Cancer Immunotherapy: Oncolytic Viruses


Oncolytic virotherapy is a promising cancer treatment that uses a replication-competent virus to selectively infect cancer cells, cause cytotoxicity, and generate anti-tumor immunity.  This approach has seen major advances in recent years using both wildtype and genetically engineered viruses.

Analyzing cancer cell killing with high sensitivity and without the need for labels/modifications, the xCELLigence Real-Time Cell Analysis (RTCA) instruments allow the interaction between viruses and target cells to be studied under conditions that approximate human physiology more closely than other in vitro techniques.  By monitoring target cell killing continuously, these instruments also do away with laborious endpoints and thereby readily yield cell killing data under many different conditions simultaneously.

Application Highlight

In the below example xCELLigence RTCA was used to monitor killing of A549 lung cancer cells by a chimeric adenovirus (Enadenotucirev, EnAd) which infects cells by binding to CD46 and/or desmoglein, both widely expressed on many carcinoma cells.  In a potency analysis, the cytotoxicity (i.e. killing kinetics) of EnAd at a range of concentrations was compared with wild-type adenoviruses Ad11p and Ad5.  At the highest concentration (red, 500 PPC (particles per cells)), EnAd and Ad11p caused complete cell killing (Cell Index decreasing to zero) between 36-48 hours post-infection.  However, at lower virus concentrations (0.8-20 PPC) EnAd is substantially more potent than Ad11p, displaying both an earlier onset of cytotoxicity and a more rapid completion of cytolysis.  When compared with EnAd and Ad11p, wildtype Ad5 is much less efficient at killing the cancer cells, requiring 5 days to achieve full cell killing even at the highest virus concentration.

This data highlights the ability of xCELLigence RTCA assays to quantitatively capture differences in the potency of different oncolytic viruses.

Killing of A549 lung cancer cells by different adenoviruses. Black arrow indicates the time of virus addition. Virus concentrations are listed as particles per cell (PPC). Figure adapted from: Mol Ther Oncolytics. 2016 Dec 10;4:18-30

Key Benefits of Using xCELLigence to Monitor Oncolytic Viruses:
  1. Label-Free: Allowing for more physiological assay conditions; labeling or secondary assays aren’t required.
  2. Real-Time: Quantitative monitoring of both fast (hours) and slow (days) killing kinetics.
  3. Sensitive: Capable of evaluating low effector cell to target cell ratios that are physiologically relevant.
  4. Simple Workflow: Requires only the addition of effector cells to target cells (in the presence or absence of antibodies); homogeneous assay without additional sample handling.
  5. Automatic Data Plotting: RTCA software enables facile data display and objective analysis, precluding the subjective data vetting that is common to imaging-based assays.

IMT Handbook

Explore Functional Potency Assays for Cancer Immunotherapy Research

  • Antibody-Dependent Cell-Mediated Cytolysis (ADCC)
  • BiTEs and Bispecific Antibodies
  • Checkpoint Inhibitors
  • CAR-T Cells

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Cancer Immunotherapy Research Grant

The research grant winner will be provided access to the xCELLigence Real Time Cell Analysis (RTCA) SP instrument, consumables, and consultation for up to 6 months. Apply by September 15, 2018.

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Webinar: Utilizing a Label-Free Real-Time Cell Analysis Technology for Immuno-Oncology Applications

The xCELLigence potency assay is a label-free, non-destructive assay that enables the measurement of complete immune cell killing kinetics from single samples. 

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Oncolytic Viruses Supporting Information:

  1. Oncolytic adenovirus expressing bispecific antibody targets T-cell cytotoxicity in cancer biopsies.  Freedman JD, Hagel J, Scott EM, Psallidas I, Gupta A, Spiers L, Miller P, Kanellakis N, Ashfield R, Fisher KD, Duffy MR, Seymour LW. EMBO Mol Med. 2017 Aug;9(8):1067-1087.
  2. Oncolytic Adenoviral Delivery of an EGFR-Targeting T-cell Engager Improves Antitumor Efficacy.  Fajardo CA, Guedan S, Rojas LA, Moreno R, Arias-Badia M, de Sostoa J, June CH, Alemany R.  Cancer Res. 2017 Apr 15;77(8):2052-2063.
  3. Oncolytic Group B Adenovirus Enadenotucirev Mediates Non-apoptotic Cell Death with Membrane Disruption and Release of Inflammatory Mediators.  Dyer A, Di Y, Calderon H, Illingworth S, Kueberuwa G, Tedcastle A, Jakeman P, Chia SL, Brown A, Silva MA, Barlow D, Beadle J, Hermiston T, Ferguson DJ, Champion B, Fisher KD, Seymour LW.  Mol Ther Oncolytics. 2016 Dec 10;4:18-30.
  4. Novel epi-virotherapeutic treatment of pancreatic cancer combining the oral histone deacetylase inhibitor resminostat with oncolytic measles vaccine virus.  Ellerhoff TP, Berchtold S, Venturelli S, Burkard M, Smirnow I, Wulff T, Lauer UM.  Int J Oncol. 2016 Nov;49(5):1931-1944.
  5. Oncolytic effects of parvovirus H-1 in medulloblastoma are associated with repression of master regulators of early neurogenesis.  Lacroix J, Schlund F, Leuchs B, Adolph K, Sturm D, Bender S, Hielscher T, Pfister SM, Witt O, Rommelaere J, Schlehofer JR, Witt H.  Int J Cancer. 2014 Feb 1;134(3):703-16.
  6. Generation of an adenovirus-parvovirus chimera with enhanced oncolytic potential.  El-Andaloussi N, Bonifati S, Kaufmann JK, Mailly L, Daeffler L, Deryckère F, Nettelbeck DM, Rommelaere J, Marchini A.  J Virol. 2012 Oct;86(19):10418-31.
  7. Efficient and selective tumor cell lysis and induction of apoptosis in melanoma cells by a conditional replication-competent CD95L adenovirus.  Fecker LF, Schmude M, Jost S, Hossini AM, Picó AH, Wang X, Schwarz C, Fechner H, Eberle J.  Exp Dermatol. 2010 Aug;19(8):e56-66.