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1. WCN24-360 The use of convalescent plasma in a kidney transplant recipient COVID-19 patient: A case report from the perspective of viral load evolution and immune response

Author

Ibañez, Natalia, primary, Aldunate, Fabian, additional, Fajardo, Alvaro, additional, Rammauro, Florencia, additional, Daghero, Hellen, additional, Ferla, Diego, additional, Pereira, Marianoel, additional, Salazar, Cecilia, additional, Pritsch, Otto, additional, Hurtado, Joaquín, additional, Tenzi, Jordan, additional, Bollati-Fogolín, Mariela, additional, Bianchi, Sergio, additional, Nin, Nicolas, additional, Moratorio, Gonzalo, additional, and Moreno, Pilar, additional

Published
2024
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3. Low switched memory B cells are associated with no humoral response after SARS-CoV-2 vaccine boosters in kidney transplant recipients

Author

Seija, Mariana, primary, García-Luna, Joaquin, additional, Rammauro, Florencia, additional, Brugnini, Andreína, additional, Trías, Natalia, additional, Astesiano, Rossana, additional, Santiago, José, additional, Orihuela, Natalia, additional, Zulberti, Catherine, additional, Machado, Danilo, additional, Recalde, Cecilia, additional, Yandián, Federico, additional, Guerisoli, Ana, additional, Noboa, Javier, additional, Orihuela, Sergio, additional, Curi, Lilian, additional, Bugstaller, Emma, additional, Noboa, Oscar, additional, Nin, Marcelo, additional, Bianchi, Sergio, additional, Tiscornia, Adriana, additional, and Lens, Daniela, additional

Published
2023
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5. PD-1/PD-L1 blockade abrogates a dysfunctional innate-adaptive immune axis in critical β-coronavirus disease

Author

Duhalde Vega, Maite, primary, Olivera, Daniela, additional, Gastão Davanzo, Gustavo, additional, Bertullo, Mauricio, additional, Noya, Verónica, additional, Fabiano de Souza, Gabriela, additional, Primon Muraro, Stéfanie, additional, Castro, Icaro, additional, Arévalo, Ana Paula, additional, Crispo, Martina, additional, Galliussi, Germán, additional, Russo, Sofía, additional, Charbonnier, David, additional, Rammauro, Florencia, additional, Jeldres, Mathías, additional, Alamón, Catalina, additional, Varela, Valentina, additional, Batthyany, Carlos, additional, Bollati-Fogolín, Mariela, additional, Oppezzo, Pablo, additional, Pritsch, Otto, additional, Proença-Módena, José Luiz, additional, Nakaya, Helder I., additional, Trias, Emiliano, additional, Barbeito, Luis, additional, Anegon, Ignacio, additional, Cuturi, María Cristina, additional, Moraes-Vieira, Pedro, additional, Segovia, Mercedes, additional, and Hill, Marcelo, additional

Published
2022
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6. ¿What did we learn from Convalescent plasma treatment in a COVID-19 patient two-time kidney transplanted? A case report from the viral and immune response evolution perspective

Author

Aldunate, Fabian, primary, Fajardo, Alvaro, additional, Rammauro, Florencia, additional, Daghero, Hellen, additional, Arce, Rodrigo, additional, Ferla, Diego, additional, Pereira-Gomez, Marianoel, additional, Ibañez, Natalia, additional, Salazar, Cecilia, additional, Iraola, Gregorio, additional, Pritsch, Otto, additional, Bollati, Mariella, additional, Hurtado, Javier, additional, Tenzi, Jordan, additional, Bianchi, Sergio, additional, Nin, Nicolas, additional, Moratorio, Gonzalo, additional, and Moreno, Pilar, additional

Published
2022
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7. Humoral Response to Heterologous SARS-CoV-2 Vaccination in Kidney Transplant Patients Is Heterogeneous and Dose Dependent

