CAMILA MALTA ROMANO
Projetos de Pesquisa
Unidades Organizacionais
LIM/52 - Laboratório de Virologia, Hospital das Clínicas, Faculdade de Medicina
11 resultados
Resultados de Busca
Agora exibindo 1 - 10 de 11
- Production of a Recombinant Dengue Virus 2 NS5 Protein and Potential Use as a Vaccine Antigen(2016) ALVES, Rbens Prince dos Santos; PEREIRA, Lennon Ramos; FABRIS, Denicar Lina Nascimento; SALVADOR, Felipe Scassi; SANTOS, Robert Andreata; ZANOTTO, Paolo Marinho de Andrade; ROMANO, Camila Malta; AMORIM, Jaime Henrique; FERREIRAA, Luis Carlos de SouzaDengue fever is caused by any of the four known dengue virus serotypes (DENV1 to DENV4) that affect millions of people worldwide, causing a significant number of deaths. There are vaccines based on chimeric viruses, but they still are not in clinical use. Anti-DENV vaccine strategies based on nonstructural proteins are promising alternatives to those based on whole virus or structural proteins. The DENV nonstructural protein 5 (NS5) is the main target of anti-DENV T cell-based immune responses in humans. In this study, we purified a soluble recombinant form of DENV2 NS5 expressed in Escherichia coli at large amounts and high purity after optimization of expression conditions and purification steps. The purified DENV2 NS5 was recognized by serum from DENV1-, DENV2-, DENV3-, or DENV4-infected patients in an epitope-conformation-dependent manner. In addition, immunization of BALB/c mice with NS5 induced high levels of NS5-specific antibodies and expansion of gamma interferon-and tumor necrosis factor alpha-producing T cells. Moreover, mice immunized with purified NS5 were partially protected from lethal challenges with the DENV2 NGC strain and with a clinical isolate (JHA1). These results indicate that the recombinant NS5 protein preserves immunological determinants of the native protein and is a promising vaccine antigen capable of inducing protective immune responses.
- Genotypic distribution of HHV-8 in AIDS individuals without and with Kaposi sarcoma Is genotype B associated with better prognosis of AIDS-KS?(2016) TOZETTO-MENDOZA, Tania Regina; IBRAHIM, Karim Yaqub; TATENO, Adriana Fumie; OLIVEIRA, Cristiane Mendes de; SUMITA, Laura Massami; SANCHEZ, Maria Carmem Arroyo; LUNA, Expedito Jose; PIERROTTI, Ligia Camara; DREXLER, Jan Felix; BRAZ-SILVA, Paulo Henrique; PANNUTI, Claudio Sergio; ROMANO, Camila MaltaAIDS-associated Kaposi's sarcoma (AIDS-KS) caused by human herpes virus 8 (HHV-8) is the most severe and resistant form of KS tumor. Our aim was to verify whether there is an association between HHV-8 variability and development of AIDS-KS in Brazil by comparing the HHV-8 variability between individuals without and with KS. Saliva samples and blood, when available, were analyzed by PCR techniques for detection of the fragments of ORF K1 of HHV-8, which were then genotyped and analyzed regarding the genetic variability. Our study described 106 positive cases for HHV-8 in the saliva from 751 AIDS patients without previous KS. In addition, we performed a phylogenetic analysis of HHV-8 in 34 of the 106 AIDS patients without KS and in 33 of the 37 patients with active KS. The distribution of HHV-8 genotypes A, B, C, and F in AIDS individuals was indistinguishable by comparing non-KS and KS groups, as well as regarding ethnicity. Considering the KS group, genotype B was associated with better prognosis of KS tumor. Interestingly. we found a particular profile of diversity within Glade C and 2 recombinant patterns of HHV-8 in the saliva of AIDS individuals without KS. We emphasize the need to achieve standard genotyping protocol for ORF K1 amplification, thus allowing for substantial detection of HHV-8 variants. Our findings can shed light on the role of HHV-8 variability in the pathogenesis of AIDS-KS.
- Serotype influences on dengue severity: a cross-sectional study on 485 confirmed dengue cases in Vitoria, Brazil(2016) VICENTE, Creuza Rachel; HERBINGER, Karl-Heinz; FROESCHL, Guenter; ROMANO, Camila Malta; CABIDELLE, Aline de Souza Areias; CERUTTI JUNIOR, CrispimBackground: Dengue is caused by a RNA virus of the family Flaviviridae, which presents four serotypes (DENV-1 to DENV-4) capable of inducing hemorrhage. The purpose of this study was to evaluate the influence of serotype on the outcome of dengue. Methods: This cross-sectional study included data from dengue cases with serotyping results that occurred between 2009 and 2013 in Vitoria, Espirito Santo, Brazil. Data were accessed through the Information System for Notifiable Diseases. Chi-square test, Fisher exact test, Mann-Whitney U test, and logistic regression were performed to assess associations between different serotypes and dengue severity, while considering gender and age. Results: The sample consisted of 485 laboratory confirmed dengue cases, of which 46.4 % were females, with median age of 26 years. Regarding overall samples, 77.3 % were caused by DENV-1, 16.1 % by DENV-4, 6.4 % by DENV-2, and 0.2 % by DENV-3. Severe dengue affected 6.6 % of all cases, of which 32.3 % of the cases caused by DENV-2, 6.4 % of those caused by DENV-4, 4.5 % of those caused by DENV-1, and none of those caused by DENV-3. Severe dengue was found to be seven times more frequent among cases of DENV-2 than among those of the other serotypes. Conclusions: The present study found that cases of DENV-2 had a higher proportion of severe dengue than among those of DENV-1 and DENV-4. Consequently, early detection of serotypes circulating in the territory could be an important approach to prevent increasing numbers of severe outcomes during dengue outbreaks by predicting the health support needed for early diagnoses and treatment of dengue cases.
