MARCIA DALASTRA LAURENTI

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Departamento de Patologia, Faculdade de Medicina - Docente
LIM/50 - Laboratório de Patologia das Moléstias Infecciosas, Hospital das Clínicas, Faculdade de Medicina - Líder

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  • article 0 Citação(ões) na Scopus
    Leishmania enriettii visceralises in the trachea, lungs, and spleen of Cavia porcellus
    (2022) ALVES-SOBRINHO, Edneia Venancio; PINHEIRO, Lucelia de Jesus; PARANAIBA, Larissa Ferreira; FONTES, Igor Campos; PARREIRAS, Patricia Martins; GONTIJO, Nelder Figueiredo; TAFURI, Wagner Luiz; LAURENTI, Marcia Dalastra; SOARES, Rodrigo Pedro
    BACKGROUND Leishmania (Mundinia) enriettii is a species commonly found in the guinea pig, Cavia porcellus. Although it is a dermotropic species, there is still an uncertainty regarding its ability to visceralise during Leishmania life cycle. OBJECTIVE Here, we investigated the ability of L. enriettii (strain L88) to visceralise in lungs, trachea, spleen, and liver of C. porcellus, its natural vertebrate host. METHODS Animals were infected sub-cutaneously in the nose and followed for 12 weeks using histological (hematoxilin-eosin) and molecular tools (polymerase chain reaction-restriction fragment length polymorphism - PCR-RFLP). To isolate parasite from C. porcellus, animals were experimentally infected for viscera removal and PCR typing targeting hsp70 gene. FINDINGS Histological analysis revealed intense and diffuse inflammation with the presence of amastigotes in the trachea, lung, and spleen up to 12 weeks post-infection (PI). Molecular analysis of paraffin-embedded tissues detected parasite DNA in the trachea and spleen between the 4th and 8th weeks PI. At the 12th PI, no parasite DNA was detected in any of the organs. To confirm that the spleen could serve as a temporary site for L. enriettii, we performed additional in vivo experiments. During 6th week PI, the parasite was isolated from the spleen confirming previous histopathological and PCR observations. MAIN CONCLUSION Leishmania enriettii (strain L88) was able to visceralise in the trachea, lung, and spleen of C. porcellus.
  • article 8 Citação(ões) na Scopus
    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 Tobias
    This 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.
  • article 5 Citação(ões) na Scopus
    Evaluation of Leishmania (Leishmania) infantum excreted-secreted antigens for detection of canine leishmaniasis
    (2016) PINEDO-CANCINO, Viviana; LAURENTI, Marcia Dalastra; KESPER, Norival; UMEZAWA, Eufrosina Setsu
    The efficacy of tests with L (L.) infantum excreted-secreted antigens (ESA) to detect canine leishmaniasis (CanL) was evaluated using immunoblotting (ESA-blot), ELISA (ESA-ELISA) and ELISA with alkaline extract from promastigotes (PAE). Of one hundred fifty-five domestic dogs tested, 100 were suspected of CanL, 23 had other diseases and 32 were healthy. Sera from the dogs suspected of CanL were tested by immunohistochemistry (IHC), and 54% were confirmed to be infected by L (L.) infantum (38 symptomatic and 16 asymptomatic). Of these, 100% were positive by ESA-blot, ESA-ELISA and PAE-ELISA. In the ESA blot their sera recognized polypeptides in the 26.5-31.5 kDa region. Of the 46% of dogs with negative IHC, 44-53% tested positive in all three tests irrespective of clinical status. The twenty-three dogs with other diseases were negative by ESA-blot, but sera from 9% and 26% of them reacted with ESA-ELISA and PAE-ELISA, respectively. The 32 healthy dogs were negative in all the tests. ESA-blot showed good correlation with IHC in the detection of CanL and a high specificity index.
