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1.
Glycoconj J ; 38(5): 539-549, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34515909

RESUMO

Recent changes in the epidemiology of meningococcal have been reported and meningococcal group W (MenW) has become the third most prevalent group isolated in Brazil in the last 10 years. In this study we have developed a conjugate vaccine for MenW using a modified reductive amination conjugation method through a covalent linkage between periodate-oxidized MenW non-O-acetylated polysaccharide and hydrazide-activated monomeric tetanus toxoid. Process control of bulks was done by physicochemical analysis including polysaccharide and protein quantification, high performance liquid chromatography - size exclusion chromatography, capillary electrophoresis, and hydrogen nuclear magnetic resonance. Conjugate bulks were best produced with concentration of polysaccharide twice as high as protein, at room temperature, and pH approximately 6.0. A scaled-up bulk (100 mg scale) was formulated and inoculated intramuscularly in mice in a dose-response study (0.1, 0.5, 1.0 and 10.0 µg of polysaccharide/dose). The immunogenicity of conjugate bulks was determined by serum bactericidal assay and ELISA assays of serum from immunized mice. ELISA and SBA titers revealed high titers of IgG and demonstrated the functionality of the antibodies produced in all doses studied 15 days after the third dose. However, significant differences were observed among them by ELISA. In conclusion, this study established the best conditions to produce MenW conjugate bulks and showed the efficacy of the obtained conjugate bulk in induce a good immune response in mice. Further experiments will need to be done to scale up the conjugation reaction and then allow the use of this conjugate in clinical trials.


Assuntos
Infecções Meningocócicas/epidemiologia , Infecções Meningocócicas/prevenção & controle , Vacinas Meningocócicas/imunologia , Neisseria meningitidis/classificação , Animais , Anticorpos Antibacterianos , Atividade Bactericida do Sangue , Brasil/epidemiologia , Feminino , Glicoconjugados , Humanos , Masculino , Camundongos , Projetos Piloto , Toxoide Tetânico/imunologia , Vacinas Conjugadas/imunologia
2.
J. pediatr. (Rio J.) ; J. pediatr. (Rio J.);97(supl.1): 84-90, Mar.-Apr. 2021. tab
Artigo em Inglês | LILACS | ID: biblio-1250221

RESUMO

Abstract Objectives: Inborn Errors of Immunity are characterized by infectious conditions and manifestations of immune dysregulation. The diversity of clinical phenotypes can make it difficult to direct the laboratory investigation. This article aims to update the investigation of immunological competence in the context of primary defects of the immune system. Source of data: Searches were carried out on Pubmed to review articles published in the last five years, in English, French or Spanish, using the terms "diagnosis" OR "investigation" AND "immunodeficiency" or "primary immunodeficiency" or "inborn errors of immunity" NOT "HIV". Recent textbook editions have also been consulted. Summary of findings: The immune system competence investigation should be started based on clinical phenotypes. Relevant data are: characterization of infectious conditions (location, recurrence, types of infectious agents, response to treatment), age during symptom onset and associated manifestations (growth impairment, allergy, autoimmunity, malignancies, fever and signs of inflammation without the identification of infection or autoimmunity) and family history. These data contribute to the selection of tests to be performed. Conclusions: The diagnostic investigation of Inborn Errors of Immunity should be guided by the clinical characterization of patients, aiming to optimize the use of complementary tests. Many diagnoses are attained only through genetic tests, which are not always available. However, the absence of a diagnosis of certainty should never delay the implementation of therapeutic measures that preserve patient life and health.


Assuntos
Humanos , Síndromes de Imunodeficiência/diagnóstico , Neoplasias , Fenótipo , Recidiva , Inflamação
3.
J Pediatr (Rio J) ; 97 Suppl 1: S84-S90, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33400918

RESUMO

OBJECTIVES: Inborn Errors of Immunity are characterized by infectious conditions and manifestations of immune dysregulation. The diversity of clinical phenotypes can make it difficult to direct the laboratory investigation. This article aims to update the investigation of immunological competence in the context of primary defects of the immune system. SOURCE OF DATA: Searches were carried out on Pubmed to review articles published in the last five years, in English, French or Spanish, using the terms "diagnosis" OR "investigation" AND "immunodeficiency" or "primary immunodeficiency" or "inborn errors of immunity" NOT "HIV". Recent textbook editions have also been consulted. SUMMARY OF FINDINGS: The immune system competence investigation should be started based on clinical phenotypes. Relevant data are: characterization of infectious conditions (location, recurrence, types of infectious agents, response to treatment), age during symptom onset and associated manifestations (growth impairment, allergy, autoimmunity, malignancies, fever and signs of inflammation without the identification of infection or autoimmunity) and family history. These data contribute to the selection of tests to be performed. CONCLUSIONS: The diagnostic investigation of Inborn Errors of Immunity should be guided by the clinical characterization of patients, aiming to optimize the use of complementary tests. Many diagnoses are attained only through genetic tests, which are not always available. However, the absence of a diagnosis of certainty should never delay the implementation of therapeutic measures that preserve patient life and health.


