Gene, Immune and Cellular Responses to Single and Combined Space Flight Conditions - A (TripleLux-A) - 09.19.18

Overview | Description | Applications | Operations | Results | Publications | Imagery

ISS Science for Everyone

Science Objectives for Everyone
Long-term space missions present a number of risks for astronauts. Some effects of the space environment level appear to act at the cellular level and it is important to understand the underlying mechanisms of these effects. This project uses a rat macrophage cell line to focus on two aspects of cellular function which may have medical importance: i) The synergy between the effects of the space radiation environment and microgravity on cellular function and, ii) The impairment of immune functions under spaceflight conditions.
Science Results for Everyone
Cells can literally blow off bacteria. Oxidative burst reaction refers to rapid release of reactive oxygen, an immune response to prevent microbes from invading the body. When rat white blood cells were exposed to either microgravity or 1 g and the gravitational force then gradually increased or decreased, oxidative burst reaction rapidly declined after transition from 1 g to 0 g but was completely restored within 42 seconds in microgravity. These results show that mammalian white blood cells can adapt to microgravity very quickly, suggesting the cells have adaption mechanisms for surviving low gravity conditions. In turn, this implies that multicellular organisms can adapt to life in space.

The following content was provided by Oliver Ullrich, Ph.D., M.D, and is maintained in a database by the ISS Program Science Office.
Information provided courtesy of the Erasmus Experiment Archive.
Experiment Details


Principal Investigator(s)
Oliver Ullrich, Ph.D., M.D, University of Zurich, Zurich, Switzerland

Cora Thiel, Ph.D., University of Zurich, Zurich, Switzerland

Information Pending

Sponsoring Space Agency
European Space Agency (ESA)

Sponsoring Organization
Information Pending

Research Benefits
Earth Benefits, Scientific Discovery

ISS Expedition Duration
September 2014 - September 2015

Expeditions Assigned

Previous Missions
Information Pending

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Experiment Description

Research Overview
The aim of Triplelux-A is to understand the mechanisms at the cellular level which underlie the following phenomena previously observed in spaceflight:

  • Impairment of immune functions under spaceflight conditions.
  • Enhancement of responses to radiation in microgravity.
  • Clearly separate the effects of microgravity from other spaceflight factors by use of an onboard 1g centrifuge.
Specifically, the ability of rat macrophages to phagocytize zymosan (as an analogue of bacteria) is assessed. This process is the first line of defense against microbial infection. Phagocytosis is quantified using luminol as a detector for reactive oxygen species produced during phagocytosis of zymosan.

The Triplelux-A experiment uses the reactive oxygen burst as a measure of the phagocytic activity of macrophages and hemocytes under spaceflight conditions.

The reactive oxygen burst is measured by a chemiluminescent assay where Oxygen (O2)-radicals convert luminol to 3-Aminophthalate, which results in the emission of light at a wavelength of 475 nm. Light emission is enhanced by the addition of exogenous hydrogen peroxide. The luminol reaction is catalyzed by peroxidase.

The TRIPLELUX biosensor test is used to analyze cellular responses by a bioluminescent or chemiluminescent reporter. This is used for in-flight measurements of gene expression (SOS lux reporter) and phagocytosis. Using an onboard 1g centrifuge and a stepwise changing of g-level from 1g to microgravity, and vice versa, it is possible to determine whether changes in these processes are caused by microgravity, radiation, or a combination of both- as well as detecting a threshold at which immune cells can sense microgravity. Furthermore, the cellular adaptation to microgravity as well as the re-adaptation to 1g is to be analyzed.

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Space Applications
Conducting studies of the immune system during spaceflight provides knowledge and understanding of the effects of space habitation on the immune system. The data from these studies is used in assessing the cellular mechanisms underlying the aggravation of radiation responses, and impairment of immune functions, during spaceflight. Understanding such risks is essential in maintaining the health and performance of crew members during long-duration missions.

Earth Applications
With greater understanding of the immune system in space, scientists can determine new countermeasures for people suffering from weakened immune systems.

