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. 2007 Mar 21;81(13):7249–7268. doi: 10.1128/JVI.02754-06

FIG. 3.

FIG. 3.

EBV M-RT-PCR of pre-LC isolated from blood. CD1a+ pre-LC were isolated from the blood of eight healthy subjects and six with AIDS, each with fewer than 200 CD4+ cells/ml of blood (mean, 70; range, 4 to 164). Cells were studied as newly isolated, after culture for 3 days, or after culture for 3 days and treatment with chemical inducers of EBV replication for 2 more days. RNA extracted from the cells was studied by EBV M-RT-PCR amplification and specific probe hybridization. (+) Control = B958 lymphoblastoid cell line DNA and Akata Burkitt's lymphoma cell line DNA; (−) Control = no DNA. (A) Newly isolated pre-LC. EBER-1 expression was demonstrated in healthy subjects 3, 4, 5, 6, 7, and 10 and in AIDS subjects 11, 12, 14, 15, and 16. With a band in the reverse transcriptase-negative (RT −) reaction mixture, it is uncertain if the result for subject 6 represents true EBER-1 expression or detection of viral genomic DNA sequences in the RNA preparation. Despite a band in the reverse transcriptase-negative reaction mixture for subject 13, the absence of a band in the corresponding reverse transcriptase-positive (RT +) reaction mixture was interpreted as an absence of EBER expression for this subject. (B) Cultured pre-LC. EBER-1 expression was demonstrated in all three subjects, including subject 8, in whom EBER-1 expression was not detected in newly isolated pre-LC. (C) Control cells. The EBV-positive Namalwa Burkitt's lymphoma cell line expresses the least EBER-1 of the known EBV-positive cell lines. The sensitivity of detection of the EBV M-RT-PCR assay for EBER-1 expression was demonstrated to be a single Namalwa cell diluted into 105 cells of the EBV-negative RHEK-1 cell line. (D) Newly isolated pre-LC. BZLF1 expression was demonstrated in healthy subject 4. (E) Cultured pre-LC. New weak BZLF1 expression was demonstrated in all three healthy subjects in whom BZLF1 expression was not detected in newly isolated pre-LC. (F) Cultured and induced pre-LC. Newly induced strong BZLF1 expression was demonstrated in all three healthy subjects in whom BZLF1 expression was not detected in newly isolated pre-LC.