title: A hollow-fiber bioreactor for expanding HIV-1 in human lymphocytes used in preparing an inactivated vaccine candidate.
abstract: An inactivated HIV vaccine intended to elicit broadly neutralizing antibodies is designed to use a pool of population-prevalent HIV-1 from plasma (PHIV), isolated before evolution of antibody-mediated genetic mutations. A suitable cell substrate (CS) for isolating such PHIV is peripheral blood mononuclear cells (PBMC) after stimulating with phytohemagglutinin (PHA) and interleukin-2 (IL-2). Feasibility of employing a hollow-fiber bioreactor under optimized conditions was investigated for large-scale expansion and efficient recovery of concentrated PHIV. Each CS batch was infected in vitro with a prototype PHIV, the infected cells were introduced into the bioreactor for 7-10 days in co-culture, and the cell-free supernatants were assayed for p24 antigen as an index of HIV synthesis. PBMC versus CD8-depleted (CD8D) CS, 20kDa versus 5kDa molecular weight cut-off (MWCO) bioreactor cartridges, 7- versus 10-day culture periods, and varying concentrations of IL-2, fetal bovine serum (FBS) and glucose content in the medium were functionally evaluated for p24 yield. PBMC cultures in 20kDa MWCO cartridges with 15% FBS, 80IU/mL IL-2 and 2.0g/L glucose produced the highest p24 yield; however, CD8D-CS, 20-30% FBS and 80 IU/mL IL-2 within 5kDa cartridges and 2.0 g/L glucose in the circulating medium was more cost-effective for synthesis of virion p24.
authors.affiliation: Department of Laboratory Medicine, University of California, San Francisco, CA 94143-0100, USA.
authors.forename: Maggie
authors.initials: M
authors.lastname: Leong
authors.forename: William
authors.initials: W
authors.lastname: Babbitt
authors.forename: Girish
authors.initials: G
authors.lastname: Vyas
mesh_terms: D016915:AIDS Vaccines / Q000302:isolation & purification*; D019149:Bioreactors / Q000821:virology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology; D003470:Culture Media; D004867:Equipment Design; D005947:Glucose / Q000378:metabolism; D015497:HIV-1 / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology*; D006801:Humans; D007376:Interleukin-2 / Q000008:administration & dosage; D019344:Lactic Acid / Q000378:metabolism; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000821:virology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D011994:Recombinant Proteins / Q000008:administration & dosage; D015164:Vaccines, Inactivated / Q000302:isolation & purification; D014776:Virus Cultivation / Q000295:instrumentation / Q000379:methods*; D014779:Virus Replication
chemical_list: D016915:AIDS Vaccines; D003470:Culture Media; C508594:IL2 protein, human; D007376:Interleukin-2; D010835:Phytohemagglutinins; D011994:Recombinant Proteins; D015164:Vaccines, Inactivated; D019344:Lactic Acid; D005947:Glucose