INVESTIGADORES
LUCCHESI paula Maria Alejandra
artículos
Título:
Antibodies anti-Shiga toxin 2 B subunit from chicken egg yolk: isolation, purification and neutralization efficacy
Autor/es:
PARMA, Y. R.; CHACANA, P. A.; ROGÉ, A.; KAHL, A.; CANGELOSI, A.; GEOGHEGAN, P.; LUCCHESI, P. M. A.; FERNÁNDEZ-MIYAKAWA, M. E.
Revista:
TOXICON
Editorial:
PERGAMON-ELSEVIER SCIENCE LTD
Referencias:
Año: 2011 vol. 58 p. 380 - 388
ISSN:
0041-0101
Resumen:
Shiga toxins (Stx1 and Stx2) are the main virulence factors of enterohemorrhagic Escherichia
coli (EHEC), a foodborne pathogen associated with diarrhea, hemorrhagic colitis and
hemolytic uremic syndrome. The aim of this study was to evaluate the antibodies against
Stx2 obtained from egg yolks of laying hens immunized with a recombinant Stx2B subunit.
A high specific response in serum was observed 25 days after the first immunization and
IgY antibodies were extracted from day 47th and purified from egg yolk. A concentration of
0.84 mg of total IgY/ml of egg yolk was obtained, of which 8% were antigen specific. The
ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution
were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and
ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivoEscherichia
coli (EHEC), a foodborne pathogen associated with diarrhea, hemorrhagic colitis and
hemolytic uremic syndrome. The aim of this study was to evaluate the antibodies against
Stx2 obtained from egg yolks of laying hens immunized with a recombinant Stx2B subunit.
A high specific response in serum was observed 25 days after the first immunization and
IgY antibodies were extracted from day 47th and purified from egg yolk. A concentration of
0.84 mg of total IgY/ml of egg yolk was obtained, of which 8% were antigen specific. The
ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution
were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and
ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivo(EHEC), a foodborne pathogen associated with diarrhea, hemorrhagic colitis and
hemolytic uremic syndrome. The aim of this study was to evaluate the antibodies against
Stx2 obtained from egg yolks of laying hens immunized with a recombinant Stx2B subunit.
A high specific response in serum was observed 25 days after the first immunization and
IgY antibodies were extracted from day 47th and purified from egg yolk. A concentration of
0.84 mg of total IgY/ml of egg yolk was obtained, of which 8% were antigen specific. The
ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution
were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and
ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivofic response in serum was observed 25 days after the first immunization and
IgY antibodies were extracted from day 47th and purified from egg yolk. A concentration of
0.84 mg of total IgY/ml of egg yolk was obtained, of which 8% were antigen specific. The
ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution
were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and
ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivofied from egg yolk. A concentration of
0.84 mg of total IgY/ml of egg yolk was obtained, of which 8% were antigen specific. The
ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution
were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and
ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivofic. The
ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution
were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and
ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivoficacy of IgY against Stx2 was determined by in vitro and in vivo
experiments. The results show that IgY was able to recognize Stx2B and Stx2 in denatured
conditions, attached to a solid-phase and free in solution. The anti-Stx2B IgY could
effectively block the biological activity of Stx2 on Vero cells and protect mice from Stx2
challenge.
The data suggest that immunization of hens with Stx2B could be a strategy to obtain at low
cost a relatively high concentration of anti-Stx2 egg yolk IgY, able to neutralize Stx2 lethal
activity. IgY technology could be an useful tool for research, diagnosis and therapy of EHEC
infection.