Bacterial expression of a designed single-chain IL-10 prevents severe lung inflammation

Montero-Blay, Ariadna, 1994-; Delgado Blanco, Javier; Rodriguez-Arce, Irene; Lastrucci, Claire; Piñero-Lambea, Carlos; Lluch-Senar, Maria 1982-; Serrano Pubull, Luis, 1982-

doi:http://dx.doi.org/10.15252/msb.202211037

Bacterial expression of a designed single-chain IL-10 prevents severe lung inflammation

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Montero-Blay A, Blanco JD, Rodriguez-Arce I, Lastrucci C, Piñero-Lambea C, Lluch-Senar M, Serrano L. Bacterial expression of a designed single-chain IL-10 prevents severe lung inflammation. Mol Syst Biol. 2023 Jan;19(1):e11037. DOI: 10.15252/msb.202211037

Abstract

Interleukin-10 (IL-10) is an anti-inflammatory cytokine that is active as a swapped domain dimer and is used in bacterial therapy of gut inflammation. IL-10 can be used as treatment of a wide range of pulmonary diseases. Here we have developed a non-pathogenic chassis (CV8) of the human lung bacterium Mycoplasma pneumoniae (MPN) to treat lung diseases. We find that IL-10 expression by MPN has a limited impact on the lung inflammatory response in mice. To solve these issues, we rationally designed a single-chain IL-10 (SC-IL10) with or without surface mutations, using our protein design software (ModelX and FoldX). As compared to the IL-10 WT, the designed SC-IL10 molecules increase the effective expression in MPN four-fold, and the activity in mouse and human cell lines between 10 and 60 times, depending on the cell line. The SC-IL10 molecules expressed in the mouse lung by CV8 in vivo have a powerful anti-inflammatory effect on Pseudomonas aeruginosa lung infection. This rational design strategy could be used to other molecules with immunomodulatory properties used in bacterial therapy.