Extracorporeal immune cell therapy of sepsis: ex vivo results

Gerd Klinkmann; Thomas Wild; Benjamin Heskamp; Fanny Doss; Sandra Doss; Lubomir Arseniev; Krasimira Aleksandrova; Martin Sauer; Daniel A Reuter; Steffen Mitzner; Jens Altrichter

doi:10.1186/s40635-022-00453-8

Extracorporeal immune cell therapy of sepsis: ex vivo results

Intensive Care Med Exp. 2022 Jun 16;10(1):26. doi: 10.1186/s40635-022-00453-8.

Authors

Gerd Klinkmann^{1

2}, Thomas Wild³, Benjamin Heskamp³, Fanny Doss³, Sandra Doss^{3

4}, Lubomir Arseniev⁵, Krasimira Aleksandrova⁵, Martin Sauer⁶, Daniel A Reuter⁷, Steffen Mitzner^{8

4}, Jens Altrichter³

Affiliations

¹ Department of Anaesthesiology and Intensive Care Medicine, University of Rostock, Schillingallee 35, 18055, Rostock, Germany. gerd.klinkmann@med.uni-rostock.de.
² Department of Anesthesiology and Intensive Care Medicine, University Medical Center Rostock, Schillingallee 35, 18057, Rostock, Germany. gerd.klinkmann@med.uni-rostock.de.
³ ARTCLINE GmbH, Schillingallee 68, 18057, Rostock, Germany.
⁴ Department of Extracorporeal Therapy Systems, Fraunhofer Institute for Cell Therapy and Immunology, Schillingallee 68, 18057, Rostock, Germany.
⁵ Cellular Therapy Centre (CTC), Medizinische Hochschule Hannover, Feodor-Lynen-Str. 21, 30625, Hannover, Germany.
⁶ Center for Anesthesiology and Intensive Care Medicine, Hospital of Magdeburg, Birkenallee 34, 39130, Magdeburg, Germany.
⁷ Department of Anaesthesiology and Intensive Care Medicine, University of Rostock, Schillingallee 35, 18055, Rostock, Germany.
⁸ Division of Nephrology, Department of Medicine, Medical Faculty, University of Rostock, Ernst-Heydemann-Str. 6, 18057, Rostock, Germany.

Abstract

Background: Immune cell dysfunction plays a central role in sepsis-associated immune paralysis. The transfusion of healthy donor immune cells, i.e., granulocyte concentrates (GC) potentially induces tissue damage via local effects of neutrophils. Initial clinical trials using standard donor GC in a strictly extracorporeal bioreactor system for treatment of septic shock patients already provided evidence for beneficial effects with fewer side effects, by separating patient and donor immune cells using plasma filters. In this ex vivo study, we demonstrate the functional characteristics of a simplified extracorporeal therapy system using purified granulocyte preparations.

Methods: Purified GC were used in an immune cell perfusion model prefilled with human donor plasma simulating a 6-h treatment. The extracorporeal circuit consisted of a blood circuit and a plasma circuit with 3 plasma filters (PF). PF1 is separating the plasma from the patient's blood. Plasma is then perfused through PF2 containing donor immune cells and used in a dead-end mode. The filtrated plasma is finally retransfused to the blood circuit. PF3 is included in the plasma backflow as a redundant safety measure. The donor immune cells are retained in the extracorporeal system and discarded after treatment. Phagocytosis activity, oxidative burst and cell viability as well as cytokine release and metabolic parameters of purified GCs were assessed.

Results: Cells were viable throughout the study period and exhibited well-preserved functionality and efficient metabolic activity. Course of lactate dehydrogenase and free hemoglobin concentration yielded no indication of cell impairment. The capability of the cells to secret various cytokines was preserved. Of particular interest is equivalence in performance of the cells on day 1 and day 3, demonstrating the sustained shelf life and performance of the immune cells in the purified GCs.

Conclusion: Results demonstrate the suitability of a simplified extracorporeal system. Furthermore, granulocytes remain viable and highly active during a 6-h treatment even after storage for 3 days supporting the treatment of septic patients with this system in advanced clinical trials.

Keywords: Clinical Use; Concentrate; Extracorporeal; Granulocyte; Sepsis; Therapy.