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Objective: To compare the clinical outcomes of patients infected with SARS-CoV-2 variants with patients infected with the original strain. Methods: This is a case control study comparing cases of COVID-19 patients infected with SARS-CoV-2 variants of concern identified by genomic sequencing, with a control group of 62 patients randomly selected from a COVID-19 database of patients diagnosed prior to the emergence of these variants. Findings: In the 40 patients infected with variants, the predominant (52.5%) was P.1 variant. The variant group presented more arthralgia (p=0.015), hyporexia (p=0.006), nausea/vomiting (p=0.048), and mental confusion (p=0.029), the last not identified in the control group. There were no significant differences in comorbidities or demographic data between the groups. Severe disease, according to the WHO Clinical Progression Scale, was identified only in those infected with the variants, as well as high-flow oxygen therapy and ICU admission (p=0.05). The two deaths reported in the study were in patients infected with the variants. Conclusions: The worst outcomes were observed in the group infected with SARS-CoV-2 variants, although no significant differences in comorbidities or demographics date were observed between the groups. Objetivo: Comparar los resultados clínicos de los pacientes infectados con variantes del SARS-CoV-2 con los pacientes infectados con la cepa original. Métodos: Se trata de un estudio de casos y controles que comprende casos de pacientes con COVID-19 infectados con variantes de la preocupación por SARS-CoV-2, identificados por secuenciación genómica, con un grupo control de 62 pacientes seleccionados aleatoriamente de una base de datos de pacientes con COVID-19 diagnosticados antes de la aparición de estas variantes de preocupación. Resultados: En los 40 pacientes infectados con las variantes, la forma predominante fue la variante P.1 (52,5%). El grupo infectado con variantes presentó más artralgia (p=0,015), hiporexia (p=0,006), náuseas/vómitos (p=0,048) y confusión mental (p=0,029), no identificándose esta última en el grupo control. No hubo diferencias significativas entre los grupos cuando se compararon las comorbilidades o los datos geográficos. La forma grave de la enfermedad, según la Escala de Progresión de la OMS, se identificó solo en aquellos infectados con formas variantes, así como el uso de oxigenoterapia de alto flujo y el ingreso en la UCI (p = 0,05). Las dos muertes reportadas en el estudio fueron en pacientes infectados con formas variantes del virus. Conclusiones: Los peores resultados se observaron en el grupo infectado con variantes del SARS-CoV-2, aunque no hubo diferencias significativas en las comorbilidades ni en los datos demográficos entre los grupos. Objetivo: Comparar os desfechos clínicos de pacientes infectados com variantes dos SARS-CoV-2 com pacientes infectados com a cepa original. Métodos: Este é um estudo caso controle que compara casos de pacientes com COVID-19 infectados com variantes de preocupação do SARS-CoV-2, identificadas por sequenciamento genômico, com um grupo controle de 62 pacientes selecionados aleatoriamente de um banco de dados de pacientes com COVID-19 diagnosticados previamente ao surgimento dessas variantes de preocupação. Resultados: Nos 40 pacientes infectados com as variantes, a forma predominante foi a variante P.1 (52,5%). O grupo infectado por variantes apresentou mais artralgia (p=0,015), hiporexia (p=0,006), náusea/vômito (p=0,048) e confusão mental (p=0,029), sendo esta última não identificada no grupo controle. Não houve diferenças significativas entre os grupos quando comparados comorbidades ou dados geográficos. Forma grave da doença, de acordo com a Escala de Progressão da OMS, foi identificada somente naqueles infectados com as formas variantes, assim como o uso de oxigenoterapia de alto fluxo e admissão na UTI (p=0,05). As duas mortes reportadas no estudo foram em pacientes infectados com formas variantes do vírus. Conclusões: Os piores desfechos foram observados no grupo infectado com variantes do SARS-CoV-2, apesar de não haver diferenças significativas nas comorbidades ou dados demográficos entre os grupos.

Texto completo: acesso restrito. p. 4460-4467 Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2014-08-11T13:43:08Z No. of bitstreams: 1 Nívea F. Luz.pdf: 1584833 bytes, checksum: aeb0961f7c6201b1a38868d69549901f (MD5) Rejected by Delba Rosa (delba@ufba.br), reason: Por gentileza, Cite todos os autores; O endereço eletrônico não confere. Grata on 2014-10-09T16:03:14Z (GMT) Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2014-10-14T11:18:15Z No. of bitstreams: 1 Nívea F. Luz.pdf: 1584833 bytes, checksum: aeb0961f7c6201b1a38868d69549901f (MD5) Approved for entry into archive by Delba Rosa (delba@ufba.br) on 2014-10-14T12:13:10Z (GMT) No. of bitstreams: 1 Nívea F. Luz.pdf: 1584833 bytes, checksum: aeb0961f7c6201b1a38868d69549901f (MD5) Made available in DSpace on 2014-10-14T12:13:10Z (GMT). No. of bitstreams: 1 Nívea F. Luz.pdf: 1584833 bytes, checksum: aeb0961f7c6201b1a38868d69549901f (MD5) Previous issue date: 2012 Visceral leishmaniasis (VL) remains a major public health problem worldwide. This disease is highly associated with chronic inflammation and a lack of the cellular immune responses against Leishmania. It is important to identify major factors driving the successful establishment of the Leishmania infection to develop better tools for the disease control. Heme oxygenase-1 (HO-1) is a key enzyme triggered by cellular stress, and its role in VL has not been investigated. In this study, we evaluated the role of HO-1 in the infection by Leishmania infantum chagasi, the causative agent of VL cases in Brazil. We found that L. chagasi infection or lipophosphoglycan isolated from promastigotes triggered HO-1 production by murine macrophages. Interestingly, cobalt protoporphyrin IX, an HO-1 inductor, increased the parasite burden in both mouse and human-derived macrophages. Upon L. chagasi infection, macrophages from Hmox1 knockout mice presented significantly lower parasite loads when compared with those from wild-type mice. Furthermore, upregulation of HO-1 by cobalt protoporphyrin IX diminished the production of TNF-α and reactive oxygen species by infected murine macrophages and increased Cu/Zn superoxide dismutase expression in human monocytes. Finally, patients with VL presented higher systemic concentrations of HO-1 than healthy individuals, and this increase of HO-1 was reduced after antileishmanial treatment, suggesting that HO-1 is associated with disease susceptibility. Our data argue that HO-1 has a critical role in the L. chagasi infection and is strongly associated with the inflammatory imbalance during VL. Manipulation of HO-1 pathways during VL could serve as an adjunctive therapeutic approach.

11 p. Submitted by Ana Valéria de Jesus Moura (anavaleria_131@hotmail.com) on 2012-03-05T18:37:25Z No. of bitstreams: 1 journal.pntd.0000965.pdf: 1202285 bytes, checksum: 5c2616df5399151d5b3c2dfc7bdb6a25 (MD5) Made available in DSpace on 2012-03-05T18:37:25Z (GMT). No. of bitstreams: 1 journal.pntd.0000965.pdf: 1202285 bytes, checksum: 5c2616df5399151d5b3c2dfc7bdb6a25 (MD5) Previous issue date: 2011-02 Background: Leishmania braziliensis is the main causative agent of cutaneous leishmaniasis in Brazil. Protection against infection is related to development of Th1 responses, but the mechanisms that mediate susceptibility are still poorly understood. Murine models have been the most important tools in understanding the immunopathogenesis of L. major infection and have shown that Th2 responses favor parasite survival. In contrast, L. braziliensis–infected mice develop strong Th1 responses and easily resolve the infection, thus making the study of factors affecting susceptibility to this parasite difficult. Methodology/Principal Findings: Here, we describe an experimental model for the evaluation of the mechanisms mediating susceptibility to L. braziliensis infection. BALB/c mice were inoculated with stationary phase promastigotes of L. braziliensis, isolates LTCP393(R) and LTCP15171(S), which are resistant and susceptible to antimony and nitric oxide (NO), respectively. Mice inoculated with LTCP393(R) presented larger lesions that healed more slowly and contained higher parasite loads than lesions caused by LTCP15171(S). Inflammatory infiltrates in the lesions and production of IFN-c, TNF-a, IL-10 and TGF-b were similar in mice inoculated with either isolate, indicating that these factors did not contribute to the different disease manifestations observed. In contrast, IL-4 production was strongly increased in LTCP393(R)-inoculated animals and also arginase I (Arg I) expression. Moreover, anti-IL-4 monoclonal antibody (mAb) treatment resulted in decreased lesion thickness and parasite burden in animals inoculated with LTCP393(R), but not in those inoculated with LTCP15171(S). Conclusion/Significance: We conclude that the ability of L. braziliensis isolates to induce Th2 responses affects the susceptibility to infection with these isolates and contributes to the increased virulence and severity of disease associated with them. Since these data reflect what happens in human infection, this model could be useful to study the pathogenesis of the L. braziliensis infection, as well as to design new strategies of therapeutic intervention.

BACKGROUND: Nitric oxide (NO) produced in macrophages plays a pivotal role as a leishmanicidal agent. A previous study has demonstrated that 20% of the L. (V.) braziliensis isolated from initial cutaneous lesions of patients from the endemic area of Corte de Pedra, Bahia, Brazil, were NO resistant. Additionally, 5 to 11% of the patients did not respond to three or more antimony treatments" (refractory patients). The aim of this study is to investigate if there is an association between the resistance of L. (V.) braziliensis to NO and nonresponsiveness to antimony therapy and cytokine production. METHODS: We evaluated the in vitro toxicity of NO against the promastigotes stages of L. (V.) braziliensis isolated from responsive and refractory patients, and the infectivity of the amastigote forms of these isolates against human macrophages. The supernatants from Leishmania infected macrophage were used to measure TNF-α and IL-10 levels. RESULTS: Using NaNO2 (pH 5.0) as the NO source, L. (V.) braziliensis isolated from refractory patients were more NO resistant (IC50 = 5.8 ± 4.8) than L. (V.) braziliensis isolated from responsive patients (IC50 = 2.0 ± 1.4). Four isolates were selected to infect human macrophages: NO-susceptible and NO-resistant L. (V.) braziliensis isolated from responsive and refractory patients. NO-resistant L. (V.) braziliensis isolated from refractory patients infected more macrophages stimulated with LPS and IFN-γ at 120 hours than NO-susceptible L. (V.) braziliensis isolated from refractory patients. Also, lower levels of TNF-α were detected in supernatants of macrophages infected with NO-resistant L. (V.) braziliensis as compared to macrophages infected with NO-susceptible L. (V.) braziliensis (p < 0.05 at 2, 24 and 120 hours), while no differences were detected in IL-10 levels. CONCLUSION: These data suggest that NO resistance could be related to the nonresponsiveness to antimony therapy seen in American Tegumentary Leishmaniasis.

Texto completo: acesso restrito. p. 607-613 Submitted by Suelen Reis (suziy.ellen@gmail.com) on 2013-08-27T11:24:37Z No. of bitstreams: 1 1-s2.0-S1567134810001048-main.pdf: 190164 bytes, checksum: 653f3fd37a385fc9938a5a0dd06157ac (MD5) Approved for entry into archive by Rodrigo Meirelles (rodrigomei@ufba.br) on 2013-11-16T12:54:26Z (GMT) No. of bitstreams: 1 1-s2.0-S1567134810001048-main.pdf: 190164 bytes, checksum: 653f3fd37a385fc9938a5a0dd06157ac (MD5) Made available in DSpace on 2013-11-16T12:54:26Z (GMT). No. of bitstreams: 1 1-s2.0-S1567134810001048-main.pdf: 190164 bytes, checksum: 653f3fd37a385fc9938a5a0dd06157ac (MD5) Previous issue date: 2010 Mucosal leishmaniasis (ML) follows localized cutaneous leishmaniasis (CL) caused by Leishmania braziliensis. Proinflammatory responses mediate CL self-healing but are exaggerated in ML. Proinflammatory monocyte chemoattractant protein 1 (MCP-1; encoded by CCL2) is associated with CL. We explore its role in CL/ML through analysis of the regulatory CCL2 −2518 bp promoter polymorphism in CL/ML population samples and families from Brazil. Genotype frequencies were compared among ML/CL cases and control groups using logistic regression and the family-based association test (FBAT). MCP-1 was measured in plasma and macrophages. The GG recessive genotype at CCL2 −2518 bp was more common in patients with ML (N = 67) than in neighborhood control (NC; N = 60) subjects (OR 1.78; 95% CI 1.01–3.14; P = 0.045), than in NC combined with leishmanin skin-test positive (N = 60) controls (OR 4.40; 95% CI 1.42–13.65; P = 0.010), and than in controls combined with CL (N = 60) patients (OR 2.78; 95% CI 1.13–6.85; P = 0.045). No associations were observed for CL compared to any groups. FBAT (91 ML and 223 CL cases in families) confirmed recessive association of ML with allele G (Z = 2.679; P = 0.007). Higher levels of MCP-1 occurred in plasma (P = 0.03) and macrophages (P < 0.0001) from GG compared to AA individuals. These results suggest that high MCP-1 increases risk of ML.

