MARCIA DALASTRA LAURENTI
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Patologia, Faculdade de Medicina - Docente
LIM/50 - Laboratório de Patologia das Moléstias Infecciosas, Hospital das Clínicas, Faculdade de Medicina - Líder
LIM/50 - Laboratório de Patologia das Moléstias Infecciosas, Hospital das Clínicas, Faculdade de Medicina - Líder
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- SUSCEPTIBILITY OF PERITONEAL MACROPHAGE FROM DIFFERENT SPECIES OF NEOTROPICAL PRIMATES TO EX VIVO Leishmania (L.) infantum chagasi-INFECTION(2012) CARNEIRO, Liliane Almeida; LAURENTI, Marcia Dalastra; CAMPOS, Marliane Batista; GOMES, Claudia Maria de Castro; CORBETT, Carlos Eduardo Pereira; SILVEIRA, Fernando TobiasThis study examined the susceptibility of peritoneal macrophage (PM) from the Neotropical primates: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus and Callimico goeldii to ex vivo Leishmania (L.) infantum chagasi-infection, the etiological agent of American visceral leishmaniasis (AVL), as a screening assay for evaluating the potential of these non-human primates as experimental models for studying AVL. The PM-susceptibility to infection was accessed by the PM-infection index (PMI) at 24, 72 h and by the mean of these rates (FPMI), as well as by the TNF-alpha, IL-12 (Capture ELISA) and Nitric oxide (NO) responses (Griess method). At 24h, the PMI of A. azarae infulatus (128) was higher than those of C. penicillata (83), C. goeldii (78), S. sciureus (77) and C. jacchus (55). At 72h, there was a significant PMI decrease in four monkeys: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) and C. jacchus (55/12), with exception of C. goeldii (78/54). The FPMI of A. azarae infulatus (82.5) and C. goeldii (66) were higher than C. jacchus (33.5), but not higher than those of C. penicillata (60.5) and S. sciureus (57.5). The TNF-alpha response was more regular in those four primates which decreased their PMI at 24/72 h: C. jacchus (145/122 pg/mu L), C. penicillata (154/130 pg/mu L), S. sciureus (164/104 pg/mu L) and A. azarae infulatus (154/104 pg/mu L), with exception of C. goeldii (38/83 pg/mu L). The IL-12 response was mainly prominent in A. infulatus and C. goeldii which presented the highest FPMI and, the NO response was higher in C. goeldii, mainly at 72 h. These findings strongly suggest that these New World primates have developed a resistant innate immune response mechanism capable of controlling the macrophage intracellular growth of L. (L.) i. chagasi-infection, which do not encourage their use as animal model for studying AVL.
- Human cutaneous leishmaniasis: interferon-dependent expression of double-stranded RNA-dependent protein kinase (PKR) via TLR2(2011) VIVARINI, Aislan de Carvalho; PEREIRA, Renata de Meirelles Santos; TEIXEIRA, Karina Luiza Dias; CALEGARI-SILVA, Teresa Cristina; BELLIO, Maria; LAURENTI, Marcia Dalastra; CORBETT, Carlos Eduardo Pereira; GOMES, Claudia Maria de Castro; SOARES, Rodrigo Pedro; SILVA, Aristobolo Mendes; SILVEIRA, Fernando Tobias; LOPES, Ulisses GazosWe investigated the type I interferon (IFN-1)/PKR axis in the outcome of the Leishmania (Leishmania) amazonensis infection, along with the underlying mechanisms that trigger and sustain this signaling pathway. Reporter assays of cell extracts from RAW-264.7 macrophages infected with L. (L.) amazonensis or HEK-293T cells cotransfected with TLR2 and PKR promoter constructions were employed. Primary macrophages of TLR2-knockout (KO) or IFNR-KO mice were infected, and the levels of PKR, IFN-1, and superoxide dismutase 1 (SOD1) transcript levels were investigated and compared. Immunohistochemical analysis of human biopsy lesions was evaluated for IFN-1 and PKR-positive cells. Leishmania infection increased the expression of PKR and IFN-beta on induction of PKR-promoter activity. The observed effects required the engagement of TLR2. TLR2-KO macrophages expressed low IFN-beta and PKR levels postinfection with a reduced parasite load. We also revealed the requirement of PKR signaling for Leishmania-induced IFN-1 expression, responsible for sustaining PKR expression and enhancing infection. Moreover, during infection, SOD1 transcripts increased and were also enhanced when IFN-1 was added to the cultures. Remarkably, SOD1 expression was abrogated in infected, dominant-negative PKR-expressing cells. Finally, lesions of patients with anergic diffuse cutaneous leishmaniasis exhibited higher levels of PKR/IFN1-expressing cells compared to those with single cutaneous leishmaniasis. In summary, we demonstrated the mechanisms and relevance of the IFN-1/PKR axis in the Leishmania infection.