CASSIA GISELE TERRASSANI SILVEIRA
Projetos de Pesquisa
Unidades Organizacionais
Instituto Central, Hospital das Clínicas, Faculdade de Medicina
17 resultados
Resultados de Busca
Agora exibindo 1 - 10 de 17
- Serum Level of YB-1 and CA12-5 in the Diagnostics of Endometriosis(2015) AHRENS, T.; SILVEIRA, C.; AGIC, A.; RODY, A.; HORNUNG, D.
- Generation of Zika virus - specific T cells from seropositive and virus-naive donors for potential use as an autologous or ""off-the-shelf"" immunotherapeutic(2019) HANAJIRI, Ryo; SANI, Gelina M.; HANLEY, Patrick J.; SILVEIRA, Cassia G.; KALLAS, Esper G.; KELLER, Michael D.; BOLLARD, Catherine M.Background: Zika virus (ZIKV) infection can cause severe birth defects in newborns with no effective currently available treatment. Adoptive transfer of virus-specific T cells has proven to be safe and effective for the prevention or treatment of many viral infections, and could represent a novel treatment approach for patients with ZIKV infection. However, extending this strategy to the ZIKV setting has been hampered by limited data on immunogenic T-cell antigens within ZIKV. Hence, we have generated ZIKV-specific T cells and characterized the cellular immune responses against ZIKV antigens. Methods: T-cell products were generated from peripheral blood of ZIKV-exposed donors, ZIKV-naive adult donors and umbilical cord blood by stimulation with pentadecamer (15mer) overlapping peptide libraries spanning four ZIKV polyproteins (C, M, E and NS1) using a Good Manufacturing Practice compliant protocol. Results: We successfully generated T cells targeting ZIKV antigens with clinically relevant numbers. The ex vivo-expanded T cells comprised both CD4(+) and CD8(+) T cells that were able to produce Th1-polarized effector cytokines and kill ZIKV-infected HLA-matched monocytes, confirming functionality of this unique T-cell product as a potential ""off-the-shelf"" therapeutic. Epitope mapping using peptide arrays identified several novel HLA class I and class II restricted epitopes within NS1 antigen, which is essential for viral replication and immune evasion. Discussion: Our findings demonstrate that it is feasible to generate potent ZIKV-specific T cells from a variety of cell sources including virus naive donors for future clinical use in an ""off-the-shelf"" setting.
- Plasmablast Expansion Following the Tetravalent, Live-Attenuated Dengue Vaccine Butantan-DV in DENV-Naive and DENV-Exposed Individuals in a Brazilian Cohort(2022) SILVEIRA, Cassia G. T.; MAGNANI, Diogo M.; COSTA, Priscilla R.; AVELINO-SILVA, Vivian I.; RICCIARDI, Michael J.; TIMENETSKY, Maria do Carmo S. T.; GOULART, Raphaella; CORREIA, Carolina A.; MARMORATO, Mariana P.; FERRARI, Lilian; NAKAGAWA, Zelinda B.; TOMIYAMA, Claudia; TOMIYAMA, Helena; KALIL, Jorge; PALACIOS, Ricardo; PRECIOSO, Alexander R.; WATKINS, David I.; KALLAS, Esper G.An effective vaccine against the dengue virus (DENV) should induce a balanced, long-lasting antibody (Ab) response against all four viral serotypes. The burst of plasmablasts in the peripheral blood after vaccination may reflect enriched vaccine-specific Ab secreting cells. Here we characterize the acute plasmablast responses from naive and DENV-exposed individuals following immunization with the live attenuated tetravalent (LAT) Butantan DENV vaccine (Butantan-DV). The frequency of circulating plasmablasts was determined by flow cytometric analysis of fresh whole blood specimens collected from 40 participants enrolled in the Phase II Butantan-DV clinical trial (NCT01696422) before and after (days 6, 12, 15 and 22) vaccination. We observed a peak in the number of circulating plasmablast at day 15 after vaccination in both the DENV naive and the DENV-exposed vaccinees. DENV-exposed vaccinees experienced a significantly higher plasmablast expansion. In the DENV-naive vaccinees, plasmablasts persisted for approximately three weeks longer than among DENV-exposed volunteers. Our findings indicate that the Butantan-DV can induce plasmablast responses in both DENV-naive and DENV-exposed individuals and demonstrate the influence of pre-existing DENV immunity on Butantan DV-induced B-cell responses.