11 p. Submitted by Ana Valéria de Jesus Moura (anavaleria_131@hotmail.com) on 2012-01-03T18:13:34Z No. of bitstreams: 1 1471-2334-10-209.pdf: 831689 bytes, checksum: 81210b7bed68f57e3ff85e018c57b8a5 (MD5) Made available in DSpace on 2012-01-03T18:13:34Z (GMT). No. of bitstreams: 1 1471-2334-10-209.pdf: 831689 bytes, checksum: 81210b7bed68f57e3ff85e018c57b8a5 (MD5) Previous issue date: 2010 Background: Nitric oxide (NO) produced in macrophages plays a pivotal role as a leishmanicidal agent. A previous study has demonstrated that 20% of the L. (V.) braziliensis isolated from initial cutaneous lesions of patients from the endemic area of Corte de Pedra, Bahia, Brazil, were NO resistant. Additionally, 5 to 11% of the patients did not respond to three or more antimony treatments” (refractory patients). The aim of this study is to investigate if there is an association between the resistance of L. (V.) braziliensis to NO and nonresponsiveness to antimony therapy and cytokine production. Methods: We evaluated the in vitro toxicity of NO against the promastigotes stages of L. (V.) braziliensis isolated from responsive and refractory patients, and the infectivity of the amastigote forms of these isolates against human macrophages. The supernatants from Leishmania infected macrophage were used to measure TNF-a and IL-10 levels. Results: Using NaNO2 (pH 5.0) as the NO source, L. (V.) braziliensis isolated from refractory patients were more NO resistant (IC50 = 5.8 ± 4.8) than L. (V.) braziliensis isolated from responsive patients (IC50 = 2.0 ± 1.4). Four isolates were selected to infect human macrophages: NO-susceptible and NO-resistant L. (V.) braziliensis isolated from responsive and refractory patients. NO-resistant L. (V.) braziliensis isolated from refractory patients infected more macrophages stimulated with LPS and IFN-g at 120 hours than NO-susceptible L. (V.) braziliensis isolated from refractory patients. Also, lower levels of TNF-a were detected in supernatants of macrophages infected with NO-resistant L. (V.) braziliensis as compared to macrophages infected with NO-susceptible L. (V.) braziliensis (p < 0.05 at 2, 24 and 120 hours), while no differences were detected in IL-10 levels. Conclusion: These data suggest that NO resistance could be related to the nonresponsiveness to antimony therapy seen in American Tegumentary Leishmaniasis.

BACKGROUND: Nitric oxide (NO•) plays a pivotal role as a leishmanicidal agent in mouse macrophages. NO• resistant Escherichia coli and Mycobacterium tuberculosis have been associated with a severe outcome of these diseases. METHODS: In this study we evaluated the in vitro toxicity of nitric oxide for the promastigote stages of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis parasites, and the infectivity of the amastigote stage for human macrophages. Parasites were isolated from patients with cutaneous, mucosal or disseminated leishmaniasis, and NO• resistance was correlated with clinical presentation. RESULTS: Seventeen isolates of L. (L.) amazonensis or L. (V.) braziliensis promastigotes were killed by up to 8 mM of more of NaNO2 (pH 5.0) and therefore were defined as nitric oxide-susceptible. In contrast, eleven isolates that survived exposure to 16 mM NaNO2 were defined as nitric oxide-resistant. Patients infected with nitric oxide-resistant Leishmania had significantly larger lesions than patients infected with nitric oxide-susceptible isolates. Furthermore, nitric oxide-resistant L. (L.) amazonensis and L. (V.) braziliensis multiplied significantly better in human macrophages than nitric oxide-susceptible isolates. CONCLUSION: These data suggest that nitric oxide-resistance of Leishmania isolates confers a survival benefit for the parasites inside the macrophage, and possibly exacerbates the clinical course of human leishmaniasis.

Texto completo: acesso restrito. p. 478-482 Submitted by Suelen Reis (suziy.ellen@gmail.com) on 2014-01-21T11:51:20Z No. of bitstreams: 1 10.1086-520017.pdf: 284401 bytes, checksum: a82c59fcd9d227388c9c10c70bb4f53d (MD5) Rejected by Flávia Ferreira (flaviaccf@yahoo.com.br), reason: Por favor faça a inclusão dos autores de todos autores do artigo. Grata, on 2014-01-28T12:59:33Z (GMT) Submitted by Suelen Reis (suziy.ellen@gmail.com) on 2014-01-28T16:04:49Z No. of bitstreams: 1 10.1086-520017.pdf: 284401 bytes, checksum: a82c59fcd9d227388c9c10c70bb4f53d (MD5) Approved for entry into archive by Flávia Ferreira (flaviaccf@yahoo.com.br) on 2014-01-30T13:28:59Z (GMT) No. of bitstreams: 1 10.1086-520017.pdf: 284401 bytes, checksum: a82c59fcd9d227388c9c10c70bb4f53d (MD5) Made available in DSpace on 2014-01-30T13:28:59Z (GMT). No. of bitstreams: 1 10.1086-520017.pdf: 284401 bytes, checksum: a82c59fcd9d227388c9c10c70bb4f53d (MD5) Previous issue date: 2007 Cutaneous leishmaniasis affects millions of people worldwide. After observations of atypical lesions in pregnant women at the health centers of Corte de Pedra, Brazil, 9 years of records were reviewed, and 26 pregnant patients were identified. A retrospective case-control study revealed that lesions in pregnant women were much larger than those in nonpregnant patients in an age- and sex-matched group (mean area, 6.08 cm2 vs. 1.46 cm2; P = .008), and many lesions had an exophytic nature. Despite foregoing treatment until after delivery, response to pentavalent antimony therapy was favorable (rate of cure with 1 course of treatment, 85%). High rates of preterm births (10.5%) and stillbirths (10.5%) were reported. Cutaneous leishmaniasis during pregnancy produces distinct lesions and may have adverse fetal effects.

Texto completo: acesso restrito. p. 251-258 Submitted by Suelen Reis (suziy.ellen@gmail.com) on 2014-01-24T15:57:07Z No. of bitstreams: 1 j.1365-3024.2007.00940.x.pdf: 240676 bytes, checksum: fcbb7c920988e7f9f6a2ee087c48d762 (MD5) Approved for entry into archive by Rodrigo Meirelles (rodrigomei@ufba.br) on 2014-09-09T16:14:33Z (GMT) No. of bitstreams: 1 j.1365-3024.2007.00940.x.pdf: 240676 bytes, checksum: fcbb7c920988e7f9f6a2ee087c48d762 (MD5) Made available in DSpace on 2014-09-09T16:14:33Z (GMT). No. of bitstreams: 1 j.1365-3024.2007.00940.x.pdf: 240676 bytes, checksum: fcbb7c920988e7f9f6a2ee087c48d762 (MD5) Previous issue date: 2007 Cutaneous (CL) and mucosal leishmaniasis (ML) are characterized by a predominant type 1 immune response (IFN-γ and TNF-α production) and strong inflammatory response in the lesions with few parasites. This exacerbated type 1 response is more evident in ML as compared to CL. Our main hypothesis is that a differential immune regulation of T cell activation leads to over reactive T cells in ML. In the present study, we investigated immunological factors that could explain the mechanisms behind it by comparing some immune regulatory mechanisms between ML and CL patients: frequency of cells expressing co-stimulatory molecules, apoptotic markers, T cell activation markers; and ability of neutralizing antibodies to IL-2, IL-12 and IL-15 do down-regulate IFN-γ production in leishmania antigen-stimulated peripheral blood mononuclear cells (PBMC). Interestingly, in CL anti-IL-2 and anti-IL-15 significantly suppressed antigen-specific IFN-γ production, while in ML only anti-IL-2 suppressed IFN-γ production. Finally, higher frequency of CD4+ T cells expressing CD28−, CD69+ and CD62Llow were observed in ML as compared to CL. These data indicate that an exacerbated type 1 response in ML is differentially regulated and not appropriately down modulated, with increased frequencies of activated effectors T cells, maintaining the persistent inflammatory response and tissue damage observed in ML.

Texto completo: acesso restrito. p. 1017–1024 Submitted by Suelen Reis (suziy.ellen@gmail.com) on 2014-03-11T15:14:47Z No. of bitstreams: 1 10.1128IAI.00914-06.pdf: 554649 bytes, checksum: e4ffaf2b2ca32eca83ae35e61b28e6e0 (MD5) Approved for entry into archive by Flávia Ferreira (flaviaccf@yahoo.com.br) on 2014-04-09T13:04:35Z (GMT) No. of bitstreams: 1 10.1128IAI.00914-06.pdf: 554649 bytes, checksum: e4ffaf2b2ca32eca83ae35e61b28e6e0 (MD5) Made available in DSpace on 2014-04-09T13:04:35Z (GMT). No. of bitstreams: 1 10.1128IAI.00914-06.pdf: 554649 bytes, checksum: e4ffaf2b2ca32eca83ae35e61b28e6e0 (MD5) Previous issue date: 2007 Leishmania spp. are intracellular protozoa residing in mononuclear phagocytes. Leishmania organisms are susceptible to microbicidal responses generated in response to phagocytosis. Assuming that both phagocyte and parasite populations are heterogeneous, it is advantageous to examine the response of individual cells phagocytosing living parasites. Because Leishmania spp. lose virulence during the raising of transfectants, we developed a method to label live Leishmania chagasi short-term with fluorescent dyes. Up to six parasite divisions were detected by flow cytometry after labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE), dioctadecyl-tetramethylindo carbocyanine perchlorate, or chloromethyl tetramethylrhodamine. Labeled parasites entered mononuclear phagocytes as determined by confocal and time-lapse microscopy. Dihydroethidium (DHE) was used to detect macrophage-derived oxidants generated during phagocytosis. Presumably Leishmania organisms are opsonized with host serum/tissue components such as complement prior to phagocytosis. Therefore, we investigated the effects of opsonization and found that this increased the efficiency of CFSE-labeled parasite entry into monocytes (84.6% ± 8.8% versus 20.2% ± 3.8% monocytes infected; P < 0.001). Opsonization also increased the percentage of phagocytes undergoing a respiratory burst (66.0% ± 6.3% versus 41.0% ± 8.3% of monocytes containing CFSE-labeled parasites; P < 0.001) and the magnitude of oxidant generation by each infected monocyte. Inhibitor data indicated that DHE was oxidized by products of the NADPH oxidase. These data suggest that opsonized serum components such as complement lead to more efficient entry of Leishmania into their target cells but at the same time activate the phagocyte oxidase to generate microbicidal products in infected cells. The parasite must balance these positive and negative survival effects in order to initiate a viable infection.