Author

Seija, Mariana, primary, Rammauro, Florencia, additional, Noboa, Javier, additional, Santiago, José, additional, Orihuela, Natalia, additional, Zulberti, Catherine, additional, Machado, Danilo, additional, Recalde, Cecilia, additional, Astesiano, Rossana, additional, Yandián, Federico, additional, Frantchez, Victoria, additional, Guerisoli, Ana, additional, Morra, Álvaro, additional, Cassinelli, Daniela, additional, Coelho, Cecilia, additional, de Aramburu, Belén, additional, González-Severgnini, Paulina, additional, Moreno, Romina, additional, Pippolo, Aldana, additional, López, Gabriela, additional, Lemos, Mónica, additional, Somariva, Lorena, additional, López, Eliana, additional, Fumero, Soledad, additional, Orihuela, Carla, additional, Suárez, Ana Laura, additional, Rodríguez, Rosalía, additional, Acuña, Gonzalo, additional, Rabaza, Victoria, additional, Perg, Nancy, additional, Cordero, Rossana, additional, Reisfeld, Cristina, additional, Olivera, Paula, additional, Montero, Paola, additional, Nogueira, Cecilia, additional, Nalerio, Catheryn, additional, Orihuela, Sergio, additional, Curi, Lilián, additional, Bugstaller, Ema, additional, Pritsch, Otto, additional, Nin, Marcelo, additional, Noboa, Oscar, additional, and Bianchi, Sergio, additional

Published
2022
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10. What have we learned from a case of convalescent plasma treatment in a two-time kidney transplant recipient COVID-19 patient? A case report from the perspective of viral load evolution and immune response.

Author

Aldunate, Fabian, Fajardo, Alvaro, Ibañez, Natalia, Rammauro, Florencia, Daghero, Hellen, Arce, Rodrigo, Ferla, Diego, Pereira-Gomez, Marianoel, Salazar, Cecilia, Iraola, Gregorio, Pritsch, Otto, Hurtado, Javier, Tenzi, Jordan, Bollati-Fogolín, Mariela, Bianchi, Sergio, Nin, Nicolas, Moratorio, Gonzalo, and Moreno, Pilar

Published
2023
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12. Comparison of antibody response to SARS-CoV-2 after two doses of inactivated virus and BNT162b2 mRNA vaccines in kidney transplant

Author

Seija, Mariana, primary, Rammauro, Florencia, additional, Santiago, José, additional, Orihuela, Natalia, additional, Zulberti, Catherine, additional, Machado, Danilo, additional, Recalde, Cecilia, additional, Noboa, Javier, additional, Frantchez, Victoria, additional, Astesiano, Rossana, additional, Yandián, Federico, additional, Guerisoli, Ana, additional, Morra, Álvaro, additional, Cassinelli, Daniela, additional, Coelho, Cecilia, additional, de Aramburu, Belén, additional, González-Severgnini, Paulina, additional, Moreno, Romina, additional, Pippolo, Aldana, additional, López, Gabriela, additional, Lemos, Mónica, additional, Somariva, Lorena, additional, López, Eliana, additional, Fumero, Soledad, additional, Orihuela, Carla, additional, Rodríguez, Rosalía, additional, Acuña, Gonzalo, additional, Rabaza, Victoria, additional, Perg, Nancy, additional, Cordero, Rossana, additional, Reisfeld, Cristina, additional, Olivera, Paula, additional, Montero, Paola, additional, Nogueira, Cecilia, additional, Nalerio, Catheryn, additional, Orihuela, Sergio, additional, Curi, Lilián, additional, Burgstaller, Ema, additional, Noboa, Oscar, additional, Pritsch, Otto, additional, Nin, Marcelo, additional, and Bianchi, Sergio, additional

Published
2021
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13. Expression, Purification, and Characterization of Bovine Leukemia Virus-Like Particles Produced in Drosophila S2 Cells

Author

Olivero-Deibe, Natalia, primary, Tomé-Poderti, Lorena, additional, Carrión, Federico, additional, Bianchi, Sergio, additional, Fló, Martín, additional, Prieto, Daniel, additional, Rammauro, Florencia, additional, Addiego, Andrés, additional, Ibañez, Natalia, additional, Portela, Madelón, additional, Duran, Rosario, additional, Berois, Mabel, additional, and Pritsch, Otto, additional

Published
2021
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14. Comparison of antibody response to SARS-CoV-2 after two doses of inactivated virus and BNT162b2 mRNA vaccines in kidney transplant.