-De Carvalho Vivarini, A., Pereira, R. M. S., Dias Teixeira, K. L., Calegari-Silva, T. C., Bellio, M., Laurenti, M. D., Corbett, C. E. P., de Castro Gomes, C. M., Soares, R. P., Mendes Silva, A., Silveira, F. T., Lopes, U. G. Human cutaneous leish-maniasis: interferon-dependent expression of double-stranded RNA-kinase (PKR) via TLR2. FASEB J. 25, 4162-4173 (2011). www.fasebj.org
- Severe visceral leishmaniasis in children: the relationship between cytokine patterns and clinical features(2013) GAMA, Monica Elinor Alves; GOMES, Claudia Maria de Castro; SILVEIRA, Fernando Tobias; LAURENTI, Marcia Dalastra; GONCALVES, Eloisa da Graca; SILVA, Antonio Rafael da; CORBETT, Carlos Eduardo PereiraIntroduction: The relationship between severe clinical manifestations of visceral leishmaniasis (VL) and immune response profiles has not yet been clarified, despite numerous studies on the subject. This study aimed to investigate the relationship between cytokine profiles and the presence of immunological markers associated with clinical manifestations and, particularly, signs of severity, as defined in a protocol drafted by the Ministry of Health ( Brazil). Methods: We conducted a prospective, descriptive study between May 2008 and December 2009. This study was based on an assessment of all pediatric patients with VL who were observed in a reference hospital in Maranhao. Results: Among 27 children, 55.5% presented with more than one sign of severity or warning sign. Patients without signs of severity or warning signs and patients with only one warning sign had the highest interferon-gamma (IFN-gamma) levels, although their interleukin 10 (IL-10) levels were also elevated. In contrast, patients with the features of severe disease had the lowest IFN-gamma levels. Three patients who presented with more than two signs of severe disease died; these patients had undetectable interleukin 2 (IL-2) and IFN-gamma levels and low IL-10 levels, which varied between 0 and 36.8pg/mL. Conclusions: Our results showed that disease severity was associated with low IFN-gamma levels and elevated IL-10 levels. However, further studies with larger samples are needed to better characterize the relationship between disease severity and cytokine levels, with the aim of identifying immunological markers of active-disease severity.
- Preclinical diagnosis of American visceral leishmaniasis during early onset of human Leishmania (L.) infantum chagasi-infection(2014) LIMA, Luciana Vieira do Rego; RAMOS, Patricia Karla Santos; CAMPOS, Marliane Batista; SANTOS, Thiago Vasconcelos dos; GOMES, Claudia Maria de Castro; LAURENTI, Marcia Dalastra; CORBETT, Carlos Eduardo Pereira; SILVEIRA, Fernando TobiasAmerican visceral leishmaniasis (AVL) is an infectious disease, often with long-duration evolution, caused by Leishmania (L.) infantum chagasi. However, although the disease is considered the major clinical manifestation of the link between L. (L.) i. chagasi and the human immune response, we have recently identified five clinical-immunological profiles of infection in the Brazilian Amazon: three asymptomatic (Asymptomatic Infection - AI, Sub-clinical Resistant Infection - SRI, and Indeterminate Initial Infection III), and two symptomatic ones [Symptomatic Infection - SI (=AVL) and Sub-clinical Oligosymptomatic Infection - SOI]. We confirm here the preclinical diagnosis of AVL through the IgM-antibody response in a case of an early infection (profile III) that evolved to the full disease after 6 weeks.`
