Preparation of Tumor Antigen-loaded Mature Dendritic Cells for Immunotherapy

Sabado, Rachel Lubong; Miller, Elizabeth A.; Spadaccia, Meredith; Vengco, Isabelita; Hasan, Farah; Bhardwaj, Nina

doi:10.3791/50085

Cited by 19 publications

(15 citation statements)

References 24 publications

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“…Human MDDCs were differentiated from monocyte fractions of human PBMCs obtained from healthy volunteer blood donors, as described previously [8,29,30]. Human buffy coats served as the source of monocytes and were obtained from the New York Blood Center (New York, NY, USA).…”

Section: Generation Of Mddcs and Primary DC Isolationmentioning

confidence: 99%

Ion efflux and influenza infection trigger NLRP3 inflammasome signaling in human dendritic cells

Fernandez

Miller

Krammer

et al. 2015

Journal of Leukocyte Biology

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The nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome, a multiprotein complex, is an essential intracellular mediator of antiviral immunity. In murine dendritic cells, this complex responds to a wide array of signals, including ion efflux and influenza A virus infection, to activate caspase-1-mediated proteolysis of IL-1β and IL-18 into biologically active cytokines. However, the presence and function of the nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome in human dendritic cells, in response to various triggers, including viral infection, has not been defined clearly. Here, we delineate the contribution of the nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome to the secretion of IL-1β, IL-18, and IL-1α by human dendritic cells (monocyte-derived and primary conventional dendritic cells). Activation of the nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome in human dendritic cells by various synthetic activators resulted in the secretion of bioactive IL-1β, IL-18, and IL-1α and induction of pyroptotic cell death. Cellular IL-1β release depended on potassium efflux and the activity of proteins nucleotide-binding oligomerization domain-like receptor protein 3 and caspase-1. Likewise, influenza A virus infection of dendritic cells resulted in priming and activation of the nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome and secretion of IL-1β and IL-18 in an M2- and nucleotide-binding oligomerization domain-like receptor protein 3-dependent manner. The magnitude of priming by influenza A virus varied among different strains and inversely corresponded to type I IFN production. To our knowledge, this is the first report describing the existence and function of the nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome in human dendritic cells and the ability of influenza A virus to prime and activate this pathway in human dendritic cells, with important implications for antiviral immunity and pathogenesis.

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Section: Generation Of Mddcs and Primary DC Isolationmentioning

confidence: 99%

Ion efflux and influenza infection trigger NLRP3 inflammasome signaling in human dendritic cells

Fernandez

Miller

Krammer

et al. 2015

Journal of Leukocyte Biology

Self Cite

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“…(11,(23)(24)(25). Previous studies have revealed that Tα1 exerts immunomodulatory effects on the maturation, differentiation and function of DCs from murine bone marrow (18,26) and healthy human PB (21).…”

Section: Discussionmentioning

confidence: 99%

Effect of thymosin α1 on the phenotypic and functional maturation of dendritic cells from children with acute lymphoblastic leukemia

Liu

Zhao

et al. 2015

Molecular Medicine Reports

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Abstract.To determine the effect of thymosin α 1 (Tα1) on the phenotypic and functional maturation of HL-60 cells, freeze-thaw antigen-loaded dendritic cells (DCs) were derived from peripheral blood mononuclear cells (PBMCs) of children with acute lymphoblastic leukemia (ALL). The DCs were generated from the PBMC samples that were collected from the PB of 10 consecutive ALL children. On day 3 of culturing, the cells in the antigen + no Tα1 (AN) and antigen + Tα1 (AT) groups were incubated with 100 µl lysates obtained from freeze-thaw cycling. After 5 days of incubation, the AT group was administered with 100 ng/ml Tα1. On day 8, the DCs were stained with fluorescein isothiocyanate-conjugated cluster of differentiation (CD)1a, CD83 and HLA-DR antibodies and analyzed by flow cytometry. In addition, the killing activity of cytotoxic T lymphocytes (CTLs) from the different groups on wild-type leukemia cells was measured. The DCs in the AT group exhibited more apparent, characteristic dendritic morphologies than the control and AN group DCs. Furthermore, the lowest expression level of CD1a, and the highest expression of CD83 and HLA-DR were observed in the AT group when compared with the AN and control groups (P<0.05). The lactate dehydrogenase release assay demonstrated that the killing rate of CTL in the AT group was significantly higher than that in the control and AN groups (P<0.01). Thus, Tα1 may markedly promote the phenotypic and functional maturation of DCs, and may serve as a suitable immunomodulator of DC-based immunotherapy for treatment of hematological malignancies.