  • article 9 Citação(ões) na Scopus
    Leishmania infantum INFECTION IN DOGS FROM THE SOUTHERN REGION OF MINAS GERAIS STATE, BRAZIL
    (2016) NUNES, Juliana Barbosa; LAURENTI, Marcia Dalastra; KANAMURA, Herminia Yohko; PEREIRA, Alessandro Antonio Costa; COLOMBO, Fabio Antonio; MARQUES, Marcos Jose
    Visceral leishmaniasis is a systemic and chronic disease and dogs are the main reservoir of the etiologic agent, Leishmania infantum (syn L. chagasi). A serological and molecular investigation of canine visceral leishmaniasis (CVL) was performed in the municipality of Alfenas, located in the southern region of Minas Gerais, where the disease is not endemic. Samples from 87 dogs were submitted to serological tests including the Dual Path Platform (DPP (R)) CVL Bio-Manguinhos rapid test, an in-house enzyme-linked immunosorbent assay (ELISA) and an immunofluorescence antibody test (IFAT), as well as molecular techniques such as a conventional polymerase chain reaction (PCR) with the RV1/RV2 primers and a quantitative PCR (qPCR) with the LinJ31, Ldon and DNApol primers. Of the 87 serum samples, eight (9.2%) were positive for Leishmania using the DPP rapid test, but only four (4.6%) were confirmed by ELISA and two (2.3%) by IFAT. In these two serologically confirmed cases, spleen and liver samples were positive by all the employed molecular and parasitological procedures performed on spleen samples. When whole blood samples were used in the molecular assays, two samples (2.3%) were positive only by qPCR. DNA extracted and amplified from the spleens of seropositive dogs was sequenced, showing 100% of similarity with the Leishmania infantum (syn L. chagasi) sequence. Thus, the first cases of CVL have been confirmed in the Alfenas region, suggesting the importance of canine surveys in non-endemic municipalities for CVL to monitor disease progression and to prevent outbreaks.
  • article 12 Citação(ões) na Scopus
    Effect of phospholipase A(2) inhibitors during infection caused by Leishmania (Leishmania) amazonensis
    (2018) BORDON, Maria L. A. C.; LAURENTI, Marcia D.; RIBEIRO, Susan Pereira; TOYAMA, Marcos H.; TOYAMA, Daniela De O.; PASSERO, Luiz Felipe D.
    Background: Lipid metabolites play an important role in parasite differentiation and virulence. Studies have revealed that Leishmania sp. uses prostaglandins to evade innate barriers, thus enabling the parasites to survive inside immune cells. Despite the role of the enzyme Phospholipase A(2) (PLA(2)) in prostaglandins production, few studies have investigated the role of parasite PLA(2) during the interaction between L. (L.) amazonensis and the host (in vitro and in vivo) immune cells. Methods: In the present work, the leishmanicidal effect of PLA(2) inhibitors, methyl arachidonyl fluorophosphonate (MAFP), bromoenol lactone (BEL) and aristolochic acid (AA) were investigated in vitro (promastigote and intracellular amastigote forms of L. (L.) amazonensis) and during in vivo infection using BALB/c mice. Results: The aforementioned inhibitors were deleterious to promastigote and amastigote forms of the L. (L.) amazonensis and were non-toxic to peritoneal macrophages from BALB/c mice. L. (L.) amazonensis-infected BALB/c mice treated with the inhibitor BEL presented decreased lesion size and skin parasitism; however, BEL treatment induced hepatotoxicity in BALB/c mice. Conclusions: Results presented herein suggested that PLA(2) inhibitors altered L. (L.) amazonensis viability. In spite of liver toxicity, treatment with BEL was the most selective compound in vitro, as well in vivo, resulting in lower skin parasitism in the infected mice. These findings corroborate the role of PLA(2) in parasite virulence and maintenance in vertebrate hosts, and suggest that molecules structurally related to BEL should be considered when planning compounds against Leishmania sp.
