Using Impedance-Based Approaches for Measuring Antigen-Specific Cytotoxic T cell Activity

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Keith Knutson photoDr. Keith L. Knutson

Mayo Clinic, Jacksonville, FL
The Vaccine & Gene Therapy Institute of Florida, Port Saint Lucie, FL

Immunotherapy, where the immune system is harnessed to treat and/or prevent disease, holds great promise in the fight against cancer. One noteworthy facet of cancer immunotherapy is the development of cancer-specific vaccines. In this webinar Dr. Keith L. Knutson from the Mayo Clinic (Jacksonville, FL) and VGTI discusses the use of ACEA’s xCELLigence® system in his efforts to discover a novel epitope for use in breast cancer vaccination. Specific topics covered include:

  • The concepts of cancer vaccine and immune therapy
  • Development of an impedance-based assay using the xCELLigence® system to monitor cytotoxic T-cell activity
  • The advantages of dynamic monitoring of cytotoxic T-cell activity using impedance-based assays vs. the traditional endpoint Chromium-51 Release Assay (CRA)

Dr. Keith L. Knutson is currently the director of the Cancer Vaccine and Immune Therapies program at VGTI and is an affiliate associate professor in the Department of Immunology at the Mayo Clinic. He received his Ph.D. in physiology and pharmacology from the University of Georgia, Athens in 1995, and completed two postdoctoral fellowships in immunology at the University of British Columbia and the University of Washington. He was a 2004 recipient of the Howard Temin Award from the National Cancer Institute.

Dr. Knutson’s current research focuses on the immunology and immunotherapy of breast and ovarian cancers. His interests are in both the basic immunobiology and clinical translation, including clinical trials. His laboratory is currently conducting research on cancer vaccines that augment CD4 helper T-cell immunity using peptide epitopes. These vaccine strategies are aimed at preventing patient relapse after conventional chemotherapies. Adoptive T-cell therapy, a strategy that involves infusing high numbers of T-cells into patients with active bulky malignancy, is also being examined in Knutson’s lab. Specific adoptive T-cell therapy projects include methods for T-cell expansion and in vitro generation of memory T-cells.

Lastly, Dr. Knutson’s lab also seeks to understand how tumors evade the immune system and immune therapies. Several different mechanisms of immune escape are being investigated, including cellular reprogramming (e.g., epithelial to mesenchymal transition) and recruitment of regulatory T-cells and myeloid-derived suppressor cells into the tumor microenvironment. In addition to his research, Dr. Knutson participates in and chairs several study sections. He is currently a full member of the Department of Defense’s Breast Cancer Research Program Integration Panel and a full member of the Tumor Microenvironment Study Section at the National Institutes of Health Center for Scientific Review.

REFERENCES:

  1. Asiedu MK, Ingle JN, Behrens MD, Radisky DC, Knutson KL. TGFbeta/TNF(alpha)-mediated epithelial-mesenchymal transition generates breast cancer stem cells with a claudin-low phenotype. Cancer Res. 2011 Jul 1;71(13):4707-19.
  2. Erskine CL, Henle AM, Knutson KL. Determining optimal cytotoxic activity of human Her2neu specific CD8 T cells by comparing the Cr51 release assay to the xCELLigence system. J Vis Exp. 2012 Aug 8;(66):e3683.
  3. Henle AM, Erskine CL, Benson LM, Clynes R, Knutson KL. Enzymatic discovery of a HER-2/neu epitope that generates cross-reactive T cells.  J Immunol. 2013 Jan 1;190(1):479-88.
  4. Mader EK, Butler G, Dowdy SC, Mariani A, Knutson KL, Federspiel MJ, Russell SJ, Galanis E, Dietz AB, Peng KW. Optimizing patient derived mesenchymal stem cells as virus carriers for a phase I clinical trial in ovarian cancer. J Transl Med. 2013 Jan 24;11:20