BACKGROUND: Disseminated leishmaniasis is an emerging infectious disease, mostly due to L. braziliensis, which has clinical and histopathological features distinct from cutaneous leishmaniasis. METHODS: In the current study we evaluated the in vitro production of the cytokines IFN-γ, TNF-α, IL-5 and IL-10 by peripheral blood mononuclear cells (PBMC) from 15 disseminated leishmaniasis and 24 cutaneous leishmaniasis patients upon stimulation with L. braziliensis antigens genotyped as disseminated leishmaniasis or cutaneous leishmaniasis isolates. RESULTS: Regardless of the source of L. braziliensis antigens, PBMC from cutaneous leishmaniasis patients produced significantly higher IFN-γ than PBMC from disseminated leishmaniasis patients. Levels of TNF-α by PBMC from cutaneous leishmaniasis patients were significantly higher than disseminated leishmaniasis patients only when stimulated by genotyped cutaneous leishmaniasis antigens. The levels of IL-5 and IL-10 production by PBMC were very low and similar in PBMCs from both disseminated leishmaniasis and cutaneous leishmaniasis patients. The immune response of each patient evaluated by the two L. braziliensis antigens was assessed in a paired analysis in which we showed that L. braziliensis genotyped as disseminated leishmaniasis isolate was more potent than L. braziliensis genotyped as cutaneous leishmaniasis isolate in triggering IFN-γ and TNF-α production in both diseases and IL-5 only in cutaneous leishmaniasis patients. CONCLUSION: This study provides evidence that antigens prepared from genotypically distinct strains of L. braziliensis induce different degrees of immune response. It also indicates that both parasite and host play a role in the outcome of L. braziliensis infection.

p.1-6 Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2013-08-27T11:36:10Z No. of bitstreams: 1 1471-2334-6-75.pdf: 289112 bytes, checksum: 456a013c7aad83ff1a9b0fa90711a95d (MD5) Approved for entry into archive by Patricia Barroso(pbarroso@ufba.br) on 2013-08-27T17:37:57Z (GMT) No. of bitstreams: 1 1471-2334-6-75.pdf: 289112 bytes, checksum: 456a013c7aad83ff1a9b0fa90711a95d (MD5) Made available in DSpace on 2013-08-27T17:37:57Z (GMT). No. of bitstreams: 1 1471-2334-6-75.pdf: 289112 bytes, checksum: 456a013c7aad83ff1a9b0fa90711a95d (MD5) Previous issue date: 2006 Background: Disseminated leishmaniasis is an emerging infectious disease, mostly due to L. braziliensis, which has clinical and histopathological features distinct from cutaneous leishmaniasis. Methods: In the current study we evaluated the in vitro production of the cytokines IFN-γ, TNF-α, IL-5 and IL-10 by peripheral blood mononuclear cells (PBMC) from 15 disseminated leishmaniasis and 24 cutaneous leishmaniasis patients upon stimulation with L. braziliensis antigens genotyped as disseminated leishmaniasis or cutaneous leishmaniasis isolates. Results: Regardless of the source of L. braziliensis antigens, PBMC from cutaneous leishmaniasis patients produced significantly higher IFN-γ than PBMC from disseminated leishmaniasis patients. Levels of TNF-α by PBMC from cutaneous leishmaniasis patients were significantly higher than disseminated leishmaniasis patients only when stimulated by genotyped cutaneous leishmaniasis antigens. The levels of IL-5 and IL-10 production by PBMC were very low and similar in PBMCs from both disseminated leishmaniasis and cutaneous leishmaniasis patients. The immune response of each patient evaluated by the two L. braziliensis antigens was assessed in a paired analysis in which we showed that L. braziliensis genotyped as disseminated leishmaniasis isolate was more potent than L. braziliensis genotyped as cutaneous leishmaniasis isolate in triggering IFN-γ and TNF-α production in both diseases and IL-5 only in cutaneous leishmaniasis patients. Conclusion: This study provides evidence that antigens prepared from genotypically distinct strains of L. braziliensis induce different degrees of immune response. It also indicates that both parasite and host play a role in the outcome of L. braziliensis infection.

Texto completo:acesso restrito. p. 341-348. Submitted by JURANDI DE SOUZA SILVA (jssufba@hotmail.com) on 2012-12-13T17:47:37Z No. of bitstreams: 1 j.1365-2249.2004.02426.x.pdf: 154075 bytes, checksum: b96d36a6be1a7e1d84e73a6a964ddf6a (MD5) Made available in DSpace on 2012-12-13T17:47:37Z (GMT). No. of bitstreams: 1 j.1365-2249.2004.02426.x.pdf: 154075 bytes, checksum: b96d36a6be1a7e1d84e73a6a964ddf6a (MD5) Previous issue date: 2004 Regulation of the immune response directed against Leishmaniais critical for the establishment of effective control of the disease. It is likely that some types of immune responses directed against Leishmania can lead to more severe clinical forms of leishmaniasis causing a poor control of the pathogen and/or pathology, while others lead to resolution of the infection with little pathology as in cutaneous leishmaniasis. To gain a better understanding of the possible role that subpopulations of T cells, and their associated cytokines have on disease progression and/or protective immune responses to L. braziliensis infection, a detailed study of the frequency of activated and memory T cells, as well as antigen specific, cytokine producing T cells was carried out. Following the determination of cytokine producing mononuclear cell populations in response to total Leishmania antigen (SLA), and to the recombinant antigen LACK, correlation analysis were performed between specific cytokine producing populations to identify models for cellular mechanisms of immunoregulation in human cutaneous leishmaniasis.These studies have shown: (1) a positive correlation between ex vivo CD45RO frequencies and antigen specific cytokine (IFN-gamma or IL-10) producing cells; (2) a negative correlation between ex vivo CD69 expression and the frequency of IFN-gamma producing cells; (3) a positive correlation amongst SLA specific, IFN-gamma or TNF-alpha and IL-10 producing lymphocytes with one another; and (4) a higher frequency of IL-10 producing, parasite specific (anti-SLA or anti-LACK), lymphocytes are correlated with a lower frequency of TNF-alpha producing monocytes, demonstrating an antigen specific delivery of IL-10 inducing negative regulation of monocyte activity.

p. 508–514 Submitted by Ana Valéria de Jesus Moura (anavaleria_131@hotmail.com) on 2011-12-11T17:03:46Z No. of bitstreams: 1 1077.pdf: 1159272 bytes, checksum: 42eaf0f40906bf9b35739de038ee64f2 (MD5) Made available in DSpace on 2011-12-11T17:03:46Z (GMT). No. of bitstreams: 1 1077.pdf: 1159272 bytes, checksum: 42eaf0f40906bf9b35739de038ee64f2 (MD5) Previous issue date: 2004-01 In Corte de Pedra (CP), northeastern Brazil, Leishmania braziliensis causes three distinct forms of American tegumentary leishmaniasis (ATL). To test the hypothesis that strain polymorphism may be involved in this disease spectrum and accurately characterize the parasite population structure in CP, we compared one L. major, two non-CP L. braziliensis, one CP L. amazonensis, and 45 CP L. braziliensis isolates, obtained over a 10-year period from localized cutaneous, mucosal, and disseminated leishmaniasis patients, with randomly amplified polymorphic DNA (RAPD). Electrophoretic profiles were mostly unique across species. All typing protocols revealed polymorphism among the 45 CP L. braziliensis isolates, which displayed eight different RAPD patterns and greater than 80% overall fingerprint identity, attesting to the adequacy of the tools to assess strain variability in CP’s geographically limited population of parasites. The dendrogram based on the sum of RAPD profiles of each isolate unveiled nine discrete typing units clustered into five clades. Global positioning showed extensive overlap of these clades in CP, precluding geographic sequestration as the mechanism of the observed structuralization. Finally, all forms of ATL presented a statistically significant difference in their frequencies among the clades, suggesting that L. braziliensis genotypes may be accompanied by specific disease manifestation after infection.

Texto completo: acesso restrito. p. 101-104 Submitted by Suelen Reis (suelen_suzane@hotmail.com) on 2012-12-17T14:07:20Z No. of bitstreams: 1 101.full.pdf: 157103 bytes, checksum: efc876856ea0c49fcc9a0e237a93cbcd (MD5) Made available in DSpace on 2012-12-17T14:07:21Z (GMT). No. of bitstreams: 1 101.full.pdf: 157103 bytes, checksum: efc876856ea0c49fcc9a0e237a93cbcd (MD5) Previous issue date: 2002-01 High levels of antileishmanial immunoglobulin E (IgE) antibodies are associated with disease activity in visceral leishmaniasis. Herein, we report our observations about the relationship between antileishmanial IgE antibodies and clinical aspects of cutaneous leishmaniasis. This study was carried out with 45 patients (29 male and 16 female), with ages ranging from 11 to 48 years. All subjects were from an area to which leishmaniasis is endemic, Corte de Pedra (Bahia, Brazil), and the duration of the illness was ≤30 days. The patients were classified as positive or negative for IgE serology in enzyme-linked immunosorbent assay with leishmanial antigens. IgE antibodies were detected in 18 patients (optical density, 0.421 ± 0.30; 95% confidence interval, 0.27 to 0.57), and only 3 (17%) had more than one ulcer. In this group the diameter of Montenegro’s reaction was 18 ± 12.2 mm. In the group with negative IgE serology, 11 of 27 patients (48%) presented two or more cutaneous ulcers, and the mean of the skin test result was 9 ± 6.9 mm. There was a positive correlation between IgE antibody levels and Montenegro’s reaction size and an inverse correlation between IgE antileishmanial antibodies and the number of skin ulcers. The presence of antileishmanial IgE antibodies in cutaneous leishmaniasis may be a result of immunoregulatory events with clinical implications.

P. 7-10,Jan-Fev. Submitted by JURANDI DE SOUZA SILVA (jssufba@hotmail.com) on 2011-09-12T12:56:37Z No. of bitstreams: 1 13164.pdf: 82743 bytes, checksum: e021d3f31a9761bc2665706a1081739e (MD5) Made available in DSpace on 2011-09-12T12:56:37Z (GMT). No. of bitstreams: 1 13164.pdf: 82743 bytes, checksum: e021d3f31a9761bc2665706a1081739e (MD5) Previous issue date: 2002 Para avaliar se os níveis séricos de IFN-g e TNF-a podem ser usados como marcadores da resposta terapêutica na leishmaniose cutânea, 54 pacientes com história de uma única lesão ulcerada com duração de até 30 dias foram arrolados para o estudo. IFN-g e TNF-a foram mensurados por ELISA no sobrenadante de cultura de linfócitos, antes e 60 dias após início da terapia. Cicatrização completa da lesão 60 dias após o tratamento foi considerada critério de cura. Os níveis de IFN-g e TNF-a foram similares em ambos os grupos antes da terapia. Houve tendência de aumento dos níveis de IFN-g nos pacientes curados aos 60 dias, os valores não alcançam significância estatística. Em ambos os grupos de pacientes, os níveis iniciais de TNF-a foram semelhantes antes da terapia e caíram significantemente após tratamento independente da cura ou manutenção da lesão ativa. Uberaba

Texto completo: acesso restrito. p.98-105 Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2014-02-03T15:22:14Z No. of bitstreams: 1 Angela Silva.pdf: 150155 bytes, checksum: b258ecad79a09b6802c5000bc6cf812a (MD5) Approved for entry into archive by Patricia Barroso (pbarroso@ufba.br) on 2014-02-06T19:37:41Z (GMT) No. of bitstreams: 1 Angela Silva.pdf: 150155 bytes, checksum: b258ecad79a09b6802c5000bc6cf812a (MD5) Made available in DSpace on 2014-02-06T19:37:41Z (GMT). No. of bitstreams: 1 Angela Silva.pdf: 150155 bytes, checksum: b258ecad79a09b6802c5000bc6cf812a (MD5) Previous issue date: 2002 Thirty-one patients with acute schistosomiasis were evaluated clinically and immunologically. Cytokine levels were determined in peripheral blood mononuclear cell (PBMC) supernatants. Levels of total and antigen-specific IgE, tumor necrosis factor (TNF)–α, and immune complexes were measured in serum samples. Clinical findings included general symptoms, liver damage, pulmonary involvement, and pericarditis. All patients had eosinophilia. Immune complexes were detected in 55% of the patients (mean±SD, 7.8±7.6 μg Eq/mL) and were associated with cough, dyspnea, and abnormal chest radiographic findings. Levels (mean ± SD) of TNF-α (1349.3±767.6 pg/mL), interleukin (IL)–1 (2683±1270 pg/mL), and IL-6 (382 ± 52.3 pg/mL) were elevated in PBMC. Serum TNF-α levels were elevated in 87% of the patients and were associated with abdominal pain. Higher interferon-γ levels were detected in PBMC of patients with acute disease than in those of patients with chronic schistosomiasis; IL-5 levels were higher in those with chronic disease. Low IL-5 levels were associated with weight loss. Proinflammatory cytokines and immune complexes with low Th2 responses might explain the immunopathogenesis of acute schistosomiasis.

