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Immunotherapy-on-Chip Against an Experimental Sepsis Model

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Abstract—

Lipopolysaccharide (LPS) is commonly used in murine sepsis models, which are largely associated with immunosuppression and collapse of the immune system. After adapting the LPS treatment to the needs of locally bred BALB/c mice, the present study explored the protective role of Micrococcus luteus peptidoglycan (PG)–pre-activated vaccine-on-chip technology in endotoxemia. The established protocol consisted of five daily intraperitoneal injections of 0.2 μg/g LPS, allowing longer survival, necessary for a therapeutic treatment application. A novel immunotherapy technology, the so-called vaccine-on-chip, consists of a 3-dimensional laser micro-textured silicon (Si) scaffold loaded with macrophages and activated in vitro with 1 μg/ml PG, which has been previously shown to exert a mild immunostimulatory activity upon subcutaneous implantation. The LPS treatment significantly decreased CD4 + and CD8 + cells, while increasing CD11b + , Gr1 + , CD25 + , Foxp3 + , and class II + cells. These results were accompanied by increased arginase-1 activity in spleen cell lysates and C-reactive protein (CRP), procalcitonin (PCT), IL-6, TNF-a, IL-10, and IL-18 in the serum, while acquiring severe sepsis phenotype as defined by the murine sepsis scoring. The in vivo application of PG pre-activated implant significantly increased the percentage of CD4 + and CD8 + cells, while decreasing the percentage of Gr1 + , CD25 + , CD11b + , Foxp3 + cells, and arginase-1 activity in the spleen of LPS-treated animals, as well as all serum markers tested, allowing survival and rescuing the severity of sepsis phenotype. In conclusion, these results reveal a novel immunotherapy technology based on PG pre-activated micro-texture Si scaffolds in LPS endotoxemia, supporting thus its potential use in the treatment of septic patients.

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Acknowledgements

We thank Dr Emmanuel Stratakis, Research Director at the Institute of Electronic Structure and Laser (IESL-FORTH), and Anna Karayiannaki for providing Si scaffolds.

Funding

This work was co-financed by Greece and the European Union (European Social Fund- ESF) through the Operational Programme «Human Resources Development, Education and Lifelong Learning 2014–2020» in the context of the project “Vaccine-on-chip against sepsis” (MIS 5048426).

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Authors and Affiliations

  1. Department of Biology, University of Crete, Heraklion, Crete, Greece

    Zerva Ioanna, Bakela Katerina & Athanassakis Irene

  2. Department of Biology, University of Crete, Heraklion, Crete, Greece

    Athanassakis Irene

Authors
  1. Zerva Ioanna
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  2. Bakela Katerina
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  3. Athanassakis Irene
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Contributions

All authors contributed to the study. Conceptualization, data analysis, writing, funding acquisition, and supervision were performed by Irene Athanassakis. Material preparation and data collection were performed by Ioanna Zerva and Katerina Bekela (equal contribution). All the authors read and approved the final manuscript.

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Correspondence to Athanassakis Irene.

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The applied protocol was approved by the national Bioethical Committee (Approval # 292314, ADA ΨΗΘ47ΛΚ-Γ5Ψ).

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The authors declare no competing interests.

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Ioanna, Z., Katerina, B. & Irene, A. Immunotherapy-on-Chip Against an Experimental Sepsis Model. Inflammation 44, 2333–2345 (2021). https://doi.org/10.1007/s10753-021-01506-y

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