Logo image
Back
Temporal Changes in Innate and Adaptive Immunity During Sepsis as Determined by ELISpot
Preprint   Open access

Temporal Changes in Innate and Adaptive Immunity During Sepsis as Determined by ELISpot

J Unsinger, D Osborne, A H Walton, E Han, L Sheets, M B Mazer, K E Remy, T S Griffith, M Rao, V P Badovinac, …
bioRxiv : the preprint server for biology
12/14/2023
DOI: 10.1101/2023.12.14.571668
PMCID: PMC10760123
PMID: 38168302
url
https://doi.org/10.1101/2023.12.14.571668 View
Preprint (Author's original)This preprint has not been evaluated by subject experts through peer review. Preprints may undergo extensive changes and/or become peer-reviewed journal articles. Open Access

Abstract

The inability to evaluate host immunity in a rapid quantitative manner in patients with sepsis has severely hampered development of novel immune therapies. The ELISpot assay is a bioassay that measures the number of cytokine-secreting cells and the relative amount of cytokine produced at the single-cell level. A key advantage of ELISpot is its excellent dynamic range enabling a more precise quantifiable assessment of host immunity. Herein, we tested the hypothesis on whether the ELISpot assay can detect dynamic changes in both innate and adaptive immunity as they often occur during sepsis. We also tested whether ELISpot could detect the effect of immune drug therapies to modulate innate and adaptive immunity. Mice were made septic using sublethal cecal ligation and puncture (CLP). Blood and spleens were harvested serially and IFN-γ and TNF-α production were compared by ELISpot and ELISA. The capability of ELISpot to detect changes in innate and adaptive immunity due to immune therapy with dexamethasone, IL-7, and arginine was also evaluated. ELISpot confirmed a decreased innate and adaptive immunity responsiveness during sepsis progression. More importantly, ELISpot was also able to detect changes in adaptive and innate immunity in response to immune-modulatory reagents, for example dexamethasone, arginine, and IL-7 in a readily quantifiable manner, as predicted by the reagents known mechanisms of action. ELISpot and ELISA results tended to parallel one another although some differences were noted. ELISpot offers a unique capability to assess the functional status of both adaptive and innate immunity over time. The results presented herein demonstrate that ELISpot can also be used to detect and follow the effects of drugs to ameliorate sepsis-induced immune dysfunction. This capability would be a major advance in guiding new immune therapies in sepsis.

Details

Metrics

15 Record Views
Logo image