Texto completo:acesso restrito. p. 1829-1834 Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2014-01-24T15:40:13Z No. of bitstreams: 1 Paulo R. Machado.pdf: 224392 bytes, checksum: 67109cc2ca55ab2deef4204989e276ef (MD5) Approved for entry into archive by Flávia Ferreira (flaviaccf@yahoo.com.br) on 2015-10-27T15:11:16Z (GMT) No. of bitstreams: 1 Paulo R. Machado.pdf: 224392 bytes, checksum: 67109cc2ca55ab2deef4204989e276ef (MD5) Made available in DSpace on 2015-10-27T15:11:16Z (GMT). No. of bitstreams: 1 Paulo R. Machado.pdf: 224392 bytes, checksum: 67109cc2ca55ab2deef4204989e276ef (MD5) Previous issue date: 2002 During the past decade, there has been an increase in the number of patients with disseminated leishmaniasis (DL), which is characterized by a large number of acneiform and papular skin lesions, with very few or no parasites in the skin tissue. The present report describes 42 cases of DL identified between 1992 and 1998 in an area where Leishmania braziliensis transmission is endemic; 8 of the patients were prospectively diagnosed. In a contrast to localized cutaneous leishmaniasis (LCL), acquisition of DL was associated with age >19 years (P

p. 101-103 Submitted by Suelen Reis (suelen_suzane@hotmail.com) on 2012-12-17T15:34:45Z No. of bitstreams: 1 4291.pdf: 74717 bytes, checksum: 58e675ae0f403132ba5858e62a9b23c7 (MD5) Made available in DSpace on 2012-12-17T15:34:45Z (GMT). No. of bitstreams: 1 4291.pdf: 74717 bytes, checksum: 58e675ae0f403132ba5858e62a9b23c7 (MD5) Previous issue date: 2002-01 Procedures for IgG depletion in visceral leishmaniasis (VL) and schistosomiasis sera using Sepharose-protein G beads also deplete IgE. In this study, the presence of IgG anti-IgE autoantibodies in sera from patients with VL (n = 10), and hepatic-intestinal schistosomiasis (n = 10) and from healthy individuals (n = 10) was investigated. A sandwich ELISA using goat IgG anti-human IgE to capture serum IgE and goat anti-human IgG peroxidase conjugate to demonstrate the binding of IgG to the IgE captured was performed. VL sera had higher titers (p < 0.05) of IgG anti-IgE autoantibodies (OD = 2.01 ± 0.43) than sera from healthy individuals (OD = 1.35 ± 0.16) or persons infected with Schistosoma mansoni (OD = 1.34 ± 0.18). The immunoblotting carried out with eluates from Sepharose-protein G beads used to deplete IgG from these sera and goat anti-human IgE peroxidase conjugate, showed a similar pattern of bands, predominating the 75 kDa e-heavy chain and also polypeptides resulting from physiological enzymatic digestion of IgE. A frequent additional band immediately above 75 kDa was observed only in VL sera.

Texto completo: acesso restrito. p. 69-73 Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2014-01-20T12:55:22Z No. of bitstreams: 1 Paulo Machado.pdf: 283941 bytes, checksum: 8a820425f50c84a4911718fdbc9636f6 (MD5) Approved for entry into archive by Flávia Ferreira (flaviaccf@yahoo.com.br) on 2014-11-21T13:50:43Z (GMT) No. of bitstreams: 1 Paulo Machado.pdf: 283941 bytes, checksum: 8a820425f50c84a4911718fdbc9636f6 (MD5) Made available in DSpace on 2014-11-21T13:50:43Z (GMT). No. of bitstreams: 1 Paulo Machado.pdf: 283941 bytes, checksum: 8a820425f50c84a4911718fdbc9636f6 (MD5) Previous issue date: 2002 The clinical characteristics and treatment outcome were determined for 26 patients who presented with early-stage cutaneous leishmaniasis. Illness duration ranged from 8 to 20 days, and the commonest clinical presentation was the presence of a papule with small central crust on a lower extremity. Prominent regional adenopathy was found in 22 (85%) of 26 patients. The results of an intradermal skin test for Leishmania were positive for 96% of those patients, and results of serologic testing were positive for 61% of patients tested. Ten (46%) of 22 patients for whom follow-up data were available developed enlargement and ulceration of the lesion despite early antimony therapy and required additional courses of treatment. Histopathological studies of samples from the lesions of 3 patients showed vasculitis. These data show that early therapy for cutaneous leishmaniasis does not prevent the development of an ulcer in one-half of patients. This unfavorable outcome underlines the relevance of local exacerbated inflammatory and immune response in the pathogenesis of the disease.

P. 537-542, Nov-Dez. Submitted by JURANDI DE SOUZA SILVA (jssufba@hotmail.com) on 2011-09-12T13:24:40Z No. of bitstreams: 1 13164.pdf: 66389 bytes, checksum: 8f5ed8c38cbb5140621e5d70d5a6ca0d (MD5) Made available in DSpace on 2011-09-12T13:24:40Z (GMT). No. of bitstreams: 1 13164.pdf: 66389 bytes, checksum: 8f5ed8c38cbb5140621e5d70d5a6ca0d (MD5) Previous issue date: 2001 O teste de Montenegro, utilizado no diagnóstico da leishmaniose tegumentar, tem sido recentemente considerado marcador de imunogenicidade após vacinação. Neste estudo, avaliou-se o poder sensibilizante deste teste e a produção de y-IFN in vitro, em indivíduos não expostos a Leishmania. Utilizaram-se para o teste antígenos de L. amazonensis produzido em nosso laboratório (grupo I) ou produzido pela FIOCRUZ-RJ (grupo II). No dia 30, 33% dos indivíduos do grupo I e 42% do grupo II converteram o teste para positivo, comparando-se com 67% do grupo I e 50% do grupo II no dia 90. y-IFN foi detectado em 56% dos indivíduos do grupo I e 17% do grupo II no dia 30 (169±309 e 11±36pg/ml) e em 67% do grupo I e 58% do grupo II no dia 360 (69±107 e 18±20pg/ml). Estes dados demonstram que o teste de Montenegro induz, além de hipersensibilidade tardia, a produção de y-IFN. Uberaba

Activation of Th1 or Th2 cells is associated with production of specific immunoglobulin isotypes, offering the opportunity to use antibody measurement for evaluation of T cell function. Schistosomiasis and visceral leishmaniasis are diseases associated with Th2 activation. However, an IgE response is not always detected in these patients. In the present study we evaluated specific IgE antibodies to S. mansoni and L. chagasi antigens by ELISA after depletion of serum IgG with protein G immobilized on Sepharose beads or RF-absorbent (purified sheep IgG antibodies anti-human IgG). In schistosomiasis patients, specific IgE to SWAP antigen was demonstrable in only 10 of 21 patients (48%) (mean absorbance ± SD = 0.102 ± 0.195) when unabsorbed serum was used. Depletion of IgG with protein G increased the number of specific IgE-positive tests to 13 (62%) and the use of RF-absorbent increased the number of positive results to 20 (95%) (mean absorbances ± SD = 0.303 ± 0.455 and 0.374 ± 0.477, respectively). Specific IgE anti-L. chagasi antibodies were not detected in unabsorbed serum from visceral leishmaniasis patients. When IgG was depleted with protein G, IgE antibodies were detected in only 3 (11%) of 27 patients, and the use of RF-absorbent permitted the detection of this isotype in all 27 visceral leishmaniasis sera tested (mean absorbance ± SD = 0.104 ± 0.03). These data show that the presence of IgG antibodies may prevent the detection of a specific IgE response in these parasite diseases. RF-absorbent, a reagent that blocks IgG-binding sites and also removes rheumatoid factor, was more efficient than protein G for the demonstration of specific IgE antibodies.

p. 1101-1105 Submitted by Santiago Fabio (fabio.ssantiago@hotmail.com) on 2012-10-24T18:07:09Z No. of bitstreams: 1 Souza-Atta, M.L.B..pdf: 169960 bytes, checksum: 536c503cc92dd82e815a9c8339681322 (MD5) Made available in DSpace on 2012-10-24T18:07:09Z (GMT). No. of bitstreams: 1 Souza-Atta, M.L.B..pdf: 169960 bytes, checksum: 536c503cc92dd82e815a9c8339681322 (MD5) Previous issue date: 1999 Activation of Th1 or Th2 cells is associated with production of specific immunoglobulin isotypes, offering the opportunity to use antibody measurement for evaluation of T cell function. Schistosomiasis and visceral leishmaniasis are diseases associated with Th2 activation. However, an IgE response is not always detected in these patients. In the present study we evaluated specific IgE antibodies to S. mansoni and L. chagasi antigens by ELISA after depletion of serum IgG with protein G immobilized on Sepharose beads or RF-absorbent (purified sheep IgG antibodies anti-human IgG). In schistosomiasis patients, specific IgE to SWAP antigen was demonstrable in only 10 of 21 patients (48%) (mean absorbance ± SD = 0.102 ± 0.195) when unabsorbed serum was used. Depletion of IgG with protein G increased the number of specific IgE-positive tests to 13 (62%) and the use of RF-absorbent increased the number of positive results to 20 (95%) (mean absorbances ± SD = 0.303 ± 0.455 and 0.374 ± 0.477, respectively). Specific IgE anti-L. chagasi antibodies were not detected in unabsorbed serum from visceral leishmaniasis patients. When IgG was depleted with protein G, IgE antibodies were detected in only 3 (11%) of 27 patients, and the use of RF-absorbent permitted the detection of this isotype in all 27 visceral leishmaniasis sera tested (mean absorbance ± SD = 0.104 ± 0.03). These data show that the presence of IgG antibodies may prevent the detection of a specific IgE response in these parasite diseases. RF-absorbent, a reagent that blocks IgG-binding sites and also removes rheumatoid factor, was more efficient than protein G for the demonstration of specific IgE antibodies. Ribeirão Preto

Texto completo. Acesso restrito. p.1731-1734 Submitted by Santiago Fabio (fabio.ssantiago@hotmail.com) on 2013-08-01T20:40:16Z No. of bitstreams: 1 777777777777777777777.pdf: 91255 bytes, checksum: 4f795003d50d28945a2af7d42727c258 (MD5) Approved for entry into archive by Flávia Ferreira(flaviaccf@yahoo.com.br) on 2013-08-05T13:29:47Z (GMT) No. of bitstreams: 1 777777777777777777777.pdf: 91255 bytes, checksum: 4f795003d50d28945a2af7d42727c258 (MD5) Made available in DSpace on 2013-08-05T13:29:47Z (GMT). No. of bitstreams: 1 777777777777777777777.pdf: 91255 bytes, checksum: 4f795003d50d28945a2af7d42727c258 (MD5) Previous issue date: 1999 This study examined the T cell responses in the early phase of Leishmania braziliensis infection. Cytokine profiles, lymphoproliferative responses, and skin test results in 25 patients with early cutaneous leishmaniasis (ECL; illness duration !60 days) were compared with those in persons with late cutaneous leishmaniasis (LCL; illness duration 12 months). Absent or low lymphoproliferative responses were observed in 8 (32%) of 25 patients and an absence of interferon (IFN)–g production in 9 (41%) of 22 patients prior to therapy. IFN-g production in ECL (mean 5 SD) was lower than in LCL (2935346 vs. 7475377 pg/mL, respectively; P ! .01). In contrast, interleukin (IL)–10 production in ECL (mean 5 SD) was higher than in LCL (246556 vs. 50541 pg/mL, respectively;P ! .01) . Restoration of lymphoproliferative responses and IFN-g production was achieved when monoclonal antibody to IL-10 or IL-12 was added to the cultures. These results show that T cell responses during early-phase infection are down-regulated by IL-10 and may facilitate parasite multiplication. Salvador

p. 426–430 Submitted by Santiago Fabio (fabio.ssantiago@hotmail.com) on 2013-08-20T19:04:15Z No. of bitstreams: 1 4444444444444444444444.pdf: 81049 bytes, checksum: 80b46900692f3f8459a653c83f5aca3e (MD5) Approved for entry into archive by Alda Lima da Silva(sivalda@ufba.br) on 2013-08-23T21:54:32Z (GMT) No. of bitstreams: 1 4444444444444444444444.pdf: 81049 bytes, checksum: 80b46900692f3f8459a653c83f5aca3e (MD5) Made available in DSpace on 2013-08-23T21:54:32Z (GMT). No. of bitstreams: 1 4444444444444444444444.pdf: 81049 bytes, checksum: 80b46900692f3f8459a653c83f5aca3e (MD5) Previous issue date: 1998 Abstract. Visceral leishmaniasis (VL) is characterized by a depression of the T helper cell type 1 immune response. Although mRNA expression for interleukin-4 (IL-4) is observed, evidence of the role of this cytokine in the pathogenesis of VL has been lacking. Since IL-4 is involved in IgE synthesis, we measured the total IgE and Leishmania antigen-specific IgE antibody levels in sera from patients with VL. Specific IgE antibodies detected by an ELISA technique after absorbing the sera with purified sheep IgG anti-human IgG were found in all 23 patients with VL and were not detected in subjects with subclinical Leishmania chagasi infection (n 5 10), Chagas’ disease (n 5 10), atopic patients (n 5 10), and healthy controls (n 5 10). Levels of Leishmania-specific IgE (optical density values) before and after treatment were 0.100 6 0.03 (mean 6 SD) and 0.028 6 0.002, respectively (P , 0.05). These results indicate that a specific IgE response is useful in the diagnosis of active disease and to evaluate response to treatment. Salvador