- Consumptive coagulopathy of severe yellow fever occurs independently of hepatocellular tropism and massive hepatic injury(2020) BAILEY, Adam L.; I, Liang- Kang; ZANELLA, Luiz Gonzaga Francisco de Assis Barros D'Elia; SILVEIRA, Cassia G. T.; HO, Yeh-Li; FOQUET, Lander; BIAL, Greg; MCCUNE, Broc T.; DUARTE-NETO, Amaro Nunes; THOMAS, Archana; RAUE, Hans-Peter; BYRNES, Kathleen; KALLAS, Esper G.; SLIFKA, Mark K.; DIAMOND, Michael S.Yellow fever (YF) is a mosquito-transmitted viral disease that causes tens of thousands of deaths each year despite the long-standing deployment of an effective vaccine. In its most severe form, YF manifests as a hemorrhagic fever that causes severe damage to visceral organs. Although coagulopathy is a defining feature of severe YF in humans, the mechanism by which it develops remains uncertain. Hepatocytes are a major target of yellow fever virus (YFV) infection, and the coagulopathy in severe YF has long been attributed to massive hepatocyte infection and destruction that results in a defect in clotting factor synthesis. However, when we analyzed blood from Brazilian patients with severe YF, we found high concentrations of plasma D-dimer, a fibrin split product, suggestive of a concurrent consumptive process. To define the relationship between coagulopathy and hepatocellular tropism, we compared infection and disease in Fah(-/-), Rag2(-/-), and Il2r gamma(-/-) mice engrafted with human hepatocytes (hFRG mice) and rhesus macaques using a highly pathogenic African YFV strain. YFV infection of macaques and hFRG mice caused substantial hepatocyte infection, liver damage, and coagulopathy as defined by virological, clinical, and pathological criteria. However, only macaques developed a consumptive coagulopathy whereas YFV-infected hFRG mice did not. Thus, infection of cell types other than hepatocytes likely contributes to the consumptive coagulopathy associated with severe YF in primates and humans. These findings expand our understanding of viral hemorrhagic disease and associated coagulopathy and suggest directions for clinical management of severe YF cases.
- Prior Dengue Virus Exposure Shapes T Cell Immunity to Zika Virus in Humans(2017) GRIFONI, Alba; PHAM, John; SIDNEY, John; O'ROURKE, Patrick H.; PAUL, Sinu; PETERS, Bjoern; MARTINI, Sheridan R.; SILVA, Aruna D. de; RICCIARDI, Michael J.; MAGNANI, Diogo M.; SILVEIRA, Cassia G. T.; MAESTRI, Alvino; COSTA, Priscilla R.; DE-OLIVEIRA-PINTO, Luzia Maria; AZEREDO, Elzinandes Leal de; DAMASCO, Paulo Vieira; PHILLIPS, Elizabeth; MALLAL, Simon; SILVA, Aravinda M. de; COLLINS, Matthew; DURBIN, Anna; DIEHL, Sean A.; CERPAS, Cristhiam; BALMASEDA, Angel; KUAN, Guillermina; COLOMA, Josefina; HARRIS, Eva; CROWE JR., James E.; STONE, Mars; NORRIS, Phillip J.; BUSCH, Michael; VIVANCO-CID, Hector; COX, Josephine; GRAHAM, Barney S.; LEDGERWOOD, Julie E.; TURTLE, Lance; SOLOMON, Tom; KALLAS, Esper G.; WATKINS, David I.; WEISKOPF, Daniela; SETTE, AlessandroWhile progress has been made in characterizing humoral immunity to Zika virus (ZIKV) in humans, little is known regarding the corresponding T cell responses to ZIKV. Here, we investigate the kinetics and viral epitopes targeted by T cells responding to ZIKV and address the critical question of whether preexisting dengue virus (DENV) T cell immunity modulates these responses. We find that memory T cell responses elicited by prior infection with DENV or vaccination with tetravalent dengue attenuated vaccines (TDLAV) recognize ZIKV-derived peptides. This cross-reactivity is explained by the sequence similarity of the two viruses, as the ZIKV peptides recognized by DENV-elicited memory T cells are identical or highly conserved in DENV and ZIKV. DENV exposure prior to ZIKV infection also influences the timing and magnitude of the T cell response. ZIKV-reactive T cells in the acute phase of infection are detected earlier and in greater magnitude in DENV-immune patients. Conversely, the frequency of ZIKV-reactive T cells continues to rise in the convalescent phase in DENV-naive donors but declines in DENV-preexposed donors, compatible with more efficient control of ZIKV replication and/or clearance of ZIKV antigen. The quality of responses is also influenced by previous DENV exposure, and ZIKV-specific CD8 T cells from DENV-preexposed donors selectively upregulated granzyme B and PD1, unlike DENV-naive donors. Finally, we discovered that ZIKV structural proteins (E, prM, and C) are major targets of both the CD4 and CD8 T cell responses, whereas DENV T cell epitopes are found primarily in nonstructural proteins. IMPORTANCE The issue of potential ZIKV and DENV cross-reactivity and how preexisting DENV T cell immunity modulates Zika T cell responses is of great relevance, as the two viruses often cocirculate and Zika virus has been spreading in geographical regions where DENV is endemic or hyperendemic. Our data show that memory T cell responses elicited by prior infection with DENV recognize ZIKV-derived peptides and that DENV exposure prior to ZIKV infection influences the timing, magnitude, and quality of the T cell response. Additionally, we show that ZIKV-specific responses target different proteins than DENV-specific responses, pointing toward important implications for vaccine design against this global threat.
