Identification of immunodominant Helicobacter pylori proteins with reactivity to H. pylori-specific egg-yolk immunoglobulin.
The importance of hens eggs as a source of specific antibodies (IgY) is well recognized. The protective effect of IgY obtained from hens immunized with Helicobacter pylori whole-cell lysate has been reported for the control of H. pylori infection. However, IgY produced by whole-cell lysates presents the possibility of cross-reactivity with other bacteria, including the normal human flora, and this could decrease the efficiency of IgY. In the present study, the immunodominant proteins of H. pylori with reactivity to H. pylori-specific IgY (IgY-Hp) were identified. IgY obtained from hens immunized with various fractions of H. pylori proteins was isolated and purified, titres of IgY-Hp against H. pylori were determined and cross-reactivity between IgY-Hp and normal human bacteria was examined by Western blot analysis. Finally, immunodominant H. pylori proteins were identified by LC/MS analysis. IgY obtained 2 months after immunization with H. pylori whole-cell lysate showed the highest antibody titre. Five immunodominant proteins were identified that were strongly reactive to IgY-Hp: urease beta-subunit (62 kDa), heat-shock protein 60 (60 kDa), urease alpha-subunit (26 kDa), probable peroxiredoxin (22 kDa) and probable thiol peroxidase (18 kDa). Immunization of hens with the immunodominant proteins identified would produce a more specific IgY against H. pylori.
Shin
Ji-Hyun
JH
Research Center for Gastroenterology1 and Departments of Gastroenterology2 and Surgery3, Dankook University College of Medicine, Cheonan, Korea 4Department of Biochemistry, Yonsei University College of Sciences, Seoul, Korea 5Department of Agricultural Chemistry, Korea University College of Life & Environmental Science, Seoul, Korea.
Nam
Seung-Woo
SW
Research Center for Gastroenterology1 and Departments of Gastroenterology2 and Surgery3, Dankook University College of Medicine, Cheonan, Korea 4Department of Biochemistry, Yonsei University College of Sciences, Seoul, Korea 5Department of Agricultural Chemistry, Korea University College of Life & Environmental Science, Seoul, Korea.
Kim
Jung-Taik
JT
Research Center for Gastroenterology1 and Departments of Gastroenterology2 and Surgery3, Dankook University College of Medicine, Cheonan, Korea 4Department of Biochemistry, Yonsei University College of Sciences, Seoul, Korea 5Department of Agricultural Chemistry, Korea University College of Life & Environmental Science, Seoul, Korea.
Yoon
Jong-Bok
JB
Research Center for Gastroenterology1 and Departments of Gastroenterology2 and Surgery3, Dankook University College of Medicine, Cheonan, Korea 4Department of Biochemistry, Yonsei University College of Sciences, Seoul, Korea 5Department of Agricultural Chemistry, Korea University College of Life & Environmental Science, Seoul, Korea.
Bang
Won-Gi
WG
Research Center for Gastroenterology1 and Departments of Gastroenterology2 and Surgery3, Dankook University College of Medicine, Cheonan, Korea 4Department of Biochemistry, Yonsei University College of Sciences, Seoul, Korea 5Department of Agricultural Chemistry, Korea University College of Life & Environmental Science, Seoul, Korea.
Roe
Im-Hwan
IH
Research Center for Gastroenterology1 and Departments of Gastroenterology2 and Surgery3, Dankook University College of Medicine, Cheonan, Korea 4Department of Biochemistry, Yonsei University College of Sciences, Seoul, Korea 5Department of Agricultural Chemistry, Korea University College of Life & Environmental Science, Seoul, Korea.
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology*; D002645:Chickens / Q000276:immunology*; D003429:Cross Reactions; D004527:Egg Proteins / Q000276:immunology*; D004926:Escherichia coli / Q000276:immunology; D005260:Female; D016480:Helicobacter pylori / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology*; D007136:Immunoglobulins / Q000276:immunology*; D007778:Lactobacillus / Q000276:immunology; D008970:Molecular Weight; D013211:Staphylococcus aureus / Q000276:immunology; D013295:Streptococcus mutans / Q000276:immunology
D001426:Bacterial Proteins; D004527:Egg Proteins; C016512:IgY; D016056:Immunodominant Epitopes; D007136:Immunoglobulins