A Comparative Analysis of Serum and Serum-free Media for Generation of Clinical Grade DCs

Napoletano, Chiara; Pinto, Dora; Bellati, Filippo; Taurino, Federica; Rahimi, Hassan; Tomao, Federica; Panici, Pierluigi Benedetti; Rughetti, Aurelia; Frati, Luigi; Nuti, Marianna

doi:10.1097/cji.0b013e318046f396

Cited by 26 publications

(28 citation statements)

References 47 publications

(69 reference statements)

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“…As expected, an increased dextran uptake by iDCs (day 6 culture) was observed compared with the day 8 mDCs; however, the increase was not statistically significant. Various studies have indicated that the uptake of dextran may be a measure of immune response (23)(24)(25). The present results also support earlier studies that suggest that increased antigen uptake is associated with IL-12 and IFN-γ secretion, and CTL response (17,26,27).…”

Section: Discussionsupporting

confidence: 90%

In vitro generation of cytotoxic T lymphocyte response using dendritic cell immunotherapy in osteosarcoma

Zhang

Kou

et al. 2016

Oncology Letters

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Abstract.Immunotherapy with tumor lysate-pulsed dendritic cells (DCs) is one of the breakthrough strategies used in the treatment of cancer. However, DC-based immunotherapies for osteosarcoma are limited. In the present study, preclinical studies of a C3H osteosarcoma mouse model (produced by subcutaneous injection of LM8 murine osteosarcoma cells) validated the concept that LM8 cell lysate-pulsed bone marrow-derived DCs may evoke a more potent immune response compared with DCs that have been matured using polyinosinic:polycytidylic acid (poly I:C). A cytotoxic T lymphocyte (CTL) response was established using two groups of C3H mice (n=9) with osteosarcoma; the treatment group consisted of LM8 cell lysate-pulsed DCs and the control group consisted of DCs matured using poly I:C. Each group was immunized with doses of 1x10 6 cells twice per week for 3 weeks. No difference in the expression of cluster of differentiation markers was identified in the two groups. DCs pulsed with LM8 cell lysate were associated with the increased induction of CTL activity. Serum interferon-γ levels were increased in mice that received DCs pulsed with LM8 cell lysate compared with that in the poly I:C-matured DC group (P<0.041). Serum interleukin-4 was decreased in the treatment group vs. the control group (P<0.033). A mixed lymphocyte reaction assay confirmed that LM8-DC immunotherapy may evoke a significant antigen-specific immune response in a mouse model. The present study reveals promising data on efficacy of a DC-based immunotherapy in the treatment of osteosarcoma; however, further clinical studies are warranted.

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Section: Discussionsupporting

confidence: 90%

In vitro generation of cytotoxic T lymphocyte response using dendritic cell immunotherapy in osteosarcoma

Zhang

Kou

et al. 2016

Oncology Letters

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“…These clinical-grade tolDCs are comparable to our previously characterised research-grade tolDCs24 25 in terms of cell surface phenotype, cytokine production and in vitro T cell modulatory function. CellGroDC was deemed the medium of choice23 and is currently used in DC vaccine development for cancer 38–40. MPLA, a low-toxicity alternative to LPS, is currently being tested in vaccine development 41.…”

Section: Discussionmentioning

confidence: 99%

“…Therapy with tolDCs therefore has great potential for the treatment of autoimmunity in humans, and the current challenge is to develop tolDCs for clinical application. Others have addressed the development of clinical-grade DCs for cancer immunotherapy 21–23. However, criteria for the development of safe tolDCs for the treatment of autoimmune disease remain to be evaluated.…”

Section: Introductionmentioning

confidence: 99%

Generation and characterisation of therapeutic tolerogenic dendritic cells for rheumatoid arthritis

et al. 2010

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ObjectivesTolerogenic dendritic cells (tolDCs) constitute a promising experimental treatment for targeting autoreactive T cells in autoimmune diseases, including rheumatoid arthritis (RA). The authors' goal is to bring tolDC therapy for RA to the clinic. Here the authors address key translational issues related to the manufacturing of tolDCs from RA patients with current good manufacturing practice (cGMP)-compliant reagents, the stability of tolDCs, and the selection of suitable quality control markers.MethodsHuman monocyte-derived tolDCs were established from RA patients and healthy controls (HCs) using the immunosuppressive drugs dexamethasone and vitamin D3, and the cGMP-grade immunomodulator, monophosphoryl lipid A, in the cGMP-compliant medium, CellGroDC. The functionality of tolDCs and tolDC-modulated autologous CD4 T cells was determined by flow cytometry, [3H]thymidine incorporation and ELISA.ResultsClinical-grade tolDCs established from patients with RA exhibit a typical tolerogenic phenotype of reduced costimulatory molecules, low production of proinflammatory cytokines and impaired stimulation of autologous antigen-specific T cells, comparable to HC tolDCs. Toll-like receptor 2 (TLR-2) was highly expressed by tolDCs but not mature DCs. Furthermore, tolDCs suppressed mature DC-induced T cell proliferation, interferon γ and interleukin 17 production, and rendered T cells hyporesponsive to further stimulation. Importantly, tolDCs were phenotypically stable in the absence of immunosuppressive drugs and were refractory to further challenge with proinflammatory mediators.ConclusionstolDCs established from patients with RA are comparable to those derived from healthy donors. TLR-2 was identified as an ideal marker for quality control of tolDCs. Potently tolerogenic and highly stable, these tolDCs are a promising cellular therapeutic for tailored immunomodulation in the treatment of RA.

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“…One of the most recent avenues that are being pursued is the use of DCs. Cell therapy remains extremely complex and the quality of cells produce strictly depends on methods and materials adopted 52.…”

Section: Novel Strategiesmentioning

confidence: 99%

Immune Effects of Trastuzumab

Nuti¹,

Bellati²,

Visconti³

et al. 2011

J. Cancer

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Trastuzumab's targeted therapy has become a stronghold for human epidermal growth factor receptor 2 positive breast cancer patients. This humanized monoclonal antibody binds to the extracellular juxta-membrane domain of HER2 and inhibits the proliferation and survival of HER2 dependent cancer cells. The ways by which this molecule exerts its action have been partially elucidated but several new mechanisms are being constantly identified. Several new agents are being introduced that interfere with HER2. Several new immunotherapy strategies are being introduced in order to direct the immune system against cells and tissues that aberrantly overexpressed HER2. We review the strategies currently adopted and those suggested against HER2 expressing tumors.

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A Comparative Analysis of Serum and Serum-free Media for Generation of Clinical Grade DCs

Cited by 26 publications

References 47 publications

In vitro generation of cytotoxic T lymphocyte response using dendritic cell immunotherapy in osteosarcoma

In vitro generation of cytotoxic T lymphocyte response using dendritic cell immunotherapy in osteosarcoma

Generation and characterisation of therapeutic tolerogenic dendritic cells for rheumatoid arthritis

Immune Effects of Trastuzumab

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