Lymphocyte phenotype, EBV DNA levels with latent gene expression, and cytokine levels in stimulated PBMCs from the patient, his parents, and a healthy blood bank donor. (A) CD4/CD8 and (B) CD8/CD56 expression of PBMCs stimulated with PHA and IL-2 for 2 days followed by IL-2 alone for 2 weeks. (C) EBV DNA levels in stimulated PBMCs. DNA from the EBV BamHI W region DNA was quantified using real-time PCR. The EBV BamHI W fragment copy number per cell was calculated by the formula N = 2 × (W/B), where N is the EBV BamHI W copy number/cell, W is the EBV BamHI W copy number, and B is the bcl-2 copy number. Copy numbers of EBV-BamHI W gene per 1×10⁶ cells are indicated. U indicates undetectable. (D) RT-PCR analysis of EBV latency genes in stimulated PBMCs. Southern blots of PCR products were hybridized with [³²P]-labeled probes. (E) Cytokine levels were measured in culture supernatants of stimulated PBMCs from a healthy donor (1), an unrelated patient with CAEBV (2), the patient with the perforin mutations (3), the patient's mother (4), and the patient's father (5). Cells were stimulated with anti-CD3 antibody (black bars) or isotype control antibody (white bars). The experiment was performed 3 times with similar results.