PAULA ORDONHEZ RIGATO
Projetos de Pesquisa
Unidades Organizacionais
LIM/56 - Laboratório de Investigação em Dermatologia e Imunodeficiências, Hospital das Clínicas, Faculdade de Medicina
7 resultados
Resultados de Busca
Agora exibindo 1 - 7 de 7
- Mucosal and systemic anti-GAG immunity induced by neonatal immunization with HIV LAMP/gag DNA vaccine in mice(2011) GOLDONI, Adriana Leticia; MACIEL JR., Milton; RIGATO, Paula Ordonhez; PIUBELLI, Orlando; BRITO, Cyro Alves de; MELO, Andrea; MARQUES, Ernesto Torres; AUGUST, Joseph Thomas; DUARTE, Alberto Jose da Silva; SATO, Maria NotomiVaccines capable of inducing mucosal immunity in early postnatal life until adulthood, protecting early sexual initiation, should be considered as strategies to vaccination against HIV. The HIV-1 GAG protein as a chimera with the lysosome-associated membrane protein (LAMP/gag), encoded by a DNA vaccine, is targeted to the endosomal/lysosomal compartment that contains class II MHC molecules and has been shown to be immunogenic in adult mice. Assuming that one such strategy could help to overcome the immunological immaturity in the early postnatal period, we have evaluated the systemic and mucosal immunogenicity of LAMP/gag immunization in neonatal mice. Intranasal immunization with LAMP/gag vaccine induced higher levels of sIgA and IgG anti-GAG antibodies in intestinal washes than did the gag vaccine. The combination of ID injections and the IN protocol with the chimeric vaccine promoted the increase of Ab levels in sera. Both vaccines induced splenic IFN-gamma- secreting cells against GAG peptide pools, as well as in vivo cytotoxic T lymphocyte (CTL) function, and increased the percentage of CD8+ T cells to the immunodominant class I peptide in gut and spleen. However, only the chimeric vaccine was able to enhance Th1/Th2 cytokine secretion in response to class II GAG peptide and to enhance IL-4-secreting cells against GAG peptides and p24 protein stimuli. Long-lasting humoral and cellular responses were detected until adult age, following neonatal immunization with the chimeric vaccine. The LAMP/gag vaccination was able to induce potent GAG-specific T and B cell immune responses in early life which are essential to elicit sustained and long-lasting mucosal and systemic humoral response.
- Development of potent class II transactivator gene delivery systems capable of inducing de novo MHC II expression in human cells, in vitro and ex vivo(2017) PALMA, M. L.; DUANGKHAE, P.; DOURADINHA, B.; VIANA, I. F. T.; RIGATO, P. O.; DHALIA, R.; MAILLIARD, R. B.; BARRATT-BOYES, S. M.; NASCIMENTO, E. J. M.; OSHIRO, T. M.; DUARTE, A. J. da Silva; MARQUES, E. T. A.Class II transactivator (CIITA) induces transcription of major histocompatibility complex (MHC) II genes and can potentially be used to improve genetic immunotherapies by converting non-immune cells into cells capable of presenting antigens to CD4(+) T cells. However, CIITA expression is tightly controlled and it remains unclear whether distinct non-immune cells differ in this transactivator regulation. Here we describe the development of gene delivery systems capable of promoting the efficient CIITA expression in non-immune cell lines and in primary human cells of an ex vivo skin explant model. Different human cell types undergoing CIITA overexpression presented high-level de novo expression of MHC II, validating the delivery systems as suitable tools for the CIITA evaluation as a molecular adjuvant for gene therapies.
conferenceObject Is the Functionality of Monocyte Derived Dendritic Cell from Chronic Mucocutaneous Candidiasis Patients Altered?(2014) RIGATO, P. Ordonhez; DIAS, A. Santos; VASCONCELOS, D. Moraes; DUARTE, A. J.conferenceObject MCM4 DEFICIENCY: A RARE VARIANT OF IMMUNODEFICIENCY OF NK CELLS ASSOCIATED TO PROPORTIONATE NANISM AND ADRENAL INSUFFICIENCY. DESCRIPTION OF THE FIRST CASE IN BRAZIL(2016) MORAES-VASCONCELOS, Dewton; RIBEIRO, Roberto; RIGATO, Paula Ordonhez; PINICHI, Paula; AOKI, Valeria; TAKAOKA, Roberto; DUARTE, Alberto Jose da Silva; SABINO, Ester Cerdeira- Regulation of HIV-Gag Expression and Targeting to the Endolysosomal/Secretory Pathway by the Luminal Domain of Lysosomal-Associated Membrane Protein (LAMP-1) Enhance Gag-Specific Immune Response(2014) GODINHO, Rodrigo Maciel da Costa; MATASSOLI, Flavio Lemos; LUCAS, Carolina Goncalves de Oliveira; RIGATO, Paula Ordonhez; GONCALVES, Jorge Luiz Santos; SATO, Maria Notomi; MACIEL JR., Milton; PECANHA, Ligia Maria Torres; AUGUST, J. Thomas; MARQUES JR., Ernesto Torres de Azevedo; ARRUDA, Luciana Barros deWe have previously demonstrated that a DNA vaccine encoding HIV-p55gag in association with the lysosomal associated membrane protein-1 (LAMP-1) elicited a greater Gag-specific immune response, in comparison to a DNA encoding the native gag. In vitro studies have also demonstrated that LAMP/Gag was highly expressed and was present in MHCII containing compartments in transfected cells. In this study, the mechanisms involved in these processes and the relative contributions of the increased expression and altered traffic for the enhanced immune response were addressed. Cells transfected with plasmid DNA constructs containing p55gag attached to truncated sequences of LAMP-1 showed that the increased expression of gag mRNA required p55gag in frame with at least 741 bp of the LAMP-1 luminal domain. LAMP luminal domain also showed to be essential for Gag traffic through lysosomes and, in this case, the whole sequence was required. Further analysis of the trafficking pathway of the intact LAMP/Gag chimera demonstrated that it was secreted, at least in part, associated with exosome-like vesicles. Immunization of mice with LAMP/gag chimeric plasmids demonstrated that high expression level alone can induce a substantial transient antibody response, but targeting of the antigen to the endolysosomal/secretory pathways was required for establishment of cellular and memory response. The intact LAMP/gag construct induced polyfunctional CD4(+) T cell response, which presence at the time of immunization was required for CD8(+) T cell priming. LAMP-mediated targeting to endolysosomal/secretory pathway is an important new mechanistic element in LAMP-mediated enhanced immunity with applications to the development of novel anti-HIV vaccines and to general vaccinology field.
