Patentes e registro sanitário de vacinas antimeningocócicas sorogrupo B no Brasil: aspectos importantes para a regulação e o controle / Patents and sanitary registration of serogroup B meningococcal / vaccine in Brazil: basic aspects for biological products regulation and control from rice bran

As vacinas são um dos instrumentos mais importantes para a saúde pública. O registro sanitário é uma das ferramentas que a vigilância sanitária dispõe para controlar a entrada de todos os medicamentos e constitui a base das informações sobre eles para o Sistema de Vigilância Sanitária. No caso da meningite meningocócica, doença causada pela bactéria Neisseria meningitidis (Nm), várias vacinas foram desenvolvidas inicialmente contra os sorogrupos A, C, W135 e Y, utilizando-se os polissacarídeos capsulares. Pelo fato de o antígeno polissacarídico do sorogrupo B não induzir boa resposta imunológica em seres humanos, outras estratégias vacinais foram testadas. O objetivo deste trabalho foi mapear as patentes de produtos vacinais contra o sorogrupo B, depositadas no período de 1990 a 2005, e comparar esses pedidos com as vacinas registradas. O levantamento dos pedidos de patentes foi realizado utilizando-se as bases de dados eletrônicos do Instituto Nacional de Propriedade Industrial (INPI) e do Escritório de Patentes Europeu (EPO), a partir de janeiro de 1995. Várias patentes solicitaram proteção dos antígenos presentes em vacina já registrada, o que não caracteriza inovação. Os maiores depositantes de patentes são as multinacionais, o que mostra a necessidade de incremento dessa pesquisa aplicada por instituições brasileiras.(AU)

Vaccines are important instruments for public health. Sanitary registration is an important tool for the national regulatory authorities to control the medicines use and it is the basic source of information for the National Sanitary Surveillance System NRAs. For meningococcal meningitis, caused by Neisseria meningitidis (Nm), many vaccines were developed against serogroups A, C. W135, and Y using the capsular polysaccharides. Considering that the serogroup B polysaccharide is poorly immunogenic in human, other vaccines strategies were tried. This study aimed at identifying the applied patents for meningitis B antigens during the period from 1990 to 2005, and these were compared to the registered products. The European Patent Office and the National Institute for Industrial Propriety data basis were used to survey the patents request. Several patents requested the antigens safekeeping, which are included into the registered vaccine, and it does not characterize an innovation. The majority of the patents depositories belong to multinational companies and the Brazilian institutions should put in practice the process of requesting the patent for their research products.(AU)

Imunização nasal em coelhos com Neisseria lactamica: importância dos antígenos de reatividade cruzada / Nasal immunization in rabbits with Neisseria lactamica: the importance of the cross reactive antigens

Neisseria lactamica, a commensal bacterium nonpathogenic to human beings and usually found in the upper respiratory tract of children, is closely related to pathogenic Neisseria meningitidis. Colonization with N. lactamica can be responsible for evolving natural immunity to meningococcal infection in childhood, when rates of meningococcus carriers are low. These features lead to suggest that N. lactamica components can be key-elements in the production of a new vaccine for N. meningitidis. As little is known about dynamic carriers and N. lactamica population diversity in children, it has been difficult choosing a representative for preparing an adequate immunogenic product. A protocol was proposed to study immunogenicity of whole cells of N. lactamica, N. meningitidis, N. sicca or N. meningitidis c (carrier-isolated) by i.n. immunization in rabbits considering the natural pathogen entry route. Oropharinx-isolated N. lactamica, N. meningitidis N. sicca, or N. meningitidis c were i.n. inoculated into adult rabbits, in a concentration of optical density 1.0 at 650nm in a volume of 1000 µL. The rabbits were immunized four times at seven-day intervals. N. subflava, N. elongata, N. sicca, N. perflava, N. mucosa strains isolated from CSF and blood were also used. The rabbits developed levels of specific lgG antibodies in serum, as determined by ELISA using whole cells of homologous an

