Background White blood cells have been shown in animal studies to play a central role in the pathogenesis of diabetic retinopathy. Lymphoblastoid cells are immortalized EBV-transformed primary B-cell leukocytes that have been extensively used as a model for conditions in which white blood cells play a primary role. The purpose of this study was to investigate whether lymphoblastoid cell lines, by retaining many of the key features of primary leukocytes, can be induced with glucose to demonstrate relevant biological responses to those found in diabetic retinopathy. Methods Lymphoblastoid cell lines were obtained from twenty-three human subjects. Differences between high and standard glucose conditions were assessed for expression, endothelial adhesion, and reactive oxygen species. Results Collectively, stimulation of the lymphoblastoid cell lines with high glucose demonstrated corresponding changes on molecular, cellular and functional levels. Lymphoblastoid cell lines up-regulated expression of a panel of genes associated with the leukocyte-mediated inflammation found in diabetic retinopathy that include: a cytokine (IL-1B fold change = 2.11, p-value = 0.02), an enzyme (PKCB fold change = 2.30, p-value = 0.01), transcription factors (NFKB-p50 fold change = 2.05, p-value = 0.01), (NFKB-p65 fold change = 2.82, p-value = 0.003), and an adhesion molecule (CD18 fold change = 2.59, 0.02). Protein expression of CD18 was also increased (p-value = 2.14x10-5 ). The lymphoblastoid cell lines demonstrated increased adhesiveness to endothelial cells (p = 1.28x10-5 ). Reactive oxygen species were increased (p = 2.56x10-6 ). Significant inter-individual variation among the lymphoblastoid cell lines in these responses was evident (F = 18.70, p < 0.0001). Conclusions Exposure of lymphoblastoid cell lines derived from different human subjects to high glucose demonstrated differential and heterogeneous gene expression, adhesion, and cellular effects that recapitulated features found in the diabetic state. Lymphoblastoid cells may represent a useful tool to guide an individualized understanding of the development and potential treatment of diabetic complications like retinopathy.
Bibliographical noteFunding Information:
The authors gratefully acknowledge financial support for this research. Funding for this work was provided by: the NIDDK Diabetic Complications Consortium (DiaComp, www.diacomp. org), grant DK076169 (MAG); Search for Vision (http://www.searchforvision.org/home.html)(MAG); National Eye Institute (https://nei.nih.gov/) R01 EY023644-01A1 (MAG);National Eye Institute Core Grant EY001792 for Vision Research; an unrestricted grant from Research to Prevent Blindness (http:// www.rpbusa.org/rpb), Inc., New York, NY; the Juvenile Diabetes Research Foundation (http://jdrf. org/) (JDRF 17-2012-480) (RN); the National Institutes of Health R01 DK065073 (RN); RSH received support from NIH/NIGMS (http://www.nigms. nih.gov) grant K08GM089941, Circle of Service Foundation (http://www.cosfoundation.org/) Early Career Investigator award, NIH/NCI (http://www. cancer.gov/) grant R21 CA139278, NIH/NIGMS grant UO1GM61393, University of Chicago Cancer Center Support Grant (#P30 CA14599), Breast Cancer SPORE Career Development Award [CA125183], a Conquer Cancer Foundation of American Society of Clinical Oncology (http://www.asco.org/) Translational Research Professorship award In Memory of Merrill J. Egorin, MD (awarded to Dr. MJ Ratain) and a pilot grant from NIH/NCATS (https://ncats.nih.gov/) grant UL1RR024999. Funding for EDIC is provided by cooperative agreement grants (1982-1993, and 2012-2017) and contracts (1982-2012) with the NIDDK (http://www.niddk.nih.gov), Division of Diabetes, Endocrinology, and Metabolic Diseases (grant numbers U01 DK094176 and U01 DK094157), as well as the National Eye Institute, National Institute of Neurological Disorders and Stroke, the General Clinical Research Center Program (1993-2007), and the Clinical Translational Science Center Awards Program (2006- present). Industry contributors have had no role in the DCCT/EDIC study but have provided free or discounted supplies or equipment to support participants' adherence to the study: Abbott Diabetes Care (Alameda, CA), Animas (Westchester, PA), Bayer Diabetes Care (North America Headquarters, Tarrytown, NY), Becton Dickinson (Franklin Lakes, NJ), Eli Lilly (Indianapolis, IN), Extend Nutrition (St. Louis, MO), Insulet Corporation (Bedford, MA), Lifescan (Milpitas, CA), Medtronic Diabetes (Minneapolis, MN), Nipro Home Diagnostics (Ft. Lauderdale, FL), Nova Diabetes Care (Billerica, MA), Omron (Shelton, CT), Perrigo Diabetes Care (Allegan, MI), Roche Diabetes Care (Indianapolis, IN), and Sanofi-Aventis (Bridgewater, NJ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.