Effect of the Heat Shock Protein 70 on the Adjuvanticity Induced by a Bacterial Flagellin of Vibrio ulnificus.
- Author:
Shee Eun LEE
1
;
Soo Jang BAE
;
Soo Young KIM
;
Young Ran KIM
;
Joon Haeng RHEE
Author Information
1. Research Institute of Vibrio Infection and Genome Research Center for Enteropathogenic Bacteria, Chonnam National University, Gwangju 501-746, South Korea. selee@chonnam.ac.kr
- Publication Type:Original Article
- Keywords:
Flagellin;
Vibrio vulnificus;
HSP70;
Adjuvanticity
- MeSH:
Animals;
Blotting, Western;
Chromatography;
Flagellin*;
Heat-Shock Proteins*;
Hot Temperature*;
HSP70 Heat-Shock Proteins*;
Immunization;
Inteins;
Mice;
Staphylococcal Protein A;
Tetanus Toxin;
Tetanus Toxoid;
Vibrio vulnificus;
Vibrio*
- From:Journal of Bacteriology and Virology
2005;35(4):299-306
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Recently we have shown that a bacterial flagellin, Vibrio vulnifiucs FlaB (Vv-FlaB), has a strong adjuvant activity to induce protective immune response. In order to investigate the adjuvanticity of Vv-FlaB, we prepared highly purified recombinant protein by using an intein fusion protein purification system. However, in the process of the purification, we unexpectedly encountered a contamination with a 70 kDa protein. We proved the 70 kDa protein as the heat shock protein 70 (HSP70) by Western blotting. Unfortunately, it was reported that the HSP70 has a strong adjuvanticity. In this study we investigated the role of contaminating HSP70 on the Vv-FlaB-mediated adjuvanticity. We separated Vv-FlaB and HSP70 by using a high performance protein purification chromatography and compared adjuvant activities of Vv-FlaB, HSP70 and Vv-FlaB/HSP70 mixture. Using an intranasal immunization mouse model, we observed that co-administration of the flagellin with tetanus toxoid (TT) induced significantly enhanced TT-specific antibody (Ig) responses. However contaminating doses of HSP70 did not affect the adjuvanticity of Vv-FlaB and furthermore HSP70 alone did not enhance TT-specific Ig response and protective immunity against lethal challenge with tetanus toxin. These results show that the HSP70 contaminating Vv-FlaB preparations did not affect the adjuvanticity of Vv-FlaB.