- Antibodies are not required to a protective immune response against dengue virus elicited in a mouse encephalitis model(2016) AMORIM, Jaime Henrique; ALVES, Rubens Prince dos Santos; BIZERRA, Raiza; PEREIRA, Sara Araujo; PEREIRA, Lennon Ramos; FABRIS, Denicar Lina Nascimento; SANTOS, Robert Andreata; ROMANO, Camila Malta; FERREIRA, Luis Carlos de SouzaGenerating neutralizing antibodies have been considered a prerequisite to control dengue virus (DENV) infection. However, T lymphocytes have also been shown to be important in a protective immune state. In order to investigate the contribution of both humoral and cellular immune responses in DENV immunity, we used an experimental model in which a non-lethal DENV2 strain (ACS46) is used to intracranially prime Balb/C mice which develop protective immunity against a lethal DENV2 strain (JHA1). Primed mice generated envelope-specific antibodies and CD8(+) T cell responses targeting mainly non-structural proteins. Immune sera from protected mice did not confer passive protection to naive mice challenged with the JHA1 strain. In contrast, depletion of CD4(+) and CD8(+) T lymphocytes significantly reduced survival of ACS46-primed mice challenged with the JHA1 strain. Collectively, results presented in this study show that a cellular immune response targeting non-structural proteins are a promising way in vaccine development against dengue.
- New findings about trichodysplasia spinulosa-associated polyomavirus (TSPyV)-novel qPCR detects TSPyV-DNA in blood samples(2016) URBANO, Paulo R.; NALI, Luiz H. S.; BICALHO, Camila S.; PIERROTTI, Ligia C.; DAVID-NETO, Elias; PANNUTI, Claudio S.; ROMANO, Camila M.A new real-time PCR assay for trichodysplasia spinulosa associated polyomavirus (TSPyV) DNA detection was designed, and blood samples from kidney transplant recipients and healthy individuals were screened. TSPyV-DNA was not detected in blood from healthy individuals, but 26.8% of kidney recipients presented TSPyV-DNA. This is the first report of TSPyV viremia.
conferenceObject Profile of HERV-W expression in multiple sclerosis patients(2016) NALI, L. H.; SILVA, I. T. da; OLIVAL, G. S. do; DIAS-NETO, E.; SILVA, D. F.; URBANO, P. R.; TILBERY, C. P.; PENALVA-DE-OLIVEIRA, A. C.; ROMANO, C. M.- Resistance-associated variants in HCV subtypes 1a and 1b detected by Ion Torrent sequencing platform(2016) GASPARETO, Karine V.; RIBEIRO, Roberto M.; MALTA, Fernanda de Mello; GOMES-GOUVEA, Michele S.; MUTO, Nair H.; ROMANO, Camila M.; MENDES-CORREA, Maria C.; CARRILHO, Flair J.; SABINO, Ester C.; PINHO, Joao R. RebelloBackground: As a result of increased understanding of the HCV life cycle, a new generation of drugs known as direct-acting antivirals (DAAs) was developed and is constantly being improved. At baseline, HCV variants resistant to DAA therapy may pre-exist, increasing the likelihood of treatment failure. The aim of this study was to investigate the presence of resistance-associated variants (RAVs) in treatment-naive patients infected with HCV subtypes 1a and 1b. Methods: Next-generation sequencing was used to assess the frequencies of NS3-4A, NS5A and NS5B RAVs in 100 HCV monoinfected DAA-naive patients (HCV-1a: n= 51; HCV-1b: n= 49). Results: Complete HCV sequence information was obtained for most samples. RAVs were detected in the NS3-4A (T54S, V55A, Q80K and R155K), NS5A (Q30H/R, H58P and Y93C/H/N) and NS5B (A421V) regions in 10%, 22% and 8%, respectively, of patients infected with HCV subtype-1a. Among the patients infected with HCV subtype-1b, mutations in the NS3-4A (F43I, T54S, Q80H, D168E and M175L), NS5A (L28M, R30Q, L31M, Q54H, A92T and Y93H) and NS5B (L159F, C316N, A421V and S556G) regions were observed in 12%, 53% and 31% of patients, respectively. Conclusions: High-throughput DNA sequencing allows an easier and more complete analysis of DAA RAVs, including mutations that represent only a minor variant of the whole viral population. RAVs to the three different classes of DAAs were found in our population. The characterization of their profile in the circulating virus is relevant to determine the better treatment option for infected individuals or to guide the implementation of treatment policies.