  • article 70 Citação(ões) na Scopus
    Serological cross-reactivity of Trypanosoma cruzi, Ehrlichia canis, Toxoplasma gondii, Neospora caninum and Babesia canis to Leishmania infantum chagasi tests in dogs
    (2014) ZANETTE, Mauricio Franco; LIMA, Valeria Marcal Felix de; LAURENTI, Marcia Dalastra; ROSSI, Claudio Nazaretian; VIDES, Juliana Peloi; VIEIRA, Rafael Felipe da Costa; BIONDO, Alexander Welker; MARCONDES, Mary
    Introduction: The aim of this study was to evaluate the serological cross-reactivity between Leishmania sp. and other canine pathogens. Methods: Positive serum samples for Ehrlichia canis, Babesia canis, Toxoplasma gondii, Neospora caninum and Trypanosoma cruzi were tested using three serological methods enzyme linked immunosorbent assay (ELISA), indirect immunofluorescent antibody test (IFAT) and Kalazar Detect™, for canine visceral leishmaniasis. Results: Of the 57 dog samples tested, 24 (42.1%) tested positive using one of the three serological methods: 10/57 (17.5%) for ELISA, 11/57 (19.3%) for IFAT and 3/57 (5.3%) for Kalazar Detect™. Conclusions: Our results demonstrated that the presence of other infectious agents may lead to cross-reactivity on leishmaniasis serological tests.
  • article 19 Citação(ões) na Scopus
    Amazonian Phlebovirus (Bunyaviridae) potentiates the infection of Leishmania (Leishmania) amazonensis: Role of the PKR/IFN1/IL-10 axis
    (2019) RATH, Carolina Torturella; SCHNELLRATH, Laila Castro; DAMASO, Clarissa R.; ARRUDA, Luciana Barros de; VASCONCELOS, Pedro Fernando da Costa; GOMES, Claudia; LAURENTI, Marcia Dalastra; SILVA, Teresa Cristina Calegari; VIVARINI, Aislan de Carvalho; FASEL, Nicolas; PEREIRA, Renata Meirelles Santos; LOPES, Ulisses Gazos
    Background Leishmania parasites are transmitted to vertebrate hosts by phlebotomine sandflies and, in humans, may cause tegumentary or visceral leishmaniasis. The role of PKR (dsRNA activated kinase) and Toll-like receptor 3 (TLR3) activation in the control of Leishmania infection highlights the importance of the engagement of RNA sensors, which are usually involved in the antiviral cell response, in the fate of parasitism by Leishmania. We tested the hypothesis that Phlebovirus, a subgroup of the Bunyaviridae, transmitted by sandflies, would interfere with Leishmania infection. Methodology/Principal findings We tested two Phlebovirus isolates, Icoaraci and Pacui, from the rodents Nectomys sp. and Oryzomys sp., respectively, both natural sylvatic reservoir of Leishmania (Leishmania) amazonensis from the Amazon region. Phlebovirus coinfection with L. (L.) amazonensis in murine macrophages led to increased intracellular growth of L. (L.) amazonensis. Further studies with Icoaraci coinfection revealed the requirement of the PKR/IFN1 axis on the exacerbation of the parasite infection. L. (L.) amazonensis and Phlebovirus coinfection potentiated PKR activation and synergistically induced the expression of IFN beta and IL-10. Importantly, in vivo coinfection of C57BL/6 mice corroborated the in vitro data. The exacerbation effect of RNA virus on parasite infection may be specific because coinfection with dengue virus (DENV2) exerted the opposite effect on parasite load. Conclusions Altogether, our data suggest that coinfections with specific RNA viruses shared by vectors or reservoirs of Leishmania may enhance and sustain the activation of host cellular RNA sensors, resulting in aggravation of the parasite infection. The present work highlights new perspectives for the investigation of antiviral pathways as important modulators of protozoan infections. Author summary Cutaneous Leishmaniasis is highly prevalent worldwide, affecting millions of people in undeveloped countries. Leishmania parasites are transmitted to humans by phlebotomine flies. The transmission occurs mainly in rural areas, although the urbanization of the disease is in process. Phlebotomine vectors may also transmit RNA virus (arboviruses) to humans. Our group has previously demonstrated the induction of the host anti-viral pathways by L. (L.) amazonensis, Since some anti-viral response pathways potentiate L. (L.) amazonensis infection and the same group of Leishmania arthropod vectors transmits RNA viruses belonging to the genus Phlebovirus, we hypothesized that coinfection could aggravate the parasitism by L. (L.) amazonensis. Using a phlebovirus isolated from L. amazonensis rodents reservoirs (Icoaraci), we demonstrated that Icoaraci coinfection enhanced the parasite load, both in vivo and in vitro. These observations were partialy corroborated with Pacui, another Phlebovirus isolate. Increased macrophage infection required IFN type I expression induced by the parasite and amplified during coinfection. Importantly, the infection by another prevalent arbovirus, DENV2, did not result in the exacerbation of the parasite infection. Overall our results support the importance of complex and multifactorial analysis of infectious processes, especially in tropical regions that frequently have endemic areas of overlap.
