IntroductionThe use of anti-retroviral therapy (ART) has dramatically reduced HIV-1 associated morbidity and mortality. However, HIV-1 infected individuals have increased rates of morbidity and mortality compared to the non-HIV-1 infected population and this appears to be related to end-organ diseases collectively referred to as Serious Non-AIDS Events (SNAEs). Circulating miRNAs are reported as promising biomarkers for a number of human disease conditions including those that constitute SNAEs. Our study sought to investigate the potential of selected miRNAs in predicting mortality in HIV-1 infected ART treated individuals.Materials and MethodsA set of miRNAs was chosen based on published associations with human disease conditions that constitute SNAEs. This case: control study compared 126 cases (individuals who died whilst on therapy), and 247 matched controls (individuals who remained alive). Cases and controls were ART treated participants of two pivotal HIV-1 trials. The relative abundance of each miRNA in serum was measured, by RTqPCR. Associations with mortality (all-cause, cardiovascular and malignancy) were assessed by logistic regression analysis. Correlations between miRNAs and CD4+ T cell count, hs-CRP, IL-6 and D-dimer were also assessed.ResultsNone of the selected miRNAs was associated with all-cause, cardiovascular or malignancy mortality. The levels of three miRNAs (miRs -21, -122 and -200a) correlated with IL-6 while miR-21 also correlated with D-dimer. Additionally, the abundance of miRs -31, -150 and -223, correlated with baseline CD4+ T cell count while the same three miRNAs plus miR-145 correlated with nadir CD4+ T cell count.DiscussionNo associations with mortality were found with any circulating miRNA studied. These results cast doubt onto the effectiveness of circulating miRNA as early predictors of mortality or the major underlying diseases that contribute to mortality in participants treated for HIV-1 infection.

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We have developed a cancer vaccine in which patient derived myeloma cells are fused with autologous dendritic cells (DCs). In this way, multiple tumor antigens are presented in the context of the immune stimulating machinery of the DC. We are conducting a phase I trial in which patients with multiple myeloma (MM) undergo serial vaccination with DC/myeloma fusions in conjunction with GM-CSF. Eligibility criteria included ≥20% plasma cells in the bone marrow and measurable paraprotein or light chain. To date, 16 patients have been enrolled and 15 have undergone therapy. Patients underwent leukapheresis collection and DCs were generated from adherent mononuclear cells cultured for 5 days with GM-CSF and IL-4 and matured by exposure to TNFa for 48–72 hours. The mean yield of DCs was 1.23 x108 cells with a mean viability of 88%. Patient derived myeloma cells were isolated from bone marrow aspirates. The mean yield of myeloma cells was 92.5 x 106 cells with a mean viability of 90%. Fusion cells were generated by coculture of DCs with myeloma cells in the presence of 50% polyethylene glycol. Fusion cells were quantified by determining the percentage of cells that co-expressed unique DC and myeloma antigens. Mean fusion efficiency and viability was 40% and 84%, respectively. As a measure of immunologic potency, DC/MM fusions stimulated allogeneic T cell proliferation, with mean stimulation indexes of 48, 36, and 9 for the DC, fusion and myeloma cell populations, respectively. To date, 15 patients have completed vaccination at a dose of 1–5x106 fusion cells. Adverse events potentially related to vaccine included injection site reactions, edema, rash, fever (infection), chills, fatigue, muscle aches, pruritis, and diarrhea. One patient with a history of prior deep venous thrombosis (DVT) developed a DVT and pulmonary embolus of uncertain relation to the vaccine. To date, 6/9 evaluable patients have demonstrated immunologic response, defined by at least 2 fold increase in IFNγ expression by CD4 and/or CD8 T cells in response to ex vivo exposure to autologous tumor lysate. Vaccination was associated with an increase in circulating tumor specific T cells as evidenced by an increase in CD8+ cells binding the MUC1 tetramer. The impact of vaccination on circulating levels of regulatory T cells is being examined. Humoral immune responses are being assessed by SERAX analysis. Three patients have ongoing stable disease at 3.5, 6, and 19 months following their initial vaccination. Six patients demonstrated disease stability with subsequent progression ranging from 9 weeks to 8 months following their first vaccine. Vaccination with DC/MM fusions has been well tolerated, associated with immunologic response, and disease stabilization in a majority of patients with multiple myeloma.