- ALTERNATIVE METHODS FOR SEQUENCING FULL TSPyV GENOMES USING SANGER OR NGS(2016) URBANO, Paulo Roberto; OLIVEIRA, Ana Carolina Soares de; ROMANO, Camila Malta
- Occurrence, genotypic characterization, and patterns of shedding of human polyomavirus JCPyV and BKPyV in urine samples of healthy individuals in SAo Paulo, Brazil(2016) URBANO, Paulo Roberto Palma; OLIVEIRA, Renato Reis; ROMANO, Camila Malta; PANNUTI, Claudio Sergio; FINK, Maria Cristina Domingues da SilvaThe objective of this study was to evaluate the prevalence, genotypic characterization, and determination of the patterns of shedding of human polyomavirus JC (JCPyV) and BK (BKPyV) in consecutive urine samples collected from healthy adults. Urine samples collected monthly over a 6 month period were screened by polymerase chain reaction (PCR) with two sets of primers complementary to the VP1 protein region specific for the JCPyV or BKPyV genome. The viral load of JCPyV and BKPyV in positive samples was determined by quantitative real time PCR. Seventy-one healthy individuals (ages between 18 and 65) were included in the study. Polyomavirus DNA urinary shedding was identified in 44 (62%) of the 71 individuals evaluated: BKPyV only in 16 (22.5%); JCPyV only in 19 (26.7%); and both in 9 (12.7%). Among the 28 individuals shedding JCPyV, the shedding was nearly continuous in 13 (46.4%) and sporadic in 15 (53.6%), whereas all BKPyV shedding was sporadic. A total of 45 (19 BKPyV and 26 JCPyV) strains were identified. Of the BKPyV strains, individuals were observed that excreted all genotypes except genotype 3 and the JCPyV strains, excretion of 5 different genotypes. Evaluating the age of individuals who excrete JCPyV and BKPyV, mostly are young adults, with a slight increase with increasing age and observing the viral load can not draw any parallel between the increase or decrease of age or excreted genotype as there was a wide variation both in the excretion of BKPyV and JCPyV. The high occurrence of isolated or simultaneous urinary shedding of JCPyV and BKPyV in healthy individuals merits further study. J. Med. Virol. 88:153-158, 2016. (c) 2015 Wiley Periodicals, Inc.
- Detection of Culex flavivirus and Aedes flavivirus nucleotide sequences in mosquitoes from parks in the city of Sao Paulo, Brazil(2016) FERNANDES, Licia Natal; PAULA, Marcia Bicudo de; ARAUJO, Alessandra Bergamo; GONCALVES, Elisabeth Fernandes Bertoletti; ROMANO, Camila Malta; NATAL, Delsio; MALAFRONTE, Rosely dos Santos; MARRELLI, Mauro Toledo; LEVI, Jose EduardoThe dengue viruses are widespread in Brazil and are a major public health concern. Other flaviviruses also cause diseases in humans, although on a smaller scale. The city of Sao Paulo is in a highly urbanized area with few green spaces apart from its parks, which are used for recreation and where potential vertebrate hosts and mosquito vectors of pathogenic Flavivirus species can be found. Although this scenario can contribute to the transmission of Flavivirus to humans, little is known about the circulation of members of this genus in these areas. In light of this, the present study sought to identify Flavivirus infection in mosquitoes (Diptera: Culicidae) collected in parks in the city of Sao Paulo. Seven parks in different sectors of the city were selected. Monthly mosquito collections were carried out in each park from March 2011 to February 2012 using aspiration and traps (Shannon and CD C-CO2). Nucleic acids were extracted from the mosquitoes collected and used for reverse-transcriptase and real-time polymerase chain reactions with genus-specific primers targeting a 200-nucleotide region in the Flavivirus NS5 gene. Positive samples were sequenced, and phylogenetic analyses were performed. Culex and Aedes were the most frequent genera of Culicidae collected. Culex flavivirus (CxFV)-related and Aedes flavivirus (AEFV)- related nucleotide sequences were detected in 17 pools of Culex and two pools of Aedes mosquitoes, respectively, among the 818 pools of non-engorged females analyzed. To the best of our knowledge, this is the first report of CxFV and AEFV in the city of Sao Paulo and Latin America, respectively. Both viruses are insect- specific flaviviruses, a group known to replicate only in mosquito cells and induce a cytopathic effect in some situations. Hence, our data suggests that CxFV and AEFV are present in Culex and Aedes mosquitoes, respectively, in parks in the city of Sao Paulo. Even though Flavivirus species of medical importance were not detected, surveillance is recommended in the study areas because of the presence of vertebrates and mosquitoes that could act as amplifying hosts and vectors of flaviviruses, providing the required conditions for circulation of these viruses.