Neisseria lactamica, a commensal bacterium nonpathogenic to human beings and usually found in the upper respiratory tract of children, is closely related to pathogenic Neisseria meningitidis. Colonization with N. lactamica can be responsible for evolving natural immunity to meningococcal infection in childhood, when rates of meningococcus carriers are low. These features lead to suggest that N. lactamica components can be key-elements in the production of a new vaccine for N. meningitidis. As little is known about dynamic carriers and N. lactamica population diversity in children, it has been difficult choosing a representative for preparing an adequate immunogenic product. A protocol was proposed to study immunogenicity of whole cells of N. lactamica, N. meningitidis, N. sicca or N. meningitidis c (carrier-isolated) by i.n. immunization in rabbits considering the natural pathogen entry route. Oropharinx-isolated N. lactamica, N. meningitidis N. sicca, or N. meningitidis c were i.n. inoculated into adult rabbits, in a concentration of optical density 1.0 at 650nm in a volume of 1000 µL. The rabbits were immunized four times at seven-day intervals. N. subflava, N. elongata, N. sicca, N. perflava, N. mucosa strains isolated from CSF and blood were also used. The rabbits developed levels of specific lgG antibodies in serum, as determined by ELISA using whole cells of homologous an

Production of outer membrane vesicles (OMV) in batch cultivation of Neisseria meningitidis serogroup B

Meningococcal disease is an important cause of death and morbidity throughout the world. Nearly 330,000 cases and 35,000 deaths occur yearly. Neisseria meningitidis, serogroup B strain N.44/89, is prevalent in Brazil. Its outer membrane vesicles (OMV) with iron regulated proteins (IRP) are released to the culture medium and are used as antigen for vaccine production. In order to have knowledge about the kinetic parameters, especially the final OMV concentration values, 20-h batch cultivations were carried out in Catlin medium with iron restriction. Process conditions comprised: 7 L bioreactor, 36ºC, 0.5 atm, overlay air flowrate of 1 L/min, agitation varying from 250 rpm to 850 rpm and dissolved oxygen control set at 10% of saturation condition. Biomass was determined by optical density at 540 nm and dry weight. Glycerol, lactate, pH and dissolved oxygen were measured from samples taken during cultivation. Outer membrane vesicle (OMV) concentration was determined by Lowry's method after ultracentrifugation. IRP presence was verified by SDS-PAGE. Highest biomass value, corresponding to the highest initial lactate concentration (7.84 g/L) was achieved at the 9th hour process time corresponding to 1.0 g/L dry biomass and 2.3 optical density at 540 nm. Lactate consumption was directly related to cell growth (yield factor: 0.24 g dry biomass / g lactate). Glycerol concentration in the medium did not change significantly during the process. OMV concentration reached the highest value of 80 mg/L at end cultivation time. The obtained results suggest that lactate is a main limiting growth factor and the maximum amount of antigen is obtained during stationary growth and cell death phases.

A doença meningocócica é uma causa importante de morte a nível mundial. Aproximadamente 330.000 casos e 35.000 mortes ocorrem anualmente. A cepa N.44/89 do sorogrupo B de Neisseria meningitidis é prevalente no Brasil. Suas vesículas de membrana externa (OMV - "outer membrane vesicles"), com proteínas reguladoras de ferro (IRP - "iron regulated proteins") liberadas no meio de cultura, são empregadas como antígeno para a produção da vacina. A fim ter o conhecimento sobre os parâmetros cinéticos, especialmente os valores finais da concentração de OMV, cultivos batelada de 20 hs foram realizados no meio de Catlin com limitação do ferro. As condições de processo compreenderam: biorreator de 7 litros, 36ºC, 0,5 atm, vazão de ar de 1 L/min, agitação variando entre 250 a 850 rpm, controle do oxigênio dissolvido em 10% da condição de saturação. A biomassa foi determinada pela densidade ótica em 540 nm e peso seco. Glicerol, lactato, pH e oxigênio dissolvido foram medidos das amostras retiradas durante o cultivo. A concentração de OMV foi determinada pelo método de Lowry após ultracentrifugação. A presença de IRP foi verificada por SDS-PAGE. O valor mais elevado de biomassa, correspondendo à concentração inicial mais elevada de lactato (7,84 g/L) foi obtido no tempo de processo de 9 horas, o qual corresponde a biomassa seca de 1,0 g/L e a densidade ótica de 2,3 em 540 nm. O consumo de lactato. foi relacionado diretamente ao crescimento celular (fator de conversão de 0,24 biomassa por lactato g/g). A concentração do glicerol no meio não se alterou significativamente ao longo do processo. A concentração de OMV alcançou o valor mais elevado de 80 mg/L no tempo final de cultivo. Os resultados obtidos sugerem que o lactato é o principal fator limitante do crescimento e o máximo do antígeno é obtido durante a fase estacionária de crescimento e de morte celular.

