A simple in vitro method for evaluating dendritic cell-based vaccinations

Abstract

BackgroundDendritic cell (DC) therapy is a promising therapy for cancer-targeting treatments. Recently, DCs have been used for treatment of some cancers. We aimed to develop an in vitro assay to evaluate DC therapy in cancer treatment using a breast cancer model.MethodsDCs were induced from murine bone marrow mononuclear cells in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with GM-CSF (20 ng/mL) and IL-4 (20 ng/mL). Immature DCs were primed with breast cancer stem cell (BCSC)-derived antigens. BCSCs were sorted from 4T1 cell lines based on aldehyde dehydrogenase expression. A mixture of DCs and cytotoxic T lymphocytes (CTLs) were used to evaluate the inhibitory effect of antigen-primed DCs on BCSCs. BCSC proliferation and doubling time were recorded based on impedance-based cell analysis using the xCELLigence system. The specification of inhibitory effects of DCs and CTLs was also evaluated using the same system.ResultsThe results showed that impedance-based analysis of BCSCs reflected cytotoxicity and inhibitory effects of DCs and CTLs at 72 hours. Differences in ratios of DC:CTL changed the cytotoxicity of DCs and CTLs.ConclusionThis study successfully used impedance-based cell analysis as a new in vitro assay to evaluate DC efficacy in cancer immunotherapy. We hope this technique will contribute to the development and improvement of immunotherapies in the near future.

Application of xCELLigence RTCA Biosensor Technology for Revealing the Profile and Window of Drug Responsiveness in Real Time

“…Advantages, disadvantages and concluding remarks: It is now evident that xCELLigence biosensor technology has numerous applications for basic research and for clinical drug development. These range from the basics of good cell culture techniques to more advanced applications, including high throughput drug screening [ 31 ] (e.g., cardiotoxicity [ 32 , 33 ]), cell mediated killing [ 7 ], toxicology [ 34 ] and immunology [ 8 , 35 ]). As our experience grows, we realise more and more applications where xCELLigence technology is valuable.…”

In vitro immunotherapy potency assays using real-time cell analysis

“…In summary, the real-time impedance-based platform and reagents described in this paper provides a sensitive, robust and reproducible assay system that can be used for quantitative assessment of different immunotherapy approaches in vitro [ 38 – 41 ]. Despite the fact that various groups have already incorporated this technology for assessment of BiTEs and bispecific antibodies [ 25 , 26 ] oncolytic viruses [ 42 , 43 ], NK-mediated cytolysis [ 44 , 45 ] and for assessment of engineered T cells such as CAR-T [ 10 , 46 , 47 ], the new adaptation to suspension cancer cells and the companion immunotherapy analysis software promises to further extend and accelerate the adoption of the technology for immunotherapy approaches.…”

Targeting breast cancer stem cells by dendritic cell vaccination in humanized mice with breast tumor: preliminary results

“…T-lymphocyte proliferation stimulated by DCs and measurement of IL-12 were evaluated as described in a previously published study. 44 There were five experimental groups with different ratios of DCs:lymphocytes (0.25:100, 0.5:100, 1:100, 2:100, and 8:100) and three control groups with DCs + phytohemagglutinin (PHA), PHA alone, or PHA + lymphocytes. The T-lymphocyte concentration was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay kit according to the manufacturer’s instructions (Sigma-Aldrich Co.).…”