Antibodies anti-Shiga toxin 2 B subunit from chicken egg yolk: Isolation, purification and neutralization efficacy

被引：29

作者：

Parma, Y. R. ^{[1
]}

Chacana, P. A. ^{[2
]}

Roge, A. ^{[3
]}

Kahl, A. ^{[2
]}

Cangelosi, A. ^{[4
]}

Geoghegan, P. ^{[4
]}

Lucchesi, P. M. A. ^{[5
]}

Fernandez-Miyakawa, M. E. ^{[1
]}

机构：

[1] Inst Nacl Tecnol Agropecuaria, Ctr Nacl Invest Agropecuarias, Inst Patobiol, RA-1712 Buenos Aires, DF, Argentina

[2] Inst Nacl Tecnol Agropecuaria, Ctr Nacl Invest Agropecuarias, Inst Virol, IncuINTA, RA-1712 Buenos Aires, DF, Argentina

[3] Inst Nacl Prod Biol, Serv Sueros & Antigenos, Buenos Aires, DF, Argentina

[4] Ctr Nacl Control Calidad Biol, Serv Immunoterapeut, Buenos Aires, DF, Argentina

[5] Univ Natl Ctr Pcia, Depto Sanidad Anim & Med Prevent, Lab Inmunoquim & Biotecnol, Buenos Aires, Tandil, Argentina

来源：

TOXICON | 2011年 / 58卷 / 04期

关键词：

Shiga toxin; Antitoxin; IgY technology; Chicken; Neutralization; Stx2B; ESCHERICHIA-COLI O157-H7; IN-VITRO; IMMUNOGLOBULIN; PROTECTION; IGY; MICE; INFECTION; BOVINE; ASSAY;

D O I：

10.1016/j.toxicon.2011.07.009

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

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 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. (C) 2011 Elsevier Ltd. All rights reserved.

引用

页码：380 / 388

页数：9