- SUSCEPTIBILITY OF Cebus apella MONKEY (PRIMATES: CEBIDAE) TO EXPERIMENTAL Leishmania (L.) infantum chagasi-INFECTION(2011) CARNEIRO, Liliane Almeida; SILVEIRA, Fernando Tobias; CAMPOS, Marliane Batista; BRIGIDO, Maria do Carmo de Oliveira; GOMES, Claudia Maria C.; CORBETT, Carlos E. P.; LAURENTI, Marcia D.In Amazonian Brazil, the Cebus apella monkey (Primates: Cebidae) has been associated with the enzootic cycle of Leishmania (V.) shawi, a dermotropic parasite causing American cutaneous leishmaniasis (ACL). It has also been successfully used as animal model for studying cutaneous leishmaniasis. In this work, there has been investigated its susceptibility to experimental Leishmania (L.) infantum chagasi-infection, the etiologic agent of American visceral leishmaniasis (AVL). There were used ten C. apella specimens, eight adult and two young, four males and six females, all born and raised in captivity. Two experimental infection protocols were performed: i) six monkeys were inoculated, intra-dermal via (ID), into the base of the tail with 2 x 10(6) promastigotes forms from the stationary phase culture medium; ii) other four monkeys were inoculated with 3 x 10(7) amastigotes forms from the visceral infection of infected hamsters by two different via: a) two by intravenous via (IV) and, b) other two by intra-peritoneal via (IP). The parameters of infection evaluation included: a) clinical: physical exam of abdomen, weigh and body temperature; b) parasitological: needle aspiration of the bone-marrow for searching of amastigotes (Giemsa-stained smears) and promastigotes forms (culture medium); c) immunological: Indirect fluorescence antibody test (IFAT) and, Delayed-type hypersensitivity (DTH). In the six monkeys ID inoculated (promastigotes forms) all parameters of infection evaluation were negative during the 12 months period of follow-up. Among the four monkeys inoculated with amastigotes forms, two IV inoculated showed the parasite in the bone-marrow from the first toward to the sixth month p. i. and following that they cleared the infection, whereas the other two IP inoculated were totally negative. These four monkeys showed specific IgG-antibody response since the third month p. i. (IP: 1/80 and IV: 1/320 IgG) toward to the 12(th) month (IP: 1/160 and IV: 1/5120). The DTH-conversion occurred in only one IV inoculated monkey with a strong (30 mm) skin reaction. Considering these results, we do not encourage the use of C. apella monkey as animal model for studying the AVL.
- Evaluation of systemic immunity in atypical cutaneous leishmaniasis caused by Leishmania (L.) infantum chagasi(2022) LAURENTI, Marcia Dalastra; SOSA-OCHOA, Wilfredo; FLORES, Gabriela Venicia Araujo; PACHECO, Carmen Maria Sandoval; TOMOKANE, Thaise Yumie; OLIVEIRA, Luanda Mara da Silva; ZUNIGA, Concepcion; SILVEIRA, Fernando Tobias; CORBETT, Carlos Eduardo PereiraIn some central-American countries, Leishmania (L.) infantum chagasi infection can cause non-ulcerated or atypical cutaneous leishmaniasis (NUCL) in addition to the classic clinical form, visceral leishmaniasis (VL). Little is known about the host-parasite relationship that can contribute to the determination of one or another clinical form. The present study had the objective to evaluate the humoral and cellular immunity in the sera of individuals affected by NUCL to improve the comprehension of this atypical host-parasite interaction. Based on clinical and laboratory diagnosis, serum of 80 individuals was collected to evaluate the cytokines and immunoglobulins profile of NUCL (n = 47), VL patients (n = 5), and negative controls (n = 28). Cytokines were detected using Cytokine Bead Array (CBA) Human Th1/Th2/Th17 kit according to the manufacturer's instructions; class (IgG and IgM), and subclass of (IgG1 and IgG2) immunoglobulins was evaluated by ELISA using specific antigens. The concentration of TNF-alpha, IFN-gamma, IL-2 and IL-4 cytokines in NUCL, VL and control was present below the detection threshold of CBA kit. IL-6, IL-10 and IL-17A cytokines was lower in NUCL compared to LV patients. Regarding to immunoglobulins, NUCL patients produced 4.0 times more IgG than the control, while VL patients produced 6.6 times more; and IgM level was 1.6 times higher in NUCL and 2.6 times in VL patients compared to the control. Concerning the immunoglobulins subclass, only VL patients showed positive reaction for IgG1, and IgG2 did not show positive reaction among the groups. The results showed a weak cellular and humoral systemic immune response in NUCL patients.
