RT-PCR amplification products of gene MAGE-A4.

Products were obtained with lymph node samples from patients with HD (HD11, HD5, HD3) or NHL and obtained with samples of normal lymphoid cells (N1, lymph node; N2, PBMC; N3, PHA-activated peripheral T cells). C+, positive control sarcoma line LB23-SARC. C−, negative control (no RNA present in the RT reaction). Amplification of MAGE-A4transcripts gives a band of 443 base pairs, distinct from that of the amplified genomic sequence of 517 base pairs (faintly visible in line N1). The levels of gene expression were evaluated by comparing band intensities with those obtained for serial dilutions (1:1, 1:3, 1:9, 1:27) from the positive and scored as follows : ++++, 100%; +++, 30% to 100%; ++, 10% to 30%; +, 3% to 10%; or ±, less than 3%. Absence of RT-PCR product is indicated as −. RNA integrity was assessed by RT-PCR amplification of the β-actin mRNA (data not shown).