The clinical spectrum of leishmaniasis and control of the infection are influenced by the parasite-host relationship. The role of cellular immune responses of the Th1 type in the protection against disease in experimental and human leishmaniasis is well established. In humans, production of IFN-g is associated with the control of infection in children infected by Leishmania chagasi. In visceral leishmaniasis, an impairment in IFN-g production and high IL-4 and IL-10 levels (Th2 cytokines) are observed in antigen-stimulated peripheral blood mononuclear cells (PBMC). Moreover, IL-12 restores IFN-g production and enhances the cytotoxic response. IL-10 is the cytokine involved in down-regulation of IFN-g production, since anti-IL-10 monoclonal antibody (mAb) restores in vitro IFN-g production and lymphoproliferative responses, and IL-10 abrogates the effect of IL-12. In cutaneous and mucosal leishmaniasis, high levels of IFN-g are found in L. amazonensis-stimulated PBMC. However, low or absent IFN-g levels were observed in antigen-stimulated PBMC from 50% of subjects with less than 60 days of disease (24 ± 26 pg/ml). This response was restored by IL-12 (308 ± 342 pg/ml) and anti-IL-10 mAb (380 ± 245 pg/ml) (P

p.143-148 Submitted by Texeira Ana (atanateixeira@gmail.com) on 2012-10-23T18:40:11Z No. of bitstreams: 1 Ribeiro-de-Jesus, A.pdf: 226801 bytes, checksum: 7a9bc9775d64eb82a1d00e52369e01f5 (MD5) Made available in DSpace on 2012-10-23T18:40:11Z (GMT). No. of bitstreams: 1 Ribeiro-de-Jesus, A.pdf: 226801 bytes, checksum: 7a9bc9775d64eb82a1d00e52369e01f5 (MD5) Previous issue date: 1998-01 The clinical spectrum of leishmaniasis and control of the infection are influenced by the parasite-host relationship. The role of cellular immune responses of the Th1 type in the protection against disease in experimental and human leishmaniasis is well established. In humans, production of IFN- is associated with the control of infection in children infected by Leishmania chagasi. In visceral leishmaniasis, an impairment in IFN- production and high IL-4 and IL-10 levels (Th2 cytokines) are observed in antigen-stimulated peripheral blood mononuclear cells (PBMC). Moreover, IL-12 restores IFN- production and enhances the cytotoxic response. IL-10 is the cytokine involved in down-regulation of IFN- production, since anti-IL-10 monoclonal antibody (mAb) restores in vitro IFN- production and lymphoproliferative responses, and IL-10 abrogates the effect of IL-12. In cutaneous and mucosal leishmaniasis, high levels of IFN- are found in L. amazonensis-stimulated PBMC. However, low or absent IFN- levels were observed in antigen-stimulated PBMC from 50% of subjects with less than 60 days of disease (24 ± 26 pg/ml). This response was restored by IL-12 (308 ± 342 pg/ml) and anti-IL-10 mAb (380 ± 245 pg/ml) (P

Texto completo. Acesso restrito. p. 355–366 Submitted by Santiago Fabio (fabio.ssantiago@hotmail.com) on 2013-09-09T18:03:41Z No. of bitstreams: 1 11111111.pdf: 374080 bytes, checksum: 95febf9cb2121d9cf26a5318582ba2d8 (MD5) Approved for entry into archive by Rodrigo Meirelles (rodrigomei@ufba.br) on 2013-11-16T13:15:44Z (GMT) No. of bitstreams: 1 11111111.pdf: 374080 bytes, checksum: 95febf9cb2121d9cf26a5318582ba2d8 (MD5) Made available in DSpace on 2013-11-16T13:15:44Z (GMT). No. of bitstreams: 1 11111111.pdf: 374080 bytes, checksum: 95febf9cb2121d9cf26a5318582ba2d8 (MD5) Previous issue date: 1996 The azurophil granules of human PMN contain four antibiotic proteins, the serprocidins, which have extensive homology to one another and to serine proteases. Azurocidin, a member of this family, is a 29-kDa glycoprotein with broad spectrum antimicrobial activity and chemotactic activity toward monocytes. Insect cells transfected with a baculovirus vector carrying azurocidin cDNA produced a recombinant azurocidin protein. We purified the recombinant azurocidin protein from the culture medium of the infected cells and showed that it retained the antimicrobial activity of the native neutrophil-derived molecule. In addition, we present evidence that a 49-amino-acid region of the recombinant azurocidin protein is required for its secretion from insect cells. Salvador

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES Introduction: Arboviruses are diseases of viral etiology transmitted to hosts through hematophagous arthropods. Of great importance, the viruses infection Zika (ZIKV) and Chikungunya (CHIKV) may be asymptomatic or present with symptoms that range from mild to severe neurological disorders, such as Guillain-Barré Syndrome and Congenital Syndrome of the Zika virus. There is also cases reports describing coagulation disorders. To date, the vascular involvement triggered by the infections caused by ZIKV and CHIKV is poorly described. Objective: To describe 2 cases of deep venous thrombosis (DVT) associated with ZIKV and to evaluate serum D-dimer levels in patients infected with ZIKV and CHIKV virus. Methodology: This is a descriptive study, where 2 cases of DVT associated with ZIKV were reported, one from the University Hospital of Sergipe and the other from the University Hospital of Paraíba. In addition, plasma samples from 172 individuals infected by ZIKV (31) and CHIKV (141) were used, from the services offered by the University Hospital of the Federal University of Sergipe, from which clinical-epidemiological information was collected through a questionnaire, molecular diagnosis using real-time reverse transcription polymerase chain reaction (RT-PCR) and D-dimer dosing by enzyme-linked immunosorbent assay (ELISA). Results: Individuals infected with ZIKV and CHIKV had a mean age of 37.5 ± 13.35 years and 41.8 ± 17.3, respectively. The most affected gender was male (64%) by ZIKV and female (86.8%) by CHIKV. D-dimer levels were increased in 19.35% of the 31 ZIKV-positive individuals and in 63.83% of the 141 CHIKV-positive individuals. Conclusion: This study describes for the first time high levels of D-dimer in people infected with ZIKV and CHIKV, providing data relevant to the scientific community about the possibility of the association of these arboviruses and the development of venous thromboembolism (VTE). Introdução: As arboviroses são doenças de etiologia viral transmitidas aos hospedeiros através de artrópodes hematófagos. Entre essas arboviroses, destacam-se atualmente as infecções pelos vírus Zika (ZIKV) e Chikungunya (CHIKV), as quais podem ser assintomáticas ou apresentarem quadros clínicos variados desde sintomas leves a graves, como os distúrbios neurológicos, a exemplo da Síndrome de Guillain-Barré e a Síndrome Congênita do Zika vírus. Foram relatados também alguns casos de complicações relacionadas aos distúrbios da coagulação. Até o momento, o acometimento vascular desencadeado pelas infecções causadas pelos ZIKV e CHIKV é pouco descrito. Objetivo: Descrever 2 casos de trombose venosa profunda (TVP) associados ao ZIKV e avaliar os níveis séricos do dímero D em pacientes infectados pelos vírus ZIKV e CHIKV. Metodologia: Trata-se de um estudo descritivo onde foram relatados 2 casos de TVP associados ao ZIKV, um proveniente do Hospital Universitário de Sergipe e o outro do Hospital Universitário da Paraíba. Além disso, foram utilizadas amostras de plasmas de 172 indivíduos infectados pelos ZIKV (31) e CHIKV (141), oriundos dos serviços ofertados pelo Hospital Universitário da Universidade Federal de Sergipe, dos quais foram coletadas informações clínico-epidemiológicas através de questionário e em suas amostras foram realizados o diagnóstico molecular pela técnica da transcrição reversa da reação em cadeia da polimerase (RT-PCR) em tempo real e a dosagem do dímero D por ensaio imunoenzimático (ELISA). Resultados: Os indivíduos infectados pelos ZIKV e CHIKV tinham idade média de 37,5±13,35 anos e 41,8±17,3, respectivamente. O gênero mais afetado foi o masculino (64%) pelo ZIKV e o feminino (86,8%) pelo CHIKV. Os níveis de dímero D foram aumentados em relação ao valor de referência (0,5μg/mL) em 19,35% dos 31 indivíduos positivos para o ZIKV e em 63,83% dos 141 indivíduos positivos para CHIKV. Conclusão: Este estudo descreve pela primeira vez níveis elevados de dímero D em pessoas infectadas pelos ZIKV e CHIKV, fornecendo dados relevantes para a comunidade científica sobre a possibilidade da associação desses arbovírus e o desenvolvimento do tromboembolismo venoso (TEV). Aracaju

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES Trypanosomatid protozoa of Leishmania genus are vector-borne parasites that infect humans causing a wide range of diseases known as Leishmaniasis. Severity of clinical forms generally depends on infecting species, thus its correct identification becomes critical to diagnosis, prognosis and treatment. The current biochemical and molecular methods for typing strains of Leishmania species are mostly effective. Although these tools remain important in the diagnosis and identification of Leishmania, they are insufficient for definitive identification. After isolate 7 clinical parasites strains, some of them belonging to atypical cases of cutaneous/mucocutaneous and visceral Leishmaniasis, we aimed to identify the specie of Leishmania involved in these cases. By combining structural analysis of the body shape, such as presence of flagellum and kinetoplast, we saw that these parasites shared morphologic characteristics with others Tripanosomatidae, however, they were not conclusive for species identification. Since PCR and isozymes assays were unsuccessful to identify 6 out of 7 clinical isolates reported here, we proceeded to determine their genome sequence. Through whole-genome sequencing analysis of parasite strains, we showed that a new pathogenic Trypanosomatid was the etiological agent in clinical cases diagnosed as Leishmaniasis in Brazil. By comparing coding sequences of over orthologous genes within 36 Trypanosomatidae organisms, we found that this new parasite is closely related to Crithidia fasciculata, which parasites exclusively mosquitoes and is considered non-infective to humans. Phenotypic characterization of 2 of these isolates from the skin and bone marrow of the same patient showed that skin isolate was attenuated compared to the bone marrow isolate for survival in the spleen in BALB/c mice. However, both of them were able to infect and induce inflammation in the liver after 4 and 6 weeks post infection. Conversely, only skin isolate was able to survive in dermis, leading to ear swelling, accompanied of presence of parasites in the ear and lymph nodes. Besides that, bone marrow isolate infection in murine macrophages showed higher number of infected cells and also increased number of parasites within macrophages comparing to the skin isolate infection. Furthermore, bone marrow isolate negatively modulated the nitric oxide production in murine macrophages, corroborating with increased infection. Our findings raise concerns about an emerging infectious disease easily confused with Leishmaniasis, opening a research path towards epidemiological questions about identification of vectors, reservoirs, distribution and reassessment of Leishmaniasis cases in Brazil. Due to its proximity to the monoxenous Crithidia genus and the geographical location of atypical Leishmaniasis cases reported here, we proposed naming this new parasite species as Cridia sergipensis. Overall, this research provides a unique knowledge on phenotipic differences associated with diverse pathologies caused by Cridia infection. Protozoários Tripanossomatídeos do gênero Leishmania são parasitos transmitidos por vetores que infectam hospedeiros vertebrados, causando um amplo espectro de doenças denominadas leishmanioses. A gravidade das formas clínicas depende, dentre outros fatores, da espécie, assim, a identificação correta se torna crítica para o diagnóstico, prognóstico e tratamento. As ferramentas bioquímicas e moleculares disponíveis para caracterizar espécies de Leishmania são, na maioria das vezes, eficientes. Embora essas metodologias sejam importantes para o diagnóstico e identificação de Leishmania, são insuficientes para identificação conclusiva. Após isolar 7 amostras clínicas de parasitos, sendo alguns deles de casos atípicos de leishmaniose cutânea/mucocutânea e visceral, o principal objetivo foi identificar a espécie de Leishmania envolvida. Através da análise estrutural do citoesqueleto, que envolve a presença de flagelo e cinetoplasto, por exemplo, foi observado que esses parasitos compartilham características morfológicas com outros Tripanossomatídeos, entretanto, não se pode afirmar qual a espécie em questão. Uma vez que os resultados da identificação por PCR e eletroforese de isoenzimas foram inconclusivos, foi realizado o sequenciamento total do genoma. O sequenciamento dos isolados clínicos dos pacientes diagnosticados com leishmanioses mostrou que se trata de uma nova espécie patogênica dentro do grupo dos Tripanossomatídeos. Ao comparar as sequências codificantes dos genes ortólogos de 36 Tripanossomatídeos, foi observado que esses parasitos são intimamente relacionados com Crithidia fasciculata, parasito exclusivo de mosquitos, e considerado não patogênico para humanos. A caracterização fenotípica de 2 desses isolados clínicos, sendo um deles isolado da pele e outro da medula óssea do mesmo paciente mostrou que o isolado da pele não foi capaz de estabelecer doença no baço de camundongos BALB/c infectados. Por outro lado, ambos os isolados infectaram e induziram reação inflamatória no fígado dos animais após 4 e 6 semanas de infecção. No entanto, apenas o isolado da pele foi capaz de sobreviver na derme dos camundongos, induzindo inflamação local com presença de parasitos na orelha e linfonodos. Além disso, a infecção com o isolado da medula óssea em macrófagos murinos exibiu um maior número de células infectadas e maior número de amastigotas dentro das células, comparada ao da pele. Adicionalmente, o isolado da medula óssea foi capaz de modular negativamente a produção de óxido nítrico em macrófagos murinos, corroborando com o aumento da infecção. Essas descobertas levantam preocupações sobre uma doença infecciosa emergente facilmente confundida com leishmaniose, abrindo caminhos para a pesquisa nos campos epidemiológicos, de vetores, reservatórios, distribuição e reavaliação de casos de leishmaniose. Devido a proximidade com os parasitos do gênero Crithidia e a localização geográfica de onde foram isolados, a nomenclatura de Cridia sergipensis foi proposta para agrupar essa nova espécie. O presente estudo contribui também para o conhecimento das diferenças fenotípicas associadas com as diversas patologias causadas pela infecção com Cridia. Aracaju

