Alveolar Macrophage based personalized therapy for the delivery of IL-37

Alveolar Macrophages (AM) are self-renewing, tissue resident immune cells in the lung. Interleukin 37 has anti-inflammatory properties to prevent hyper-inflammation and tissue damage. IL-37 has been shown to counteract asthma or cigarette smoke induced inflammation. The cytokine downregulates allergic airway inflammation by compensating for disease-enhancing effects of IL-1β and IL-33. The aim of this work is the development of a cell-based personalized therapy with AMs that have been modified by adeno-associated virus (AAV) to express IL-37. They will be re-transplanted into the lung in different disease models of asthma and COPD to mitigate the inflammatory phenotype. Via in silico methods primers were designed to clone the open reading frame of IL-37 by PCR. The DNA-Fragment was sequenced and inserted into an expression plasmid. The gene expression was confirmed by real-time PCR. By western blot protein production was verified to prove the function of the plasmid. AAV serotype 2, 6 and 9 were built, first with GFP to test the function of the virus, and later with the IL-37 expressing plasmid. By real-time PCR, microscopy and ELISA successful transduction and AAV function was proven as a first step in a cell line. AMs were isolated from mice and the cell type was verified by FACS. The cells were cultured to increase the cell number. By confocal microscopy was shown that AM can be successfully transduced by AAV. In summary AMs are an easily recovered cell type. The cells can stably express a transgene and produce a prolonged therapeutic effect. The advantage of the therapy is the absence of an immune response, as the therapy is carried out with the patient's own cells.