PRISCILLA RAMOS COSTA
Projetos de Pesquisa
Unidades Organizacionais
LIM/60 - Laboratório de Imunologia Clínica e Alergia, Hospital das Clínicas, Faculdade de Medicina
4 resultados
Resultados de Busca
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- Performance comparison of the Maxim and Sedia Limiting Antigen Avidity assays for HIV incidence surveillance(2019) SEMPA, Joseph B.; WELTE, Alex; BUSCH, Michael P.; HALL, Jake; HAMPTON, Dylan; FACENTE, Shelley N.; KEATING, Sheila M.; MARSON, Kara; PARKIN, Neil; PILCHER, Christopher D.; MURPHY, Gary; GREBE, Eduard; SEMP, Joseph; MATTEN, David; BRAND, Hilmarie; CHIBAWARA, Trust; MCKINNEY, Elaine; FACENTE, Shelley; KEATING, Sheila; LEBEDEVA, Mila; KASSANJEE, Reshma; LAEYENDECKER, Oliver; QUINN, Thomas; BURNS, David; LITTLE, Susan; SANDS, Anita; HALLETT, Tim; OWEN, Sherry Michele; PAREKH, Bharat; SEXTON, Connie; PRICE, Matthew; KAMALI, Anatoli; LOEB, Lisa; MARTIN, Jeffrey; DEEKS, Steven G.; HOH, Rebecca; BARTOLOMEI, Zelinda; CERQUEIRA, Natalia; SANTOS, Breno; ZABTOSKI, Kellin; LIRA, Rita de Cassia Alves; SPERHACKE, Rosa Dea; MOTTA, Leonardo R.; PAGANELLA, Machline; KALLAS, Esper; TOMIYAMA, Helena; TOMIYAMA, Claudia; COSTA, Priscilla; NUNES, Maria A.; REIS, Gisele; SAUER, Mariana M.; NAKAGAWA, Zelinda; FERRARI, Lilian; AMARAL, Ana P.; MILANI, Karine; KARIM, Salim S. Abdool; KARIM, Quarraisha Abdool; NDUNGU, Thumbi; MAJOLA, Nelisile; SAMSUNDER, Natasha; NANICHE, Denise; MANDOMANDO, Inacio; V, Eusebio Macete; SANCHEZ, Jorge; LAMA, Javier; DUERR, Ann; CAPOBIANCHI, Maria R.; SULIGOI, Barbara; STRAMER, Susan; WILLIAMSON, Phillip; VERMEULEN, Marion; SABINO, EsterBackground Two manufacturers, Maxim Biomedical and Sedia Biosciences Corporation, supply CDC-approved versions of the HIV-1 Limiting Antigen Avidity EIA (LAg) for detecting 'recent' HIV infection in cross-sectional incidence estimation. This study assesses and compares the performance of the two assays for incidence surveillance. Methods We ran both assays on a panel of 2,500 well-characterized HIV-1-infected specimens. We analysed concordance of assay results, assessed reproducibility using repeat testing and estimated mean durations of recent infection (MDRIs) and false-recent rates (FRRs) for a range of normalized optical density (ODn) thresholds, alone and in combination with viral load thresholds. We defined three hypothetical surveillance scenarios, similar to the Kenyan and South African epidemics, and a concentrated epidemic. These scenarios allowed us to evaluate the precision of incidence estimates obtained by means of various recent infection testing algorithms (RITAs) based on each of the two assays. Results The Maxim assay produced lower ODn values than the Sedia assay on average, largely as a result of higher calibrator readings (mean OD of 0.749 vs. 0.643), with correlation of normalized readings lower (R-2 = 0.908 vs. R-2 = 0.938). Reproducibility on blinded control specimens was slightly better for Maxim. The MDRI of a Maxim-based algorithm at the 'standard' threshold (ODn <= 1.5 & VL > 1,000) was 201 days (95% CI: 180,223) and for Sedia 171 (152,191). The difference Differences in MDRI were estimated at 32.7 (22.9,42.8) and 30.9 days (21.7,40.7) for the two algorithms, respectively. Commensurately, the Maxim algorithm had a higher FRR in treatment-naive subjects (1.7% vs. 1.1%). The two assays produced similar precision of incidence estimates in the three surveillance scenarios. Conclusions Differences between the assays can be primarily attributed to the calibrators supplied by the manufacturers. Performance for surveillance was extremely similar, although different thresholds were optimal (i.e. produced the lowest variance of incidence estimates) and at any given ODn threshold, different estimates of MDRI and FRR were obtained. The two assays cannot be treated as interchangeable: assay and algorithm-specific performance characteristic estimates must be used for survey planning and incidence estimation.
