Harnessing dendritic cells (DC) to treat HIV infection is considered a key strategy to improve anti-HIV treatment and promote the discovery of functional or sterilizing cures. Although this strategy represents a promising approach, the results of currently published trials suggest that opportunities to optimize its performance still exist. In addition to the genetic and clinical characteristics of patients, the efficacy of DC-based immunotherapy depends on the quality of the vaccine product, which is composed of precursor-derived DC and an antigen for pulsing. Here, we focus on some factors that can interfere with vaccine production and should thus be considered to improve DC-based immunotherapy for HIV infection.
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.
BackgroundNLRP3-inflammasome activation was evaluated in monocyte-derived dendritic cells (DC) obtained through IL-4 (IL4-DC) or IFN-α (IFN-DC) protocols and pulsed with chemically inactivated HIV-1. Inflammasome’ genes expression and IL-1β secretion were compared in DC isolated from 15 healthy subjects (HC) and 10 HIV-1 infected individuals (HIV+).FindingsWhether HIV was able to increased NLRP3-inflammasome genes expression and IL-1β secretion in IL4-DC from HC, the induction of inflammasome appeared significantly reduced in IFN-DC from HC, suggesting a different responsive state of IFN-DC compared to IL4-DC. No inflammasome activation was observed in IL4-DC as well as in IFN-DC derived from HIV + subjects, confirming previous findings on “unresponsive” state of DC derived from HIV + possibly due to chronic inflammatory state of these individuals.ConclusionsOur results showed that IFN-α differently modulates inflammasome expression during monocytes-DC in vitro differentiation. These findings could be of interest considering the on-going research about DC manipulation and therapeutic strategies for HIV + involving DC-based immune-vaccines.
Dendritic cell (DC)-based immunotherapy is a promising strategy for the treatment of HIV-infected individuals. Different from the conventional protocol for DC differentiation based on the cytokine IL-4 (IL4-DCs), several studies have suggested obtaining DCs by culturing monocytes with type I IFN (IFN-α) to yield IFN-DCs, as performed in cancer therapy. To evaluate the phenotypic and functional characteristics, monocytes from HIV-infected subjects were differentiated into IFN-DCs or IL4-DCs, pulsed with chemically inactivated HIV and stimulated with pro-inflammatory cytokines. A comparative analysis between both types of monocyte-derived DCs (MoDCs) showed that immature IFN-DCs were phenotypically distinct from immature IL4-DCs at the baseline of differentiation, presenting a pre-activated profile. From the functional profile, we determined that IFN-DCs were capable of producing the cytokine IL-12 p70 and of inducing the production of IFN-γ by CD4 + T lymphocytes but not by TCD8+ lymphocytes. Our results suggest that IFN-DCs derived from HIV-infected individuals are able to recognize and present viral antigens to induce TCD4+ cellular immunity to HIV.
The decline in number and function of T cells is a hallmark of HIV infection, and preservation or restoration of HIV-specific cellular immune response is a major goal of AIDS treatment. Dendritic cells (DCs) play a key role in the initiation and maintenance of the immune response, and their use as a vaccine vehicle is a promising strategy for enhancing vaccine efficacy. We evaluated the potential of DC-mediated immunization with a DNA vaccine consisting of HIV-1-p55gag (gag, group-specific antigen) associated to lysosomal associated protein (LAMP) sequence (LAMP/ gag vaccine). Immunization of mice with mouse DCs transfected with LAMP/gag (Lg-mDCs) stimulated more potent B-and T-cell responses than naked DNA or DCs pulsed with inactivated HIV. Anti-Gag antibody levels were sustained for at least 3 mo after immunization, and recall T-cell responses were also strongly detected at this time point. Human DCs transfected with LAMP/gag (Lg-hDCs) were also activated and able to stimulate greater T-cell response than native gag-transfected DCs. Coculture between Lg-hDCs and T lymphocytes obtained from patients with HIV resulted in upregulation of CD38, CD69, HLA-DR, and granzyme B by CD4 + and CD8 + T cells, and increased IFN-g and TNF-a production. These results indicate that the use of LAMP/gag-DC may be an efficient strategy for enhancing immune function in patients with HIV
Dedico este trabalho à minha avó Maria do Pranto, minha rainha e guerreira por ser a minha fonte de inspiração e aprendizado.
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AgradecimentosAgradeço primeiramente a Deus por ser meu guia protetor, fonte de força e pelo acompanhamento a cada dia desta trajetória.Agradeço aos amigos e familiares, especialmente aos meus pais Guida Santillo e Roberto Santillo, pelo amor e por terem me proporcionado o estudo; ao meu irmãoWillian Santillo e ao meu noivo Thiago Bonatti pelo carinho, confiança e por acreditarem no meu potencial! Agradeço a vocês pelas palavras amigas, pelo carinho, amor, sorriso e reconhecimento em todos os momentos. Obrigada por toda admiração demonstrada. On day 5 of culture, DCs were pulsed with chemically Aldrithiol-2-inactivated HIV and stimulated for 48 hours with a cocktail of proinflammatory cytokines. Subsequently, we assessed IFN-DC surface markers' expression and the ability of IFN-DCs present antigens and induce proliferation and IFN-γ production by allogeneic and autologous CD4+ and CD8+ lymphocytes, respectively, by flow cytometry. Moreover, we available the IL-10 and IL-12p70 secretion by ELISA. RESULTS: IFN-DCs isolated from HIV-infected individuals showed morphological and phenotypic characteristics of semi-activated cells already in the basal state of maturity, and exhibit features of plasmacytoid DCs and NK cells, different from that seen in the IL4-DCs. Furthermore, the IFN-DCs were able to produce IL-12p70 after activate/viral pulse stimuli accompanied of lower IL-10 secretion, like observed in IL4-DCs. Finally, the IFN-DCs showed capable of stimulate CD4+ and CD8+ T lymphocytes proliferation and IFN-γ production in comparable levels to IL4-DCs. CONCLUSIONS: Although IFN-DCs and IL4-DCs are distinct populations of MDDCs presenting distinct morphology and phenotype, we observed that the ability to recognize and present viral antigens and stimulate T lymphocytes response were similar between the two populations.
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