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
13 resultados
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- In situ study of cellular immune response in human cutaneous lesions caused by Leishmania (Viannia) panamensis in Panama(2021) GONZALEZ, Kadir; CALZADA, Jose Eduardo; TOMOKANE, Thaise Yumie; PACHECO, Carmen Maria Sandoval; FLORES, Gabriela Venicia Araujo; GOMES, Claudia Maria Castro; CORBETT, Carlos Eduardo Pereira; SALDANA, Azael; LAURENTI, Marcia DalastraAims: Leishmaniasis is considered a disease with multiple clinical/immunopathological characteristics, depending on the immunity of the host and the species of the parasite. In Panama, the most prevalent species that causes localized cutaneous leishmaniasis (LCL) is Leishmania (Viannia) panamensis, and its immune response is poorly studied. Therefore, we evaluated by immunohistochemistry, the in situ immune response during this infection. Methods and Results: Biopsies from Panamanian patients with LCL were collected and processed by histological techniques. Infection by L. (V.) panamensis was demonstrated by isolation in culture and molecular characterization by Hsp70-RFLP. The in situ immune response was assessed by immunohistochemistry. The immune response was characterized by predominance of T cells, mainly CD8 cells that showed positive correlation with IFN-gamma and Granzyme B. CD4 cells presented positive correlation with both IFN-gamma and IL-13, pointed by mixed cellular immune response. Regulatory response was characterized by FoxP3 cells, which showed positive correlation to IL-10 but not with TGF-beta. Conclusions: L. (V.) panamensis infection triggers a mixed cellular immune response, characterized by the presence of pro-inflammatory, anti-inflammatory and regulatory elements in the skin lesion of Panamanian patients. These data contribute to a better understanding of the immunopathogenesis of Leishmania Viannia infection in Panama.
- Salivary gland homogenates from wild-caught sand flies Lutzomyia flaviscutellata and Lutzomyia (Psychodopygus) complexus showed inhibitory effects on Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis infection in BALB/c mice(2014) FRANCESQUINI, Fernanda C.; SILVEIRA, Fernando T.; PASSERO, Luiz Felipe D.; TOMOKANE, Thaise Y.; CARVALHO, Ana Kely; CORBETT, Carlos Eduardo P.; LAURENTI, Marcia D.During the natural transmission of Leishmania parasites, the infected sand fly female regurgitates promastigotes into the host's skin together with its saliva. It has been reported that vector saliva contains immunomodulatory molecules that facilitate the establishment of infection. Thus, the main objective of this study was to evaluate the specificity of Lutzomyia (Lu.) flaviscutellata and Lu.(Psychodopygus) complexus salivas on the infectivity of Leishmania (L.) (Leishmania) amazonensis and L.(Viannia) braziliensis, respectively. BALB/c mice were inoculated into the skin of hind footpad with L.(L.) amazonensis and L.(V.) braziliensis promastigotes in the absence or presence of Lu.flaviscutellata and Lu.(P.) complexus salivary gland homogenates (SGHs). The evolution of the infection was evaluated by lesion size, histopathological analysis and determination of the parasite load in the skin biopsies collected from the site of infection at 4 and 8weeks PI. The lesion size and the parasite load of both groups of mice infected in the presence of SGHs were smaller than the control groups. The histopathological features showed that the inflammatory reaction was less prominent in the groups of mice infected in the presence of both SGHs when compared to the control group. The results showed that the presence of SGHs of Lu.flaviscutellata and Lu.(P.) complexus led to induction of processes that were disadvantageous to parasite establishment during infection by L.(L.) amazonensis and L.(V.) braziliensis. An inhibitory effect on Leishmania infection could be observed in both groups inoculated with SGHs, especially when the SGH from Lu.(P.) complexus was used.
