Characterization of Monocyte-Derived Dendritic Cells used in Immunotherapy for Hiv-1-Infected Individuals

Abstract

Aims: A therapeutic vaccine based on monocyte-derived dendritic cells (MDDCs) has been shown to represent a promising strategy for the treatment of cancer and viral infections. Here, we characterized the MDDCs used as an immunogen in a clinical trial for an anti-HIV-1 therapeutic vaccine. Patients & Methods: Monocytes obtained from 17 HIV-infected individuals were differentiated into MDDCs and, after loading with autologous HIV, the cells were characterized concerning surface molecule expression, migratory and phagocytosis capacity, cytokine production and the induction of an effective cell-mediated immune response. Results: The MDDCs were able to induce antigen-specific responses in autologous CD4+ and CD8+ T lymphocytes. Conclusions: Despite a large interindividual variability, the results suggested that MDDCs present the potential to promote immune responses in vaccinated patients.

Activation of Latent HIV-1 T Cell Reservoirs with a Combination of Innate Immune and Epigenetic Regulators

“…cGAMP induces modest reactivation in vitro without affecting cell death. Recent achievements in cancer immunotherapy (24)(25)(26) have raised the possibility that the application of immunotherapeutic approaches to other fields, including HIV research, could promote the clearance of the latent HIV-1 reservoir (6,27,28). To evaluate the abilities of immunostimulatory biologic agents to induce the reactivation of HIV-1 provirus and to selectively induce the death of latently HIV-1-infected cells, we first analyzed the effects of three different agonists of innate immunity-the RIG-I agonist M8, previously characterized by our group (29), and the STING agonists cGAMP and c-di-GMP (the cyclic dinucleotide of GMP)-on HIV-1 reactivation in J-Lat 10.6 cells, an in vitro model of HIV-1 latency that contains a green fluorescent protein (GFP) gene substitution in the Nef open reading frame (ORF) (30).…”

Flagellin/NLRC4 Pathway Rescues NLRP3-Inflammasome Defect in Dendritic Cells From HIV-Infected Patients: Perspective for New Adjuvant in Immunocompromised Individuals

“…CD4 + T lymphocytes activation was measured by the meaning of intracellular staining of IFN-γ. Briefly, autologous co-cultures of MDDC and lymphocytes were treated with lymphocyte mitogens Ionomycin (1 μg/mL; Sigma-Aldrich, Merck) and Phorbol Myristate Acetate (10 ng/mL; Sigma-Aldrich, Merck) for 96 h (31). Twenty microgram per milliliter Brefeldin A (Sigma-Aldrich, Merck) was added 6 h before the end of co-culture to block Golgi secretory pathway.…”

Caracterização genética e funcional de células dendríticas de indivíduos HIV+ estimuladas com flagelina

“…CD4 + T lymphocytes activation was measured by the meaning of intracellular staining of IFN-γ. Briefly, autologous co-cultures of MDDC and lymphocytes were treated with lymphocyte mitogens Ionomycin (1 µg/mL; Sigma-Aldrich, Merck) and Phorbol Myristate Acetate (10 ng/mL; Sigma-Aldrich, Merck) for 96 h (31). Twenty microgram per milliliter Brefeldin A (Sigma-Aldrich, Merck) was added 6 h before the end of co-culture to block Golgi secretory pathway.…”

“…MDDC were treated with 5 µg/mL FLG for 24 h and phenotypic profile, TNF and IL-12 secretion as well as CD4 + T lymphocytes activation in co-culture experiments were assessed based on previously published protocols (31,32).…”

“…To test whether FLG-treated MDDC are able to induce a properly adaptive immunity response, a MDDC/lymphocytes coculture assay was performed using autologous cells as previously described (31,32). Lymphocytes alone were used as a negative control (Neg) (Figures 1D,E).…”