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.