ADCC of adherent target cells: How to improve prediction of treatment efficacy

In a new paper (Anabel Zwick, et al., Journal of Immunological Methods,, the autors report results from granulocyte-mediated ADCC obtained using real-time monitoring with xCELLigence (ACEA Biosciences) .

While the impedance assay overcame limitations of end-point assays, such as the requirement of target cells in suspension or unspecific release of probes, results at short-term were comparable to 51-chromium or Calcein release assays. 

However, in the long run, real-time monitoring revealed additionally cell-line specific kinetics and long-term efficacy. Monitoring IgA-dependent neutrophil-mediated ADCC for more than four hours allowed the documentation of residual non-killed target cells that grew out again, although short-term kinetics indicated efficient killing.

The authors discuss additional critical parameters for the prediction of treatment efficacy appearing only after prolonged monitoring of ADCC.

In conclusion, xCELLigence provides additional valuable information on kinetics of ADCC against target cells, effector cell performance, efficacy and additional critical parameters of cytotoxicity.



Figure: Monitoring target cell killing in the presence of granulocytes and IgA2.0 (blue line).
Short-term endpoint assays (<28 h) indicate effective cell killing.
Long-term kinetics however revealed that  target cell growth is progressing again
after approx. 32h.

Dr. Stefan Lohse, Universitätsklinikum des Saarlandes und Medizinische Fakultät der Universität des Saarlandes, Institut für Virologie, Germany:

"We are working with xCELLigence SP and DP. Running assays is very convenient: after seeding the cells, plates are placed in the incubator and automatic measurement is initiated.

Monitoring of cellular behavior is feasible at any time online in real-time. However, due to the high sensitivity of the system, frequent usage of the incubator should be avoided as disturbances might get visible in the result plots.

Main advantages of the system are the capability of long-term measurements and online cell
monitoring without disturbance of the cells and the avoidance of any dyes using adherently growing target cells - distinct advantages to traditional endpoint assays, what finally led us to the results discussed in the recent paper."



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