Nasopharyngeal carcinoma (NPC) is an epithelial malignancy which develops with high incidence in Southern China, Northern Africa and among Eskimos. We have analyzed EBV transcription in NPC. These studies have revealed that the latent membrane protein (LMP) is expressed in all NPC specimens and in the C15 NPC tumor may be encoded by two forms of LMP mRNA. These studies have also identified a new RNA which contains a previously unidentified open reading frame within the BamHI A fragment. To further characterize EBV transcription in NPC, cDNA libraries will be prepared to three additional tumors which have been successfully established in nude mice. The EBV specific cDNAs will be identified and sequenced. Transcription representing LMP, LMP2, and BARFO will be analyzed in the nude mice tumors by cDNA cloning using both oligodeoxythymidine and specific synthetic oligonucleotides for priming of cDNA synthesis. The polymerase chain reaction (PCR) will be utilized to identify RNAs, containing the 171 kb intron within BARFO, identified in C15, which is not present in a related mRNA transcribed in the B95-8 cell line. PCR analysis will be used to identify expression of this specific mRNA in RNA obtained from biopsy specimens and in cell lines. To identify the authentic protein encoded by the BARFO ORF, a fusion protein will be synthesized with glutathione in transferase. The fusion protein will be used to produce antisera to identify the protein as expressed in NPC tissue. The fusion protein will also be used to screen human sera for reactivity to the putative protein. Based on our previous findings we will continue to identify EBV-associated hyperplasias by identifying and characterizing the EBV termini and identification of the EBERs in in situ hybridizations.