main L. chagasi-parasitized tissue. Wehave discussed the importance of the association between parasite density, immunological/ histopathological aspects and clinical status of the CVL, their current applications, challenges for the future and potential opportunities in CVL research.Item Cellular immunophenotypic profile in the splenic compartment during canine visceral leishmaniasis.(2014) Reis, Alexandre Barbosa; Carvalho, Andréa Teixeira de; Giunchetti, Rodolfo Cordeiro; Roatt, Bruno Mendes; Vital, Wendel Coura; Nicolato, Roney Luiz de Carvalho; Lemos, Denise da Silveira; Oliveira, Rodrigo Corrêa de; Martins Filho, Olindo AssisTo determine the role of the spleen in the pathogenesis of canine visceral leishmaniasis (CVL), we analyzed cellular immunophenotypic profiles of 52 dogs naturally infected with Leishmania infantum, clinically classified as follows: asymptomatic dogs-I (AD-I), seroneg-ative/PCR+; asymptomatic dogs-II (AD-II), seropositive/PCR+; oligosymptomatic dogs (OD) and symptomatic dogs (SD). Seven non-infected dogs (CD) were included as a control group. AD-II presented higher levels of CD8+ T splenocytes and lower TCD4+/TCD8+ ratio in com-parison with CD. OD and SD showed lower percentages of CD21+ as compared with AD-II. All seropositive dogs presented lower levels of CD45RA+ than CD. Regardless of the stimuli used, the proliferation index from splenocytes in vitro was inversely correlated with clini-cal status. After LSA stimulation, there was a higher percentage of specific CD8+ T in AD-II than CD and non-stimulated culture. In contrast, splenocytes from SD under in vitro LSA stimulation induced decreased MHC-II+ expression in comparison with all groups, and non-stimulated culture. In conclusion, the role of CD8+ T splenocytes seems to be important for an effective immunological response, a hallmark of asymptomatic CVL, whereas the pro-nounced loss of MHC-II expression upon LSA stimulation is a biomarker of symptomatic CVL.Item Isotype patterns of immunoglobulins : hallmarks for clinical status and tissue parasite density in brazilian dogs naturally infected by Leishmania (Leishmania) chagasi.(2006) Reis, Alexandre Barbosa; Carvalho, Andréa Teixeira de; Vale, André Macedo; Marques, Marcos José; Giunchetti, Rodolfo Cordeiro; Mayrink, Wilson; Guerra, Luanda Liboreiro; Andrade, Renata Aline de; Oliveira, Rodrigo Corrêa de; Martins Filho, Olindo AssisThe role of anti-leishmanial immune response underlying the susceptibility/resistance during canine visceral leishmaniasis (CVL) has been recognized throughout ex vivo and in vitro investigations. Recently, we demonstrated that immunoglobulin levels (Igs), as well as the parasite load are relevant hallmarks of distinct clinical status of CVL. To further characterize and upgrade the background on this issue, herein, we have evaluated, inLeishmania ( Leishmania ) chagasinaturally infected dogs, the relationship between tissue parasitism (skin, bone marrow, spleen, liver and lymph node), the CVL clinical status (asymptomatic (AD), with no suggestive signs of the disease; oligosymptomatic (OD), with maximum three clinical signs—opaque bristles; localized alopecia and moderate loss of weight; symptomatic (SD), serologically positive with severe clinical signs of visceral leishmaniasis), and the humoral immunological profile of anti-Leishmania immunoglobulins (IgG, IgG1, IgG2, IgM, IgA and IgE). Our major statistically significant findings revealed distinct patterns of tissue parasite density within L. chagasi-infected dogs despite their clinical status, pointing out the spleen and skin as the most relevant sites of high parasitism during ongoing CVL. Parasite density of bone marrow and spleen were the most reliable parasitological markers to decode the clinical status of CVL. Moreover, the parasite density of bone marrow better correlates with most anti- Leishmania Igs reactivity. Additionally, a prognostic hallmark for canine visceral leishmaniasis