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Operational Requirements and Protocols
This experiment should be performed using the same batch of reagents and cells as the flight experiment, following the actual flight sequence. In case it is impossible to perform a near simultaneous ground reference, a postflight ground control duplicating the flight sequence is acceptable.
- Upload: Culture Tubes (with cells) and Luminol/Peroxidase/Zymosan Liquid reservoirs in NASA Cold Stowage (CS) (at -80°C). HM Interface plates (with culture medium) at ambient, max 7 days;
- On-orbit stowage in BIOLAB Temperature Control Unit at -20°C for the HM I/F plates (with culture medium) and the Liquid reservoirs (Back-up scenario: NASA Cold Stowage (CS));
- On-orbit stowage in NASA CS at -80°C for the Culture Tubes; Max stowage time of Culture Tubes in frozen stowage below -78°C on-orbit 4 months;
- The cells are thawed at ambient temperature (+18-28°C) for ~50 minutes, then the experiment is run at +37°C in BIOLAB. Following a preincubation (reconstitution) period of ~30 minutes, the cells are incubated for ~3 hours with a mixture of zymosan, peroxidase, and luminol. The chemiluminescence associated with the phagocytosis of the zymosan particles is measured. One set of samples is run under Zero-g and a second set under 1-g.

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Decadal Survey Recommendations

Information Pending

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Results/More Information

The TripleLux-A investigation studies the effects of microgravity on macrophage function. Macrophages are immune cells that engulf and break down microorganisms, cellular debris, and foreign particles. Macrophages are essential for cellular repair and overall healthy immune function, mainly due to their ability to perform a process called oxidative burst reaction, which causes chemical degradation of internalized particles, such as bacteria, that are a threat to the immune system. Results of this study showed that during the transition from a 1-g centrifuge to microgravity on the space station, the macrophages ability to generate oxidative burst reactions was reduced immediately, but subsequently recovered to their normal capabilities in less than a minute.
The “ultra-fast” adaptation indicates that mammalian macrophages are highly adaptive to a low-gravity environment; therefore, key cellular functions of multicellular life could adapt to and exist in a low-gravity environment, enabling long-duration space exploration beyond low-Earth orbit.

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Results Publications

    Thiel CS, de Zelicourt D, Tauber S, Adrian A, Franz M, Simmet DM, Schoppmann K, Hauschild S, Krammer S, Christen M, Bradacs G, Paulsen K, Wolf SA, Braun M, Hatton JP, Kurtcuoglu V, Franke S, Tanner S, Cristoforetti S, Sick B, Hock B, Ullrich O.  Rapid adaptation to microgravity in mammalian macrophage cells. Scientific Reports. 2017 February 27; 7(1): 13 pp. DOI: 10.1038/s41598-017-00119-6.

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Ground Based Results Publications

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ISS Patents

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Related Publications

    Rabbow E, Rettberg P, Baumstark-Khan C, Horneck G.  The SOS-LUX-LAC-FLUORO-Toxicity-test on the International Space Station (ISS). Advances in Space Research. 2003; 31(6): 1513-1524. DOI: 10.1016/S0273-1177(03)00086-3.

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Related Websites

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image The Advanced Experimental Containment (AEC) hardware for the TripleLux experiments. Image courtesy of ESA.
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image NASA Image: ISS043E167915 - View of the Triplelux-A experiment hardware.
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image NASA Image: ISS043E167919 - View of the Triplelux-A experiment hardware.
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image NASA Image: ISS043E152043 - Photos documenting the Gene, Immune and Cellular Responses to Single and Combined Space Flight Conditions – A (Triplelux-A) experiment.
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image NASA Image: ISS043E181041 - European Space Agency (ESA) astronaut Samantha Cristoforetti prepares the TripleLux-A experiment for return on SpaceX’s Dragon cargo craft.
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image NASA Image: ISS043E181042 - European Space Agency (ESA) astronaut Samantha Cristoforetti prepares the TripleLux-A experiment for return on SpaceX’s Dragon cargo craft.
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image NASA Image: ISS043E181153 - Photographic documentation of closeout activities for the Triplelux-A experiment. The Experiment Container (EC) was unstowed and removed from Biolab, then the EC Handling Mechanism (HM) Interface was also unstowed and documented. Triplelux-A uses a rat macrophage cell line to investigate and compare the ability of macrophages to kill pathogens under normal gravity and microgravity conditions. The goal is to gain a better understanding of immune suppression in spaceflight.
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