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Analysis of Pro-inflammatory Cytokine and Type II Interferon Induction by Nanoparticles

  • Timothy M. Potter
  • Barry W. Neun
  • Jamie C. Rodriguez
  • Anna N. Ilinskaya
  • Marina A. DobrovolskaiaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1682)

Abstract

Cytokines, chemokines, and interferons are released by the immune cells in response to cellular stress, damage and/or pathogens, and are widely used as biomarkers of inflammation. Certain levels of cytokines are needed to stimulate an immune response in applications such as vaccines or immunotherapy where immune stimulation is desired. However, undesirable elevation of cytokine levels, as may occur in response to a drug or a device, may lead to severe side effects such as systemic inflammatory response syndrome or cytokine storm. Therefore, preclinical evaluation of a test material’s propensity to cause cytokine secretion by healthy immune cells is an important parameter for establishing its safety profile. Herein, we describe in vitro methods for analysis of cytokines, chemokines, and type II interferon in whole blood cultures derived from healthy donor volunteers. First, whole blood is incubated with controls and tested nanomaterials for 24 h. Then, culture supernatants are analyzed by ELISA to detect IL-1β, TNFα, IL-8, and IFNγ. The culture supernatants can also be analyzed for the presence of other biomarkers secreted by the immune cells. Such testing would require additional assays not covered in this chapter and/or optimization of the test procedure to include relevant positive controls and/or cell types.

Key words

Cytokines Chemokines Interferons Inflammation Whole blood Immunostimulation 

Notes

Acknowledgment

This project has been funded in whole or in part by federal funds from the National Cancer Institute, National Institutes of Health, under contract HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Timothy M. Potter
    • 1
  • Barry W. Neun
    • 1
  • Jamie C. Rodriguez
    • 1
  • Anna N. Ilinskaya
    • 1
  • Marina A. Dobrovolskaia
    • 1
    Email author
  1. 1.Cancer Research Technology Program, Nanotechnology Characterization LaboratoryLeidos Biomedical Research, Inc., Frederick National Laboratory for Cancer ResearchFrederickUSA

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