Abstract
Studying innate immunity in humans is crucial for understanding its role in the pathophysiology of systemic inflammation, particularly in the complex setting of sepsis. Therefore, we standardized a step-by-step process from the venipuncture to the transfer in a human model system, while closely monitoring the inflammatory response for up to three hours. We designed an animal-free, human whole blood sepsis model using a commercially available, simple to use, tubing system. First, we analyzed routine clinical parameters, including cell count and blood gas analysis. Second, we demonstrated that extracellular activation markers (e.g., CD11b and CD62l) as well as intracellular metabolic (intracellular pH) and functional (generation of radical oxygen species) features remained stable after incubation in the whole blood model. Third, we mimicked systemic inflammation during early sepsis by exposure of whole blood to pathogen-associated molecular patterns. Stimulation with lipopolysaccharide revealed the capability of the model system to evoke a sepsis-like inflammatory phenotype of innate immunity. In summary, the presented model serves as a convenient, economic, and reliable platform to study innate immunity in human whole blood, which may yield clinically important insights.
Highlights
Innate immunity plays a vital role as the vanguard against numerous harmful carriers of damage- and pathogen associated molecular patterns (DAMPs and PAMPs, respectively)
The blood system is subjected to various alterations because the interaction with other organ systems is interrupted while in parallel the continuous contact and interaction with the intact endothelium is lost
The ex vivo blood is exposed to artificial surfaces, resulting in activation of thromboinflammation [32]
Summary
Innate immunity plays a vital role as the vanguard against numerous harmful carriers of damage- and pathogen associated molecular patterns (DAMPs and PAMPs, respectively). To study the complex innate immune response to PAMP exposure, various small or large, simple or complex animal models are used [4,5,6]. The exact exposure time and the amount of pathogens within the blood during sepsis cannot be determined with certainty. As another alternative, injection of immune-stimulants, including lipopolysaccharide (LPS), has been performed and investigated in healthy human volunteers [8,9,10]. Because excessive inflammation during sepsis remains a major clinical and scientific challenge, there is an unabated research need to elucidate the underlying immunological pathomechanisms, for example, by utilizing human whole blood in a standardized and reliable manner
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