Development of a Mixed Lymphocyte Reaction (MLR) assay for evaluating the allogenicity of cell therapy

Cell therapies represent a promising class of treatments with the potential to address many incurable diseases through unique and powerful mechanisms of action.

Despite recent successes, these therapies still encounter significant challenges that impede their widespread application and commercialisation. Key obstacles include the identification of suitable cell sources and the generation of a sufficiently viable product.

In contrast to autologous cell therapies, which utilise cells derived from the patient, allogeneic cell therapies offer the advantage of being «off-the-shelf» treatments. These therapies, including induced pluripotent stem cells (iPSCs), mesenchymal stem cell-based therapies (MSCs), and CAR T cells, can be produced from a single donor or a limited pool of donors, making them accessible to multiple patients. However, the use of allogeneic products necessitates HLA-compatible donors and immune suppression, and they carry significant risks such as graft-versus-host disease. Consequently, both the FDA and EMA mandate specific tests to demonstrate the allo-compatibility of allogeneic treatments.

This application note describes several methods for efficiently assessing lymphocyte proliferation upon non-specific T cell stimulation and allogeneic recognition between non-HLA-matched, randomly selected PBMCs from healthy donors.