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES Visceral Leishmaniasis (VL) is a serious infectious disease and is increasing geographical expansion and urbanization. patterns of transmission of the disease are altered due to human actions. The dog is considered the main reservoir of the parasite. The development of the disease depends in part on the host immune system. Neutrophils are considered the first line of defense against pathogens and important agents in the control of LV. These cells have an additional mechanism to eliminate microorganisms that occur with the release of extracellular networks (Neutrophil Extracellular Traps -NETs). This work has two overall objectives, the first being an epidemiological study of LV in Sergipe and the second evaluation of NETs formation in human neutrophils and dogs with LV stimulated with L. infantum, so this thesis was divided into chapters, for better understanding. Epidemiological evaluation were used SINAN data and serosurveys canines Zoonosis Aracaju-SE. A descriptive analysis of the data and construction of distribution maps of disease was performed. Sergipe Aracaju and from 2008 to 2014 the incidence rate in humans with LV increased and the percentage of infected dogs has doubled in the capital. The percentage of cases positive for human VL by sex, according to the age of the patient group, showed that over 15 years the disease affects more males. The spatial distribution analysis allowed to view areas of the city with the highest concentration of human and canine VL. The neighborhoods located in poor areas or growing areas were those with the highest incidence of the disease in humans and dogs. The results show that presents endemicity for human and canine VL. In order to evaluate the formation of NETs were used human groups: Control; LV treated and positive DTH. The dogs were divided into control; Asymptomatic and symptomatic. Neutrophils were isolated from patients with LV University Hospital / UFS and dogs diagnosed with VT by Zoonoses were incubated with and without stimulation of parasite and NETs measures in the supernatant of the cultures after 90 minutes. The parasitic load determined after 24 and 48 hours of interaction with neutrophils treated by the limiting dilution technique. In the evaluation of NETs induction of human neutrophils and release dogs with LV greater amount of NETs when stimulated with L. infantum. When compared between the groups individuals positive DTH release fewer NETs stimulated or not with the parasite. At 24 and 48 hours LV treated subjects have a higher parasite load and showed positive DTH fewer parasite than the other groups tested. Dogs with LV release signals greater amount of NETs when stimulated with L. infantum and have higher parasitic load after 48 hours. It is observed that the release of NETs and control of parasitic load by neutrophils varies according to the clinical form of the LV, in humans and dogs. A Leishmaniose Visceral (LV) é uma doença infecciosa grave e está em crescente expansão geográfica e urbanização. Os padrões de transmissão da doença são alterados devido às ações antrópicas. O cão é considerado o principal reservatório do parasito. O desenvolvimento da doença depende em parte do sistema imune do hospedeiro. Os neutrófilos são considerados a primeira linha de defesa do organismo contra agentes patogênicos e importantes no controle da LV. Estas células têm um mecanismo adicional para eliminar microorganismos que ocorre com a liberação de redes extracelulares (Neutrophil Extracelular Traps -NETs). Este trabalho teve dois objetivos gerais, sendo o primeiro um estudo epidemiológico da LV em Sergipe e o segundo a avaliação da formação de NETs em neutrófilos de humanos e cães com LV estimulados com L. infantum, assim essa tese foi dividida em capítulos, para melhor compreensão. Na avaliação epidemiológica foram utilizados dados do SINAN e inquéritos sorológicos caninos do Zoonoses de Aracaju-SE. Foi realizada uma análise descritiva dos dados e construção de mapas de distribuição da doença. Em Sergipe e Aracaju de 2008 a 2014 o coeficiente de incidência em humanos com LV aumentou e o percentual de cães infectados dobrou na capital. A porcentagem de casos positivos para LV humana por sexo, de acordo com a faixa etária do paciente, mostrou que acima de 15 anos a doença acomete mais o sexo masculino. A análise de distribuição espacial permitiu visualizar áreas da cidade com maior concentração de LV humana e canina. Os bairros situados em áreas com situação econômica desfavorável ou em zonas de expansão foram os que apresentaram maior incidência da doença em humanos e cães. Os resultados demonstram que apresenta caráter endêmico para a LV humana e canina. Com a finalidade de avaliar a formação de NETs foram utilizados grupos humanos: Controle; Tratados LV e DTH positivos. Os cães foram divididos em: Controle; Assintomáticos e sintomáticos. Foram isolados neutrófilos de pacientes com LV do Hospital Universitário/UFS e de cães diagnosticados com LV pelo Zoonoses foram incubadas, com e sem estímulo do parasito e NETs medidas no sobrenadante das culturas após 90 minutos. A Carga parasitária determinada após 24 e 48 horas de interação com neutrófilos tratados, através da técnica de diluição limitante. Na avaliação da indução de NETs, os neutrófilos de humanos e cães com LV liberam maior quantidade de NETs quando estimulados com L. infantum. Quando comparado entre os grupos indivíduos DTH positivos liberam menor quantidade de NETs estimulados ou não com o parasito. Em 24 e 48 horas indivíduos tratados com LV apresentam maior carga parasitária e os DTH positivos demonstraram menor quantidade do parasito do que os outros grupos testados. Os cães com sinais de LV liberam maior quantidade de NETs quando estimulados com L. infantum e apresentam maior carga parasitária após 48 horas. Observa-se que a liberação de NETs e o controle da carga parasitária através dos neutrófilos variam de acordo com a forma clínica da LV, em humanos e cães.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES Visceral leishmaniasis is a neglected disease caused by Leishmania infantum in Brazil. The Ecto-NTPDase activities, described in various parasites including Leishmania, is involved in adhesion, infection and virulence of these parasites, functioning as a molecular mechanism able to modulate the host immune response and promoting the disease progression. E-NTPDases are enzymes that hydrolyze nucleotides tri and/or di-phosphate into monophosphate products. The extracellular metabolism of these nucleotides has an important role in infection by L. infantum and other intracellular pathogens. These nucleotides are powerful extracellular signals involved in various cellular processes, which are part of the purinergic signalling that modulates the immune response. The objective of this study was to evaluate the role of ecto-NTPDase activities in adherence and infection of L. infantum in human macrophages. The ecto-NTPDase activities of parasites were evaluated in four different media or conditions: (1) purine-depleted, (2) adenosine-enriched, (3) with inhibitor of ecto-NTPDase activities, and (4) with anti-E antibody NTPDase 1. The effects of these treatments on the adhesion and infection of these parasites in human macrophages were investigated. Immunolocalization of the E-NTPDase in the membrane of L. infantum was also evaluated. The ecto-nucleotidase activity is modulated according to the availability of adenosine in the medium. Parasites cultivated in a purine-depleted medium showed increased hydrolytic capacity of nucleotides and higher infectivity. Parasites grown in adenosine-enriched medium had lower ecto-nucleotidase activity and infectivity. Growing parasites with an inhibitor of Ecto-NTPDase activities led to an increase in the adherence of Leishmania to non-phagocytic cells. E-NTPDase enzymes responsible for the hydrolysis of nucleotides are present in the membrane of L. infantum. The higher hydrolysis capacity is associated with increased adhesion capacity and infection of these parasites in human macrophages. A Leishmaniose visceral é uma doença negligenciada causada, no Brasil, pelo protozoário Leishmania infantum. A atividade ecto-nucleotidásica descrita em vários parasitos incluindo Leishmania sp está associada à nutrição, adesão, infecção e virulência destes, funcionando como um mecanismo molecular capaz de modular a resposta imune do hospedeiro, favorecendo a progressão da doença. As ecto-nucleotidases da família E-NTPDases são enzimas capazes de hidrolisar nucleotídeos tri e/ou di-fosfatados em seus produtos monofosfatados. O metabolismo extracelular destes nucleotídeos tem um papel importante na infecção por L. infantum e outros patógenos intracelulares. Estes nucleotídeos extracelulares são potentes sinalizadores envolvidos em vários processos celulares, incluindo a sinalização purinérgica que está relacionada à modulação da resposta imune. O objetivo deste trabalho foi avaliar o papel da atividade ecto-nucleotidásica em L. infantum na adesão e infecção de macrófagos humanos. Avaliação da atividade ecto-nucleotidásica de parasitos crescidos em meio depletado de purina, meio enriquecido com adenosina, incubados previamente com inibidor da atividade NTPDásica, ou na presença do anticorpo Anti E-NTPDase 1 e investigar os efeitos desses tratamentos na adesão e infecção desses parasitos em macrófagos humanos. A atividade ecto-nucleotidásica dos parasitas foram avaliadas em quatro condições diferentes: (1) na meio depletado de purina, (2) meio enriquecido com adenosina, (3) com o inibidor da atividade ecto-nucleotidásica, e (4) incubada com anticorpo anti-E NTPDase 1. foram investigados os efeitos desses tratamentos sobre a adesão e infecção destes parasitas em macrófagos humanos. Imunodetectamos a presença de E-NTPDases presentes na membrana de L. infantum, e comparamos a atividade ecto-nucleotidásica de diferentes cepas de L. infantum. A atividade ecto-nucleotidásica, é modulada de acordo com a disponibilidade de adenosina do meio, parasitos cultivados em meio depletado de purina apresentaram o aumento da capacidade hidrolítica de nucleotídeos, e maior capacidade infectiva, os parasitos cultivados em meio suplementado com adenosina apresentaram uma menor atividade ecto-nucleotidásica e uma redução de 30% na capacidade infectiva. O aumento da atividade ecto-nucleotidásica levou a um aumento na adesão em células não fagocíticas. As enzimas E-NTPDases estão presentes na membrana de L. infantum. Concluímos que atividade ecto-nucleotidásica está associada a maior capacidade de adesão e infecção destes parasitos in vitro.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Search for new therapeutic alternatives for leishmaniasis is considered essential by the World Health Organization, due to the high toxicity of the drugs currently used, its high cost and the risk of resistance. In Brazil, it is used three drugs to treat leishmaniasis: meglumine antimoniate and two formulations of amphotericin B (deoxycholate and liposomal). This study is a piggy-back evaluation linked to a multicentric clinical trial with the objective to perform a decision analysis between the treatment regimens for visceral leishmaniasis, for both adults and children. For this end, it was performed analysis of the data obtained in the state of Sergipe from 62 patients randomized to treatment with meglumine antimoniate (group A), considered the first line treatment in Brazil, amphotericin B deoxycholate (group B), liposomal amphotericin B (group C) and the combined regimen of meglumine antimoniate with liposomal amphotericin B (group D), in order to conduct a decision analysis from among these therapeutic regimens for both adults and children. To this end, two scenarios were considered: scenario 1 is the currently prevailing in Brazil, where patients are always treated as possible on an outpatient basis with meglumine antimoniate, although with a smaller monitoring on its potentially lethal side effects and a possible lower adherence to treatment. In the scenario 2, patients are hospitalized during the treatment period, with a better monitoring of clinical and laboratory parameters of the patient, although with a higher risk of nosocomial infections and increasing treatment costs. In terms of security, there is an emphasis on amphotericin B deoxycholate as the one that presents a greater amount of serious and potentially lethal adverse reactions, with an average of 2 severe reactions per patient. From the viewpoint of cost-efficacy, for scenario 1 the meglumine antimoniate remains the treatment of choice for both adults and children. However, in scenario 2, the combination of drugs (group D) presented itself as the most costeffective for both adults and children. From this study it became clear, from a safety point of view for the patient, the need to evaluate both therapeutic regimens, particularly amphotericin B deoxycholate, such as the treatment scenarios for visceral leishmaniasis. A busca por novas alternativas terapêuticas para as leishmanioses é considerada essencial pela Organização Mundial da Saúde, em virtude da elevada toxicidade dos medicamentos atualmente utilizados, seu alto custo e o risco de resistência. No Brasil, utilizam-se três drogas para o tratamento da leishmaniose: o antimoniato de meglumina e as formulações de anfotericina B (desoxicolato e lipossomal). Este estudo é uma análise econômica atrelada a um ensaio clínico multicêntrico, com o objetivo de se realizar uma análise de decisão entre os esquemas terapêuticos para a leishmaniose visceral, tanto para adultos como para crianças. Para tal, foram utilizados os dados obtidos do estado de Sergipe, onde foram acompanhados 62 pacientes, randomizados para tratamento com antimoniato de meglumina (grupo A), considerado o esquema de primeira escolha no Brasil, anfotericina B desoxicolato (grupo B), anfotericina B lipossomal (grupo C) e o esquema terapêutico combinado de antimoniato de meglumina com anfotericina B lipossomal (grupo D). Na análise de decisão, foram considerados dois cenários: o cenário 1, atualmente existente no Brasil, no qual os pacientes são tratados sempre que possível ambulatorialmente com o antimoniato de meglumina, porém com um menor acompanhamento de suas reações adversas potencialmente letais e uma possível menor adesão ao tratamento. Já no cenário 2, os pacientes são internados durante todo o período do tratamento, com um melhor monitoramento dos parâmetros clínicos e laboratoriais do paciente, porém com um risco maior de infecções hospitalares e aumento de custos do tratamento. No quesito segurança, há um destaque para a anfotericina B desoxicolato como sendo a que apresenta uma maior quantidade de reações adversas graves e potencialmente letais, com uma média de 2 reações graves/paciente. Já no ponto de vista de custo-eficácia, para o cenário 1 o antimoniato de meglumina continua sendo o tratamento de primeira escolha, tanto para adultos como para crianças. Porém, no cenário 2, a combinação de medicamentos (grupo D) se apresentou como o mais custo-eficaz tanto para adultos como para crianças. Deste estudo evidenciou-se, do ponto de vista de segurança ao paciente, a necessidade de reavaliação tanto dos esquemas terapêuticos, particularmente a anfotericina B desoxicolato, como dos cenários de tratamento para a leishmaniose visceral.

