Abstract

The immediate goal of this research is to construct an effective, nontoxic acellular vaccine against infection by Bordetella pertussis. Pertussis toxin is an extracellular protein produced by B. Pertussis that is both a determinant of virulence and an antigen that elicits protective immunity. Conditions will be established to produce a toxoid of pertussis toxin that lacks cytotoxicity but retains immunogenicity. The genetically engineered toxoid of pertussis toxin will be tested alone and with other antigens of B. pertussis as acellular vaccine candidates. The long-term goals are to develop strategies for the construction of an acellular vaccine against B. pertussis that are applicable for the generation of recombinant DNA-derived acellular vaccines against other bacterial !pathogens. The research project described will define the catalytic properties of the Sl subunit of pertussis toxin.
The aim i s to define the function of specific residues within the Sl subunit that reduce toxicity but retain immunogenicity. This research will also focus on determining the mechanism of action of pertussis toxin by defining how pertussis toxin is activated, enters sensitive cells, and interacts with G-proteins.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Modified Research Career Development Award (K04)
Project #
5K04AI001087-02
Application #
3071050
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Project Start
1992-07-01
Project End
1997-06-30
Budget Start
1993-07-01
Budget End
1994-06-30
Support Year
2
Fiscal Year
1993
Total Cost
Indirect Cost

  Institution

Name
Medical College of Wisconsin
Department
Type
Schools of Medicine
DUNS #
073134603
City
Milwaukee
State
WI
Country
United States
Zip Code
53226

  Publications

Ganesan, A K; Frank, D W; Misra, R P et al. (1998) Pseudomonas aeruginosa exoenzyme S ADP-ribosylates Ras at multiple sites. J Biol Chem 273:7332-7
Liu, S; Yahr, T L; Frank, D W et al. (1997) Biochemical relationships between the 53-kilodalton (Exo53) and 49-kilodalton (ExoS) forms of exoenzyme S of Pseudomonas aeruginosa. J Bacteriol 179:1609-13
Knight, D A; Barbieri, J T (1997) Ecto-ADP-ribosyltransferase activity of Pseudomonas aeruginosa exoenzyme S. Infect Immun 65:3304-9
Liu, S; Kulich, S M; Barbieri, J T (1996) Identification of glutamic acid 381 as a candidate active site residue of Pseudomonas aeruginosa exoenzyme S. Biochemistry 35:2754-8
Finck-Barbancon, V; Barbieri, J T (1996) Preferential processing of the S1 subunit of pertussis toxin that is bound to eukaryotic cells. Mol Microbiol 22:87-95
Xu, Y; Barbieri, J T (1996) Pertussis toxin-catalyzed ADP-ribosylation of Gi-2 and Gi-3 in CHO cells is modulated by inhibitors of intracellular trafficking. Infect Immun 64:593-9
Kulich, S M; Frank, D W; Barbieri, J T (1995) Expression of recombinant exoenzyme S of Pseudomonas aeruginosa. Infect Immun 63:1-8
Krueger, K M; Barbieri, J T (1995) The family of bacterial ADP-ribosylating exotoxins. Clin Microbiol Rev 8:34-47
Knight, D A; Finck-Barbancon, V; Kulich, S M et al. (1995) Functional domains of Pseudomonas aeruginosa exoenzyme S. Infect Immun 63:3182-6
Finck-Barbancon, V; Barbieri, J T (1995) ADP-ribosylation of alpha i3C20 by the S1 subunit and deletion peptides of S1 of pertussis toxin. Biochemistry 34:1070-5

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