The polymerase chain reaction (PCR) has become a powerful tool in the world of molecular biology (1). Using specific oligonucleotides complimentary to a known sequence of DNA in conjunction with Taq DNA polymerase, it is possible to synthesize billions of copies of that DNA from only one starting ...

Epstein-Barr virus (EBV) is a lymphotropic γ herpes virus. Infection of human B cells with EBV in vitro results in their immortalization and the resulting cell lines are named lymphoblastoid cell lines (LCLs) (1). In these cells, EBV establishes mainly a latent infection, characterized by the ex ...

There have been no reports of the direct entry of Epstein-Barr virus (EBV) into a fully permissive lytic cycle in any cell in vitro. Virus does, however spontaneously move from latency into a lytic cycle of replication in a very small percentage of the population of most B-cell lines in culture and this num ...

Epstein-Barr virus (EBV) is able to override the mechanisms that normally regulate the proliferation of human B-lymphocytes. In the absence of other extracellular signals, EBV infects resting B-lymphocytes and drives the infected cells into the cell-division cycle. Recently, there ...

Epstein-Barr virus (EBV) is associated with both lymphoma (for example, Burkitt’s lymphoma) and epithelial carcinoma (for example, nasopharyngeal carcinoma). Among about 90 genes encoded by the virus (1), seven latent genes—(Epstein-Barr viral nuclear antigen 1 (EBNA1), EBNA2, EBNA ...

When studying the contributions of Epstein-Barr virus (EBV) to tumorogenesis, it is often advantageous to analyze the effects of the expression of one viral protein on the host cell separately from the effects of other viral proteins that are being concurrently expressed during a normal inf ...

The analysis of the effects of expression of one viral protein on the host cell separately from the effect of other viral proteins is often limited by the efficiency of the transfection method. Lymphoid cells are effectively transfected by electroporation (1–4). However, the transfection e ...

The development of in vitro systems for the culture of human epithelial cells has aided the study of epithelial cell transformation by the epitheliotrophic papilloma-viruses (1). Although long-term proliferation of keratinocytes in vitro can now be achieved, the tissue-culture env ...

The inability to directly infect epithelial cells in vitro with Epstein-Barr virus (EBV) (1) has led to the development of epithelial cell-culture models to examine virus-induced growth transformation and productive infection.

Flow cytometry allows rapid multi-parameter analyses of individual cells. Cells are illuminated with incident laser light of a specific wavelength. Resultant light-scatter signals and fluorescence-emission signals from fluorochromes contained at the surface of the cell or w ...

In Epstein-Barr virus (EBV)-seropositive individuals, cell-mediated immunity against EBV can be monitored in vitro as the capacity of T-cell lymphocytes to inhibit virusinduced proliferation of autologous B cells. In vitro infection of peripheral blood mononuclear cells (PBM ...

The techniques presented in this chapter describe the experimental procedure for the identification of the nonrandom DNA fragmentation associated with apoptosis. The major benefits of this method are its ability to detect a low level of DNA fragmentation and its ability to detect large D ...

Currently, the antigen specificity of Epstein-Barr virus (EBV)-specific cytotoxic T cells (CTLs) can be determined using a set of recombinant vaccinia viruses expressing individual EBV proteins. These recombinant viruses are used to infect EBV-negative target cells, which are then ...

Although Epstein-Barr virus (EBV) infection elicits both CD4- and CD8-positive T-cell responses, a considerable body of experimental evidence indicates that the latter responses play a major role in controlling the number of EBV-infected cells and proliferation of EBV-transform ...

Limiting dilution assays (LDA) are designed to define an unknown frequency of effector cells in a population. LDA are dose-response assays that allow detection of an all-or-none (positive or negative) immunoresponse in each individual culture within replicates that vary in the number of r ...

Antibodies to many Epstein-Barr virus (EBV) gene products are present in sera from infected human carriers. Historically, the ready demonstration of antibodies to lytic cycle proteins in particular allowed the seroepidemiological studies, which implicated EBV as a factor in vario ...

Western blotting is a well established technique for identifying Epstein-Barr virus (EBV)-encoded proteins in lysates from cell lines or biopsy material. The basic technique involves separation of proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (S ...

There are many different approaches to a biochemical analysis of virus glycoproteins. However, because Epstein-Barr virus (EBV) does not replicate to high titer in any cell line, several of these are compromised by the relatively small amounts of glycoprotein that are made. For example, unl ...

The yeast two-hybrid assay invented by Stanley Fields and Stephen Elledge (1–4) is a powerful tool to investigate protein-protein interactions, the essence of many biological processes. Owing to the adaptability of the assay procedure, the ease of manipulating and cultivating Saccha ...

The Epstein-Barr viral nuclear antigen 2 (EBNA2) latency protein is a transcriptional activator that plays a critical role in regulation of Epstein-Barr virus (EBV) latency gene expression and in EBV-induced B-cell immortalization. GAL4-fusion constructions have been instrume ...