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Infection and Immunity, April 2003, p. 2239-2243, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.2239-2243.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Enhanced Murine Antigen-Specific Gamma Interferon and Immunoglobulin G2a Responses by Using Mycobacterial ESAT-6 Sequences in DNA Vaccines

F. Chris Minion,1* Sreekumar A. Menon,1,{dagger} Gregory G. Mahairas,2,{ddagger} and M. J. Wannemuehler1

Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011,1 Institute for Systems Biology, Seattle, Washington 981092

Received 23 August 2002/ Returned for modification 11 November 2002/ Accepted 3 January 2003

The Mycobacterium tuberculosis protein ESAT-6 has unusual immune stimulating activities, has been implicated in the recall of long-lived immunity, and induces protection against tuberculosis in mice. For many diseases caused by bacterial or viral pathogens, a strong cell-mediated immune (i.e., type 1) response is often required for recovery or protection. Therefore, it is important to design immunization regimens that induce agent-specific type 1 immunity. We have shown in previous studies that ESAT-6 could enhance antigen-specific type 1 immune responses in BALB/c mice against a second antigen when presented as a purified fusion protein. It was also of interest to determine if ESAT-6 could enhance the type 1 response against a second antigen beyond that afforded by DNA vaccination through CpG motifs. This was tested by using gene fusions of ESAT-6 and the Mycoplasma hyopneumoniae surface antigen P71. Modified P71 gene sequences were cloned with or without ESAT-6 sequences into a DNA vaccine vector and were used to immunize mice. Splenic lymphocytes from vaccinated mice were tested for gamma interferon (IFN-{gamma}) and interleukin-10 (IL-10) secretion. Serum antibodies were examined for P71 antigen-specific isotype responses. When stimulated in vitro with purified P71 antigen, splenocytes from the ESAT-6:P71 vaccinates secreted higher levels of IFN-{gamma} and lower levels of IL-10 compared to those of vaccinates receiving the P71 construct alone. Furthermore, the immunoglobulin G2a serum antibody levels were significantly higher in the ESAT-6:P71 vaccinates compared to those of the vaccinates receiving P71 alone. In conclusion, ESAT-6 was shown to enhance antigen-specific type 1 immune responses in BALB/c mice when used in DNA vaccines.

* Corresponding author. Mailing address: Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, IA 50011. Phone: (515) 294-6347. Fax: (515) 294-8500. E-mail: fcminion{at}iastate.edu.

Editor: J. D. Clements

{dagger} Present address: Wyeth Research, Pearl River, NY 10965-1299.

{ddagger} Present address: Regulone Corp., Seattle, WA 98199.

Infection and Immunity, April 2003, p. 2239-2243, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.2239-2243.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Olsen, I., Boysen, P., Kulberg, S., Hope, J. C., Jungersen, G., Storset, A. K. (2005). Bovine NK Cells Can Produce Gamma Interferon in Response to the Secreted Mycobacterial Proteins ESAT-6 and MPP14 but Not in Response to MPB70. Infect. Immun. 73: 5628-5635 [Abstract] [Full Text]  


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