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1.
PLoS One ; 13(9): e0204222, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30260981

RESUMO

Campylobacteriosis is a widespread infectious disease, leading to a major health and economic burden. Chickens are considered as the most common infection source for humans. Campylobacter mainly multiplies in the mucus layer of their caeca. No effective control measures are currently available, but passive immunisation of chickens with pathogen-specific maternal IgY antibodies, present in egg yolk of immunised chickens, reduces Campylobacter colonisation. To explore this strategy further, anti-Campylobacter nanobodies, directed against the flagella and major outer membrane proteins, were fused to the constant domains of chicken IgA and IgY, combining the benefits of nanobodies and the effector functions of the Fc-domains. The designer chimeric antibodies were effectively produced in leaves of Nicotiana benthamiana and seeds of Arabidopsis thaliana. Stable expression of the chimeric antibodies in seeds resulted in production levels between 1% and 8% of the total soluble protein. These in planta produced antibodies do not only bind to their purified antigens but also to Campylobacter bacterial cells. In addition, the anti-flagellin chimeric antibodies are reducing the motility of Campylobacter bacteria. These antibody-containing Arabidopsis seeds can be tested for oral passive immunisation of chickens and, if effective, the chimeric antibodies can be produced in crop seeds.


Assuntos
Anticorpos Antibacterianos/metabolismo , Campylobacter/imunologia , Plantas Geneticamente Modificadas/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Anticorpos de Domínio Único/metabolismo , Animais , Anticorpos Antibacterianos/imunologia , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Campylobacter/fisiologia , Infecções por Campylobacter/imunologia , Infecções por Campylobacter/prevenção & controle , Infecções por Campylobacter/veterinária , Galinhas , Flagelos/genética , Flagelos/imunologia , Flagelina/imunologia , Imunidade Materno-Adquirida , Imunoglobulina A/genética , Imunoglobulina A/metabolismo , Imunoglobulinas/genética , Imunoglobulinas/metabolismo , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/prevenção & controle , Proteínas Recombinantes de Fusão/imunologia , Anticorpos de Domínio Único/imunologia , Nicotiana/genética , Nicotiana/metabolismo
2.
Vet Res ; 48(1): 86, 2017 12 08.
Artigo em Inglês | MEDLINE | ID: mdl-29216932

RESUMO

Campylobacter infections are among the most prevalent foodborne infections in humans, resulting in a massive disease burden worldwide. Broilers have been identified as the major source of campylobacteriosis and reducing Campylobacter loads in the broiler caeca has been proposed as an effective measure to decrease the number of infections in humans. Failure of current methods to control Campylobacter in broilers stresses the urgency to develop novel mitigation measures. We obtained six nanobodies with a broad specificity, that recognize strains belonging to the two most relevant species, Campylobacter jejuni and Campylobacter coli. The target of the nanobodies was identified as the major outer membrane protein, a porin that contributes to bacterial virulence and viability. Multimerization of the nanobodies led to agglutination of C. jejuni cells, which may affect colonization in the chicken gut. These Campylobacter-specific nanobodies may be useful to develop a strategy for preserving chickens from Campylobacter colonization.


Assuntos
Anticorpos Antibacterianos/imunologia , Infecções por Campylobacter/veterinária , Campylobacter coli/imunologia , Campylobacter jejuni/imunologia , Galinhas , Doenças das Aves Domésticas/prevenção & controle , Anticorpos de Domínio Único/imunologia , Animais , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas de Bactérias/imunologia , Infecções por Campylobacter/imunologia , Infecções por Campylobacter/microbiologia , Infecções por Campylobacter/prevenção & controle , Epitopos/imunologia , Porinas/imunologia , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/microbiologia
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