Prokaryotic expression of Staphylococcus aureus Clumping factor B and evaluation of the antiserum-mediated opsonic activity.
- Author:
Lili ZHANG
;
Shanshan CAO
;
Yifeng ZHANG
;
Chunling XUAN
;
Youqiang WU
;
Songhua HU
- Publication Type:Journal Article
- MeSH:
Adhesins, Bacterial;
immunology;
Animals;
Antibodies, Bacterial;
blood;
Escherichia coli;
Flow Cytometry;
Immune Sera;
Immunoglobulin G;
blood;
Opsonin Proteins;
immunology;
Phagocytosis;
Rabbits;
Staphylococcal Infections;
immunology;
Staphylococcus aureus
- From:
Chinese Journal of Biotechnology
2015;31(11):1623-1631
- CountryChina
- Language:Chinese
-
Abstract:
Staphylococcus aureus is a major cause of hospital-acquired infection. Because the bacteria are very easy to become resistant to antibiotics, vaccination is a main method against S. aureus infection. Clumping factor B (ClfB) is an adhesion molecule essential for S. aureus to colonize in the host mucosa and is regarded as an important target antigen. In this study, we successfully used Escherichia coli to express a segment encoding the N1-N3 regions of ClfB protein (Truncated-ClfB) cloned from S. aureus. The protein was purified by affinity and ion exchange chromatographies and gel filtration. Rabbits were immunized three times with purified Truncated-ClfB. After that, blood was collected to prepare serum which were then used for measurement of antibody level. Phagocytosis of S. aureus opsonized by the serum was determined by a flow cytometry. Results show that the serum IgG titer reached 1:640 000. Phagocytosed S. aureus by polymorphonuclear leukocytes were significantly more when the bacteria were opsonized by the serum from Truncated-ClfB immunized rabbits than those from no immunized group (P < 0.01). Therefore, the results indicated that Truncated-ClfB could be a promising vaccine candidate against S. aureus infection.