- A human inferred germline antibody binds to an immunodominant epitope and neutralizes Zika virus(2017) MAGNANI, Diogo M.; SILVEIRA, Cassia G. T.; ROSEN, Brandon C.; RICCIARDI, Michael J.; PEDRENO-LOPEZ, Nuria; GUTMAN, Martin J.; BAILEY, Varian K.; MAXWELL, Helen S.; DOMINGUES, Aline; GONZALEZ-NIETO, Lucas; AVELINO-SILVA, Vivian I.; TRINDADE, Mateus; NOGUEIRA, Juliana; OLIVEIRA, Consuelo S.; MAESTRI, Alvino; FELIX, Alvina Clara; LEVI, Jose Eduardo; NOGUEIRA, Mauricio L.; MARTINS, Mauricio A.; MARTINEZ-NAVIO, Jose M.; FUCHS, Sebastian P.; WHITEHEAD, Stephen S.; BURTON, Dennis R.; DESROSIERS, Ronald C.; KALLAS, Esper G.; WATKINS, David I.The isolation of neutralizing monoclonal antibodies (nmAbs) against the Zika virus (ZIKV) might lead to novel preventative strategies for infections in at-risk individuals, primarily pregnant women. Here we describe the characterization of human mAbs from the plasmablasts of an acutely infected patient. One of the 18 mAbs had the unusual feature of binding to and neutralizing ZIKV despite not appearing to have been diversified by affinity maturation. This mAb neutralized ZIKV (Neut(50) similar to 2 mu g/ml) but did not react with any of the four dengue virus serotypes. Except for the expected junctional diversity created by the joining of the V-(D)-J genes, there was no deviation from immunoglobulin germline genes. This is a rare example of a human mAb with neutralizing activity in the absence of detectable somatic hypermutation. Importantly, binding of this mAb to ZIKV was specifically inhibited by human plasma from ZIKV-exposed individuals, suggesting that it may be of value in a diagnostic setting.
