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1.
Sci Rep ; 12(1): 3890, 2022 03 10.
Artículo en Inglés | MEDLINE | ID: mdl-35273234

RESUMEN

The new outbreak of coronavirus disease 2019 (COVID-19) has infected and caused the death of millions of people worldwide. Intensive efforts are underway around the world to establish effective treatments. Immunoglobulin from immunized animals or plasma from convalescent patients might constitute a specific treatment to guarantee the neutralization of the virus in the early stages of infection, especially in patients with risk factors and a high probability of progressing to severe disease. Worldwide, a few clinical trials using anti-SARS-CoV-2 immunoglobulins from horses immunized with the entire spike protein or fragments of it in the treatment of patients with COVID-19 are underway. Here, we describe the development of an anti-SARS-CoV-2 equine F(ab')2 immunoglobulin using a newly developed SARS-CoV-2 viral antigen that was purified and inactivated by radiation. Cell-based and preclinical assays showed that the F(ab')2 immunoglobulin successfully neutralizes the virus, is safe in animal models, and reduces the severity of the disease in a hamster model of SARS-CoV-2 infection and disease.


Asunto(s)
COVID-19/terapia , Inmunoglobulinas/uso terapéutico , Receptores Inmunológicos/uso terapéutico , SARS-CoV-2/inmunología , Animales , Ensayo de Inmunoadsorción Enzimática , Femenino , Caballos/inmunología , Humanos , Inmunoglobulinas/inmunología , Inmunoglobulinas/aislamiento & purificación , Masculino , Mesocricetus/inmunología , Plasmaféresis/veterinaria , Receptores Inmunológicos/inmunología
2.
Sci Rep, v. 12, 3890, mar. 2022
Artículo en Inglés | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4257

RESUMEN

The new outbreak of coronavirus disease 2019 (COVID-19) has infected and caused the death of millions of people worldwide. Intensive efforts are underway around the world to establish effective treatments. Immunoglobulin from immunized animals or plasma from convalescent patients might constitute a specific treatment to guarantee the neutralization of the virus in the early stages of infection, especially in patients with risk factors and a high probability of progressing to severe disease. Worldwide, a few clinical trials using anti-SARS-CoV-2 immunoglobulins from horses immunized with the entire spike protein or fragments of it in the treatment of patients with COVID-19 are underway. Here, we describe the development of an anti-SARS-CoV-2 equine F(ab′)2 immunoglobulin using a newly developed SARS-CoV-2 viral antigen that was purified and inactivated by radiation. Cell-based and preclinical assays showed that the F(ab′)2 immunoglobulin successfully neutralizes the virus, is safe in animal models, and reduces the severity of the disease in a hamster model of SARS-CoV-2 infection and disease.

3.
Biochem Biophys Res Commun ; 545: 145-149, 2021 03 19.
Artículo en Inglés | MEDLINE | ID: mdl-33550095

RESUMEN

In March 2013 it was reported by the World Health Organization (WHO) the first cases of human infections with avian influenza virus A (H7N9). From 2013 to December 2019, 1568 cases have been reported with 616 deaths. H7N9 infection has been associated with high morbidity and mortality rates, and vaccination is currently the most effective way to prevent infections and consequently flu-related severe illness. Developing and producing vaccines against pandemic influenza viruses is the main strategy for a response to a possible pandemic. This study aims to present the production of three industrial lots under current Good Manufacturing Practices (cGMP) of the active antigen used to produce the pandemic influenza vaccine candidate against A(H7N9). These batches were characterized and evaluated for quality standards and tested for immunogenicity in mice. The average yield was 173.50 ± 7.88 µg/mL of hemagglutinin and all the preparations met all the required specifications. The formulated H7N9 vaccine is poorly immunogenic and needs to be adjuvanted with an oil in water emulsion adjuvant (IB160) to achieve a best immune response, in a prime and in a boost scheme. These data are important for initial production planning and preparedness in the case of a H7N9 pandemic.


Asunto(s)
Subtipo H7N9 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/biosíntesis , Gripe Humana/prevención & control , Pandemias/prevención & control , Animales , Antígenos Virales/biosíntesis , Antígenos Virales/inmunología , Composición de Medicamentos/métodos , Composición de Medicamentos/estadística & datos numéricos , Industria Farmacéutica/normas , Femenino , Humanos , Vacunas contra la Influenza/inmunología , Vacunas contra la Influenza/aislamiento & purificación , Gripe Humana/inmunología , Gripe Humana/virología , Ratones , Ratones Endogámicos BALB C , Vacunas de Productos Inactivados/biosíntesis , Vacunas de Productos Inactivados/inmunología , Vacunas de Productos Inactivados/aislamiento & purificación
4.
Biochem Biophys Res Commun, v. 545, p. 145-149, mar. 2021
Artículo en Inglés | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-3533

