title: Autologous aldrithiol-2-inactivated HIV-1 combined with polyinosinic-polycytidylic acid-poly-L-lysine carboxymethylcellulose as a vaccine platform for therapeutic dendritic cell immunotherapy.
abstract: Therapeutic interventions for HIV-1 that successfully augment adaptive immunity to promote killing of infected cells may be a requisite component of strategies to reduce latent cellular reservoirs. Adoptive immunotherapies utilizing autologous monocyte-derived dendritic cells (DCs) that have been activated and antigen loaded ex vivo may serve to circumvent defects in DC function that are present during HIV infection in order to enhance adaptive immune responses. Here we detail the clinical preparation of DCs loaded with autologous aldrithiol-2 (AT-2)-inactivated HIV that have been potently activated with the viral mimic, Polyinosinic-polycytidylic acid-poly-l-lysine carboxymethylcellulose (Poly-ICLC). HIV is first propagated from CD4+ T cells from HIV-infected donors and then rendered non-replicative by chemical inactivation with aldrithiol-2 (AT-2), purified, and quantified. Viral inactivation is confirmed through measurement of Tat-regulated β-galactosidase reporter gene expression following infection of TZM-bl cells. In-process testing for sterility, mycoplasma, LPS, adventitious agents, and removal of AT-2 is performed on viral preparations. Autologous DCs are generated and pulsed with autologous AT-2-inactivated virus and simultaneously stimulated with Poly-ICLC to constitute the final DC vaccine product. Phenotypic identity, maturation, and induction of HIV-specific adaptive immune responses are confirmed via flow cytometric analysis of DCs and cocultured autologous CD4+ and CD8+ T cells. Lot release criteria for the DC vaccine have been defined in accordance with Good Manufacturing Practice (GMP) guidelines. The demonstrated feasibility of this approach has resulted in approval by the FDA for investigational use in antiretroviral (ART) suppressed individuals. We discuss how this optimized DC formulation may enhance the quality of anti-HIV adaptive responses beyond what has been previously observed during DC immunotherapy trials for HIV infection.
authors.affiliation: Icahn School of Medicine at Mount Sinai, Division of Infectious Diseases, New York, NY, USA. Electronic address: elizabeth.a.miller@mssm.edu.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Miller
authors.affiliation: Icahn School of Medicine at Mount Sinai, Tisch Cancer Institute, New York, NY, USA.
authors.forename: Meredith
authors.initials: M
authors.lastname: Spadaccia
authors.affiliation: Icahn School of Medicine at Mount Sinai, Tisch Cancer Institute, New York, NY, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Sabado
authors.affiliation: AIDS and Cancer Virus Program Inc., Frederick, MD, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Chertova
authors.affiliation: AIDS and Cancer Virus Program Inc., Frederick, MD, USA.
authors.forename: Julian
authors.initials: J
authors.lastname: Bess
authors.affiliation: AIDS and Cancer Virus Program Inc., Frederick, MD, USA.
authors.forename: Charles Mac
authors.initials: CM
authors.lastname: Trubey
authors.affiliation: Icahn School of Medicine at Mount Sinai, Tisch Cancer Institute, New York, NY, USA.
authors.forename: Rose Marie
authors.initials: RM
authors.lastname: Holman
authors.affiliation: Oncovir Inc., Washington, DC, USA.
authors.forename: Andres
authors.initials: A
authors.lastname: Salazar
authors.affiliation: AIDS and Cancer Virus Program Inc., Frederick, MD, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Lifson
authors.affiliation: Icahn School of Medicine at Mount Sinai, Tisch Cancer Institute, New York, NY, USA.
authors.forename: Nina
authors.initials: N
authors.lastname: Bhardwaj
mesh_terms: D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives; D016915:AIDS Vaccines / Q000737:chemistry* / Q000276:immunology*; D056704:Adaptive Immunity; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002266:Carboxymethylcellulose Sodium / Q000031:analogs & derivatives*; D002460:Cell Line; D003713:Dendritic Cells / Q000276:immunology*; D004220:Disulfides; D015658:HIV Infections / Q000276:immunology / Q000628:therapy; D015497:HIV-1 / Q000254:growth & development / Q000276:immunology* / Q000302:isolation & purification / Q000502:physiology; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D011070:Poly I-C / Q000276:immunology*; D011107:Polylysine / Q000031:analogs & derivatives* / Q000276:immunology; D038301:Virus Inactivation; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D016915:AIDS Vaccines; D004220:Disulfides; C001105:2,2'-dipyridyl disulfide; D011107:Polylysine; D015082:2,2'-Dipyridyl; C019531:poly ICLC; D001616:beta-Galactosidase; D002266:Carboxymethylcellulose Sodium; D011070:Poly I-C
keywords: Dendritic cell; HIV-1; Poly-ICLC; Therapeutic vaccine