- Ontogeny of the B- and T-cell response in a primary Zika virus infection of a dengue-naive individual during the 2016 outbreak in Miami, FL(2017) RICCIARDI, Michael J.; MAGNANI, Diogo M.; GRIFONI, Alba; KWON, Young-Chan; GUTMAN, Martin J.; GRUBAUGH, Nathan D.; GANGAVARAPU, Karthik; SHARKEY, Mark; SILVEIRA, Cassia G. T.; BAILEY, Varian K.; PEDRENO-LOPEZ, Nuria; GONZALEZ-NIETO, Lucas; MAXWELL, Helen S.; DOMINGUES, Aline; MARTINS, Mauricio A.; PHAM, John; WEISKOPF, Daniela; ALTMAN, John; KALLAS, Esper G.; ANDERSEN, Kristian G.; STEVENSON, Mario; LICHTENBERGER, Paola; CHOE, Hyeryun; WHITEHEAD, Stephen S.; SETTE, Alessandro; WATKINS, David I.Zika virus (ZIKV) is a mosquito-borne flavivirus of significant public health concern. In the summer of 2016, ZIKV was first detected in the contiguous United States. Here we present one of the first cases of a locally acquired ZIKV infection in a dengue-naive individual. We collected blood from a female with a maculopapular rash at day (D) 5 and D7 post onset of symptoms (POS) and we continued weekly blood draws out to D148 POS. To establish the ontogeny of the immune response against ZIKV, lymphocytes and plasma were analyzed in a longitudinal fashion. The plasmablast response peaked at D7 POS (19.6% of CD19(+) B-cells) and was undetectable by D15 POS. ZIKV-specific IgM was present at D5 POS, peaked between D15 and D21 POS, and subsequently decreased. The ZIKV-specific IgG response, however, was not detected until D15 POS and continued to increase after that. Interestingly, even though the patient had never been infected with dengue virus (DENV), cross-reactive IgM and IgG binding against each of the four DENV serotypes could be detected. The highest plasma neutralization activity against ZIKV peaked between D15 and D21 POS, and even though DENV binding antibodies were present in the plasma of the patient, there was neither neutralization nor antibody dependent enhancement (ADE) of DENV. Interestingly, ADE against ZIKV arose at D48 POS and continued until the end of the study. CD4(+) and CD8(+) T-cells recognized ZIKV-NS2A and ZIKV-E, respectively. The tetramer positive CD8(+) T-cell response peaked at D21 POS with elevated levels persisting for months. In summary, this is the first study to establish the timing of the ontogeny of the immune response against ZIKV.
- Global Assessment of Dengue Virus-Specific CD4(+) T Cell Responses in Dengue-Endemic Areas(2017) GRIFONI, Alba; ANGELO, Michael A.; LOPEZ, Benjamin; O'ROURKE, Patrick H.; SIDNEY, John; CERPAS, Cristhiam; BALMASEDA, Angel; SILVEIRA, Cassia G. T.; MAESTRI, Alvino; COSTA, Priscilla R.; DURBIN, Anna P.; DIEHL, Sean A.; PHILLIPS, Elizabeth; MALLAL, Simon; SILVA, Aruna D. De; NCHINDA, Godwin; NKENFOU, Celine; COLLINS, Matthew H.; SILVA, Aravinda M. de; LIM, Mei Qiu; MACARY, Paul A.; TATULLO, Filippo; SOLOMON, Tom; SATCHIDANANDAM, Vijaya; DESAI, Anita; RAVI, Vasanthapram; COLOMA, Josefina; TURTLE, Lance; RIVINO, Laura; KALLAS, Esper G.; PETERS, Bjoern; HARRIS, Eva; SETTE, Alessandro; WEISKOPF, DanielaBackground: Dengue is a major public health problem worldwide. Assessment of adaptive immunity is important to understanding immunopathology and to define correlates of protection against dengue virus (DENV). To enable global assessment of CD4(+) T cell responses, we mapped HLA-DRB1-restricted DENV-specific CD4(+) T cell epitopes in individuals previously exposed to DENV in the general population of the dengue-endemic region of Managua, Nicaragua. Methods: HLA class II epitopes in the population of Managua were identified by an in vitro IFN gamma ELISPOT assay. CD4(+) T cells purified by magnetic bead negative selection were stimulated with HLA-matched epitope pools in the presence of autologous antigen-presenting cells, followed by pool deconvolution to identify specific epitopes. The epitopes identified in this study were combined with those previously identified in the DENV endemic region of Sri Lanka, to generate a ""megapool"" (MP) consisting of 180 peptides specifically designed to achieve balanced HLA and DENV serotype coverage. The DENV CD4MP(180) was validated by intracellular cytokine staining assays. Results: We detected responses directed against a total of 431 epitopes, representing all 4 DENV serotypes, restricted by 15 different HLA-DRB1 alleles. The responses were associated with a similar pattern of protein immunodominance, overall higher magnitude of responses, as compared to what was observed previously in the Sri Lanka region. Based on these epitope mapping studies, we designed a DENV CD4 MP180 with higher and more consistent coverage, which allowed the detection of CD4(+) T cell DENV responses ex vivo in various cohorts of DENV exposed donors worldwide, including donors from Nicaragua, Brazil, Singapore, Sri Lanka, and U.S. domestic flavivirus-naive subjects immunized with Tetravalent Dengue Live-Attenuated Vaccine (TV005). This broad reactivity reflects that the 21 HLA-DRB1 alleles analyzed in this and previous studies account for more than 80% of alleles present with a phenotypic frequency >= 5% worldwide, corresponding to 92% phenotypic coverage of the general population (i.e., 92% of individuals express at least one of these alleles). Conclusion: The DENV CD4 MP180 can be utilized to measure ex vivo responses to DENV irrespective of geographical location.
