THAISE YUMIE TOMOKANE
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Patologia, Faculdade de Medicina
LIM/50 - Laboratório de Patologia das Moléstias Infecciosas, Hospital das Clínicas, Faculdade de Medicina
LIM/50 - Laboratório de Patologia das Moléstias Infecciosas, Hospital das Clínicas, Faculdade de Medicina
8 resultados
Resultados de Busca
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- 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.
- Chromosomal segments may explain the antibody response cooperation for canine leishmaniasis pathogenesis(2020) BATISTA, Luis F. S.; TORRECILHA, Rafaela B. P.; SILVA, Rafaela B.; UTSUNOMIYA, Yuri T.; SILVA, Thais B. F.; TOMOKANE, Thaise Y.; PACHECO, Acacio D.; BOSCO, Anelise M.; PAULAN, Silvana C.; ROSSI, Claudio N.; COSTA, Gustavo N. O.; MARCONDES, Mary; CIARLINI, Paulo C.; NUNES, Caris M.; MATTA, Vania L. R.; LAURENTI, Marcia D.Visceral leishmaniasis (VL) is marked by hyperactivation of a humoral response secreting high quantity of immunoglobulins (Igs) that are inaccessible to intracellular parasites. Here we investigated the contributions of the antibody response to the canine leishmaniasis pathogenesis. Using correlation and genome-wide association analysis, we investigated the relationship of anti-Leishmania infantum immunoglobulin classes levels with parasite burden, clinical response, renal/hepatic biochemical, and oxidative stress markers in dogs from endemic areas of VL. Immunoglobulin G (IgG) and IgA were positively correlated with parasite burden on lymph node and blood. Increased IgG, IgA and IgE levels were associated with severe canine leishmaniasis (CanL) whereas IgM was elevated in uninfected exposed dogs. Correlations of IgM, IgG and IgA with creatinine, urea, AST and ALT levels in the serum were suggested an involvement of those Igs with renal and hepatic changes. The correlogram of oxidative radicals and antioxidants revealed a likely relationship of IgM, IgG and IgA with oxidative stress and lipid pemxidation in the blood, suggested as mechanisms mediating tissue damage and CanL worsening. The gene mapping on chromosomal segments associated with the quantitative variation of immunoglobulin classes identified genetic signatures involved with reactive oxygen species generation, phagolysosome maturation and rupture, free iron availability, Thl/Th2 differenciation and, immunoglobulin clearance. The findings demonstrated the roles of the antibody response as resistance or susceptibility markers and mediators of CanL pathogenesis. In addition we pinpointed candidate genes as potential targets for the therapy against the damage caused by exacerbated antibody response and parasitism in VL.
- 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.
- Genome-Wide Association Study of Cell-Mediated Response in Dogs Naturally Infected by Leishmania infantum(2016) BATISTA, Luis F. S.; UTSUNOMIYA, Yuri T.; SILVA, Thais B. F.; DIAS, Raissa A.; TOMOKANE, Thaise Y.; PACHECO, Acacio D.; MATTA, Vania L. R. da; SILVEIRA, Fernando T.; MARCONDES, Mary; NUNES, Caris M.; LAURENTI, Marcia D.A genome-wide association study (GWAS) could unravel the complexity of the cell-mediated immunity (CMI) to canine leishmaniasis (CanL). Therefore, we scanned 110,165 single-nucleotide polymorphisms (SNPs), aiming to identify chromosomal regions associated with the leishmanin skin test (LST), lymphocyte proliferation assay (LPA), and cytokine responses to further understand the role played by CMI in the outcome of natural Leishmania infantum infection in 189 dogs. Based on LST and LPA, four CMI profiles were identified (LST-/LPA(-), LST+ /LPA(-), LST- /LPA(+), and LST+ /LPA(+)), which were not associated with subclinically infected or diseased dogs. LST+/LPA(+) dogs showed increased interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) levels and mild parasitism in the lymph nodes, whereas LST-/ LPA(+) dogs, in spite of increased IFN-beta, also showed increased interleukin-10 (IL-10) and transforming growth factor beta (TGF-beta) levels and the highest parasite load in lymph nodes. Low T cell proliferation under low parasite load suggested that L. infantum was not able to induce effective CMI in the early stage of infection. Altogether, genetic markers explained 87%, 16%, 15%, 11%, 0%, and 0% of phenotypic variance in TNF-alpha, TGF-beta, LST, IL-10, IFN-gamma, and LPA, respectively. GWAS showed that regions associated with TNF-alpha include the following genes: IL12RB1, JAK3, CCRL2, CCR2, CCR3, and CXCR6, involved in cytokine and chemokine signaling; regions associated with LST, including COMMD5 and SHARPIN, involved in regulation of NF-kappa B signaling; and regions associated with IL-10, including LTBP1 and RASGRP3, involved in T regulatory lymphocytes differentiation. These findings pinpoint chromosomic regions related to the cell-mediated response that potentially affect the clinical complexity and the parasite replication in canine L. infantum infection.
- 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.