- Low-Cost Ultra-Wide Genotyping Using Roche/454 Pyrosequencing for Surveillance of HIV Drug Resistance(2012) DUDLEY, Dawn M.; CHIN, Emily N.; BIMBER, Benjamin N.; SANABANI, Sabri S.; TAROSSO, Leandro F.; COSTA, Priscilla R.; SAUER, Mariana M.; KALLAS, Esper G.; O'CONNOR, David H.Background: Great efforts have been made to increase accessibility of HIV antiretroviral therapy (ART) in low and middle-income countries. The threat of wide-scale emergence of drug resistance could severely hamper ART scale-up efforts. Population-based surveillance of transmitted HIV drug resistance ensures the use of appropriate first-line regimens to maximize efficacy of ART programs where drug options are limited. However, traditional HIV genotyping is extremely expensive, providing a cost barrier to wide-scale and frequent HIV drug resistance surveillance. Methods/Results: We have developed a low-cost laboratory-scale next-generation sequencing-based genotyping method to monitor drug resistance. We designed primers specifically to amplify protease and reverse transcriptase from Brazilian HIV subtypes and developed a multiplexing scheme using multiplex identifier tags to minimize cost while providing more robust data than traditional genotyping techniques. Using this approach, we characterized drug resistance from plasma in 81 HIV infected individuals collected in Sao Paulo, Brazil. We describe the complexities of analyzing next-generation sequencing data and present a simplified open-source workflow to analyze drug resistance data. From this data, we identified drug resistance mutations in 20% of treatment naive individuals in our cohort, which is similar to frequencies identified using traditional genotyping in Brazilian patient samples. Conclusion: The developed ultra-wide sequencing approach described here allows multiplexing of at least 48 patient samples per sequencing run, 4 times more than the current genotyping method. This method is also 4-fold more sensitive (5% minimal detection frequency vs. 20%) at a cost 3-5 x less than the traditional Sanger-based genotyping method. Lastly, by using a benchtop next-generation sequencer (Roche/454 GS Junior), this approach can be more easily implemented in low-resource settings. This data provides proof-of-concept that next-generation HIV drug resistance genotyping is a feasible and low-cost alternative to current genotyping methods and may be particularly beneficial for in-country surveillance of transmitted drug resistance.
- IL-10-Producing Regulatory B Cells Are Decreased in Patients with Common Variable Immunodeficiency(2016) BARSOTTI, Nathalia Silveira; ALMEIDA, Rafael Ribeiro; COSTA, Priscilla Ramos; BARROS, Myrthes Toledo; KALIL, Jorge; KOKRON, Cristina MariaCommon variable immunodeficiency (CVID) is the most prevalent symptomatic primary immunodeficiency in adults. CVID patients often present changes in the frequency and function of B lymphocytes, reduced number of Treg cells, chronic immune activation, recurrent infections, high incidence of autoimmunity and increased risk for malignancies. We hypothesized that the frequency of B10 cells would be diminished in CVID patients because these cells play an important role in the development of Treg cells and in the control of T cell activation and autoimmunity. Therefore, we evaluated the frequency of B10 cells in CVID patients and correlated it with different clinical and immunological characteristics of this disease. Forty-two CVID patients and 17 healthy controls were recruited for this study. Cryo-preserved PBMCs were used for analysis of T cell activation, frequency of Treg cells and characterization of B10 cells by flow cytometry. IL-10 production by sorted B cells culture and plasma sCD14 were determined by ELISA. We found that CVID patients presented decreased frequency of IL-10-producing CD24(hi)CD38(hi) B cells in different cell culture conditions and decreased frequency of IL-10-producing CD24(hi)CD27(+) B cells stimulated with CpG+PIB. Moreover, we found that CVID patients presented lower secretion of IL-10 by sorting-purified B cells when compared to healthy controls. The frequency of B10 cells had no correlation with autoimmunity, immune activation and Treg cells in CVID patients. This work suggests that CVID patients have a compromised regulatory B cell compartment which is not correlated with clinical and immunological characteristics presented by these individuals.
- Subsets of Memory CD4(+) T Cell and Bactericidal Antibody Response to Neisseria meningitidis Serogroup C after Immunization of HIV-Infected Children and Adolescents(2014) MILAGRES, Lucimar G.; COSTA, Priscilla R.; SILVA, Giselle P.; CARVALHO, Karina I.; PEREIRA-MANFRO, Wania F.; FERREIRA, Bianca; BARRETO, Daniella M.; FROTA, Ana Cristina C.; HOFER, Cristina B.; KALLAS, Esper G.Meningococcal disease is endemic in Brazil, with periodic outbreaks and case fatality rates reach as high as 18 to 20% of cases. Conjugate vaccines against meningococci are immunogenic in healthy children. However, we have previously shown a poor bactericidal antibody response to a Men C conjugate vaccine in Brazilian HIV-infected children and adolescents after a single vaccine administration. The goal of the present work was to investigate associations between bactericidal antibody response induced by MenC vaccine and the frequency and activation profile (expression of CD38, HLA-DR and CCR5 molecules) of total CD4(+) memory T cell sub-populations in HIV-1-infected children and adolescents. Responders to vaccination against MenC had a predominance (about 44%) of CD4(+) T-INTERMEDIATE subset followed by T-TRANSITIONAL memory subset (23 to 26%). Importantly, CD4(+) T-INT frequency was positively associated with bactericidal antibody response induced by vaccination. The positive correlation persisted despite the observation that the frequency T-INT CD38(+)HLA-DR+ was higher in responders. In contrast, CD4(+) T-CENTRAL MEMORY (T-CM) subset negatively correlated with bactericidal antibodies. In conclusion, these data indicate that less differentiated CD+ T cells, like TCM may be constantly differentiating into intermediate and later differentiated CD4(+) T cell subsets. These include CD4 T-INT subset which showed a positive association with bactericidal antibodies.