- Dengue Virus Evades AAV-Mediated Neutralizing Antibody Prophylaxis in Rhesus Monkeys(2017) MAGNANI, Diogo M.; RICCIARDI, Michael J.; BAILEY, Varian K.; GUTMAN, Martin J.; PEDRENO-LOPEZ, Nuria; SILVEIRA, Cassia G. T.; MAXWELL, Helen S.; DOMINGUES, Aline; GONZALEZ-NIETO, Lucas; SU, Qin; NEWMAN, Ruchi M.; PACK, Melissa; MARTINS, Mauricio A.; MARTINEZ-NAVIO, Jose M.; FUCHS, Sebastian P.; RAKASZ, Eva G.; ALLEN, Todd M.; WHITEHEAD, Stephen S.; BURTON, Dennis R.; GAO, Guangping; DESROSIERS, Ronald C.; KALLAS, Esper G.; WATKINS, David I.Development of vaccines against mosquito-borne Flaviviruses is complicated by the occurrence of antibody-dependent enhancement (ADE), which can increase disease severity. Long-term delivery of neutralizing antibodies (nAbs) has the potential to effectively block infection and represents an alternative to vaccination. The risk of ADE may be avoided by using prophylactic nAbs harboring amino acid mutations L234A and L235A (LALA) in the immunoglobulin G (IgG) constant region. Here, we used recombinant adeno-associated viruses (rAAVs) to deliver the anti-dengue virus 3 (DENV3) nAb P3D05. While the administration of rAAV-P3D05-rhesus immunoglobulin GI (rhIgGl)-LALA to rhesus macaques engendered DENV3-neutralizing activity in serum, it did not prevent infection. The emergence of viremia following DENV3 challenge was delayed by 3-6 days in the rAAV-treated group, and replicating virus contained the envelope mutation K64R. This neutralization-resistant variant was also confirmed by virus outgrowth experiments in vitro. By delivering P3D05 with unmutated Fc sequences, we further demonstrated that DENV3 also evaded wild-type nAb prophylaxis, and serum viral loads appeared to be higher in the presence of low levels of unmutated P3D05-rhIgGl. Our study shows that a vectored approach for long-term delivery of nAbs with the LALA mutations is promising, but prophylaxis using a single nAb is likely insufficient at preventing DENV infection and replication.
- Humoral and cellular immune responses to CoronaVac up to one year after vaccination(2022) COSTA, Priscilla Ramos; CORREIA, Carolina Argondizo; MARMORATO, Mariana Prado; DIAS, Juliana Zanatta de Carvalho; THOMAZELLA, Mateus Vailant; SILVA, Amanda Cabral da; OLIVEIRA, Ana Carolina Soares de; GUSMAO, Arianne Fagotti; FERRARI, Lilian; FREITAS, Angela Carvalho; PATINO, Elizabeth Gonzalez; GRIFONI, Alba; WEISKOPF, Daniela; SETTE, Alessandro; SCHARF, Rami; KALLAS, Esper Georges; SILVEIRA, Cassia Gisele TerrassaniCoronavac is a widely used SARS-CoV-2 inactivated vaccine, but its long-term immune response assessment is still lacking. We evaluated SARS-CoV-2-specific immune responses, including T cell activation markers, antigen-specific cytokine production and antibody response following vaccination in 53 adult and elderly individuals participating in a phase 3 clinical trial. Activated follicular helper T (Tfh), non-Tfh and memory CD4(+) T cells were detected in almost all subjects early after the first vaccine dose. Activated memory CD4(+) T cells were predominantly of central and effector memory T cell phenotypes and were sustained for at least 6 months. We also detected a balanced Th1-, Th2- and Th17/Th22-type cytokine production that was associated with response over time, together with particular cytokine profile linked to poor responses in older vaccinees. SARS-CoV-2-specific IgG levels peaked 14 days after the second dose and were mostly stable over one year. CoronaVac was able to induce a potent and durable antiviral antigen-specific cellular response and the cytokine profiles related to the response over time and impacted by the senescence were defined.