- Leishmania sp identification by PCR associated with sequencing of target SSU rDNA in paraffin-embedded skin samples stored for more than 30 years(2011) LIMA, Ana Carolina S. de; ZAMPIERI, Ricardo A.; TOMOKANE, Thaise Y.; LAURENTI, Marcia D.; SILVEIRA, Fernando T.; CORBETT, Carlos E. P.; FLOETER-WINTER, Lucile M.; GOMES, Claudia M. C.Paraffin-embedded samples commonly stored at educational and research institutions constitute tissues banks for follow-up or epidemiological studies; however, the paraffin inclusion process involves the use of substances that can cause DNA degradation. In this study, a PCR protocol was applied to identify Leishmania strains in 33 paraffin-embedded skin samples of patients with American cutaneous leishmaniasis. DNA was obtained by the phenol-chloroform protocol following paraffin removal and then used in PCR or nested PCR based on the nucleotide sequence of the small subunit ribosomal RNA (SSU rDNA). The amplicons obtained were cloned and sequenced to determine the single nucleotide polymorphism that distinguishes between different Leishmania species or groups. This assay allowed to distinguish organisms belonging to the subgenus Viannia and identify L. (Leishmania) amazonensis and L. (L.) chagasi of the Leishmania subgenus. Of the 33 samples, PCR and nested PCR identified 91% of samples. After sequencing the PCR product of 26 samples, 16 were identified as L. (L.) amazonensis, the other 10 contain organisms belonging to the L. (Viannia) sub-genus. These results open a huge opportunity to study stored samples and promote relevant contributions to epidemiological studies.
- Serological and infection statuses of dogs from a visceral leishmaniasis-endemic area(2014) LARANJEIRA, Daniela Farias; MATTA, Vania Lucia Ribeiro da; TOMOKANE, Thaise Yumie; MARCONDES, Mary; CORBET, Carlos Eduardo Pereira; LAURENTI, Marcia DalastraOBJECTIVE: This study investigated the serological status of dogs living in a visceral leishmaniasis-endemic area and its correlation with the parasitological condition of the animals. METHODS: Canine humoral response was evaluated using the sera of 134 dogs by enzyme-linked immunosorbent assay and immunohistochemistry to detect parasites in the skin, lymph node, and spleen of the animals. The specific antibodies investigated were IgG, IgG1, IgG2, and IgE. RESULTS: According to the parasitological, laboratory, and clinical findings, the dogs were placed into one of four groups: asymptomatic with (AP+, n = 21) or without (AP-, n = 36) Leishmania tissue parasitism and symptomatic with (SP+, n = 52) or without (SP-, n = 25) parasitism. Higher IgG and IgE levels were positively correlated with the infection condition and parasite load, but not with the clinical status. In all groups, total IgG was the predominant antibody, which occurred at the expense of IgG2 instead of IgG1. Most of the infected dogs tested positive for IgG (SP+, 98.1%; AP+, 95.2%), whereas this was not observed with IgE (SP+, 80.8%; AP+, 71.2%). The most relevant finding was the high positivity of the uninfected dogs for Leishmania-specific IgG (SP-, 60.0%; AP-, 44.4%), IgE (SP-, 44.0%; AP-, 27.8%), IgG1 (SP-, 28.0%; AP-, 22.2%), and IgG2 antibodies (SP-, 56.0%; AP-, 41.7%). CONCLUSIONS: The serological status of dogs, as determined by any class or subclass of antibodies, did not accurately distinguish dogs infected with L. (L.) infantum chagasi from uninfected animals. The inaccuracy of the serological result may impair not only the diagnosis, but also epidemiological investigations and strategies for visceral leishmaniasis control. This complex serological scenario occurring in a visceral leishmaniasis-endemic area highlights the challenges associated with canine diagnosis and points out the difficulties experienced by veterinary clinicians and coordinators of control programs.
