Long-lasting specific antibodies against CETP induced by subcutaneous and mucosal administration of a 26-amino acid CETP epitope carried by heat shock protein 65 kDa in the absence of adjuvants.
The heat shock protein 65 kDa (Hsp65) of Mycobacterium tuberculosis var. bovis was fused with the linear polypeptide epitope of cholesteryl ester transfer protein C-terminal fragment (CETPC) and expressed as soluble protein in Escherichia coli. The fusion protein Hsp65-CETPC was purified by anion exchange column and eluted at 100-130 mM NaCl in 10mM phosphate buffer (pH 8.0), and then used to immunize mice via subcutaneous injection or intranasal delivery in the absence of adjuvants. Antibodies against CETPC were detected in immunized mice sera by enzyme-linked immunosorbent assay (ELISA) and verified by Western blot analysis. Specific antibodies were successfully induced and lasted for more than 12 weeks in animals immunized with the fusion protein via both subcutaneous and intranasal routes even in the absence of adjuvants. Results showed that Hsp65 could be used as a convenient carrier molecule for presenting foreign polypeptide epitopes, such as CETPC, to the immune system in vivo. Antibodies induced by Hsp65-CETPC could partially inhibit the excessive activity of CETP to normal level. Therefore, Hsp65-CETPC might be further developed to a vaccine against atherosclerosis.
Gaofu
Qi
Q
Minigene Pharmacy Laboratory, Biopharmaceutical College, China Pharmaceutical University, Tongjia Xiang 24, Nanjing 210009, PR China.
Dan
Mao
M
Jie
Wu
W
Liao
Zhongping
Z
Li
Zong
Z
Roque
Rouel S
RS
Jingjing
Liu
L
D000276:Adjuvants, Immunologic; D000281:Administration, Intranasal; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000032:analysis; D001500:BCG Vaccine / Q000276:immunology; D001426:Bacterial Proteins / Q000008:administration & dosage / Q000276:immunology*; D015153:Blotting, Western; D002352:Carrier Proteins / Q000008:administration & dosage / Q000276:immunology*; D018834:Chaperonin 60; D018833:Chaperonins / Q000008:administration & dosage / Q000276:immunology*; D053480:Cholesterol Ester Transfer Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D005822:Genetic Vectors; D006023:Glycoproteins / Q000008:administration & dosage / Q000276:immunology*; D018928:Immunity, Mucosal / Q000276:immunology*; D007279:Injections, Subcutaneous; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000097:blood / Q000276:immunology
D000276:Adjuvants, Immunologic; D000906:Antibodies; D001500:BCG Vaccine; D001426:Bacterial Proteins; D002352:Carrier Proteins; D018834:Chaperonin 60; D053480:Cholesterol Ester Transfer Proteins; D000939:Epitopes; D006023:Glycoproteins; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; C074119:heat-shock protein 65, Mycobacterium; D018833:Chaperonins