Long-lasting specific antibodies against P277 induced by mucosal administration of P277 repeat sequences carried by Hsp65 in the absence of adjuvants

Liang Jin; Yu Wang; Qiyan Xiong; Qingmei Chen; Jianping Li; Aihua Zhu; Rongyue Cao; Jie Wu; Jingjing Liu

doi:10.1016/j.vaccine.2006.11.052

Long-lasting specific antibodies against P277 induced by mucosal administration of P277 repeat sequences carried by Hsp65 in the absence of adjuvants

Vaccine. 2007 Mar 1;25(11):2043-50. doi: 10.1016/j.vaccine.2006.11.052. Epub 2006 Dec 8.

Authors

Liang Jin¹, Yu Wang, Qiyan Xiong, Qingmei Chen, Jianping Li, Aihua Zhu, Rongyue Cao, Jie Wu, Jingjing Liu

Affiliation

¹ Minigene Pharmacy Laboratory, Biopharmaceutical College, China Pharmaceutical University, Tongjia Xiang 24, Nanjing 210009, PR China.

PMID: 17224213
DOI: 10.1016/j.vaccine.2006.11.052

Abstract

To improve the weak immunogenicity of peptide P277, the recombinant expression plasmid pET28-Hsp65-6 x P277 was constructed by inserting 5 x P277 which was amplified by PCR into the vector pET28-Hsp65-P277. It was transformed into Escherichia coli BL21 (DE3) and the fusion protein (Hsp65-6 x P277) was expressed effectively as soluble protein after inducing by lactose. The fusion protein was purified by means of cell disruption, ammonium sulfate precipitation, double-distilled H2O dialysis, DEAE52-cellulose column chromatography, and then used to immunize female NOD mice with three i.n. inoculations in the absence of adjuvants. Serum samples from the immunized mice were collected at 3 weeks interval. Antibodies against P277 and HSP65 were detected in immunized mice sera by enzyme-linked immunosorbent assay (ELISA) and Western blot. Specific antibodies were successfully induced and lasted for more than 20 weeks in animals immunized with the fusion protein via intranasal route even in the absence of adjuvants.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Administration, Intranasal
Animals
Antibodies / blood
Antibodies, Bacterial / blood
Antibody Affinity
Antibody Specificity
Bacterial Proteins / genetics
Bacterial Proteins / immunology*
Blood Glucose / analysis
Blotting, Western
Chaperonin 60
Chaperonins / genetics
Chaperonins / immunology*
Diabetes Mellitus, Type 1 / prevention & control*
Disease Models, Animal
Enzyme-Linked Immunosorbent Assay
Female
Heat-Shock Proteins / genetics
Heat-Shock Proteins / immunology*
Mice
Mice, Inbred NOD
Peptide Fragments / genetics
Peptide Fragments / immunology*
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / immunology
Vaccines, Synthetic / genetics
Vaccines, Synthetic / immunology*

Substances

Antibodies
Antibodies, Bacterial
Bacterial Proteins
Blood Glucose
Chaperonin 60
Heat-Shock Proteins
Peptide Fragments
Recombinant Fusion Proteins
Vaccines, Synthetic
heat-shock protein 65, Mycobacterium
peptide 277, heat shock protein 60
Chaperonins