- Macrophage Polarization in the Skin Lesion Caused by Neotropical Species of Leishmania sp(2021) PACHECO, Carmen M. Sandoval; V, Gabriela Araujo Flores; GONZALEZ, Kadir; GOMES, Claudia M. de Castro; PASSERO, Luiz F. D.; TOMOKANE, Thaise Y.; SOSA-OCHOA, Wilfredo; ZUNIGA, Concepcion; CALZADA, Jose; SALDANA, Azael; CORBETT, Carlos E. P.; SILVEIRA, Fernando T.; LAURENTI, Marcia D.Macrophages play important roles in the innate and acquired immune responses against Leishmania parasites. Depending on the subset and activation status, macrophages may eliminate intracellular parasites; however, these host cells also can offer a safe environment for Leishmania replication. In this sense, the fate of the parasite may be influenced by the phenotype of the infected macrophage, linked to the subtype of classically activated (M1) or alternatively activated (M2) macrophages. In the present study, M1 and M2 macrophage subsets were analyzed by double-staining immunohistochemistry in skin biopsies from patients with American cutaneous leishmaniasis (ACL) caused by L. (L.) amazonensis, L. (V.) braziliensis, L. (V.) panamensis ,and L. (L.) infantum chagasi. High number of M1 macrophages was detected in nonulcerated cutaneous leishmaniasis (NUCL) caused by L. (L.) infantum chagasi (M1=112 +/- 12, M2=43 +/- 12 cells/mm(2)). On the other side, high density of M2 macrophages was observed in the skin lesions of patients with anergic diffuse cutaneous leishmaniasis (ADCL) (M1=195 +/- 25, M2=616 +/- 114), followed by cases of localized cutaneous leishmaniasis (LCL) caused by L. (L.) amazonensis (M1=97 +/- 24, M2=219 +/- 29), L. (V.) panamensis (M1=71 +/- 14, M2=164 +/- 14), and L. (V.) braziliensis (M1=50 +/- 13, M2=53 +/- 10); however, low density of M2 macrophages was observed in NUCL. The data presented herein show the polarization of macrophages in skin lesions caused by different Leishmania species that may be related with the outcome of the disease.
- Evaluation of Regulatory Immune Response in Skin Lesions of Patients Affected by Nonulcerated or Atypical Cutaneous Leishmaniasis in Honduras, Central America(2018) FLORES, Gabriela Venicia Araujo; PACHECO, Carmen Maria Sandoval; TOMOKANE, Thaise Yumie; OCHOA, Wilfredo Sosa; VALERIANO, Concepcion Zuniga; GOMES, Claudia Maria Castro; CORBETT, Carlos Eduardo Pereira; LAURENTI, Marcia DalastraIn Honduras, Leishmania (L.) infantum chagasi causes both visceral leishmaniasis (LV) and nonulcerated or atypical cutaneous leishmaniasis (NUCL). NUCL is characterized by mononuclear inflammatory infiltration of the dermis, composed mainly of lymphocytes followed by macrophages with discrete parasitism. Considering that little is known about the pathogenesis of NUCL, the aim of this study was to evaluate the regulatory response in situ in skin lesions of patients affected by NUCL. Biopsies (n = 20) from human cutaneous nonulcerative lesions were collected and processed by usual histological techniques. The in situ regulatory immune response was evaluated by immunohistochemistry using antihuman CD4, FoxP3, IL-10, and TGF-beta antibodies. CD4(+), FoxP3(+), TGF-beta(+), and IL-10(+) cells were observed in the dermis with inflammatory infiltration in all studied cases and at higher densities compared to the normal skin controls. A positive and strong correlation was observed between CD4(+) and FoxP3(+) cells, and a positive and moderate correlation was observed between FoxP3(+) and TGF-beta(+) but not with IL-10(+) cells. The data suggest that T regulatory FoxP3(+) cells and the regulatory cytokines, especially TGF-beta, play an important role in the immunopathogenesis of NUCL, modulating a cellular immune response in the skin, avoiding tissue damage, and leading to low tissue parasitic persistence.
