Pre-evaluation
Determination of toxicity of the drug in “in vitro” cell cultures to determine suitable concentrations of the drug (also adapted to the in vivo concentrations assumed to be used); evaluation of a comparator drug to be included.
Possible in vitro tests:
- Does the new drug stimulate the innate immunity of humans?
- cytokine determination after stimulation with the new drug: monocyte derived cytokines (like IL-1b, TNFα|, IL-6…)
- up-regulation of adhesion molecules (CD40, CD86) on monocytes or immature DC after drug addition for 24-48hrs
2. Does the new drug stimulate the adaptive immunity and does it stimulate via the p-i mechanism?
- stimulatory potential of the novel compound on peripheral mononuclear cells (PBMC) in lymphocyte transformation tests (LTT)
- stimulatory potential of the novel compound on CD69 up-regulation in PBMC, and determination of the involved cell population
- cytokine determination after stimulation with the new drug: T cell derived cytokines like IL-2, IFNγ, IL-13, IL-5 etc. will be tested
- stimulatory potential of the novel compound on CD71 up-regulation of T cell lines (mostly drug, some peptide specific)
- TCR-transfected hybridoma cell lines: evaluation of the new compound for its capacity to induce a partial (TCR downregulation) or complete (IL-2 production) activation.
- basophil activation test of the new compound using basophils of atopic donors: effect on CD203c up-regulation or CD63 expression. The results will be compared to various drugs often or rarely inducing drug hypersensitivity reactions; In some instances, a similar drug is already available and the reactivity can be compared