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1.
Chinese Journal of Biotechnology ; (12): 1623-1631, 2015.
Article in Chinese | WPRIM | ID: wpr-240549

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.


Subject(s)
Adhesins, Bacterial , Allergy and Immunology , Animals , Antibodies, Bacterial , Blood , Escherichia coli , Flow Cytometry , Immune Sera , Immunoglobulin G , Blood , Opsonin Proteins , Allergy and Immunology , Phagocytosis , Rabbits , Staphylococcal Infections , Allergy and Immunology , Staphylococcus aureus
2.
Protein & Cell ; (12): 761-769, 2014.
Article in English | WPRIM | ID: wpr-757468

ABSTRACT

Bacterial cell division is strictly regulated in the formation of equal daughter cells. This process is governed by a series of spatial and temporal regulators, and several new factors of interest to the field have recently been identified. Here, we report the requirement of gluconate 5-dehydrogenase (Ga5DH) in cell division of the zoonotic pathogen Streptococcus suis. Ga5DH catalyzes the reversible reduction of 5-ketogluconate to D-gluconate and was localized to the site of cell division. The deletion of Ga5DH in S. suis resulted in a plump morphology with aberrant septa joining the progeny. A significant increase was also observed in cell length. These defects were determined to be the consequence of Ga5DH deprivation in S. suis causing FtsZ delocalization. In addition, the interaction of FtsZ with Ga5DH in vitro was confirmed by protein interaction assays. These results indicate that Ga5DH may function to prevent the formation of ectopic Z rings during S. suis cell division.


Subject(s)
Bacterial Proteins , Chemistry , Genetics , Metabolism , Cell Division , Cell Shape , Cytoskeletal Proteins , Chemistry , Genetics , Metabolism , Mutation , Oxidoreductases , Genetics , Metabolism , Protein Binding , Streptococcus suis
3.
Protein & Cell ; (12): 997-1005, 2011.
Article in English | WPRIM | ID: wpr-757312

ABSTRACT

There is a great need for new vaccine development against influenza A viruses due to the drawbacks of traditional vaccines that are mainly prepared using embryonated eggs. The main component of the current split influenza A virus vaccine is viral hemagglutinin (HA) which induces a strong antibody-mediated immune response. To develop a modern vaccine against influenza A viruses, the current research has been focused on the universal vaccines targeting viral M2, NP and HA proteins. Crystallographic studies have shown that HA forms a trimer embedded on the viral envelope surface, and each monomer consists of a globular head (HA1) and a "rod-like" stalk region (HA2), the latter being more conserved among different HA subtypes and being the primary target for universal vaccines. In this study, we rationally designed the HA head based on the crystal structure of the 2009-pandemic influenza A (H1N1) virus HA as a model, tested its immunogenicity in mice, solved its crystal structure and further examined its immunological characteristics. The results show that the HA globular head can be easily prepared by in vitro refolding in an E. coli expression system, which maintains its intact structure and allows for the stimulation of a strong immune response. Together with recent reports on some similar HA globular head preparations we conclude that structure-based rational design of the HA globular head can be used for subtype-specific vaccines against influenza viruses.


Subject(s)
Animals , Antibodies, Viral , Allergy and Immunology , Crystallography, X-Ray , Drug Design , Female , Freund's Adjuvant , Hemagglutinin Glycoproteins, Influenza Virus , Genetics , Allergy and Immunology , Humans , Influenza A Virus, H1N1 Subtype , Genetics , Allergy and Immunology , Influenza Vaccines , Influenza, Human , Allergy and Immunology , Virology , Mice , Mice, Inbred BALB C , Models, Molecular , Protein Folding , Recombinant Proteins , Genetics , Allergy and Immunology , Structure-Activity Relationship , Vaccination , Vaccines, Subunit
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