Author

Seija, Mariana, Rammauro, Florencia, Santiago, José, Orihuela, Natalia, Zulberti, Catherine, Machado, Danilo, Recalde, Cecilia, Noboa, Javier, Frantchez, Victoria, Astesiano, Rossana, Yandián, Federico, Guerisoli, Ana, Morra, Álvaro, Cassinelli, Daniela, Coelho, Cecilia, Aramburu, Belén de, González-Severgnini, Paulina, Moreno, Romina, Pippolo, Aldana, and López, Gabriela

Subjects
*SARS-CoV-2, *ANTIBODY formation, *COVID-19, *COVID-19 vaccines, *KIDNEY transplantation
Abstract

Background Antibody response against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) after mRNA or adenoviral vector-based vaccines is weak in kidney transplant (KT) patients. However, few studies have focused on humoral response after inactivated virus-based vaccines in KT. Here, we compare antibody response following vaccination with inactivated virus (CoronaVac®) and BNT162b2 mRNA. Methods A national multicentre cross-sectional study was conducted. The study group was composed of patients from all KT centres in Uruguay, vaccinated between 1 and 31 May 2021 (CoronaVac®, n = 245 and BNT162b2, n = 39). The control group was constituted of 82 healthy individuals. Participants had no prior confirmed coronavirus disease 2019 (COVID-19) test. Blood samples were collected between 30 and 40 days after the second dose. Serum-specific immunoglobulin G (IgG) antibodies against the receptor-binding domain (RBD) of SARS-CoV-2 Spike protein were determined using the COVID-19 IgG QUANT ELISA Kit. Results Only 29% of KT recipients showed seroconversion (36.5% BNT162b2, 27.8% inactivated virus, P = 0.248) in comparison with 100% in healthy control with either vaccine. Antibody levels against RBD were higher with BNT162b mRNA than with inactivated virus [median (interquartile range) 173 (73–554) and 29 (11–70) binding antibody units (BAU)/mL, P < 0.034] in KT and 10 times lower than healthy control [inactivated virus: 308 (209–335) and BNT162b2: 2638 (2608–3808) BAU/mL, P < 0.034]. In multivariate analysis, variables associated with negative humoral response were age, triple immunosuppression, estimated glomerular filtration rate and time post-KT. Conclusion Seroconversion was low in KT patients after vaccination with both platforms. Antibody levels against SARS-CoV-2 were lower with inactivated virus than BNT162b mRNA. These findings support the need for strategies to improve immunogenicity in KT recipients after two doses of either vaccine. [ABSTRACT FROM AUTHOR]

Published
2022
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17. Expression, Purification, and Characterization of Bovine Leukemia Virus-Like Particles Produced in Drosophila S2 Cells

Author

Madelón Portela, Sergio Bianchi, Daniel Prieto, Andrés Addiego, Natalia Olivero-Deibe, Martín Fló, Rosario Durán, Federico Carrión, Natalia Ibañez, Lorena Tomé-Poderti, Florencia Rammauro, Otto Pritsch, Mabel Berois, Olivero-Deibe Natalia, Instituto Pasteur (Montevideo), Tomé Poderti Lorena Magalí, Instituto Pasteur (Montevideo), Carrión Federico, Instituto Pasteur (Montevideo), Bianchi Sergio, Instituto Pasteur (Montevideo), Fló Díaz Martín, Instituo Pasteur (Montevideo), Prieto Mena Daniel, IIBCE, Rammauro Florencia, Instituo Pasteur (Montevideo), Addiego Andrés, Instituo Pasteur (Montevideo), Ibañez Natalia, Instituo Pasteur (Montevideo), Portela María Magdalena, Instituo Pasteur (Montevideo), Durán Rosario, Instituo Pasteur (Montevideo), Berois Mabel, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología., and Pritsch Otto, Instituo Pasteur (Montevideo)

Subjects
Gag, Env, Furin cleavage site, Virus-like particles (VLP), Retrovirus, Bovine leukemia virus, biology, Schneider 2 cells, viruses, Bovine leukemia virus (BLV), Wild type, virus diseases, General Medicine, biology.organism_classification, Virology, Deltaretrovirus, Virus, Immunogens, Cell culture, biology.protein, Furin
Abstract