Conselho Nacional de Desenvolvimento Científico e Tecnológico A leishmaniose visceral ou calazar é uma doença negligenciada, fatal que afeta a população pobre nos países em desenvolvimento. Há falha no controle da transmissão da doença devido à demora no diagnóstico, tratamento e toxicidade das drogas utilizadas, necessitando de novas estratégias. Dentre as estratégias seria de extrema importância o desenvolvimento de uma vacina a fim de minimizar a frequência dos casos e eliminar a doença. Nesse estudo, avaliamos a resposta imune celular a 10 antígenos recombinantes (SMT, CPB, NH, A2, KMP11, VIZ92, NS, A2MAT, 8NSA e NSA) bem assim ao antígeno solúvel de Leishmania (SLA), dosando as seguintes citocinas: IL-1ß, IL-12p70, IFN-γ, IL-6, IL-10, IL-27, TNF-α. Células mononucleares do sangue periférico (CMSP) de 37 indivíduos: LV ativa, LV curado, DTH+ e controles sadios foi estimulado com os antígenos acima por 72 horas a 37ºC/5% de CO2. Ressaltamos que não houve alteração significativa de outras citocinas, além do INF-γ, TNF-α e IL-10, em resposta aos demais antígenos recombinantes não destacados nos resultados deste estudo. Assim, observamos concentração elevada de INF-γ e/ou TNF-α acompanhados de resposta ao IL-10 em indivíduos mais protegidos em resposta ao estímulo dos antígenos: CPB, VIZ92 e 8NSA; destacamos que em relação ao antígeno CPB tem-se concentração significativa de INF-γ no grupo DTH+ se comparado ao grupo LV ativa e correlação direta entre as concentrações de INF-γ e TNF-α entre estes dois grupos. Sugerimos que os antígenos CPB, VIZ92 e 8NSA possam estar associados à indução de resposta protetora e ser promissores para utilização em imunoterapia e imunoprofilaxia. A leishmaniose visceral ou calazar é uma doença negligenciada, fatal que afeta a população pobre nos países em desenvolvimento. Há falha no controle da transmissão da doença devido à demora no diagnóstico, tratamento e toxicidade das drogas utilizadas, necessitando de novas estratégias. Dentre as estratégias seria de extrema importância o desenvolvimento de uma vacina a fim de minimizar a frequência dos casos e eliminar a doença. Nesse estudo, avaliamos a resposta imune celular a 10 antígenos recombinantes (SMT, CPB, NH, A2, KMP11, VIZ92, NS, A2MAT, 8NSA e NSA) bem assim ao antígeno solúvel de Leishmania (SLA), dosando as seguintes citocinas: IL-1ß, IL-12p70, IFN-γ, IL-6, IL-10, IL-27, TNF-α. Células mononucleares do sangue periférico (CMSP) de 37 indivíduos: LV ativa, LV curado, DTH+ e controles sadios foi estimulado com os antígenos acima por 72 horas a 37ºC/5% de CO2. Ressaltamos que não houve alteração significativa de outras citocinas, além do INF-γ, TNF-α e IL-10, em resposta aos demais antígenos recombinantes não destacados nos resultados deste estudo. Assim, observamos concentração elevada de INF-γ e/ou TNF-α acompanhados de resposta ao IL-10 em indivíduos mais protegidos em resposta ao estímulo dos antígenos: CPB, VIZ92 e 8NSA; destacamos que em relação ao antígeno CPB tem-se concentração significativa de INF-γ no grupo DTH+ se comparado ao grupo LV ativa e correlação direta entre as concentrações de INF-γ e TNF-α entre estes dois grupos. Sugerimos que os antígenos CPB, VIZ92 e 8NSA possam estar associados à indução de resposta protetora e ser promissores para utilização em imunoterapia e imunoprofilaxia.

Leishmaniose visceral (LV) é uma doença grave causada pelo protozoário do gênero Leishmania. Caracterizada por uma infecção do sistema reticuloendotelial e intensa resposta inflamatória, a LV clássica apresenta como principais manifestações clínicas: hepatoesplenomegalia, febre e edema, além de neutropenia, plaquetopenia e hipergamaglobulinemia. A presença da inflamação mediada por IFN-g e IL-12, teoricamente controlaria a infecção, contudo, o que se observa é a participação de citocinas Th2 associadas a IL-10. Entretanto, não está claro porque 90% dos indivíduos infectados não desenvolvem a doença enquanto outros desenvolvem a forma clássica e alguns evoluem para forma mais severa da doença sem responder ao tratamento. Considerando esse espectro de respostas o objetivo deste trabalho foi avaliar a resposta imunológica de indivíduos infectados com Leishmaniachagasie associá-lacom o desenvolvimento da doença bem como a resposta terapêutica. Para associar perfil de citocinas com desenvolvimento da doença, soros de pacientes com diagnóstico confirmado para LV foram avaliados quanto a presença de citocinas, sCD14, MIF e LPS e então comparados com controles saudáveis e indivíduos assintomáticos. Os resultados sugerem que sCD14 pode estar iniciando a ativação da resposta inflamatória via IFN-g e estimulando a produção de IL-27, IL-10 e IL-6. Estas estão fortemente associadas com hepatoesplenomegalia, neutropenia e plaquetopenia. Foi observado também que indivíduos com IL-6 maior que 200pg/mL evoluíram para óbito. Para avaliar a regulação de macrófagos na LV, células mononucleares do sangue periférico foram isoladas de sangue venoso de indivíduos assintomáticos e sintomático, diferenciadas em macrófagos e em seguida infectados com Leishmaniachagasi nos tempos de 2, 24, 48 e 72 horas. Os sobrenadantes das infecções foram avaliados quanto a presença de citocinas e as células coradas para avaliação doíndice de infecção. Apesar de no início da infecção os indivíduos assintomáticos apresentaram maior percentual de macrófagos infectados em relação aos sintomáticos, foi observado um controle da infecção já com 24 horas. Os indivíduos sintomáticos, pelo contrário, exibiram um aumento tanto no percentual de macrófagos infectados quanto no número de amastigotas por macrófagos e mesmo após 48 horas não controlou a infecção. Macrófagos de indivíduos assintomáticos apresentaram maior produção IL-12p70 após duas horas de infecção, enquanto que macrófagos de indivíduos sintomáticos apresentam maior produção de IL-10. Esses resultados sugerem que o controle da infecção ocorre já no início da infecção pela produção de citocinas pró-inflamatórias em detrimento das moduladoras. Em adição, para identificarbiomarcadores imunológicos de bom prognóstico da LV pós tratamento, soros de pacientes foram coletados antes, durante e após o tratamento e avaliados quanto a produção de citocinas, quimiocinas e metaloproteinases. Os dados indicam que IFN-g, TNF-a, IL-10 e IL-17, FGF e VEGF associados com a clínica podem compor um painel de triagem para desenvolvimento de novos fármacos contra LV. Juntos nossos dados reforçam a estreita relação de IFN-g, IL-10, IL-6, TNF-a e IL-27 na dinâmica imunológica da LV e sugerem a participação direta de sCD14 na ativação da resposta imune frente a Leishmaniachagasi.