  • article 5 Citação(ões) na Scopus
    LEISHMANICIDAL ACTIVITY in vivo OF A MILTEFOSINE DERIVATIVE IN Mesocricetus auratus
    (2020) SILVA, Joana C. da; NUNES, Juliana B.; GONTIJO, Vanessa S.; MALAQUIAS, Luiz Cosme C.; FREITAS, Rossimiriam P. de; ALVES, Rosemeire B.; COLOMBO, Fabio A.; LAURENTI, Marcia D.; MARQUES, Marcos J.
    Visceral leishmaniasis (VL) is a chronic and systemic disease; if untreated, it can cause death in a large number of cases. The therapy is based on the use of antimonials, which have been used for over 50 years. However, cases of resistance have been reported in some countries. In this context, miltefosine (MIL) was introduced to treat antimonial unresponsive cases. Nonetheless, in recent years MIL unresponsive and relapse cases of VL have increasingly been reported. In the current study, the therapeutic potential of compound 5-(4-(3-methanesulfonatepropyl)-1H-1,2,3-triazol-1-yl)dodecyl methanesulfonate (C11), an MIL derivative, was assessed in an experimental VL hamster model. For this purpose, golden hamsters (Mesocricetus auratus) were infected with Leishmania (L.) infantum chagasi and treated daily for 10 days with C11 and MIL administered orally; in addition, Glucantime (GLU), peritoneal route, were administered at 15, 10, 50 mg/kg body weight/day, respectively. Twenty four hours after the end of treatment the animals were euthanatized; and the specimens were collected to evaluate the relative mRNA expression of cytokines IFN-gamma, TNF-alpha, IL-17, TGF-beta, IL-4 and IL-10 in fragments of the spleen and liver; moreover, the parasitism in these organs was evaluated as well as the main histopathological alterations. The C11-treated animals showed greater expression of IL-17 and TNF-alpha cytokines and reduced expression of IL-10 in the spleen in comparison to the infected untreated group (UTG) (p <0.05). The C11 and GLU groups showed a significant reduction in the IgG levels in comparison to the UTG group (p <0.05). Moreover, the C11-treated animals had fewer parasites in the spleen than the UTG animals (reduction of 95.9%), as well as a greater preservation of white pulp architecture in the spleen than the UTG, GLU and MIL groups (p <0.05). For the liver, the animals from the C11 and MIL groups showed a significant increase in TNF-alpha relative expression in comparison to the UTG animals, which would explain the increase in the number of granulomas and the reduction in the parasitic load (p <0.05). Combined, these findings indicate that C11 is an interesting compound that should be considered for the development of new drugs against VL, mainly due to its leishmanicidal effect and immunostimulating action.
