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. Epub 2023 Jan 4.


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.

Keywords: infection; interleukin; live biotherapeutics; mycoplasma; protein engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteria
  • Cytokines
  • Humans
  • Inflammation / pathology
  • Interleukin-10* / genetics
  • Lung
  • Mice
  • Pneumonia* / pathology
  • Pneumonia* / prevention & control
  • Pseudomonas aeruginosa


  • Interleukin-10
  • Cytokines

Associated data

  • PDB/1y6k
  • PDB/2ilk
  • PDB/1j7v
  • PDB/6x93
  • PDB/2wyu