Estudo da Imunogenicidade da proteína de Classe 5C de Neisseria meningitidis B em camundongos imunizados pela via nasal / Immunogenicity of intranasally administered class 5C protein of Neisseria meningitidis in mice

Neisseria meningitidis is a human pathogen for which no fully effective vaccine is avaliable. Meningococcal vaccines currently approved for use in humans are made from purified capsular polysaccharides. Such vaccines are effective against serogroups A and C, although in the United States, a quadrivalent vaccine containing four types of meningococcal serogroups A, C, Y, and W-135) is effective among people over two years of age. Unfortunately, such capsular polysaccharide vaccines present limitations as they have no effect on children younger than two years and the immune response is short-lived in older children and in adolescents. Development of a vaccine against serogroup B poses an enormous problem due to the similarity between the B capsular polysaccharide structure and a polysialic acid containing glycopeptides that are part of the human brain tissue, raising the possibility of occurrence of autoimmunity. Promising studies have been conducted on the development of a vaccine based on N. meningitidis outer membrane proteins (OMPs). The major OMPs N. meningitidis are designated from class 1 through class 5. However, OMP vaccines are weak antigens in infants. In an effort to establish efficient immunization in infants, we here tested a vaccine consisting of purified class 5C protein from one reference Neisseria meningitidis serogroup B and different adjuvants. This study aimed

Avaliação da resposta imune em coelhos pela via de imunização nasal com antígenos de Neisseria meningitis B selecionados para os imunotipos L3,7,9 e L8 por meio de anticorpos monoclonais / Evaluation of the immune response in rabbits immunized intranasally with antigens of Neisseria meningitidis B selected for the immunotypes L3,7,9 or 8 with monoclonal antibodies

Serogroup B Neisseria meningitidis is responsible for fulminant septcemia and it is a common cause of pyogenic meningitis. In addition to sporadic outbreaks, large epidemics of serogroup B meningococcal disease occur in many parts of the world. Therefore, the development of a vaccine against N.meningididis serogroup B remains a high priority worldwide. Unlike others serogroups to wich the capsular polysaccharides constitute efficacious vaccines, the serogroup B capsule is poorly immunogenic in humans. In modern vaccine development, strong emphasis has been laid on mucosal immunization system. In particular, the intranasal (i.n) holds promise for a potential induction of protective immune responses, since its able to elicit both local and strong systemic immune response. Intranasal vaccination may therefore be of particular interest against respiratory tract infections, such as those caused by N.meningitidis. Lipopolysaccharides(LPS) are complex molecules which are part of outer membrane, of Gram-negative bacteria. The advantage of mucosal immunization is that vaccines containing lipopolysaccharides may be delivered safely via mucosa route without causing adverse side effect seen in parental immunization. The New Zeland white rabbits was employed as model for representative by intranasal immunization in humans with regards to vaccine disposition. Immunizing with native outer mem

Estudo da Imunogenicidade da proteína de Classe 5C de Neisseria meningitidis B em camundongos imunizados pela via nasal / Immunogenicity of intranasally administered class 5C protein of Neisseria meningitidis in mice

Neisseria meningitidis is a human pathogen for which no fully effective vaccine is avaliable. Meningococcal vaccines currently approved for use in humans are made from purified capsular polysaccharides. Such vaccines are effective against serogroups A and C, although in the United States, a quadrivalent vaccine containing four types of meningococcal serogroups A, C, Y, and W-135) is effective among people over two years of age. Unfortunately, such capsular polysaccharide vaccines present limitations as they have no effect on children younger than two years and the immune response is short-lived in older children and in adolescents. Development of a vaccine against serogroup B poses an enormous problem due to the similarity between the B capsular polysaccharide structure and a polysialic acid containing glycopeptides that are part of the human brain tissue, raising the possibility of occurrence of autoimmunity. Promising studies have been conducted on the development of a vaccine based on N. meningitidis outer membrane proteins (OMPs). The major OMPs N. meningitidis are designated from class 1 through class 5. However, OMP vaccines are weak antigens in infants. In an effort to establish efficient immunization in infants, we here tested a vaccine consisting of purified class 5C protein from one reference Neisseria meningitidis serogroup B and different adjuvants. This study aimed