- Comparative Genomic Analyses of New and Old World Viscerotropic Leishmanine Parasites: Further Insights into the Origins of Visceral Leishmaniasis Agents(2023) SILVEIRA, Fernando Tobias; SOUSA, Edivaldo Costa; SILVESTRE, Rodrigo Vellasco Duarte; SANTOS, Thiago Vasconcelos dos; SOSA-OCHOA, Wilfredo; VALERIANO, Concepcion Zuniga; RAMOS, Patricia Karla Santos; CASSEB, Samir Mansour Moraes; LIMA, Luciana Vieira do Rego; CAMPOS, Marliane Batista; MATTA, Vania Lucia da; GOMES, Claudia Maria; FLORES, Gabriela V. Araujo; PACHECO, Carmen M. Sandoval M.; CORBETT, Carlos Eduardo; LAURENTI, Marcia DalastraVisceral leishmaniasis (VL), also known as kala-azar, is an anthropozoonotic disease affecting human populations on five continents. Aetiologic agents belong to the Leishmania (L.) donovani complex. Until the 1990s, three leishmanine parasites comprised this complex: L. (L.) donovani Laveran & Mesnil 1903, L. (L.) infantum Nicolle 1908, and L. (L.) chagasi Lainson & Shaw 1987 (=L. chagasi Cunha & Chagas 1937). The VL causal agent in the New World (NW) was previously identified as L. (L.) chagasi. After the development of molecular characterization, however, comparisons between L. (L.) chagasi and L. (L.) infantum showed high similarity, and L. (L.) chagasi was then regarded as synonymous with L. (L.) infantum. It was, therefore, suggested that L. (L.) chagasi was not native to the NW but had been introduced from the Old World by Iberian colonizers. However, in light of ecological evidence from the NW parasite's enzootic cycle involving a wild phlebotomine vector (Lutzomyia longipalpis) and a wild mammal reservoir (the fox, Cerdocyon thous), we have recently analyzed by molecular clock comparisons of the DNA polymerase alpha subunit gene the whole-genome sequence of L. (L.) infantum chagasi of the most prevalent clinical form, atypical dermal leishmaniasis (ADL), from Honduras (Central America) with that of the same parasite from Brazil (South America), as well as those of L. (L.) donovani (India) and L. (L.) infantum (Europe), which revealed that the Honduran parasite is older ancestry (382,800 ya) than the parasite from Brazil (143,300 ya), L. (L.) donovani (33,776 ya), or L. (L.) infantum (13,000 ya). In the present work, we have now amplified the genomic comparisons among these leishmanine parasites, exploring mainly the variations in the genome for each chromosome, and the number of genomic SNPs for each chromosome. Although the results of this new analysis have confirmed a high genomic similarity (similar to 99%) among these parasites [except L. (L.) donovani], the Honduran parasite revealed a single structural variation on chromosome 17, and the highest frequency of genomic SNPs (more than twice the number seen in the Brazilian one), which together to its extraordinary ancestry (382,800 ya) represent strong evidence that L. (L.) chagasi/L. (L.) infantum chagasi is, in fact, native to the NW, and therefore with valid taxonomic status. Furthermore, the Honduran parasite, the most ancestral viscerotropic leishmanine parasite, showed genomic and clinical taxonomic characteristics compatible with a new Leishmania species causing ADL in Central America.
- Whole-Genome Sequencing of Leishmania infantum chagasi Isolates from Honduras and Brazil(2021) SILVEIRA, Fernando Tobias; SOUSA JUNIOR, Edivaldo Costa; SILVESTRE, Rodrigo Vellasco; COSTA-MARTINS, Andre Guilherme; PINHEIRO, Kenny da Costa; OCHOA, Wilfredo Sosa; SANTOS, Thiago Vasconcelos dos; RAMOS, Patricia Karla; CASSEB, Samir; SILVA, Sandro Patroca da; VALERIANO, Concepcion Zuniga; LIMA, Luciana Vieira; CAMPOS, Marliane Batista; MATTA, Vania Lucia da; GOMES, Claudia Maria; FLORES, Gabriela Venicia; PACHECO, Carmen Maria; CORBETT, Carlos Eduardo; NAKAYA, Helder; LAURENTI, Marcia DalastraThis work reports on the whole-genome sequencing of Leishmania infantum chagasi from Honduras (Central America) and Brazil (South America).