RESUMEN

In March 2013 it was reported by the World Health Organization (WHO) the first cases of human infections with avian influenza virus A (H7N9). From 2013 to December 2019, 1568 cases have been reported with 616 deaths. H7N9 infection has been associated with high morbidity and mortality rates, and vaccination is currently the most effective way to prevent infections and consequently flu-related severe illness. Developing and producing vaccines against pandemic influenza viruses is the main strategy for a response to a possible pandemic. This study aims to present the production of three industrial lots under current Good Manufacturing Practices (cGMP) of the active antigen used to produce the pandemic influenza vaccine candidate against A(H7N9). These batches were characterized and evaluated for quality standards and tested for immunogenicity in mice. The average yield was 173.50 ± 7.88 μg/mL of hemagglutinin and all the preparations met all the required specifications. The formulated H7N9 vaccine is poorly immunogenic and needs to be adjuvanted with an oil in water emulsion adjuvant (IB160) to achieve a best immune response, in a prime and in a boost scheme. These data are important for initial production planning and preparedness in the case of a H7N9 pandemic.

5.
São Paulo; s.n; 2011. 134 p.
Tesis en Portugués | LILACS, Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP, SESSP-IBACERVO | ID: biblio-1080937

RESUMEN

Os Influenzavirus podem ser classificados de acordo com suas glicoproteínas externas hemaglutinina (HA) e neuraminidase (NA), ambas apresentando alta variabilidade genética e antigênica. No presente estudo foi realizada análise molecular do gene HA, do vírus influenza A (IA) em amostras colhidas de crianças e lactentes com sintomatologia respiratória atendidas no Hospital Universitário da Universidade de São Paulo (USP), durante os anos de 1995 a 2006. Um total de 3.009 amostras foram analisadas por duplex RT-PCR e 4,38% (n=132) foram positivas, sendo 12,1% (n=16) Influenza B e 87,9% (n=116) IA, das quais 9% (n=9) eram H1N1, 91% (n=91) eram H3N2 e 13,8% (n=16) não foram subtipadas. A região HA1 do gene HA de 39 amostras foi sequenciada e as sequências comparadas com as cepas vacinais e circulantes dos respectivos anos. A região de ligação ao receptor foi conservada em todas as amostras e foram verificadas alterações de aminoácidos principalmente nos sítios antigênicos e arredores. No geral, as cepas vacinais foram compatíveis com as circulantes em São Paulo.


The Influenzavirus can be classified according to their external glycoproteins hemagglutinin (HA) and neuraminidase (NA), both showing high genetic and antigenic variability. In the present study was carried out molecular analysis of the HA gene of influenza A (IA) in samples harvested from children and infants, with respiratory symptoms attended at University Hospital, University of Sao Paulo (USP), during the years 1995 to 2006. A total of 3,009 samples were analyzed by duplex RT-PCR and 4.38% (n = 132) were positive, being 12.1% (n = 16) Influenza B and 87.9% (n = 116) IA, where which 9% (n = 9) were H1N1 and 91% (n = 91) were H3N2 and 13.8% (n = 16) did not subtyped. The HA1 region of HA gene of 39 samples were sequenced and the sequences compared with vaccine strains and circulating strains in those years. The receptor-binding region was conserved in all samples and aminoacid changes were observed mainly in the antigenic sites and surroundings. Overall, the vaccine strains were consistent with those circulating in Sao Paulo.


Asunto(s)
Humanos , Niño , Genética , Alphainfluenzavirus/genética , Virología , Hemaglutininas
6.
J Virol Methods ; 148(1-2): 115-9, 2008 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-18068812

RESUMEN

Human respiratory syncytial virus (HRSV) is the main cause of acute lower respiratory tract infections in infants and children. Rapid diagnosis is required to permit appropriate care and treatment and to avoid unnecessary antibiotic use. Reverse transcriptase (RT-PCR) and indirect immunofluorescence assay (IFA) methods have been considered important tools for virus detection due to their high sensitivity and specificity. In order to maximize use-simplicity and minimize the risk of sample cross-contamination inherent in two-step techniques, a RT-PCR method using only a single tube to detect HRSV in clinical samples was developed. Nasopharyngeal aspirates from 226 patients with acute respiratory illness, ranging from infants to 5 years old, were collected at the University Hospital of the University of Sao Paulo (HU-USP), and tested using IFA, one-step RT-PCR, and semi-nested RT-PCR. One hundred and two (45.1%) samples were positive by at least one of the three methods, and 75 (33.2%) were positive by all methods: 92 (40.7%) were positive by one-step RT-PCR, 84 (37.2%) by IFA, and 96 (42.5%) by the semi-nested RT-PCR technique. One-step RT-PCR was shown to be fast, sensitive, and specific for RSV diagnosis, without the added inconvenience and risk of false positive results associated with semi-nested PCR. The combined use of these two methods enhances HRSV detection.


Asunto(s)
Infecciones por Virus Sincitial Respiratorio/diagnóstico , Virus Sincitial Respiratorio Humano/aislamiento & purificación , Infecciones del Sistema Respiratorio/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Preescolar , Humanos , Lactante , Recién Nacido , Faringe/virología , Infecciones por Virus Sincitial Respiratorio/virología , Virus Sincitial Respiratorio Humano/genética , Sensibilidad y Especificidad
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