The IMBG Test for Evaluating the Pharmacodynamic Response to Immunosuppressive Therapy in Kidney Transplant Patients: Current Evidence and Future Applications

Julio Pascual, Marta Crespo, Jose Portoles, Carlos Jimenez, Alvaro Ortega-Carrion, Teresa Diez, Isabel Portero, Julio Pascual, Marta Crespo, Jose Portoles, Carlos Jimenez, Alvaro Ortega-Carrion, Teresa Diez, Isabel Portero

Abstract

Immunosuppressive drugs are widely used to prevent rejection after kidney transplantation. However, the pharmacological response to a given immunosuppressant can vary markedly between individuals, with some showing poor treatment responses and/or experiencing serious side effects. There is an unmet need for diagnostic tools that allow clinicians to individually tailor immunosuppressive therapy to a patient's immunological profile. The Immunobiogram (IMBG) is a novel blood-based in vitro diagnostic test that provides a pharmacodynamic readout of the immune response of individual patients to a range of immunosuppressants commonly used in kidney transplant recipients. Here, we discuss the current approaches used to measure the pharmacodynamic responses of individual patients to specific immunosuppressive drugs in vitro, which can then be correlated with patient's clinical outcomes. We also describe the procedure of the IMBG assay, and summarize the results obtained using the IMBG in different kidney transplant populations. Finally, we outline future directions and other novel applications of the IMBG, both in kidney transplant patients and other autoimmune diseases.

Keywords: cellular pharmacodynamics; immune cell assay; immunosuppressive therapy; infection; transplant rejection.

Conflict of interest statement

The institutional sponsor of the research studies performed with the IMBG is BIOHOPE, a start-up Biotech company based in Madrid (Spain) that received a competitive grant from the European Commission to perform some of these studies. A.O., T.D. and I.P. are employees at Biohope. The authors J.P. (Julio Pascual), M.C., J.P. (Jose Portoles) and C.J. are independent investigators who participated in the IMBG research studies, have no shares at BIOHOPE, and do not receive financial compensation from Biohope. Only common expenses (materials, working hours) that arose during these studies were charged to EU grants awarded to these research projects. The authors declare that this review was conducted in the absence of other third-party financial or commercial relationships that could be construed as a potential conflict of interest. The research mentioned involved the use of proprietary technologies belonging to BIOHOPE: patent “method for predicting and monitoring clinical response to immunomodulatory therapy”; co-Inventors, J.D., M.d.S., V.S., I.P.; (2018) European Patent; https://patentscope.wipo.int/search/es/detail.jsf?docId=WO2019002305&_cid=P21-LEQ1AF-12273-1 (accessed on 1 January 2023).

Figures

Figure 1
Figure 1
Mechanisms of action of immunosuppressive drugs on patient’s T cells.
Figure 2
Figure 2
IMBG assay procedure. PBMCs are extracted from the patient’s blood sample and immunologically stimulated to induce their activation and proliferation. These activated PBMCs are embedded in a hydrogel substrate, which is then loaded into segregated channels in the IMBG plate. PBMCs in each channel are exposed to a concentration gradient of a distinct immunosuppressant. Next, PBMC activation and proliferation along the concentration gradient are measured using a resazurin-based immunofluorescence assay, which provides a read-out of PBMC response to each immunosuppressant. For each immunosuppressant, dose–response curves are generated based on 15 immunofluorescence readings taken at sequential points along the concentration gradient in the IMBG channel.
Figure 3
Figure 3
Measurement of metabolic activity. The measurement is performed by means of ubiquitous reducing agents in cells (NADH and NADPH) with of a vital probe (resazurin). Resazurin is reduced via the aerobic respiration of metabolically active cells, changing resazurin to resorufin, a reaction that fluoresces when exposed to green light.
Figure 4
Figure 4
Immunobiogram dose–response curves obtained for a patient. Each immunosuppressive drug is tested in triplicate in the Immunobiogram plate, and the mean value of these measurements (IMS mean) is used to obtain the final curve for each immunosuppressant.

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