Material complementario: https://www.frontiersin.org/articles/10.3389/fviro.2021.756559/full#supplementary-material Bovine leukemia virus (BLV) is an oncogenic deltaretrovirus that infects cattle worldwide. In Uruguay, it is estimated that more than 70% of dairy cattle are infected, causing serious economic losses due to decreased milk production, increased calving interval, and livestock losses due to lymphosarcoma. Several attempts to develop vaccine candidates that activate protective immune responses against BLV were performed, but up to date, there is no vaccine that ensures efficient protection and/or decreased viral transmission. The development and application of new vaccines that effectively control BLV infection represent amajor challenge for countries with a high prevalence of infection. In this study, we generated two Drosophila melanogaster S2 stable cell lines capable of producing BLV virus-like particles (BLV-VLPs). One of them, BLV-VLP1, expressed both Gag and Env wild-type (Envwt) full-length proteins, whereas BLV-VLP2 contain Gag together with a mutant form of Env non-susceptible to proteolytic maturation by cellular furin type enzymes (EnvFm).We showed that Envwt is properly cleaved by cellular furin, whereas EnvFm is produced as a full-length gp72 precursor, which undergoes some partial cleavage. We observed that said mutation does not drastically affect its expression or its entry into the secretory pathway of S2 insect cells. In addition, it is expressed on the membrane and retains significant structural motifs when expressed in S2 insect cells. Morphology and size of purified BLV-VLPs were analyzed by transmission electron microscopy and dynamic light scattering, showing numerous non-aggregated and approximately spherical particles of variable diameter (70–200 nm) as previously reported for retroviral VLPs produced using different expression systems. Furthermore, we identified two N-glycosylation patterns rich in mannose in EnvFm protein displayed on VLP2. Our results suggest that the VLPs produced in Drosophila S2 cells could be a potential immunogen to be used in the development of BLV vaccines that might contribute, in conjunction with other control strategies, to reduce the transmission of the virus. CSIC I+D 2014 ANII: ALI_1_2016_2_129851; POS_NAC_2015_1_109471 PEDECIBA-FOCEM: COF 03/11 CAP: BFPD_2020_1#28143834

Published
2021
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18. Soluble SARS-CoV-2 RBD and human ACE2 peptidase domain produced in Drosophila S2 cells show functions evoking virus-cell interface.

Author

Carrión F, Rammauro F, Olivero-Deibe N, Fló M, Portela MM, Lima A, Durán R, Pritsch O, and Bianchi S

Subjects
Animals, Humans, Antibodies, Neutralizing chemistry, Antibodies, Viral chemistry, Drosophila metabolism, Spike Glycoprotein, Coronavirus genetics, Spike Glycoprotein, Coronavirus chemistry, Angiotensin-Converting Enzyme 2, COVID-19, SARS-CoV-2 metabolism
Abstract

The interaction between the receptor-binding domain (RBD) of the spike glycoprotein of SARS-CoV-2 and the peptidase domain of the human angiotensin-converting enzyme 2 (ACE2) allows the first specific contact at the virus-cell interface making it the main target of neutralizing antibodies. Here, we show a unique and cost-effective protocol using Drosophila S2 cells to produce both RBD and soluble human ACE2 peptidase domain (shACE2) as thermostable proteins, purified via Strep-tag with yields >40 mg L -1 in a laboratory scale. Furthermore, we demonstrate its binding with K D values in the lower nanomolar range (independently of Strep-tag removal) and its capability to be blocked by serum antibodies in a competition ELISA with Strep-Tactin-HRP as a proof-of-concept. In addition, we assess the capacity of RBD to bind native dimeric ACE2 overexpressed in human cells and its antigen properties with specific serum antibodies. Finally, for completeness, we analyzed RBD microheterogeneity associated with glycosylation and negative charges, with negligible effect on binding either with antibodies or shACE2. Our system represents an accessible and reliable tool for designing in-house surrogate virus neutralization tests (sVNTs), enabling the rapid characterization of neutralizing humoral responses elicited against vaccines or infection, especially in the absence of facilities to conduct virus neutralization tests. Moreover, our biophysical and biochemical characterization of RBD and shACE2 produced in S2 cells lays the groundwork for adapting to different variants of concern (VOCs) to study humoral responses elicited against different VOCs and vaccine formulations., (© 2023 The Protein Society.)

Published
2023
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