[Preparation oral liposome-encapsulated recombinant Helicobacter pylori heat shock protein 60 vaccine for prevention of Hp infection].
OBJECTIVE
To prepare oral liposome-encapsulated recombinant Helicobacter pylori (Hp) heat shock protein 60 (Hsp60) vaccine and investigate its effect against Hp infection in mice.


METHODS
The recombinant vector PET-22(+)/Hsp60 was transformed into BL21(DE3) E.coli. The recombinant protein was purified with Ni-NTA agrose resin and the oral liposome-encapsulated vaccine was prepared with phosphatidyl choline and cholesterols using film method, with the size distribution of the folate liposomes measured by transmission electronic microscopy. BALB/c mice were divided into 5 groups and immunized by intragastric administration of PBS, liposome, rHsp60 plus choleratoxin (CT), liposome-encapsulated rHsp60, and liposome-encapsulated rHsp60 plus CT, respectively, given once a week for 4 weeks. All the mice were challenged by Hp for 3 times within two weeks following the last immunization and sacrificed 3 weeks after the last challenge. Hp detection was performed by fast urease test. Semi-quantitative assessment of the bacterial colonization density observation of the inflammation severity and gastric histopathology were carried out.


RESULTS
The soluble expression product accounted for 27% of the total bacterial protein. The purity of recombinant fusion protein was about 95% after purification. The mean size of the folate liposomes was 0.7+/-0.4 mum. PBS or liposome alone showed no immune-enhancing effect, and rHsp60 plus CT, liposome-encapsulated rHsp60 and liposome-encapsulated rHsp60 plus CT had the protective rates against Hp infection of 73.3%, 66.7% and 86.7%, respectively. The latter 3 preparations effected significantly reduced Hp infection and alleviated the inflammation in the gastric mucosa of the mice challenged with Hp.


CONCLUSIONS
The oral liposome may serve as a potential adjuvant for Hp vaccine in preventing Hp infection.
Huang
Wen
W
Institute of Digestive Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. huangsh666@263.net
Bai
Yang
Y
Wang
Ji-de
JD
Wu
Jin-bao
JB
Li
Guo-feng
GF
Zhang
Wei-ming
WM
Zhou
Dian-yuan
DY
D000818:Animals; D001428:Bacterial Vaccines / Q000008:administration & dosage* / Q000096:biosynthesis* / Q000276:immunology; D018834:Chaperonin 60 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D016481:Helicobacter Infections / Q000517:prevention & control*; D016480:Helicobacter pylori / Q000276:immunology*; D008081:Liposomes; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000276:immunology
D001428:Bacterial Vaccines; D018834:Chaperonin 60; D008081:Liposomes; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic