[Biological effects of immune serum from fusion protein of beta-amyloid peptide and HbcAg/MIR on AD transgeneic cells].
OBJECTIVE
To investigate the expression of fusion protein c-Abeta-c, a fusion protein from the beta amyloid peptide (Abeta) and the Hepatitis B core antigen/ Major immunodominant region (HbcAg/MIR), in the BL21/pET28 prokaryotic expression system and to immunize mice with the expressed fusion protein and evaluate the immunogenicity and biological effects of the serum in vitro.


METHODS
The recombinant prokaryotic expression plasmid PET-28a /c-Abeta-c was constructed by molecular cloning technique and the c-Abeta-c fusion gene expression was induced in E. coli BL21 by isopropyl-1-thio-b-Dgalactopyranoside (IPTG). The expressed fusion protein was analyzed by SDS-PAGE. Intraperitoneal injection (i.p.) of the purified c-Abeta-c fusion protein was given to the BALB/c mice. The anti-Abeta effect of the immune serum was detected by indirect ELISA. The biological effect of the immune serum on Alzheimer's disease (AD) transgeneic cells was assessed by MTT assay and flow cytometer.


RESULTS
The c-Abeta-c fusion protein was found in the sediment of the isolated bacteria. The expressed protein comprised more than 30% of the total proteins in the bacteria sediment. The anti-Abeta antibody in the serum of the immunized mice reached 1:16000. The antiserum reduced the cytotoxicity of Abeta peptide in the AD transgeneic cells and significantly decreased the apoptosis of cells.


CONCLUSIONS
The c-Abeta-c fusion protein has good Abeta immunogenicity and the animal immune serum efficiently inhibits the cytotoxicity of Abeta peptide.
Liu
Zhao-hui
ZH
Department of Anatomy, Histology, and Embryology, School of Medicine, Xi'an Jiaotong University, Key Laboratory of Environment and Genes Related to Disease, Ministry of Education, Xi'an 710061, China.
Feng
Gai-feng
GF
Li
Yue-ying
YY
Ma
Yan-bing
YB
Hu
Hai-tao
HT
D000544:Alzheimer Disease / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D016229:Amyloid beta-Peptides / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D018014:Gene Transfer Techniques; D006512:Hepatitis B Core Antigens / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D007106:Immune Sera / Q000276:immunology*; D016056:Immunodominant Epitopes / Q000276:immunology*; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D015183:Restriction Mapping
D016229:Amyloid beta-Peptides; D006512:Hepatitis B Core Antigens; D007106:Immune Sera; D016056:Immunodominant Epitopes; D011993:Recombinant Fusion Proteins