was found, highlighting strong correlation between IgG1 and asymptomatic disease, but with IgA, IgE and IgG2 displaying better association with symptomatic disease. The new aspects of this study highlighted pioneer findings that correlated the degree of tissue parasite density (low (LP), medium (MP) and high (HP) parasitism) with distinct patterns of anti- Leishmania Igs reactivity. In this scope, our data re-enforce the anti- Leishmania IgG but with IgA reactivity as the better marker for overall tissue parasitism. The association between clinical status, Ig profile and the tissue parasitism support a novel investigation on the impact of humoral immune response and susceptibility/resistance mechanism during ongoing CVLItem Parasite density and impaired biochemical/hematological status are associated with severe clinical aspects of canine visceral leishmaniasis.(2006) Reis, Alexandre Barbosa; Martins Filho, Olindo Assis; Carvalho, Andréa Teixeira de; Carvalho, Maria das Graças; Mayrink, Wilson; Silva, João Carlos França da; Giunchetti, Rodolfo Cordeiro; Genaro, Odair; Oliveira, Rodrigo Corrêa deWe have performed a detailed investigation in 40 dogs naturally infected withLeishmania infantum ( syn. chagasi), subdivided into three groups: asymptomatic (AD = 12), oligosymptomatic (OD = 12) and symptomatic (SD = 16), based on their clinical features. Twenty non-infected dogs (CD) were included as control group. Serological analysis, performed by IFAT and ELISA, demonstrated higher antibodies titers in SD in comparison to the AD. A positive correlation was found between parasite density in the spleen and skin smears as well as the bone marrow parasitism with clinical status of the infection. We observed that the progression of the disease from asymptomatic to symptomatic clinical form was accompanied by intense parasitism in the bone marrow. It is likely that this led to the impaired biochemical/hematological status observed. Finally, we believe that the follow-up of these parameters could be a relevant approach to be used as markers during therapeutic and vaccine evaluations.Item Immunotherapy against experimental canine visceral leishmaniasis with the saponin enriched-Leishmune ® vaccine.(2007) Santos, Fernanda Nunes; Borja Cabrera, Gulnara Patricia; Myashiro, L. M.; Grechi, Juliana; Reis, Alexandre Barbosa; Moreira, Márcio Antônio Batistela; Martins Filho, Olindo Assis; Luvizotto, Maria Cecília Rui; Menz, Ingrid; Pessôa, L. M.; Gonçalves, Pablo Rodrigues; Palatnik, Marcos; Souza, Clarisa Beatriz Palatnik deIn order to assess the immunotherapeutic potential on canine visceral leishmaniasis of the Leishmune ® vaccine, formulated with an increased adjuvant concentration (1 mg of saponin rather than 0.5 mg), 24 mongrel dogs were infected with Leishmania (L.) chagasi . The enriched-Leishmune ® vaccine was injected on month 6, 7 and 8 after infection, when animals were seropositive and symptomatic. The control group were injected with a saline solution. Leishmune ® -treated dogs showed significantly higher levels of anti-FML IgG antibodies (ANOVA; p < 0.0001), a higher and stable IgG2 and a decreasing IgG1 response, pointing to a TH1 T cell mediated response. The vaccine had the following effects: it led to more positive delayed type hypersensitivity reactions against Leishmania lysate in vaccinated dogs (75%) than in controls (50%), to a decreased average of CD4+ Leishmania -specific lymphocytes in saline controls (32.13%) that fell outside the 95% confidence interval of the vaccinees (41.62%, CI95% 43.93–49.80) and an increased average of the clinical scores from the saline controls (17.83) that falls outside the 95% confidence interval for the Leishmune ® immunotherapy-treated dogs (15.75, CI95% 13.97–17.53). All dogs that received the vaccine were clustered, and showed lower clinical scores and normal CD4+ counts, whereas 42% of the untreated dogs showed very diminished CD4+ and higher clinical score. The