conferenceObject DEFECTIVE ACTIVATION OF DENDRITIC CELLS IN A CASE OF LOCALLY INVASIVE ASPERGILLOSIS(2012) MORAES-VASCONCELOS, D.; RIGATO, P. O.; DIAS, A. Santos; ALVES, C.; ORII, N. M.; OGUSUKU, S.; FERREIRA, M. DominguesPrimary immunodeficiencies are rare and usually first manifest during childhood. Invasive aspergillosis is the leading cause of mortality in phagocyte defects, reflecting the key role of these cells in the host defense against opportunistic fungi. Patients with AD STAT-3 de ficiency are prone to colonization of lung cavities (pneumatoceles) by Aspergillus species leading to local invasion and rarely disseminated infection. Other phagocytic and T-cell disorders are uncommonly associated with invasive aspergillosis. We describe herein the case of a young man, 34 years old, born to a non-consanguineous family, presenting chronic sinusitis treated with multiple antibiotic schemes for more than one year without resolution. He then presented tenderness in the frontal area of the head, fo llowed by ulceration. The computed tomography of brain showed osteolytic lesion of the skull, associated to invasion of the skin and paranasal sinuses. The biopsy showed fungal structures, identified as Aspergillus fumigatus. Laboratorial investigation evidenced normal blood cell counts, Ig levels, DHR, G6PD, myeloperoxidase and lymphocyte immunophenotyping. Lymphoproliferation assays showed decreased response to tetanus toxoid and toxoplasma, but normal response to CMV and T cell mitogens. Monocyte derived dendritic cells presented decreased activation parameters after Aspergillus as well as by Candida antigen stimulation. Ag specific T cell costimulation was also severely decreased when compared to healthy controls. This is to our knowledge the first case of a dendritic cell disturbance associated to invasive Aspergillus infection. This case report highlights the complex coordination of both innate and acquired pathways mediating host defense against Aspergillus infection.- Maternal LAMP/p55gagHIV-1 DNA Immunization Induces In Utero Priming and a Long-Lasting Immune Response in Vaccinated Neonates(2012) RIGATO, Paula Ordonhez; MACIEL JR., Milton; GOLDONI, Adriana Leticia; PIUBELLI, Orlando Guerra; ORII, Noemia Mie; MARQUES, Ernesto Torres; AUGUST, Joseph Thomas; DUARTE, Alberto Jose da Silva; SATO, Maria NotomiInfants born to HIV-infected mothers are at high risk of becoming infected during gestation or the breastfeeding period. A search is thus warranted for vaccine formulations that will prevent mother-to-child HIV transmission. The LAMP/gag DNA chimeric vaccine encodes the HIV-1 p55gag fused to the lysosome-associated membrane protein-1 (LAMP-1) and has been shown to enhance anti-Gag antibody (Ab) and cellular immune responses in adult and neonatal mice; such a vaccine represents a new concept in antigen presentation. In this study, we evaluated the effect of LAMP/gag DNA immunization on neonates either before conception or during pregnancy. LAMP/gag immunization of BALB/c mice before conception by the intradermal route led to the transfer of anti-Gag IgG1 Ab through the placenta and via breastfeeding. Furthermore, there were an increased percentage of CD4+ CD25+ Foxp3+ T cells in the spleens of neonates. When offspring were immunized with LAMP/gag DNA, the anti-Gag Ab response and the Gag-specific IFN-gamma-secreting cells were decreased. Inhibition of anti-Gag Ab production and cellular responses were not observed six months after immunization, indicating that maternal immunization did not interfere with the long-lasting memory response in offspring. Injection of purified IgG in conjunction with LAMP/gag DNA immunization decreased humoral and cytotoxic T-cell responses. LAMP/gag DNA immunization by intradermal injection prior to conception promoted the transfer of Ab, leading to a diminished response to Gag without interfering with the development of anti-Gag T- and B-cell memory. Finally, we assessed responses after one intravenous injection of LAMP/gag DNA during the last five days of pregnancy. The intravenous injection led to in utero immunization. In conclusion, DNA vaccine enconding LAMP-1 with Gag and other HIV-1 antigens should be considered in the development of a protective vaccine for the maternal/fetal and newborn periods.