In situ RT-PCR detection of Epstein-Barr virus immediate-early transcripts in CD4+ and CD8+ T lymphocytes.
AIDS-related Epstein-Barr virus (EBV)-associated T cell lymphomas are emerging as a new, distinct histopathological entity. The pathway whereby EBV infects T cells as well as the initial EBV transcriptional program in T cells has not been established. In order to shed light on the early events of the EBV infection of T cells, we have used in situ reverse transcription based polymerase chain reaction (RT-PCR) to study the initial EBV transcriptional program in homogeneous CD4+ and CD8+ lymphocytes. Following EBV infection, Epstein-Barr nuclear antigen (EBNA) expression could be detected in T rosetting CD4+ and CD8+ T lymphocytes. Only a few cells showed viral capsid antigen (VCA). EBV immediate-early gene transcripts (BZLF1, BRLF1, and BMLF1) encoded in the BamHI Z, R, and M fragments could be detected by in situ RT-PCR in the EBV producer cell line B95.8. Both BZLF1 and BRLF1 immediate-early transcripts, but not BMLF1 transcript, could be detected in individual CD4+ and CD8+ T cells infected with EBV. Demonstration of EBV mRNA transcripts encoding immediate-early transcriptional transactivators in EBV-infected T cells provides the first evidence for a possible mechanism whereby EBV could contribute to T cell proliferation and EBV-associated T cell malignancies.
Guan
M
M
Department of Microbiology and Immunology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA.
Romano
G
G
Henderson
E E
EE
D000956:Antigens, Viral / Q000378:metabolism*; D002051:Burkitt Lymphoma / Q000821:virology; D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D018414:CD8-Positive T-Lymphocytes / Q000821:virology*; D002467:Cell Nucleus / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004854:Herpesvirus 4, Human / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014764:Viral Proteins
D000956:Antigens, Viral; C068382:BRLF1 protein, Human herpesvirus 4; C063676:BZLF1 protein, Herpesvirus 4, Human; D004268:DNA-Binding Proteins; D017874:Immediate-Early Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D014764:Viral Proteins