title: Ex vivo production of autologous whole inactivated HIV-1 for clinical use in therapeutic vaccines.
abstract: This study provides a detailed description and characterization of the preparation of individualized lots of autologous heat inactivated HIV-1 virions used as immunogen in a clinical trial designed to test an autologous dendritic-cell-based therapeutic HIV-1 vaccine (Clinical Trial DCV-2, NCT00402142). For each participant, ex vivo isolation and expansion of primary virus were performed by co-culturing CD4-enriched PBMCs from the HIV-1-infected patient with PBMC from HIV-seronegative unrelated healthy volunteer donors. The viral supernatants were heat-inactivated and concentrated to obtain 1 mL of autologous immunogen, which was used to load autologous dendritic cells of each patient. High sequence homology was found between the inactivated virus immunogen and the HIV-1 circulating in plasma at the time of HIV-1 isolation. Immunogens contained up to 10⁹ HIV-1 RNA copies/mL showed considerably reduced infectivity after heat inactivation (median of 5.6 log₁₀), and were free of specified adventitious agents. The production of individualized lots of immunogen based on autologous inactivated HIV-1 virus fulfilling clinical-grade good manufacturing practice proved to be feasible, consistent with predetermined specifications, and safe for use in a clinical trial designed to test autologous dendritic cell-based therapeutic HIV-1 vaccine.
authors.affiliation: AIDS Research Group-IDIBAPS-HIVACAT, Spain. cgil@clinic.ub.es
authors.forename: Cristina
authors.initials: C
authors.lastname: Gil
authors.forename: Núria
authors.initials: N
authors.lastname: Climent
authors.forename: Felipe
authors.initials: F
authors.lastname: García
authors.forename: Carmen
authors.initials: C
authors.lastname: Hurtado
authors.forename: Sara
authors.initials: S
authors.lastname: Nieto-Márquez
authors.forename: Agathe
authors.initials: A
authors.lastname: León
authors.forename: M Teresa
authors.initials: MT
authors.lastname: García
authors.forename: Cristina
authors.initials: C
authors.lastname: Rovira
authors.forename: Laia
authors.initials: L
authors.lastname: Miralles
authors.forename: Judith
authors.initials: J
authors.lastname: Dalmau
authors.forename: Tomás
authors.initials: T
authors.lastname: Pumarola
authors.forename: Manel
authors.initials: M
authors.lastname: Almela
authors.forename: Javier
authors.initials: J
authors.lastname: Martinez-Picado
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Lifson
authors.forename: Laura
authors.initials: L
authors.lastname: Zamora
authors.forename: José M
authors.initials: JM
authors.lastname: Miró
authors.forename: Christian
authors.initials: C
authors.lastname: Brander
authors.forename: Bonaventura
authors.initials: B
authors.lastname: Clotet
authors.forename: Teresa
authors.initials: T
authors.lastname: Gallart
authors.forename: José M
authors.initials: JM
authors.lastname: Gatell
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology* / Q000627:therapeutic use; D023241:Antiretroviral Therapy, Highly Active; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism / Q000821:virology; D005434:Flow Cytometry; D015658:HIV Infections / Q000517:prevention & control / Q000821:virology; D015497:HIV-1 / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D017421:Sequence Analysis; D015164:Vaccines, Inactivated / Q000276:immunology*; D019562:Viral Load; D038301:Virus Inactivation
chemical_list: D016915:AIDS Vaccines; D015164:Vaccines, Inactivated