Optimization of a dendritic cell-based assay for the in vitro priming of naïve human CD4+ T cells

Monocyte‐derived dendritic cells can induce autoreactive CD4+ T cells showing myeloid lineage directed reactivity in healthy individuals

“…This impedes not only induction of autoimmunity but also antitumor immunity, thereby weakening the concept of functional immune surveillance against malignant cells in the autologous and HLAmatched setting. However, immune responses against autologous malignant cells have frequently been observed, although the functional avidity of these T cells may often be questionable [8][9][10][11][12][13].In line with the observations of autologous tumor-reactive T-cell responses, proliferation of (naive) T cells in response to autologous monocyte-derived dendritic cells (autoDCs) has also been demonstrated in in vitro studies [14][15][16][17]. The immunological basis and the biological relevance of this autoDC reactivity, seen both in human and in mouse models, have not been clarified so far [15,16,18,19].…”

Functional and Phenotypic Analysis of Two-Day Monocyte-Derived Dendritic Cells Suitable for Immunotherapy Purposes

“…The results suggested that 2d-aDCs generated from total PBMCs (as described by Martinuzzi et al [25]) or from purified monocytes as described by Moser et al [27], [ Figure 7A and 7B, respectively] induce the expansion of tumor antigen-specific CD8+ T cells against HER2 peptide and activate the effector function of these cells, evidenced by HER2-dependent IFN-γ secretion [ Figure 7D] and the expression of CD154 in CD4 T cells obtained from breast cancer patients [ Figure 7C]. These results suggest that 2d-aDCs induce the specific response and expansion of T cells in breast cancer similar to 2d-stDCs but with an increased Th1 cytokine production.…”

“…After 24 hours the iDCs were subsequently maturated using either 2d-stDCs or 2d-aDCs maturation cocktails, with or without the addition of 5 µM of the corresponding peptide(s) for 6 days [ Figure S1A]. The second method is based on the methodology of Moser et al [27]; briefly, frozen PBMCs were sorted (FACS Aria II -BD) into three different populations, CD14+ (monocytes), CD4+ and CD8+ T cells, using the BD FACS Aria II System (BD Biosciences). The monocytes were differentiated into 2d-stDCs or 2d-aDCs, pulsed or unpulsed with corresponding peptides or proteins (5 µM each) and subsequently cultured with purified CD4+ or CD8+ T cells for 14 days at a ratio of 50:1 (T cell: DCs) in AIM-V culture media (Life Technologies), for CD8+ T cell culture 30 UI/mL IL-2 was added.…”

“…Using naïve T-cells cells from the same patient sample, we used a second in vitro system based on the methodology of Moser et al [27]. Briefly, naïve CD4+ and CD8+ T cells were sorted and cultured for 14 days with 2d-stDCs or 2d-aDCs in the presence of HER2/neu KIFGSLAFL peptide or the 20-mer HER2/neu peptide pool, followed by boosting with 2d-stDCs or 2d-aDCs for an additional 6 days [ Figure S1B].…”

Synergistic effect of methionine encephalin (MENK) combined with pidotimod(PTD) on the maturation of murine dendritic cells (DCs)

“…So it is crucial to understand the regulation of DC maturation to gain further insight into their in vivo function. [21][22][23][24][25] Both MENK and PTD are peptide molecules of TH1 modulatiors, 5,16 and each of these peptides, when used separately, promotes functional maturation of dendritic cells and have a marked effect on suppressing the progression of some tumors. 26,27 Whether these two peptides exert synergistic effect on DCs remains unclear so far.…”