  • article 22 Citação(ões) na Scopus
    Histopathological characteristics of cutaneous lesions caused by Leishmania Viannia panamensis in Panama
    (2018) GONZÁLEZ, Kadir; DIAZ, Rosendo; FERREIRA, Aurea F.; GARCÍA, Víctor; PAZ, Héctor; CALZADA, José E.; RUÍZ, Michelle; LAURENTI, Márcia; SALDAñA, Azael
    ABSTRACT Cutaneous leishmaniasis (CL) is an endemic disease in the Republic of Panama, caused by Leishmania (Viannia) parasites, whose most common clinical manifestation is the presence of ulcerated lesions on the skin. These lesions usually present a chronic inflammatory reaction, sometimes granulomatous, with the presence of lymphocytes, plasma cells and macrophages. This study describes the histopathological characteristics found in the skin lesions of patients with CL caused by Leishmania (V.) panamensis in Panama. We analyzed 49 skin biopsy samples from patients with clinical suspicion of CL, by molecular tests (PCR for subgenus Viannia and HSP-70) and by Hematoxylin-Eosin staining. Samples were characterized at the species level by PCR-HSP-70/RFLP. From the 49 samples studied, 46 (94%) were positive by PCR and were characterized as Leishmania (V.) panamensis. Of these, 48% were positive by Hematoxylin-Eosin staining with alterations being observed both, in the epidermis (85%) and in the dermis (100%) of skin biopsies. The inflammatory infiltrate was characterized according to histopathological patterns: lymphohistiocytic (50%), lymphoplasmacytic (61%) and granulomatous (46%) infiltration, being the combination of these patterns frequently found. The predominant histopathological characteristics observed in CL lesions caused by L. (V.) panamensis in Panama were: an intense inflammatory reaction in the dermis with a combination of lymphohistiocytic, lymphoplasmacytic and granulomatous presentation patterns and the presence of ulcers, acanthosis, exocytosis and spongiosis in the epidermis.
  • article 0 Citação(ões) na Scopus
    First report of putative Leishmania RNA virus 2 (LRV2) in Leishmania infantum strains from canine and human visceral leishmaniasis cases in the southeast of Brazil
    (2023) REGO, Felipe Dutra; SILVA, Eduardo Sergio da; LOPES, Valeriana Valadares; TEIXEIRA-NETO, Rafael Goncalves; BELO, Vinicius Silva; JR, Antonio Augusto Fonseca; PEREIRA, Diego Andrade; PENA, Heber Paulino; LAURENTI, Marcia Dalastra; V, Gabriela Araujo; MATTA, Vania Lucia Ribeiro da; CHOUMAN, Islam Hussein; BURRIN, Thaina Bergantin; SANDOVAL, Carmen M.; BARROUIN-MELO, Stella Maria; PINHO, Flaviane Alves de; ANDRADE, Helida Monteiro de; NUNES, Ramon Vieira; GONTIJO, Celia Maria Ferreira; SOCCOL, Vanete Thomaz; KLOCEK, Donnamae; GRYBCHUK, Danyil; MACEDO, Diego Henrique; MONTE-NETO, Rubens Lima do; YURCHENKO, Vyacheslav; SOARES, Rodrigo Pedro
    BACKGROUND Leishmania RNA virus 1 (LRV1) is commonly found in South American Leishmania parasites belonging to the subgenus Viannia, whereas Leishmania RNA virus 2 (LRV2) was previously thought to be restricted to the Old-World pathogens of the subgenus Leishmania. OBJECTIVES In this study, we investigated the presence of LRV2 in strains of Leishmania (L.) infantum, the causative agent of visceral leishmaniasis (VL), originating from different hosts, clinical forms, and geographical regions. METHODS A total of seventy-one isolates were screened for LRV2 using semi-nested reverse transcription-polymerase chain reaction (RT-PCR) targeting the RNA-dependent RNA polymerase (RdRp) gene. FINDINGS We detected LRV2 in two L. infantum isolates (CUR268 and HP-EMO) from canine and human cases, respectively. MAIN CONCLUSIONS To the best of our knowledge, this is the first detection of LRV2 in the New World.