Abstract
The accumulating knowledge about host–pathogen interactions in infectious diseases shows how the immune system interfaces with pathogens, and thus, helps us in understanding the pathogenesis of diseases and improving their treatment. Purified antigens are indispensable while investigating the immune response in both innate and acquired immunities. It is ideal to use native antigens purified from the host organisms in native conditions that sustain their biological activity completely. However, purification of native antigens, especially on a large scale, is technically difficult and generally time consuming. Purifying protein as a peptide-tagged fusion protein is an effective approach. Purification of a recombinant protein engineered to incorporate a polyhistidine tag at either the carboxyl or amino terminus is an established method, and it can be easily modified to obtain optimal results under different conditions. Heat-shock proteins were highly conserved during evolution and are highly homologous between prokaryotic and eukaryotic cells. Their molecular mimicry might have roles in the pathogenesis of chronic inflammatory diseases. We successfully generated histidine-tagged recombinant heat-shock proteins from the periodontopathogens, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. The recombinant proteins allowed us to evaluate the immune response to these antigens in periodontitis patients.
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References
Hochuli, E., Bannwarth, W., Döbeli, H., Gentz, R., and Stuber, D. (1988) Genetic approach to facilitate purification of recombinant proteins with a novel metal chelate adsorbent. Bio/Technology. 6, 1321–1325.
Hengen, P. (1995) Purification of His-Tag fusion proteins from Escherichia coli. Trends Biochem. Sci. 20, 285–286.
Ford, P. J., Gemmell, E., Hamlet, S. M., Hasan, A., Walker, P. J., West, M. J., Cullinan, M. P., and Seymour, G. J. (2005) Cross-reactivity of GroEL antibodies with human heat shock protein 60 and quantification of pathogens in atherosclerosis. Oral Microbiol. Immunol. 20, 296–302.
Kiessling, R., Gronberg, A., Ivanyi, J., Söderström, K., Ferm, M., Kleinau, S., Nilsson, E., and Klareskog, L. (1991) Role of hsp60 during autoimmune and bacterial inflammation. Immunol. Rev. 121, 91–111.
Maeda, H., Miyamoto, M., Kokeguchi, S., Kono, T., Nishimura, F., Takashiba., S., and Murayama, Y. (2000) Epitope mapping of heat shock protein 60 (GroEL) from Porphyromonas gingivalis. FEMS Immunol. Med. Microbiol. 28, 219–224.
Tabeta, K., Yamazaki, K., Hotokezaka, H., Yoshie, H., and Hara, K. (2000) Elevated humoral immune response to heat shock protein 60 (hsp60) family in periodontitis patients. Clin. Exp. Immunol. 120, 285–293.
Tabeta, K., Yoshie, H., and Yamazaki, K. (2001) Characterization of serum antibody to Actinobacillus actinomycetemcomitans GroEL-like protein in periodontitis patients and healthy subjects. Oral Microbiol. Immunol. 16, 290–295.
Yamazaki, K., Ohsawa, Y., Itoh, H., Ueki, K., Tabeta, K., Oda, T., Nakajima, T., Yoshie, H., Saito, S., Oguma, F., Kodama, M., Aizawa, Y., and Seymour, G. J. (2004) T-cell clonality to Porphyromonas gingivalis and human heat shock protein 60 s in patients with atherosclerosis and periodontitis. Oral Microbiol. Immunol. 19, 160–167.
Yamazaki, K., Ohsawa, Y., Tabeta, K., Ito, H., Ueki, K., Oda, T., Yoshie, H., and Seymour, G. J. (2002) Accumulation of human heat shock protein 60-reactive T cells in the gingival tissues of periodontitis patients. Infect. Immun. 70, 2492–2501.
Acknowledgments
This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (#19390536 and #20659325) and the Promotion of Niigata University Research Project.
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Tabeta, K., Yamazaki, K. (2010). Analysis of Immune Responses to Purified Recombinant Antigens of Periodontal Pathogens. In: Seymour, G., Cullinan, M., Heng, N. (eds) Oral Biology. Methods in Molecular Biology, vol 666. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-820-1_21
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DOI: https://doi.org/10.1007/978-1-60761-820-1_21
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