Immortalized Epsteinndash;Barr Virusndash;positive B-cell Lines Obtained by Prolonged Culture of Peripheral Blood Mononuclear Cells From Human Immunodeficiency Virus Type 1ndash;positive Patients

摘要

ObjectivesTo study the factors that determine malignant B cell growth in human immunodeficiency virus type 1 (HIV-1)ndash;infected patients.Study DesignB-cell lines (lymphocyte cell lines lsqb;LCLrsqb;) were developed after nonstimulated culture of peripheral blood mononuclear cells (PBMC) from HIV-1ndash;positive (HIV-1plus;) patients. Human immunodeficiency virus type 1 replication in culture, Epsteinndash;Barr virus (EBV) latent oncogene expression, and cell-to-cell interaction were studied after nonstimulated culture of HIV-1plus;PBMC, analyzing their contribution to LCL appearance.MethodsNonstimulated PBMC cultures of HIV-1plus;PBMC and controls (N-PBMC) were established. Lymphocyte cell lines were characterized. Epsteinndash;Barr virus latent membrane protein 1 (LMP-1) and Epsteinndash;Barr nuclear antigen 2 were detected by polymerase chain reaction (PCR). Clonality of LCL was determined by light chain restriction (flow cytometry) and immunoglobulin H chain rearrangement (semi-nested PCR). Peripheral blood mononuclear cell phenotypes were studied at different intervals of culture.ResultsLymphocyte cell lines were obtained in 73percnt; of HIV-1plus;PBMC cultures, compared with 6percnt; in N-PBMC. All LCL were EBV-positive (EBVplus;). B-cell lineage was established, and up to 12 different B-cell clones were expanded from the same individual. Occurrence of LCL was more frequent in cultures with HIV-1 replication, high LMP-1 expression in viable B cells, and high CD4colon;CD8 ratio. Human immunodeficiency virus type 1 replication persisted in 53percnt; of the LCL.ConclusionsIn vitro HIV-1 replication and persistence of viable EBVplus;lymphoblasts favor spontaneous in vitro outgrowth of LCL in HIV-1plus;patients.