- Immunopathological characterization of human cutaneous leishmaniasis lesions caused by Leishmania (Viannia) spp. in Amazonian Brazil(2017) GOMES, Claudia Maria Castro; SOUSA, Maria Gloria Teixeira; MENEZES, Joyce Prieto Bezerra; BATISTA, Marliane Campos; LIMA, Ana Carolina Stocco; BELDA JR., Walter; BRADSHAW, Daniel; GAMA, Monica Elinor Alves; LAURENTI, Marcia Dalastra; SILVEIRA, Fernando Tobias; CORBETT, Carlos Eduardo PereiraAmerican cutaneous leishmaniasis (ACL) is a chronic infectious disease caused by different protozoan species of Leishmania, and it is endemic in both tropical and subtropical countries. Using immunohistochemistry, we investigate the density of CD68(+), lysozyme(+), CD1a(+), factor XIIIa(+), CD4(+), CD8(+), CD56(+), interferon (IFN)-gamma(+), and inducible NO synthase (iNOS(+)) cells. These cells were analyzed from 22 biopsy samples obtained from the lesions of ACL patients, whose infection was caused by Leishmania (Viannia) spp. Histopathological analysis showed dense mononuclear inflammatory infiltration in the dermis, which was composed of lymphocytes, macrophages, plasma cells, and discrete tissue parasitism. Granulomatous reactions were also present in the majority of cases. The density of the activated macrophages was higher than that of inactivated macrophages in the lesions. The density of Langerhans cells (CD1a(+)) was lower than that of dermal dendrocytes (factor XIIIa(+)). The density of CD8(+) T lymphocytes was higher than that of CD4(+) T lymphocytes. The cellular density of these immunological markers in relation to the species of Leishmania demonstrated that L. (Viannia) sp. lesions had higher IFN-gamma expression than that Leishmania (Viania) braziliensis lesions. The evaluation of these markers, according to disease progression, did not reveal any significant differences. L. (Viannia) sp. infection leads to a favorable immune response in the host, as predominantly represented by lysozyme(+), factor XIIIa(+), CD8(+) T cells, and the expression of (IFN)-gamma(+) at the lesion site.
- Serum Cytokine Responses over the Entire Clinical-Immunological Spectrum of Human Leishmania (L.) infantum chagasi Infection(2016) RAMOS, Patricia Karla; CARVALHO, Karina Inacio; ROSA, Daniela Santoro; RODRIGUES, Ana Paula; LIMA, Luciana Vieira; CAMPOS, Marliane Batista; GOMES, Claudia Maria C.; LAURENTI, Marcia Dalastra; CORBETT, Carlos Eduardo; SILVEIRA, Fernando TobiasThe clinical-immunological spectrum of human Leishmania (L.) infantum chagasi infection in Amazonian Brazil was recently reviewed based on clinical, DTH, and IFAT (IgG) evaluations that identified five profiles: three asymptomatic (asymptomatic infection, AI; subclinical resistant infection, SRI; and indeterminate initial infection, III) and two symptomatic (symptomatic infection, SI; American visceral leishmaniasis, AVL; and subclinical oligosymptomatic infection, SOI). TNF-alpha, IL-4, IL-6, and IL-10 serum cytokines were analyzed using multiplexed Cytometric Bead Array in 161 samples from endemic areas in the Brazilian Amazon: SI [AVL] (21 cases), III (49), SRI (19), SOI (12), AI (36), and a control group [CG] (24). The highest IL-6 serumlevels were observed in the SI profile (AVL); higher IL-10 serum levels were observed in SI than in SOI or CG and in AI and III than in SOI; higher TNF-alpha serum levels were seen in SI than in CG. Positive correlations were found between IL-6 and IL-10 serum levels in the SI and III profiles and between IL-6 and TNF-alpha and between IL-4 and TNF-alpha in the III profile. These results provide strong evidence for associating IL-6 and IL-10 with the immunopathogenesis of AVL and help clarify the role of these cytokines in the infection spectrum.