- 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.
conferenceObject Identification of Leishmania spp. isolated from patients with American cutaneous leishmaniasis from Amazonian Brazil by hsp70 PCR-RFLP(2015) LIMA, A. C. S.; CAMPOS, M. B.; TOMOKANE, T. Y.; LAURENTI, M. D.; CORBETT, C. E. P.; SILVEIRA, F. T.; GOMES, C. M. C.- Leishmania (L.) infantum chagasi isolated from skin lesions of patients affected by non-ulcerated cutaneous leishmaniasis lead to visceral lesion in hamsters(2023) FLORES, Gabriela V. Araujo; PACHECO, Carmen M. Sandoval; FERREIRA, Aurea F.; TOMOKANE, Thaise Yumie; NUNES, Juliana B.; COLOMBO, Fabio A.; SOSA-OCHOA, Wilfredo H.; ZUNIGA, Concepcion; SILVEIRA, Fernando T.; CORBETT, Carlos E. P.; LAURENTI, Marcia D.In Central America, Leishmania (L.) infantum chagasi infection causes visceral leishmaniasis (VL) and non -ulcerated cutaneous leishmaniasis (NUCL). The aim of the present study was to evaluate the course of an experimental infection in hamsters caused by L. (L.) infantum chagasi isolated from patients affected by NUCL compared with a strain isolated from a patient with VL. Stationary phase parasites in culture were inoculated through subcutaneous and intraperitoneal routes in hamsters. Following the post-infection times, a histopath-ological study, parasite load and cytokine determination in skin from the cutaneous inoculation site and viscera were performed. Animals subcutaneously infected with the different strains did not develop macroscopic lesions at the inoculation site, and the histopathological changes in the dermis were very slight. Regarding the histo-pathological study of the viscera, we observed the portal mononuclear inflammatory infiltrate, the presence of nodules in the hepatic parenchyma and the proliferation of macrophages in the spleen, which increased over the infection course. Overall, the parasite load in the liver and spleen and in the total IgG titres in the sera of infected hamster showed an increase with the time of infection, regardless of the route of inoculation. Regarding cellular immunity, we did not observe an increase or decrease in pro-and anti-inflammatory cytokines compared to the healthy control, except for IL-10, which was evident in the infected animals. The data showed that strains iso-lated from NUCL cause visceral lesions in the hamsters regardless of the route of inoculation, and they were similar to parasites isolated from VL humans.
- Dynamic of the Cellular Immune Response at the Dermal Site of Leishmania (L.) amazonensis and Leishmania (V.) braziliensis Infection in Sapajus apella Primate(2014) LAURENTI, Marcia Dalastra; PASSERO, Luiz Felipe Domingues; TOMOKANE, Thaise Yumie; FRANCESQUINI, Fernanda de Camargo; ROCHA, Mussya Cisotto; GOMES, Claudia Maria de Castro; CORBETT, Carlos Eduardo Pereira; SILVEIRA, Fernando TobiasThe purpose of this study was to characterize the immunopathological response in the skin of S. apella infected with Leishmania (L.) amazonensis and L. (V.) braziliensis parasites, the main causative agents of localized cutaneous leishmaniasis in South America. In infected animals, amastigote forms of L. (L.) amazonensis could be detected till 120 days postinfection (PI), while, in L. (V.) braziliensis infection, parasites could be detected until 180 days PI in the skin sections. CD20(+) cells were detected throughout the experimental time in both groups as well as in CD3(+) cells, which appeared to be activated because high densities of inducible nitric oxide synthase (iNOS(+)) cells were detected at 60 and 90 days PI in both studied groups. After 60 and 120 days PI, decrease in iNOS(+) cells was observed in L. (L.) amazonensis and L. (V.) braziliensis, respectively, which was associated with parasite clearance. Increase in lysozyme(+) cells was observed during the experimental infections, which also can be associated with parasite killing.