Monitoring functional immune responses with a cytokine release assay: ISS flight hardware design and experimental protocol for whole blood cultures executed under microgravity conditions

Judith-Irina Buchheim; Matthias Feuerecker; Michele Balsamo; Marco Vukich; Merel Van Walleghem; Kevin Tabury; Roel Quintens; Randy Vermeesen; Bjorn Baselet; Sarah Baatout; Bernd Rattenbacher; Inês Antunes; Thu Jennifer Ngo-Anh; Brian Crucian; Alexander Choukér

doi:10.3389/fphys.2023.1322852

Monitoring functional immune responses with a cytokine release assay: ISS flight hardware design and experimental protocol for whole blood cultures executed under microgravity conditions

Front Physiol. 2024 Jan 15:14:1322852. doi: 10.3389/fphys.2023.1322852. eCollection 2023.

Authors

Judith-Irina Buchheim¹, Matthias Feuerecker¹, Michele Balsamo², Marco Vukich^{2

3}, Merel Van Walleghem^{4

5}, Kevin Tabury⁵, Roel Quintens⁵, Randy Vermeesen⁵, Bjorn Baselet⁵, Sarah Baatout⁵, Bernd Rattenbacher⁶, Inês Antunes³, Thu Jennifer Ngo-Anh³, Brian Crucian⁷, Alexander Choukér¹

Affiliations

¹ Laboratory of Translational Research "Stress and Immunity", Department of Anesthesiology, LMU University Hospital, LMU Munich, Munich, Germany.
² Kayser Italia S.r.l, Livorno, Italy.
³ European Space Research and Technology Centre (ESTEC), European Space Agency (ESA), Noordwijk, Netherlands.
⁴ European Astronaut Center (EAC), European Space Agency (ESA), Cologne, Germany.
⁵ Belgian Nuclear Research Centre (SCK CEN), Radiobiology Unit, Nuclear Medical Application Institute, Mol, Belgium.
⁶ Biotechnology Space Support Center (Biotesc), Lucerne University of Applied Sciences and Arts (HSLU), Luzerne, Switzerland.
⁷ Immunology Lab, NASA Johnsons Space Center, Houston, TX, United States.

Abstract

Introduction: Long-term space missions trigger a prolonged neuroendocrine stress response leading to immune system dysregulation evidenced by susceptibility to infections, viral reactivation, and skin irritations. However, due to existing technical constraints, real-time functional immune assessments are not currently available to crew inflight. The in vitro cytokine release assay (CRA) has been effectively employed to study the stimulated cytokine response of immune cells in whole blood albeit limited to pre- and post-flight sessions. A novel two-valve reaction tube (RT) has been developed to enable the execution of the CRA on the International Space Station (ISS). Methods: In a comprehensive test campaign, we assessed the suitability of three materials (silicone, C-Flex, and PVC) for the RT design in terms of biochemical compatibility, chemical stability, and final data quality analysis. Furthermore, we thoroughly examined additional quality criteria such as safety, handling, and the frozen storage of antigens within the RTs. The validation of the proposed crew procedure was conducted during a parabolic flight campaign. Results: The selected material and procedure proved to be both feasible and secure yielding consistent and dependable data outcomes. This new hardware allows for the stimulation of blood samples on board the ISS, with subsequent analysis still conducted on the ground. Discussion: The resultant data promises to offer a more accurate understanding of the stress-induced neuroendocrine modulation of immunity during space travel providing valuable insights for the scientific community. Furthermore, the versatile nature of the RT suggests its potential utility as a testing platform for various other assays or sample types.

Keywords: astronaut health; functional immune testing; immune monitoring; in vitro cytokine release assay; long-term spaceflight; neuroendocrine stress response.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work has been supported by the DLR on behalf of the Federal Ministry of Economics and Technology (BMWi 50WB0719 and 50WB0919, 50WB1319, 50WB2222, 50WB2026), the European Space Agency (ESA) (ELIPS 3 and 4, and SciSpacE programs) as well as the Belgian Science Policy Office [BELSPO/ESA PRODEX grant numbers PEA_4000140806 and CO-90-11-2801-04 (IMPULSE)] and by the Medical and Clinician Scientist Program (MCSP) of the LMU Munich, Germany. Initial development of the whole blood assay and prototype flight hardware was funded by the NASA Johnson Space Center, Human Research Program, Human Health and Countermeasures Element.