J Korean Acad Periodontol.  2008 Aug;38(Suppl):335-342. 10.5051/jkape.2008.38.Suppl.335.

Production of the polyclonal subunit C protein antibody against Aggregatibacter actinomycetemcomitans cytolethal distending toxin

• Lee SJ
• Park SY
• Ko SY
• Ryu SH
• Kim HS

• KMID: 1486868
• DOI: http://doi.org/10.5051/jkape.2008.38.Suppl.335

Abstract

PURPOSE
Cytolethal distending toxin (CDT) considered as a key factor of localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis is composed of five open reading frames (ORFs). Among of them, the individual role of CdtA and CdtC is not clear; several reports presents that CDT is an AB2 toxin and they enters the host cell via clathrin-coated pits or through the interaction with GM3 ganglioside. So, CdtA, CdtC, or both seem to be required for the delivery of the CdtB protein into the host cell. Moreover, recombinant CDT was suggested as good vaccine material and antibody against CDT can be used for neutralization or for a detection kit.
MATERIALS AND METHODS
We constructed the pET28a-cdtC plasmid from Aggregatibacter actinomycetemcomitans Y4 by genomic DNA PCR and expressed in BL21 (DE3) Escherichia coli system. We obtained the antibody against the recombinant CdtC in mice system. Using the anti-CdtC antibody, we test the native CdtC detection by ELISA and Western Blotting and confirm the expression time of native CdtC protein during the growth phase of A. actinomycetemcomitans.
RESULTS
In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and generated the anti CdtC antibody against recombinant CdtC subunit expressed in E. coli system. Our anti CdtC antibody can be interacting with recombinant CdtC and native CDT in ELISA and Western system. Also, CDT holotoxin existed at 24h but not at 48h meaning that CDT holotoxin was assembled at specific time during the bacterial growth.
CONCLUSION
In conclusion, we thought that our anti CdtC antibody could be used mucosal adjuvant or detection kit development, because it could interact with native CDT holotoxin.

Keyword

Aggregatibacter actinomycetemcomitans; cytolethal distending toxin

MeSH Terms

Aggressive Periodontitis
Animals
Bacterial Toxins
Blotting, Western
DNA
Edetic Acid
Endocarditis
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Meningitis
Mice
Open Reading Frames
Osteomyelitis
Plasmids
Polymerase Chain Reaction
Bacterial Toxins
DNA
Edetic Acid