Monocytes are major inflammatory cells in the circulation, and monocyte activity is associated with diseases such as rheumatoid arthritis, cancer and heart failure. Circulating monocytes are of two main types: classical (expressing high levels of the surface marker CD14), and non-classical (expressing CD16, and little CD14). Careful studies of monocyte subset function suggest these should not be simplistically categorized as pro- or anti-inflammatory. Intermediate monocytes have characteristics between these two. Increasing evidence suggests that the balance of classical and non-classical monocytes is altered in many disease states.
Companies are increasingly looking to study monocytes in clinical trials, and in the last year Java scientists have developed several customized monocyte assays to measure responses in clinical and preclinical studies. Among the customized monocyte assays that we have developed are:
• measurement by flow cytometry of a drug-specific target in monocytes and other blood cells in a phase I clinical trial,
• determination of monocyte phenotypic balance (proportion of classical, non-classical and intermediate monocytes) in preclinical and clinical studies,
• isolation of highly pure monocytes in a phase I study for PK analysis.
These and other assays (including proteomic and transcriptomic analysis) may be customised for use in clinical trials in rheumatology, cardiovascular disease and other areas to assess efficacy, test mechanism of action or differentiate a compound from competitors.
As part of our development process for customized monocyte assays (as well as other areas, such as platelet function and characterization), Java Clinical Research ensures that the assessments are appropriate for every study. We optimize variables such as timepoints, assay type, agonist conditions, etc. for the drug being investigated. Monocytes may be studied in whole blood, in purified peripheral blood mononuclear cells (PBMCs), or isolated as highly pure cell populations using magnetic bead selection systems, depending on assay requirements.
At Java we have also implemented some standard monocyte assays that measure how monocytes function and their interactions with other cell types, including measurement of cell-specific microparticles, monocyte-platelet aggregates and LPS-stimulation assays.