University of Washington

Other

Kurt Strand

Timothy M Rose

Marnix L Bosch

George J Todaro

This invention relates to polynucleotides encoding Glycoprotein B from the RFHV/KSHV subfamily of gamma herpes viruses, three members of which are characterized in detail. DNA extracts were obtained from Macaque nemestrina and Macaque mulatta monkeys affected with retroperitoneal fibromatosis (RF), and human AIDS patients affected with Kaposi's sarcoma (KS). The extracts were amplified using consensus-degenerate oligonucleotide probes designed from known protein and DNA sequences of gamma herpes viruses. The nucleotide sequences of a 319 base pair fragment are about 76% identical between RFHV1 and KSHV, and about 60-63% identical with the closest related gamma herpes viruses outside the RFHV/KSHV subfamily. Protein sequences encoded within these fragments are are about 91% identical between RFHV1 and KSHV, and &lt;.about.65% identical to that of other gamma herpes viruses. The full-length KSHV Glycoprotein B sequence comprises a transmembrane domain near the N-terminus, and a plurality of potentially antigenic sites in the extracellular domain. Materials and methods are provided to characterize Glycoprotein B encoding regions of members of the RFHV/KSHV subfamily, including but not limited to RFHV1, RFHV2, and KSHV Peptides, polynucleotides, and antibodies of this invention can be used for diagnosing infection, and for eliciting an immune response against Glycoprotein B.

Glycoprotein B of the RFHV/KSHV subfamily of herpes viruses

This invention provides isolated polynucleotides encoding DNA polymerases of three members of a subfamily of gamma herpes viruses. Two were obtained from macaque monkeys affected with retroperitoneal fibromatosis, the other from human AIDS patients affected with Kaposi's sarcoma. A 454-base pair fragment encoding a region near the active site of the DNA polymerase is 69-83% identical amongst the three viruses, but only 54-68% identical with other known gamma herpes sequences and &lt;55% identical with alpha and beta herpes sequences. Also provided are polynucleotides encoding DNA polymerase from related viruses in the RFHV/KSHV subfamily. Polynucleotides prepared according to the sequence data can be used as reagents to detect and characterize related sequences. Such reagents may be used to detect members of the RFHV/KSHV subfamily, including but not limited to RFHV, RFHV2, and KSHV. Corresponding polypeptides and peptide fragments may be obtained by expressing the polynucleotide or by chemical synthesis. They may be used for detecting specific antibody potentially present in the serum of infected subjects. They may also be used for designing or screening pharmaceutical compounds that limit viral replication by inhibiting DNA polymerase activity.

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Issaquah, WA, United States of America

Kurt Strand