- Canine leishmaniasis: Genome-wide analysis and antibody response to Lutzomyia longipalpis saliva(2018) BATISTA, Luis F. S.; UTSUNOMIYA, Yuri T.; SILVA, Thais B. F.; CARNEIRO, Mariana M.; PAIVA, Joyr S. F.; SILVA, Rafaela B.; TOMOKANE, Thaise Y.; ROSSI, Claudio N.; PACHECO, Acacio D.; TORRECILHA, Rafaela B. P.; SILVEIRA, Fernando T.; MARCONDES, Mary; NUNES, Caris M.; LAURENTI, Marcia D.The anti-inflammatory properties of sand fly saliva favor the establishment of the Leishmania infantum infection. In contrast, an antibody response against Lutzomyia longipalpis saliva is often associated with a protective cell-mediated response against canine visceral leishmaniasis. Genetic studies may demonstrate to what extent the ability to secrete antisaliva antibodies depends on genetic or environmental factors. However, the genetic basis of canine antibody response against sand fly saliva has not been assessed. The aim of this study was to identify chromosomal regions associated with the anti-Lu. longipalpis salivary IgG response in 189 dogs resident in endemic areas in order to provide information for prophylactic strategies. Dogs were classified into five groups based on serological and parasitological diagnosis and clinical evaluation. Anti-salivary gland homogenate (SGH) IgG levels were assessed by Enzyme-Linked Immunosorbent Assay (ELISA). Genomic DNA was isolated from blood samples and genotyped using a SNP chip with 173,662 single nucleotide polymorphism (SNP) markers. The following linear regression model was fitted: IgG level = mean + origin + sex + age + use of a repellent collar, and the residuals were assumed as pseudo-phenotypes for the association test between phenotypes and genotypes (GWA). A component of variance model that takes into account polygenic and sample structure effects (EMMAX) was employed for GWA. Phenotypic findings indicated that anti-SGH IgG levels remained higher in exposed and subclinically infected dogs than in severely diseased dogs even in regression model residuals. Five associated markers were identified on chromosomes 2, 20 and 31. The mapped genes included CD180 (RP105) and MITF related to the rapid activation of B lymphocytes and differentiation into antibody-secreting plasma cells. The findings pointed to chromosomal segments useful for functional confirmation studies and a search for adjuvant molecules of the anti-saliva response.
- Expression of inducible nitric oxide synthase in macrophages inversely correlates with parasitism of lymphoid tissues in dogs with visceral leishmaniasis(2014) SANCHES, Francoise P.; TOMOKANE, Thaise Y.; MATTA, Vania L. R. Da; MARCONDES, Mary; CORBETT, Carlos E. P.; LAURENTI, Marcia D.Background: There are only a few studies reporting the role of nitric oxide metabolites for controlling macrophage intracellular parasitism, and these are controversial. Therefore, the present study aimed to evaluate the expression of inducible nitric oxide synthase (iNOS) in the lymph nodes and spleen of dogs affected by visceral leishmaniasis through immunohistochemistry and to determine its correlation with tissue parasite burden and serum interferon (IFN)-gamma levels. Twenty-eight dogs were selected and assigned to one of two groups, symptomatic (n = 18) and asymptomatic (n = 10), according to clinical status and laboratory evaluation. A negative control group (n = 6) from a non-endemic region for visceral leishmaniasis was included as well. Results: Parasite density (amastigotes/mm(2)) was similar between clinical groups in the lymph nodes (P = 0.2401) and spleen (P = 0.8869). The density of iNOS(+) cells was higher in infected dogs compared to controls (P < 0.05), without a significant difference in lymph node (P = 0.3257) and spleen (P = 0.5940) densities between symptomatic and asymptomatic dogs. A positive correlation was found between the number of iNOS(+) cells in lymph nodes and interferon-gamma levels (r = 0.3776; P = 0.0303), and there was a negative correlation between parasites and iNOS+ cell densities both in lymph nodes (r = -0.5341; P = 0.0034) and spleen (r = -0.4669; P = 0.0329). Conclusion: The negative correlation observed between tissue parasitism and the expression of iNOS may be a reflection of NO acting on the control of parasites.
- Molecular tools confirm natural Leishmania (Viannia) guyanensis/L. (V.) shawi hybrids causing cutaneous leishmaniasis in the Amazon region of Brazil(2021) LIMA, Ana Carolina S.; GOMES, Claudia Maria C.; TOMOKANE, Thaise Y.; CAMPOS, Marliane Batista; ZAMPIERI, Ricardo A.; JORGE, Carolina L.; LAURENTI, Marcia D.; SILVEIRA, Fernando T.; CORBETT, Carlos Eduardo P.; FLOETER-WINTER, Lucile MariaSeven isolates from patients with American cutaneous leishmaniasis in the Amazon region of Brazil were phenotypically suggestive of Leishmania (Viannia) guyanensis/L. (V.) shawi hybrids. In this work, two molecular targets were employed to check the hybrid identity of the putative hybrids. Heat shock protein 70 (hsp70) gene sequences were analyzed by three different polymerase chain reaction (PCR) approaches, and two different patterns of inherited hsp70 alleles were found. Three isolates presented heterozygous L. (V.) guyanensis/L. (V.) shawi patterns, and four presented homozygous hsp70 patterns involving only L. (V.) shawi alleles. The amplicon sequences confirmed the RFLP patterns. The high-resolution melting method detected variant heterozygous and homozygous profiles. Single-nucleotide polymorphism genotyping/cleaved amplified polymorphic site analysis suggested a higher contribution from L. (V.) guyanensis in hsp70 heterozygous hybrids. Additionally, PCR-RFLP analysis targeting the enzyme mannose phosphate isomerase (mpi) gene indicated heterozygous and homozygous cleavage patterns for L. (V.) shawi and L. (V.) guyanensis, corroborating the hsp70 findings. In this communication, we present molecular findings based on partial informative regions of the coding sequences of hsp70 and mpi as markers confirming that some of the parasite strains from the Brazilian Amazon region are indeed hybrids between L. (V.) guyanensis and L. (V.) shawi.