Immunophenotyping Reveals No Significant Perturbation to PBMC Subsets When Co-cultured With Colorectal Adenocarcinoma Caco-2 Cells Exposed to X-Rays
Open Access
- 2 June 2020
- journal article
- research article
- Published by Frontiers Media SA in Frontiers in Immunology
- Vol. 11, 1077
- https://doi.org/10.3389/fimmu.2020.01077
Abstract
In vitroco-culture models between tumor cells and peripheral blood mononuclear cells (PBMCs) allow studying the interplay between these cell populations, potentially gaining insight into thein vivoresponse of the immune system to the presence of the tumor, as well as to possible other agents as radiation used for therapeutic purposes. However, great care is needed in the experimental optimization of models and choice of conditions, as some setups might offer a limited possibility to capture subtle immune perturbations. A co-culture model of PBMCs from healthy donors and colorectal adenocarcinoma Caco-2 cells was successfully adopted in a previous work to measure effects on Caco-2 and modulation of signaling when these latter are irradiated. We here tested if the same experimental setting allows to measure perturbations to the main PBMC subsets: we performed immunophenotyping by means of flow cytometry and quantified helper and cytotoxic T cells, NK cells, and B cells, when PBMCs are cultured alone (control), in presence of non-irradiated Caco-2 cells or when these latter are exposed to a 10 Gy X-ray dose from a conventional radiotherapy accelerator. To measure a baseline response in all experimental conditions, PBMCs were not further stimulated, but only followed in their time-evolution up to 72 h post-irradiation of Caco-2 and assembly of the co-culture. In this time interval PBMCs maintain a high viability (measured via the MTT assay). Caco-2 viability (MTT) is slightly affected by the presence of PBMCs and by the high radiation dose, confirming their radioresistance. Immunophenotyping results indicate a large inter-individual variability for different population subsets already at the control level. We analyzed relative population changes and we detected only a small but significant perturbation to cytotoxic T cells. We conclude that this model, as it is, is not adequate for the measurements of subtler immune perturbations (if any, not washed-out by inter-individual differences). For this purpose, the model needs to be modified and further optimized e.g., including a pre-treatment strategy for PBMCs. We also performed a pooled analysis of all experimental observations with principal component analysis, suggesting the potential of this tool to identify subpopulations of similarly-responding donors.Funding Information
- Università degli Studi di Pavia
This publication has 27 references indexed in Scilit:
- Functional Analysis via Standardized Whole-Blood Stimulation Systems Defines the Boundaries of a Healthy Immune Response to Complex StimuliImmunity, 2014
- NKG2D signaling in cancer immunosurveillanceInternational Journal of Cancer, 2014
- The resveratrol analog 4,4'-dihydroxy-trans-stilbene suppresses transformation in normal mouse fibroblasts and inhibits proliferation and invasion of human breast cancer cellsCarcinogenesis: Integrative Cancer Research, 2012
- Immunostimulatory effect of faecalBifidobacteriumspecies of breast-fed and formula-fed infants in a peripheral blood mononuclear cell/Caco-2 co-culture systemBritish Journal of Nutrition, 2011
- Colorectal cancerThe Lancet, 2010
- Effects of wheat germ agglutinin on human gastrointestinal epithelium: Insights from an experimental model of immune/epithelial cell interactionToxicology and Applied Pharmacology, 2009
- Matrix metalloproteinases in inflammatory bowel disease: Boon or a bane?Inflammatory Bowel Diseases, 2007
- Lymphocyte subsets' reference ranges in an age- and gender-balanced population of 100 healthy adults—A monocentric German studyClinical Immunology, 2005
- Modulation of human dendritic cell phenotype and function by probiotic bacteriaGut, 2004
- Disregulation in TH1 and TH2 subsets of CD4 + T cells in peripheral blood of colorectal cancer patients and involvement in cancer establishment and progressionCancer Immunology, Immunotherapy, 1996