Large-scale production of CD4+ T cells from HIV-1-infected donors after CD3/CD28 costimulation

Bruce L. Levine*, Julio Cotte, Carolynn C. Small, Richard G. Carroll, James L. Riley, Wendy B. Bernstein, Dennis E. Van Epps, R. Alan Hardwick, Carl H. June

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

107 Scopus citations


We describe a procedure for large-scale enrichment, growth, and harvesting CD4+ T cells. This method may be effective for HIV-1 immunotherapy, as the mode of stimulation, with anti-CD3 plus anti-CD28 coated beads (CD3/CD28 beads) induces a potent antiviral effect. PBMC were obtained by density gradient centrifugation of an apheresis product. Monocytes/macrophages were removed by incubating PBMC with beads coated with IgG. The cells were then magnetically depleted of B cells and CD8+ cells with mouse anti-CD20 and anti-CD8 MAbs and sheep antimouse coated beads. The remaining cells were >80% CD4+ and were transferred to gas-permeable bags containing CD3/CD28 beads and cultured in a closed system. After 14 days, the cell number increased an average of 37-fold, and cells were nearly 100% CD4+. Viral load, assessed by DNA PCR for HIV-1 gag, decreased >10-fold during culture in the absence of antiretroviral agents. Removal of CD3/CD28 beads from the cell suspension was accomplished by passing cells plus beads (3-30 × 109 cells in 2-12 L) over a MaxSep® magnetic separator using gravity-driven flow. The cells were then concentrated to 300 ml in an automated centrifuge. This process allows safe and efficient growth of large numbers of CD4+ T cells from HIV-1+ donors.

Original languageEnglish
Pages (from-to)437-448
Number of pages12
JournalJournal of Hematotherapy and Stem Cell Research
Issue number5
StatePublished - Oct 1998
Externally publishedYes


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