Avaliação da resposta imune em coelhos pela via de imunização nasal com antígenos de Neisseria meningitis B selecionados para os imunotipos L3,7,9 e L8 por meio de anticorpos monoclonais / Evaluation of the immune response in rabbits immunized intranasally with antigens of Neisseria meningitidis B selected for the immunotypes L3,7,9 or 8 with monoclonal antibodies

Serogroup B Neisseria meningitidis is responsible for fulminant septcemia and it is a common cause of pyogenic meningitis. In addition to sporadic outbreaks, large epidemics of serogroup B meningococcal disease occur in many parts of the world. Therefore, the development of a vaccine against N.meningididis serogroup B remains a high priority worldwide. Unlike others serogroups to wich the capsular polysaccharides constitute efficacious vaccines, the serogroup B capsule is poorly immunogenic in humans. In modern vaccine development, strong emphasis has been laid on mucosal immunization system. In particular, the intranasal (i.n) holds promise for a potential induction of protective immune responses, since its able to elicit both local and strong systemic immune response. Intranasal vaccination may therefore be of particular interest against respiratory tract infections, such as those caused by N.meningitidis. Lipopolysaccharides(LPS) are complex molecules which are part of outer membrane, of Gram-negative bacteria. The advantage of mucosal immunization is that vaccines containing lipopolysaccharides may be delivered safely via mucosa route without causing adverse side effect seen in parental immunization. The New Zeland white rabbits was employed as model for representative by intranasal immunization in humans with regards to vaccine disposition. Immunizing with native outer mem

Imunização nasal em coelhos com Neisseria lactamica: importância dos antígenos de reatividade cruzada / Nasal immunization in rabbits with Neisseria lactamica: the importance of the cross reactive antigens

Neisseria lactamica, a commensal bacterium nonpathogenic to human beings and usually found in the upper respiratory tract of children, is closely related to pathogenic Neisseria meningitidis. Colonization with N. lactamica can be responsible for evolving natural immunity to meningococcal infection in childhood, when rates of meningococcus carriers are low. These features lead to suggest that N. lactamica components can be key-elements in the production of a new vaccine for N. meningitidis. As little is known about dynamic carriers and N. lactamica population diversity in children, it has been difficult choosing a representative for preparing an adequate immunogenic product. A protocol was proposed to study immunogenicity of whole cells of N. lactamica, N. meningitidis, N. sicca or N. meningitidis c (carrier-isolated) by i.n. immunization in rabbits considering the natural pathogen entry route. Oropharinx-isolated N. lactamica, N. meningitidis N. sicca, or N. meningitidis c were i.n. inoculated into adult rabbits, in a concentration of optical density 1.0 at 650nm in a volume of 1000 µL. The rabbits were immunized four times at seven-day intervals. N. subflava, N. elongata, N. sicca, N. perflava, N. mucosa strains isolated from CSF and blood were also used. The rabbits developed levels of specific lgG antibodies in serum, as determined by ELISA using whole cells of homologous an

Neisseria lactamica, a commensal bacterium nonpathogenic to human beings and usually found in the upper respiratory tract of children, is closely related to pathogenic Neisseria meningitidis. Colonization with N. lactamica can be responsible for evolving natural immunity to meningococcal infection in childhood, when rates of meningococcus carriers are low. These features lead to suggest that N. lactamica components can be key-elements in the production of a new vaccine for N. meningitidis. As little is known about dynamic carriers and N. lactamica population diversity in children, it has been difficult choosing a representative for preparing an adequate immunogenic product. A protocol was proposed to study immunogenicity of whole cells of N. lactamica, N. meningitidis, N. sicca or N. meningitidis c (carrier-isolated) by i.n. immunization in rabbits considering the natural pathogen entry route. Oropharinx-isolated N. lactamica, N. meningitidis N. sicca, or N. meningitidis c were i.n. inoculated into adult rabbits, in a concentration of optical density 1.0 at 650nm in a volume of 1000 µL. The rabbits were immunized four times at seven-day intervals. N. subflava, N. elongata, N. sicca, N. perflava, N. mucosa strains isolated from CSF and blood were also used. The rabbits developed levels of specific lgG antibodies in serum, as determined by ELISA using whole cells of homologous an