SBIR-STTR Award

Pertussis is a disease of global significance that particularly affects infants during the first year of life. Currently licensed pertussis vaccines are highly efficacious, but questions regarding their safety have led to public fears of vaccine-related injuries. These investigations examine the possibility of producing a subunit vaccine composed of pertussis antigens expressed in heterologous hosts by virtue of recombinant DNA methodology.In preliminary experiments, specific antigenic components of the bacterium believed capable of eliciting or augmenting protective immune responses against disease will be isolated. These components are: (1) the S-1 subunit of the pertussis toxin; (2) the 58,000-dalton subunit of the filamentous hemagglutinin; and (3) each of three serotype-specffic fhnbrial agglutinogens. N-terminal amino acid sequences will be obtained for each polypeptide with a gas-phase microsequenator. A series of oligonucleotide probes representing predicted nucleotide sequences encoding each antigen will be synthesized and used as hybridization probes to isolate the genes for these proteins from B. pertussis genomic libraries.In later investigations, these genes will be installed in host-vector systems capable of highlevel expression of heterologous proteins. The recombinant-derived proteins subsequently produced will be assessed for immunopotency in experimental animals.National Institute of Allergy and Infectious Diseases (NIAID)

Vaccines of inactivated Bordetella pertussis have provided an effective means of controlling whooping cough. Questions regarding the safety of these vaccines have led to public fear of vaccine injuries, litigation for which has reached proportions that ultimately threaten the nation's vaccin supply and have chilling effect on new vaccine development. To meet these challenges, we have taken a recombinant DNA approach to the improvement of pertussis vaccine. Pertussis toxin is a major virulence factor of the organism; recent clinical evidence suggests that inactivated toxin alone may confer disease protection. We have molecularly cloned portions of the toxin operon into bacterial expression vectors and obtained very high levels of selected toxin subunits as non-fusion proteins in recombinant E. coli. Preliminary studies demonstrate certain of these subunits retain their biological activity, are highly immunogenic, and confer protection against disease challenge in experimental animals. We propose to express other toxin subunits, and perhaps other virulence factors, in various recombinant hosts, genetically modify these proteins to eliminate their reactogenicity and test them for immunoprotection in a number of formulations. Genetically-engineered vaccines are expected to be economical to produce, free of reactogenic components, and both highly saf and efficacious.National Institute of Allergy and Infectious Diseases (NIAID)