increase in clinical signs of the saline treated group was correlated with an increase in anti-FML antibodies (p < 0.0001), the parasitological evidence ( p = 0.038) and a decrease inLeishmania -specific CD4+ lymphocyte proportions (p = 0.035). These results confirm the immunotherapeutic potential of the enriched-Leishmune ® vaccine. The vaccine reduced the clinical symptoms and evidence of parasite, modulating the outcome of the infection and the dog’s potential infectiosity to phlebotomines. The enriched-Leishmune ® vaccine was subjected to a safety analysis and found to be well tolerated and safeItem Immunogenicity of a killed Leishmania vaccine with saponin adjuvant in dogs.(2007) Giunchetti, Rodolfo Cordeiro; Oliveira, Rodrigo Corrêa de; Martins Filho, Olindo Assis; Carvalho, Andréa Teixeira de; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Souza, Juliana Vitoriano de; Moreira, Nádia das Dores; Malaquias, Luiz Cosme Cotta; Castro, Luciana Lisboa Mota e; Lana, Marta de; Reis, Alexandre BarbosaCellular and humoral immune responses of dogs to a candidate vaccine, composed of Leishmania braziliensis promastigote protein plus saponin as adjuvant, have been investigated as a pre-requisite to understanding the mechanisms of immunogenicity against canine visceral leishmaniasis (CVL). The candidate vaccine elicited strong antigenicity related to the increases of anti- Leishmania IgG isotypes, together with higher levels of lymphocytes, particularly of circulating CD8 + T-lymphocytes and Leishmania chagasi antigen-specific CD8 + T-lymphocytes. As indicated by the intense cell proliferation and increased nitric oxide production during in vitro stimulation by L. chagasisoluble antigens, the candidate vaccine elicited an immune activation status potentially compatible with effective control of the etiological agent of CVL.Item Clinical value of anti-Leishmania (Leishmania) chagasi IgG titers detected by flow cytometry to distinguish infected from vaccinated dogs.(2007) Andrade, Renata Aline de; Reis, Alexandre Barbosa; Gontijo, Célia Maria Ferreira; Braga, Lidiane Bento; Rocha, Roberta Dias Rodrigues; Araújo, Márcio Sobreira Silva; Vianna, Leonardo Rocha; Martins Filho, Olindo AssisLeishmune® vaccination covers a broader number of endemic areas of canine visceral leishmaniasis (CVL) and therefore the development of new serological devices able to discriminate CVL from Leishmune® vaccinees becomes an urgent need considering the post-vaccine seroconversion detected throughout conventional methodologies. Herein, we have described the establishment of a flow cytometry based methodology to detect anti-fixedL. ( L. ) chagasipromastigotes antibodies (FC-AFPA-IgG, FC-AFPA-IgG1 and FC-AFPA-IgG2) in sera samples from Leishmania ( Leishmania ) chagasiinfected dogs and Leishmune® vaccinees. The results of FC-AFPA were reported along the sera titration curve (1:128–1:524,288), as percentage-of-positive-fluorescent-parasite (PPFP). The use of PPFP = 20% as a cut-off edge to segregate negative and positive results at sera dilution 1:2048 revealed outstanding performance indexes that elect FC-AFPA-IgG and IgG2 (both detected by polyclonal FITC-labeled second step reagent) applicable to the serological diagnosis of CVL, with 100% of specificity for both IgG and IgG2 and 97 and 93% of sensitivity, respectively. Moreover, FC-AFPA-IgG, applied at sera dilution 1:2048, also appeared as a useful tool to discriminate L. chagasiinfected dogs from Leishmune® vaccinees, with 76% of specificity. Outstanding likelihood indexes further support the performance of FC-AFPA-IgG for exclusion diagnosis of CVL in Leishmune® vaccinees. Analysis of FC-AFPA-IgG at sera dilution 1:8192 revealed the most outstanding indexes, demonstrating that besides the ability of PPFP 20% to exclude the diagnosis of CVL, a PPFP values higher 80%, mostly observed for infected dogs (INF) have a minimal change to come from a non-infected animal (NI) or Leishmune®.