J Bacteriol Virol.  2005 Dec;35(4):299-306.

Effect of the Heat Shock Protein 70 on the Adjuvanticity Induced by a Bacterial Flagellin of Vibrio ulnificus

Affiliations
  • 1Research Institute of Vibrio Infection and Genome Research Center for Enteropathogenic Bacteria, Chonnam National University, Gwangju 501-746, South Korea. selee@chonnam.ac.kr
  • 2Chonnam Dental Research Institute,School of Dentistry, Chonnam National University, Gwangju 500-757, South Korea.
  • 3National Research Laboratory of Molecular Microbial Pathogenesis and Department of Microbiology, Chonnam National University Medical School, Gwangju 501-746, South Korea.

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.

Keyword

Flagellin; Vibrio vulnificus; HSP70; Adjuvanticity

MeSH Terms

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*
Flagellin
HSP70 Heat-Shock Proteins
Heat-Shock Proteins
Staphylococcal Protein A
Tetanus Toxin
Tetanus Toxoid
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