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“…Patients received 5 doses of 5×10 6 DCs pulsed with combination of rPSMA and rSurvivin (ratio 1 : 1), in each dose and administered as a subcutaneous injection (see study scheme; Fig. 1 Generation of Monocyte-Derived DCs The DC vaccines were generated as previously described by Sabado et al 30) Briefly, 1-3×10 10 peripheral blood mononuclear cells (PBMCs) were collected by a 3 h leukapheresis procedure from each patient, and then placed in a tissue culture flask adherence at a density of 1×10 6 cells/cm 2 in the presence of 1% human serum albumin (Baxter, Deerfield, U.S.A.). After 2 h of incubation at 37°C in 5% CO 2 , non-adherent cells were removed by rinsing with sterile phosphate buffered saline (PBS).…”

Section: Patient Population and Study Designmentioning

confidence: 99%

Survivin and PSMA Loaded Dendritic Cell Vaccine for the Treatment of Prostate Cancer

Wang

et al. 2015

Biological & Pharmaceutical Bulletin

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Key words dendritic cell vaccine; prostate cancer; autologous cellular immunotherapy; prostate specific membrane antigen; Survivin Immunotherapy has recently emerged as a promising treatment modality for management of advanced hormone-refractory prostate cancer (HRPC), in light of new advances in discovery of cancer-specific antigens (tumor-associated antigen (TAA)), and adoptive cellular targeting. Dendritic cell (DC) based adoptive immunotherapy is one such treatment. Dendritic cells are the professional antigen presenting cells that efficiently activate T lymphocytes by presenting antigen to naïve major histocompatibility complex I restricted CD8+ cells and to major histocompatibility complex II restricted CD4+ cells. Dendritic cells generated from autologous monocytes (CD14+ cells) concentrated by aphaeresis, using a culture medium enriched with granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-4, or IL-13 1-4) and pulsed ex vivo with TAA have been found to elicit a potent T-cell-mediated immune response and also protect against further tumour challenges. [5][6][7][8] Results from various clinical studies indicate that DC-based therapy can lead to significant therapeutic benefit in cancer patients without any significant toxic side effects. [9][10][11][12] In 2010, the U.S. Food and Drugs Administration (FDA) approved Sipuleucel-T, an autologous dendritic cell vaccine for the treatment of HRPC, this has paved way for a new category of autologous cellular immunotherapy for management of prostate cancer.Prostate specific membrane antigen (PSMA) is a type II membrane protein with folate hydrolase activity expressed primarily in prostate epithelium and a limited number of other cell types. This well-defined prostate expression is significantly elevated in prostate cancer, especially in advanced stages, [12][13][14] furthermore, PSMA immunoreactivity is absent to moderate in most hyperplastic and benign tissues. These features make PSMA an excellent target for prostate cancer immunotherapy. Several PSMA-based vaccines have been developed and clinical trials indicate that these immunotherapy approaches can be safely administered and can induce immune responses in patients with advanced carcinoma of prostate. 15,16) Survivin is a member of the family of inhibitors of apoptosis which is involved in cell cycle progression and control of apoptosis.17) It is over-expressed in a number of human cancers [18][19][20][21] and is often associated with poor prognosis. [22][23][24] Because of its frequent over-expression in many cancer types, survivin constitutes an interesting tumour-associated antigen (TAA) target for T cell based immunotherapy. 21,25,26) Because of its nodal properties, and over-expression in virtually every human tumour, survivin has been intensely pursued as a drug target for novel cancer therapeutics. [27][28][29] Various studies suggest that a combination of TAA might be more efficient in dendritic cell based cancer immunotherapy than using a single TAA.These observations prom...

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Preparation of Tumor Antigen-loaded Mature Dendritic Cells for Immunotherapy

Cited by 19 publications

References 24 publications

Ion efflux and influenza infection trigger NLRP3 inflammasome signaling in human dendritic cells

Ion efflux and influenza infection trigger NLRP3 inflammasome signaling in human dendritic cells

Effect of thymosin α1 on the phenotypic and functional maturation of dendritic cells from children with acute lymphoblastic leukemia

Survivin and PSMA Loaded Dendritic Cell Vaccine for the Treatment of Prostate Cancer

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