Immortalized Epstein-Barr virus-positive B-cell lines obtained by prolonged culture of peripheral blood mononuclear cells from human immunodeficiency virus type 1-positive patients.

RUIBAL ARES B; BELMONTE L; BARÉ P; SCOLNIK M; PALACIOS MF; BAYO HANZA MC; GALMARINI C; MÉNDEZ G; BRACCO MM

Journal of Human Virology

Lippincott-Raven Publishers

Año: 2001 vol. 4 p. 200 - 213

1090-9508

Immortalized cell lines (LCL) were obtained by spontaneous outgrowth of B lymphoblastoid cells in 73% of unstimulated prolonged cultures of peripheral blood mononuclear cells (PBMC) from HIV+ hemophilic patients without lymphoma, in contrast to 6% in HIV- controls (N PBMC). The method selected for culture allows for monocyte/macrophage (M/M) differentiation and HIV replication in the absence of exogenous stimuli. CD8 cell removal or cyclosporin addition were not required for the induction of B lymphocyte immortalization. All LCL were EBV+, as judged by the presence of EBV latent oncogenes LMP 1 and EBNA 2. LMP 1-del was demonstrated in 43 % of these LCL. LCL were more frequently obtained after prolonged culture of PBMC in which active HIV replication had taken place in vitro. Low rate HIV replication persisted in 53 % of them. The morphology of LCL resembles that of AIDS diffuse large cell lymphomas. B cell lineage was established by the expression of pan-B cell markers and CD23. Different B cell clones could be expanded from the same individual, as demonstrated by light chain restriction or immunoglobulin heavy chain (IgH) expression and IgH gene rearrangement. LCL also expressed activation markers (CD38, HLA-DR, CD40 and CD30). The simple method described here may be useful to identify conditions that allow the expansion of EBV+ B cell clones in HIV+.