City of Hope

Hema Pande

John A Zaia

Arthur D Riggs

Brian R Clark

Disclosed are amino acid sequences of the late 64 kilodalton protein of human cytomegalovirus (HCMVgp64), useful in diagnosing and preventing HCMV infections.

Method for detection and prevention of human cytomegalovirus infection

Immunogens for use in human cytomegalovirus vaccines or skin tests which comprise human cytomegalovirus matrix proteins or fractions thereof are disclosed. It is also disclosed that such matrix proteins are target antigens for human cytomegalovirus specific cytotoxic T-lymphocytes.

Induction of cytolytic T-lymphocytes with cytomegalovirus polypeptides

A DNA probe has been isolated which is capable of hybridizing to an oligonucleotide sequence coding for a polypeptide from a major 64 Kilodalton protein of human cytomegalovirus (HCMVgp64). The probe has a sequence of at least seventeen (17) to as many as seven hundred twenty-one (721) nucleotides. The DNA fragments coding for the major late protein of human cytomegalovirus (HCMVgp64) may be hybridized to DNA fragments of HCMV DNA from an individual having human cytomegalovirus infection. The major late protein of human cytomegalovirus (HCMVgp64) also reacts with T-lymphocytes of an individual after natural infection of that individual with human cytomegalovirus. Thus, the HCMVgp64 protein may be used as a vaccine to prevent HCMV infection.

A DNA probe has been isolated which is capable of hybridizing to an oligonucleotide sequence coding for a polypeptide from a major 64 Kilodalton protein of human cytomegalovirus (HCMVgp64). The probe has a sequence of at least seventeen (17) to as many as seven hundred twenty-one 721) nucleotides. The probe may be labelled as by radioactivity. The probe has been used to screen DNA fragments constituting a subgenomic library of human cytomegalovirus DNA to obtain DNA fragments coding for the major late protein of human cytomegalovirus. The DNA fragments coding for the major late protein of human cytomegalovirus (HCMVgp64) may be hybridized to DNA fragments of HCMV DNA from an individual having human cytomegalovirus infection. The viral DNA can be used as whole HCMV DNA or as fragments formed by digesting the human cytomegalovirus DNA with a restriction endonuclease such as one of the restriction endonucleases EcoRI, BamHI, XbaI, HindIII and PrtI. During the screening of clinical specimens, the DNA fragment coding for HCMVgp64 of human cytomegalovirus hybridizes to whole HCMV DNA or to a particular one of the DNA fragments produced by digesting the human cytomegalovirus DNA with the restriction endonucleases. As a resuslt of such hybridization, the identity of human cytomegalovirus may be established. The particular DNA fragment may have a map location of approximately 0.50 to 0.51 units in the human cytomegalovirus (strain Towne) genome. It may have a sequence of about 721 nucleotides. The major late protein of human cytomegalovirus (HCMVgp64) also reacts with T-lymphocytes of an individual after natural infection of that individual with human cytomegalovirus. Thus, the HCMVgp64 protein may be used as a vaccine to prevent HCMV infection.

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Arcadia, CA, United States of America

Hema Pande