Visceral leishmaniasis is endemic in many Brazilian states, including Sergipe. Through a cross-sectional study done with relatives of patients with visceral leishmaniasis, we determined the frequency of asymptomatic infection by Leishmania chagasi using Montenegro skin test and/or Enzyme Linked Immunosorbent Assay (ELISA). Questionnaire containing socio-demographic data, presence of signs and symptoms, physical examination of 109 subjects were analyzed. Environmental factors such as the presence of dog, chicken and wood in the residence were also investigated. Finally, the levels of cytokines Tumor Necrosis Factor (TNF)-α, Transforming Growth Factor (TGF)-β and Interleukin (IL)-1, IL-6, IL-8, IL-10 and IL-12 were measured in 59 of these individuals. The frequency of positivity was detected in 51.8% of participants, being higher in males (61.9%). Positive testes were also more common in people over 14 years (79.7%). All positive individuals were considered asymptomatic and did not return to the hospital with complaints until the end of the study, suggesting that did not progress to other clinical forms. There was no difference between the origin: capital or the state, but more positive cases occurred at the capital, confirming the growing urbanization of the disease. Environmental factors was statistically significant when analyzed presence of dog and waste wood together (p

Visceral leishmaniasis or kala-azar is a zoonotic disease of domestic and wild canids, caused by the protozoan Leishmania chagasi and transmitted between dogs and man by the vector Lutzomyia longipalpis. The purpose of this research was to analize the spatial distribution of human and canine cases of visceral leishmaniasis in Aracaju, Sergipe from 2005 to 2012; characterize a nitric oxide resistant and susceptible isolates of L. chagasi from dogs in Aracaju, performing proteomic analysis of it by bidimensional electrophoresis and checking their interaction between dog and fox macrophages. In the analysis of the spatial distribution of the disease during the study period, there were 2060 canine and 110 human cases. It was found that the neighborhoods, Zona de Expansão and Mosqueiro, peripheral areas with lower population densities, recent occupation, poor urban infrastructure, had a number of significant human and canine cases. In Dezoito do Forte, Santos Dumont and America neighborhoods, were isolated nitric oxide resistant Leishmania in dogs and in humans, which could confirm the dog as a source of infection for humans and maybe could confirmation that disease in dogs precedes the occurrence of the disease in humans. Infection of fox macrophages by strains of L. chagasi, revealed that the macrophages control the infection rapidly regardless of whether the strain is resistant or not to nitric oxide. This result corroborates the fact that wild canids such as foxes, have an important role in the sylvatic cycle of visceral leishmaniasis, serving as a reservoir for the disease and rarely displaying clinical forms of the disease, unlike the domestic dog. Proteomic analysis was performed in promastigotes in logarithmic and stationary phase showing a difference in the expression of many proteins in both the promastigote growth phases. As an interesting fact we observed an over-expression of a cytosolic malate dehydrogenase and that it may probably be related to an intrinsic resistance of this isolate to nitric oxide. Related to resistance to nitric oxide, overexpression of trypanothione peroxidase was constant at all stages of the growth curve. Also observed was the overexpression of a calpain that may be responsible for inducing greater susceptibility to antimonials and lower susceptibility to miltefosine. A leishmaniose visceral (LV) ou calazar é uma zoonose de canídeos silvestres e domésticos, causada pelo protozoário Leishmania chagasi e transmitida entre os canídeos e destes ao homem pelo vetor Lutzomyia longipalpis. Os objetivos deste estudo são analisar a distribuição temporo-espacial de casos de leishmaniose visceral canina e humana em Aracaju, Sergipe de 2005 a 2012; avaliar e comparar a regulação de macrófagos de cão e de raposas na infecção por isolados de L. chagasi através do índice de infecção de macrófagos, e realizar análise proteômica das fases de crescimento logarítmica e estacionária de promastigotas de isolados de L. chagasi resistentes e susceptíveis ao óxido nítrico, através da técnica de eletroforese bidimensional. Na análise da distribuição espacial da doença aconteceram 2060 casos caninos e 110 casos humanos. Foi verificado que os bairros da Zona de Expansão e Mosqueiro, áreas periféricas, com mais baixas densidades populacionais, ocupação recente, baixa renda e infraestrutura urbana pouco presente, tiveram um número de casos caninos e humanos expressivos. Nos bairros Dezoito do Forte, Santos Dumont e América, foram isoladas Leishmania resistentes ao óxido nítrico de cães e humanos, o que poderia confirmar o cão como fonte de infecção para o homem e, talvez, uma confirmação que a doença em cães pode preceder a ocorrência da doença em humanos. A infecção de macrófagos de raposa por cepas de L. chagasi resistentes e susceptíveis ao óxido nítrico, revelou que esses controlam rapidamente a infecção independentemente se a cepa é resistente ou não ao óxido nítrico. Este resultado corrobora o fato de que os canídeos silvestres, como as raposas, têm um papel importante no ciclo silvestre da leishmaniose visceral, servindo de reservatório para a doença e raramente apresentando a forma clínica da doença, diferente do cão doméstico. Foi realizada a análise proteômica de promastigotas em fase logarítmica e em fase estacionária revelando uma diferença na expressão de várias proteínas em ambas as fases de crescimento do promastigota. Como fato mais interessante observou-se um aumento acentuado na expressão de uma malato desidrogenase citosólica e que provavelmente apresenta relação com a resistência intrínseca deste isolado ao óxido nítrico. Relacionada com a resistência ao óxido nítrico, um aumento na expressão da tripanotiona peroxidase foi constante em todas as fases de curva de crescimento. Também foi observado um aumenta na expressão de uma calpaína que poderia ser responsável por induzir susceptibilidade maior aos antimoniais e menor susceptibilidade à miltefosina.

Submitted by Hiolanda Rêgo (hiolandarego@gmail.com) on 2014-07-23T17:58:32Z No. of bitstreams: 1 Tese_ICS_ Cláudia Leal Macêdo.pdf: 5476501 bytes, checksum: 7e98dd0076ef773e91e0d9b2c5b6c101 (MD5) Approved for entry into archive by Flávia Ferreira (flaviaccf@yahoo.com.br) on 2014-07-26T03:20:19Z (GMT) No. of bitstreams: 1 Tese_ICS_ Cláudia Leal Macêdo.pdf: 5476501 bytes, checksum: 7e98dd0076ef773e91e0d9b2c5b6c101 (MD5) Made available in DSpace on 2014-07-26T03:20:19Z (GMT). No. of bitstreams: 1 Tese_ICS_ Cláudia Leal Macêdo.pdf: 5476501 bytes, checksum: 7e98dd0076ef773e91e0d9b2c5b6c101 (MD5) O carcinoma infiltrante da mama apresenta características clínicas, morfológicas e evolutivas diversas que parecem traduzir a heterogeneidade tumoral. Semelhanças entre padrões de expressão gênica e imunoistoquímica dos receptores hormonais, do HER2 e das citoqueratinas basais e luminais permitiram agrupar tumores com padrões morfológicos e imunofenotípicos diferentes, identificando subtipos: luminal A (RE+,RP+,HER2-); luminal B (RE+,RP+,HER2+); HER2 + (RE-,RP-,HER2+); triplonegativo (RE-,RP-,HER2-); basal-símile (RE-,RP-,HER2-CK5/6+ e/ou EFGR+) e “mama normal” (RE-,RP-,HER2-CK5/6- e/ou EFGR-). No presente estudo buscou-se correlacionar características clínico-patológicas e evolutivas com perfil imunoistoquímico dos subtipos moleculares em mulheres mastectomizadas. Nas análises estatísticas empregaram-se os métodos de Kaplan-Meier e a regressão de Cox. Utilizaram-se receptores hormonais, anti-c-erbB-2, anti-CK5/6, anti-EGFR, anti- CK8, anti-p63. Calcularam-se as associações entre variáveis pelas razões de prevalência. Observaram-se média de idade inferior a dos países desenvolvidos além de maior frequência de tumores pT2, de grau II e do ductal infiltrante e elevada frequência de metástases axilares. A sobrevida global foi de 80,1% em um período de dez anos. Comprometimento das margens cirúrgicas, metástases à distância e status menopausal foram variáveis importantes na sobrevida. Observou-se frequência elevada dos subtipos triplo-negativo e basal-símile e associações significantes entre estes subtipos e o HER2 + com tumores de maior dimensão (pT3) e grau histológico III confirmando que tem pior prognóstico. Expressão positiva de CK5/6 e EGFR foi a única diferença entre o subtipo basal-símile e o triplo-negativo, indicando que a subdivisão do triplo-negativo não seria necessária. Concluiu-se que os subtipos podem ser caracterizados pela avaliação dos fatores clínico-patológicos somados ao perfil imunoistoquímico.

Leishmaniasis is a serious public health problem worldwide, requiring effective measures for its control and treatment. Visceral leishmaniasis is the most severe form and has a lethal potential. Depending on the interaction between the parasite and the host immune response, different clinical manifestations may develop: asymptomatic, oligosymptomatic and symptomatic. Macrophages play a central role in immune response. However, their microbicidal activity may be impaired, leading to the survival and proliferation of the parasites inside them. The objective of this study was to evaluate the leishmanicidal activity of macrophages isolated from treated and asymptomatic visceral leishmaniasis subjects after infection by Leishmania chagasi. Our hypothesis was that macrophages isolated from treated subjects were more susceptible to infection by Leishmania chagasi in relation to asymptomatic patients. From a cross-sectional study, 05 symptomatic patients and 02 asymptomatic were investigated at University Hospital. Mononuclear cells were isolated from peripheral blood of 07 patients in order to obtain monocytes and further differentiation into macrophages. These were infected with the isolated LVHSE-07 from L. chagasi for 2, 72 and 96 hours in conditions with and without stimulus of LPS and IFN-. To examine whether macrophages responded differently to the symptomatic patient's own Leishmania, 03 symptomatic and 02 asymptomatic patients were reconvened, so that their macrophages would be infected with Leishmania from symptomatic patients (LVHSE-24, LVHSE-28, LVHSE-37). Leishmanicide response of macrophages was assessed by determining the percentage of infected macrophages and the number of amastigotes inside macrophages. The results suggest that the macrophages of the symptomatic patients were more infected, especially when used patient's own Leishmania, and they did not control infection, even in the presence of stimulus, unlike asymptomatic's macrophages, which controlled infection, independently of what Leishmania was used. This study demonstrated that macrophages from symptomatic patients are more susceptible to infection by Leishmania chagasi, when compared with macrophages from asymptomatic patients; macrophages from symptomatic patients are more infected with their own isolated of L. chagasi; macrophages from asymptomatic patients are less infected with L. chagasi and have a parasitic load lower than the macrophages of symptomatic patients, regardless of the Leishmania isolated used in infection. It follows that macrophages are related to susceptibility or resistance to infection by Leishmania chagasi. As leishmanioses são um grave problema de saúde pública no mundo, demandando medidas eficazes no seu controle e tratamento. A leishmaniose visceral é a forma mais grave e pode ser letal, caso não seja tratada. A depender da interação entre o parasita e a resposta imune do hospedeiro, podem-se desenvolver diferentes manifestações clínicas: forma assintomática, oligossintomática e sintomática. Os macrófagos desempenham um papel fundamental na resposta imune. Entretanto, suas atividades leishmanicidas podem estar prejudicadas, levando à sobrevivência e proliferação dos parasitas no seu interior. Assim, o objetivo geral deste trabalho foi avaliar a atividade leishmanicida de macrófagos infectados por Leishmania chagasi isolados de indivíduos tratados e assintomáticos para Leishmaniose visceral. A nossa hipótese era que macrófagos isolados de indivíduos tratados eram mais susceptíveis à infecção por Leishmania chagasi em relação aos de pacientes assintomáticos. A partir de um estudo transversal, foram investigados 05 indivíduos tratados e 02 assintomáticos do Hospital Universitário. Células mononucleares do sangue periférico foram isoladas dos 07 pacientes para obtenção de monócitos e posterior diferenciação em macrófagos. Estes foram infectados com o isolado LVHSE-07 de L. chagasi durante 2, 72 e 96 horas em condições sem e com estímulo de LPS e IFN-. Para analisar se os macrófagos respondiam de maneira diferenciada com a Leishmania do próprio indivíduo tratado, foram reconvocados 03 tratados e 02 assintomáticos para infectar seus macrófagos com as Leishmanias dos tratados (LVHSE-24, LVHSE-28, LVHSE-37). A resposta leishmanicida dos macrófagos foi analisada determinando-se o percentual de macrófagos infectados e o número de amastigotas no interior dos macrófagos. Os resultados sugerem que os macrófagos dos indivíduos tratados foram mais infectados, principalmente quando utilizada a Leishmania do próprio indivíduo, e não controlaram a infecção mesmo na presença do estímulo, ao contrário dos macrófagos dos assintomáticos que sugerem um controle da infecção independente da Leishmania utilizada. Este estudo mostrou que macrófagos de indivíduos tratados são mais susceptíveis à infecção por Leishmania chagasi quando comparados com macrófagos de pacientes assintomáticos; macrófagos indivíduos tratados são mais infectados com seus próprios isolados de L. chagasi; macrófagos de pacientes assintomáticos são menos infectados por L. chagasi e apresentam uma carga parasitária menor que os macrófagos de indivíduos tratados independente do isolado de Leishmania utilizado na infecção. Conclui-se que os macrófagos estão relacionados com a susceptibilidade ou resistência à infecção por Leishmania chagasi.