RECOMBINANT DNA-DERIVED PERTUSSIS SUBUNIT VACCINE

PERTUSSIS IS A DISEASE OF GLOBAL SIGNIFICANCE THAT PARTICULARLY AFFECTS INFANTS DURING THE FIRST YEAR OF LIFE. CURRENTLY LICENSED PERTUSSIS VACCINES ARE HIGHLY EFFICACIOUSBUT 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 THEPERTUSSIS TOXIN; (2) THE 58,000-DALTON SUBUNIT OF THE FILAMENTOUS HEMAGGLUTININ; AND (3) EACH OF THREE SEROTYPE- SPECIFIC FIMBRIAL 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, THESEGENES WILL BE INSTALLED IN HOST-VECTOR SYSTEMS CAPABLE OF HIGH-LEVEL EXPRESSION OF HETEROLOGOUS PROTEINS. THE RECOMBINANT-DERIVED PROTEINS SUBSEQUENTLY PRODUCED WILL BE ASSESSED FOR IMMUNOPOTENCY IN EXPERIMENTAL ANIMALS.