J Korean Acad Periodontol.  2008 Dec;38(4):565-578. 10.5051/jkape.2008.38.4.565.

Production and characterization of cross-reactive anti-Porphyromonas gingivalis heat shock protein 60 monoclonal antibody

• Choi JI
• Kim SJ
• Lee JY
• Lee JY

• KMID: 1999588
• DOI: http://doi.org/10.5051/jkape.2008.38.4.565

Abstract

PURPOSE
Porphyromonas gingivalis (P. gingivalis) heat shock protein (HSP)60 may play a role in the immunopathogenesis of periodontitis as well as atherosclerosis by modulating autoimmune reaction due to its high level of sequence homology between bacteria and human counterpart. The purpose of this study was to identify immunodomiant epitope of P. gingivalis HSP60 that is reactive exclusively to the homologous bacteria without reacting with human HSP.
MATERIALS AND METHODS
The present study was performed to identify the peptide specifically recognized by anti-P. gingivalis HSP60 monoclonal antibodies mono-reactive to P. gingivalis HSP60.
RESULTS
Four different hybridomas were cloned producing monoclonal IgG antibodies exclusively to P. gingivalis HSP60. Thirty seven synthetic peptides (20-mer with 5-amino acid overlapping) were synthesized. All of these peptide were subject to SDS-PAGE for immunblot analysis. One peptide (TVPGGGTTYIRAIAALEGLK) and the other peptide (TLVVNRLRGSLKICAVKAPG) were recognized by all and one of the four monoclonal antibodies, respectively, that reacted solely with P. gingivalis HSP60. Immunohistochemistry to identify the localization of the HSP60 in the diseased gingival tissues revealed that all of the four monoclonal antibodies were highly reacted with the diseased gingival tissue than normal gingival tissue.
CONCLUSION
The P. gingivalis HSP60 peptides (TVPGGGTTYIRAIAALEGLK and TLVVNRLRGSLKICAVKAPG, respectively) are positively involved in the immunopathologic process of periodontal disease. The peptide may potentially be developed as vaccine candidates. Further investigations are under way to identify more clones producing monoclonal antibodies reactive to P. gingivalis HSP and to other periodontopathogenic bacteria as well, while maintaining specificities to human counterpart.

Keyword

epitope; anti-P.gingivalis HSP60; monoclonal antibody; periodontitis

MeSH Terms

Antibodies
Antibodies, Monoclonal
Atherosclerosis
Bacteria
Chaperonin 60
Clone Cells
Electrophoresis, Polyacrylamide Gel
Heat-Shock Proteins
Hot Temperature
Humans
Hybridomas
Immunoglobulin G
Immunohistochemistry
Peptides
Periodontal Diseases
Periodontitis
Porphyromonas gingivalis
Sequence Homology
Antibodies
Antibodies, Monoclonal
Chaperonin 60
Heat-Shock Proteins
Immunoglobulin G
Peptides