Assuntos
Síndromes de Imunodeficiência , Neoplasias , Humanos , Síndromes de Imunodeficiência/diagnóstico , Inflamação , Fenótipo , Recidiva
4.
Glycoconj J ; 35(1): 3-13, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-28929266

RESUMO

High temperature is known to cause some instability in polysaccharide-protein conjugated vaccines and studies under stress conditions may be useful in determining whether short-term accidental exposure to undesired conditions can compromise product quality. In this study, we examined the structural stability of three industrial batches of Brazilian Meningococcal C conjugate bulk (MPCT) incubated at 4, 37, and 55 °C for 5 weeks. The effect of exposure to the storage temperatures was monitored by HPLC-SEC, CZE, CD and NMR techniques. The immunological significance of any physicochemical changes observed in MPCT was determined by SBA and ELISA assays of serum from immunized mice. Fluorescence emission spectra at 4 and 37 °C were similar among all samples and compatible with the native fold of the carrier protein. Fluorescence spectra of MPCT stored at 55 °C decreased in intensity and had a significant red-shift, indicating conformational changes. Far-UV CD spectra revealed a trend toward loss of structural conformation as storage temperature was increased to 55 °C. The NMR data showed modified signal intensity of the aromatic and aliphatic residues, mainly for samples incubated at 55 °C, suggesting a partial loss of tertiary structure. About 50% free saccharide content was found in bulks stored at 55 °C, but no difference was observed in the IgG or SBA titers. The present study showed physicochemical methods alone are insufficient to predict the biological activity of a MPCT conjugate vaccine without extensive validation against immunological data. However, they provide a sensitive means of detecting changes induced in a vaccine exposed to adverse environmental condition.


Assuntos
Vacinas Meningocócicas/imunologia , Absorção de Radiação , Animais , Imunogenicidade da Vacina , Vacinas Meningocócicas/química , Camundongos , Neisseria meningitidis Sorogrupo C/imunologia , Estabilidade Proteica
5.
Clin Oral Implants Res ; 24(9): 963-8, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22697457

RESUMO

OBJECTIVES: To present some immunological aspects of fresh-frozen allogeneic bone grafting for lateral bone augmentation, based on the quantitative evaluation of IL-10, IL-1ß, IFN- γ and TNF- α in patients sera. MATERIAL AND METHODS: Thirty-three partially or totally edentulous patients received fresh-frozen allogeneic bone (AL - 20 patients) or autologous bone onlay block grafts (AT - 13 patients) prior to oral implant placement. Blood samples were collected from each patient at various time-points during a 6 month-period (baseline, 14, 30, 90 and 180 days postoperatively). Quantitative evaluation of IL-10, IL-1ß, IFN- γ and TNF- α was performed by enzyme linked immunosorbent assay (ELISA). RESULTS: For all evaluated markers and at all evaluated periods, inter-group comparisons showed no statistically significant differences between the groups, while the observed values were within normal levels. For AL-treated patients, intra-group evaluation showed statistically significant increase of TNF-α from baseline to 90 (P < 0.001) and 180 (P < 0.01) days, and from 14 to 90 (P < 0.01) and 180 (P < 0.05) days. IFN- γ showed intercalated results, with a decrease from baseline to 14 days (P < 0.05), and increase from 14 to 90 days (P < 0.001) and 180 (P < 0.05) days. No differences between the periods of evaluation were found for the AT group. CONCLUSIONS: AL grafting for lateral bone augmentation, similar to AT grafting, does not seem to challenge the immune system significantly.


Assuntos
Aumento do Rebordo Alveolar/métodos , Biomarcadores/sangue , Transplante Ósseo/métodos , Implantação Dentária Endóssea/métodos , Arcada Edêntula/cirurgia , Adulto , Idoso , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interferon gama/sangue , Interleucina-10/sangue , Interleucina-1beta/sangue , Masculino , Pessoa de Meia-Idade , Retalhos Cirúrgicos , Resultado do Tratamento , Fator de Necrose Tumoral alfa/sangue
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