Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibody
Abstract Background Dysregulated production of interleukin (IL)-6 is implicated in the pathology of inflammatory bowel disease (IBD). Neutralization of IL-6 in the gut by safe probiotic bacteria may help alleviate intestinal inflammation. Here, we developed Lactococcus lactis with potent and selecti...
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| Format: | Article |
| Language: | English |
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BMC
2022-07-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-022-01873-7 |
| _version_ | 1811218289324457984 |
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| author | Abida Zahirović Aleš Berlec |
| author_facet | Abida Zahirović Aleš Berlec |
| author_sort | Abida Zahirović |
| collection | DOAJ |
| description | Abstract Background Dysregulated production of interleukin (IL)-6 is implicated in the pathology of inflammatory bowel disease (IBD). Neutralization of IL-6 in the gut by safe probiotic bacteria may help alleviate intestinal inflammation. Here, we developed Lactococcus lactis with potent and selective IL-6 binding activity by displaying IL-6-specific affibody on its surface. Results Anti-IL-6 affibody (designated as ZIL) was expressed in fusion with lactococcal secretion peptide Usp45 and anchoring protein AcmA. A high amount of ZIL fusion protein was detected on bacterial surface, and its functionality was validated by confocal microscopy and flow cytometry. Removal of IL-6 from the surrounding medium by the engineered L. lactis was evaluated using enzyme-linked immunosorbent assay. ZIL-displaying L. lactis sequestered recombinant human IL-6 from the solution in a concentration-dependent manner by up to 99% and showed no binding to other pro-inflammatory cytokines, thus proving to be highly specific for IL-6. The removal was equally efficient across different IL-6 concentrations (150–1200 pg/mL) that were found to be clinically relevant in IBD patients. The ability of engineered bacteria to capture IL-6 from cell culture supernatant was assessed using immunostimulated human monocytic cell lines (THP-1 and U-937) differentiated into macrophage-like cells. ZIL-displaying L. lactis reduced the content of IL-6 in the supernatants of both cell lines in a concentration-dependent manner by up to 94%. Dose response analysis showed that bacterial cell concentrations of 107 and 109 CFU/mL (colony forming units per mL) were required for half-maximal removal of recombinant and macrophage-derived IL-6, respectively. Conclusion The ability of ZIL-displaying L. lactis to bind pathological concentrations of IL-6 at common bacterial doses suggests physiological significance. |
| first_indexed | 2024-04-12T07:07:18Z |
| format | Article |
| id | doaj.art-7b0ee63e1bb94de39a71fef6ecb00694 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-12T07:07:18Z |
| publishDate | 2022-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-7b0ee63e1bb94de39a71fef6ecb006942022-12-22T03:42:45ZengBMCMicrobial Cell Factories1475-28592022-07-0121111510.1186/s12934-022-01873-7Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibodyAbida Zahirović0Aleš Berlec1Department of Biotechnology, Jožef Stefan InstituteDepartment of Biotechnology, Jožef Stefan InstituteAbstract Background Dysregulated production of interleukin (IL)-6 is implicated in the pathology of inflammatory bowel disease (IBD). Neutralization of IL-6 in the gut by safe probiotic bacteria may help alleviate intestinal inflammation. Here, we developed Lactococcus lactis with potent and selective IL-6 binding activity by displaying IL-6-specific affibody on its surface. Results Anti-IL-6 affibody (designated as ZIL) was expressed in fusion with lactococcal secretion peptide Usp45 and anchoring protein AcmA. A high amount of ZIL fusion protein was detected on bacterial surface, and its functionality was validated by confocal microscopy and flow cytometry. Removal of IL-6 from the surrounding medium by the engineered L. lactis was evaluated using enzyme-linked immunosorbent assay. ZIL-displaying L. lactis sequestered recombinant human IL-6 from the solution in a concentration-dependent manner by up to 99% and showed no binding to other pro-inflammatory cytokines, thus proving to be highly specific for IL-6. The removal was equally efficient across different IL-6 concentrations (150–1200 pg/mL) that were found to be clinically relevant in IBD patients. The ability of engineered bacteria to capture IL-6 from cell culture supernatant was assessed using immunostimulated human monocytic cell lines (THP-1 and U-937) differentiated into macrophage-like cells. ZIL-displaying L. lactis reduced the content of IL-6 in the supernatants of both cell lines in a concentration-dependent manner by up to 94%. Dose response analysis showed that bacterial cell concentrations of 107 and 109 CFU/mL (colony forming units per mL) were required for half-maximal removal of recombinant and macrophage-derived IL-6, respectively. Conclusion The ability of ZIL-displaying L. lactis to bind pathological concentrations of IL-6 at common bacterial doses suggests physiological significance.https://doi.org/10.1186/s12934-022-01873-7Inflammatory bowel diseaseIL-6MicrobiotaLactococcus lactisDelivery system |
| spellingShingle | Abida Zahirović Aleš Berlec Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibody Microbial Cell Factories Inflammatory bowel disease IL-6 Microbiota Lactococcus lactis Delivery system |
| title | Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibody |
| title_full | Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibody |
| title_fullStr | Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibody |
| title_full_unstemmed | Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibody |
| title_short | Targeting IL-6 by engineered Lactococcus lactis via surface-displayed affibody |
| title_sort | targeting il 6 by engineered lactococcus lactis via surface displayed affibody |
| topic | Inflammatory bowel disease IL-6 Microbiota Lactococcus lactis Delivery system |
| url | https://doi.org/10.1186/s12934-022-01873-7 |
| work_keys_str_mv | AT abidazahirovic targetingil6byengineeredlactococcuslactisviasurfacedisplayedaffibody AT alesberlec targetingil6byengineeredlactococcuslactisviasurfacedisplayedaffibody |