“Multiple cytokine secreting cells”: central players in pharmaco-dynamics of immuno-modulatory therapies?

In a workshop hosted by the European Medicine Agency the possible role of in vitro cytokine release assays for prediction of unwanted side effects of novel (immuno-) therapeutics has been discussed. With respect to the latter the “cytokine release syndrome” and “cytokine storm” are severe and sometimes fatal adverse clinico-pathological events for which predictive assay systems are urgently needed.

In the workshop the central role of so-called “multiple cytokine secreting cells” in the pharmaco-dynamics of immuno-modulatory compounds and associated adverse events was pointed out.

As the name implies, these cells are characterized by the production of several cytokines in any single cell and thus contrast cells in which only one or two cytokines may be detected. In other words: “Multiple Cytokine Secretion” is a functional characteristic of a given cell and pretty much every T cell may become such a multiple cytokine secreting cell upon activation. Multiple cytokine secretion is a temporary state, only as long as the cell is being activated.

Physiological stimulation of T cells to multiple cytokine secretion is required for host defense and tissue remodeling. However, aberrant or un-intentional stimulation under pathological conditions or by (over-dosing of) immuno-modulatory compounds may result in severe pathologies, such as the above-mentioned “Cytokine Release Syndrome“.

The analysis of multiple cytokine secreting cells requires sophisticated multi-parameter assays, like multiplex immunological assays for measuring cytokine profiles in cellular secretions under various stimulation conditions. Ultimately, it requires multi-color intracellular cytokine staining by flow cytometry, which allows multiple cytokine determinations at the single level.

FOCUS Immunology is experienced in providing state-of-the-science analyses for multiple cytokine secreting cells under GLP conditions. If you are interested in learning more about FOCUS Immunology’s experience and offers or if you want to discuss your specific experimental needs, please feel free to contact
Dr. Eddy Bruyns, Head of FOCUS Immunology Laboratory via e-mail eddy.bruyns@focus-cdd.com or via telephone +49 6221 64935124.

EMEA workshop: “In vitro cytokine release assays to predict Cytokine Release Syndrome: the current-state-of-the science”

Biologicals Testing Package, esp. for higher risk NBEs

The advent of biologicals has brought new challenges to drug development. Proteins with immunomodulatory activity such as monoclonal antibodies directed against major switches of the immune system or proteins such as receptor antagonists or agonists may have dramatic effects on immune cell function.

The recent past has shown just how dramatic these effects may be for the well-being of – in this case – volunteers in a clinical study.

The ensuing investigation of this incident resulted in committee recommendations among which the use of human ex vivo / in vitro cultures had a prominent place. This is based on the rationale that biologicals are very often species-specific and thus traditional animal models are in all likelihood not adequate to investigate effects on the immune system.

FOCUS Immunology offers a range of assays to assess the effects of biologicals on different aspects of immune cell activity, which allow a comprehensive analysis of possible immunomodulatory activities.

Specifically, the following assays are provided by FOCUS Immunology : (i) cytokine release, (ii) cell proliferation, (iii) expression of activation markers, (iv) oxidative burst reaction, (v) chemotaxis, (vi) NK cell activity, (vii) cytotoxicity (CDC, ADCC), (viii) NO production, (ix) histamine release, (x) changes in the frequency of defined cell populations.

It may be advisable to test not only resting cells but also cells after suboptimal stimulation in order to account for activation processes which may not suffice by themselves to activate resting cells but may strongly augment activation processes occurring independently.

It also may be advisable to test different means of presenting the tested biological to the cells, from soluble to different types of surface-bound presentation.