CYTOMEGALOVIRUS GLYCOPROTEIN B RECOMBINANT ANTIGENS

THIS RESEARCH WILL EXPRESS RECOMBINANT FORMS OF HUMAN CYTOMEGALOVIRUS (CMV) GLYCOPROTEIN B (GB) FOR USE IN THE DEVELOPMENT OF A SUBUNIT VACCINE AGAINST CMV. IN VITRO MUTAGENESIS AND OTHER RECOMBINANT DNA TECHNIQUES WILL BE USED TO GENERATE TWO FORMS OF GB: (1) AN UNCLEAVED GB MOLECULE LACKING THE C-TERMINAL TRANSMEMBRANE AND CYTOPLASMIC DOMAIN, AND (2) A C-TERMINAL GB FRAGMENT FUSED TO A HETEROLOGOUS SECRETORY SIGNAL SEQUENCE. EXPRESSION PLASMIDS ENCODING THESE CONSTRUCTS WILL BE TESTED IN MAMMALIAN CELLS FOR SECRETION AND REACTIVITY WITH CMV-NEUTRALIZING MONOCLONAL ANTIBODIES. THE AIM IS TO DEVELOP STABLE CELL LINES SECRETING HIGH LEVELS OF A SINGLE GB POLYPEPTIDE THAT RETAINS THE MAJOR GB-NEUTRALIZING EPITOPES. LONG-TERM OBJECTIVES WILL INVOLVEPURIFICATION OF RECOMBINANT FORMS OF GB PRODUCED BY MAMMALIAN CELLS. THIS MATERIAL WILL BE TESTED IN ANIMALS FOR THE ABILITY TO GENERATE NEUTRALIZING ANTIBODIES. THOSE RECOMBINANT ANTIGENS THAT ELICIT A STRONG NEUTRALIZINGRESPONSE WILL BE USED TO DEVELOP A SUBUNIT VACCINE CAPABLE OF REDUCING THE SERIOUS CONSEQUENCES OF CMV DISEASE IN NEONATES AND TRANSPLANT RECIPIENTS. THIS VACCINE WILL BE INITIALLY TESTED IN SERONEGATIVE TRANSPLANT RECIPIENTS. FURTHER CLINICAL TRIALS WILL ALSO BE DONE IN FEMALES OF CHILDBEARING AGE AT RISK FOR CMV INFECTION, SUCH AS PARENTS OF CHILDREN ATTENDING DAYCARE CENTERS.