Jean-Pol Frippiat

Could spaceflight-associated immune system weakening preclude the expansion of human presence beyond Earth’s orbit?

This year, we celebrate the 40th birthday of the first landing of humans on the moon. By 2020, astronauts should return to the lunar surface and establish an outpost there that will provide a technical basis for future manned missions to Mars. This paper summarizes major constraints associated with a trip to Mars, presents immunological hazards associated with this type of mission, and shows that our current understanding of the immunosuppressive effects of spaceflight is limited. Weakening of the immune system associated with spaceflight is therefore an area that should be considered more thoroughly before we undertake prolonged space voyages.

Stress response and humoral immune system alterations related to chronic hypergravity in mice

Spaceflights are known to induce stress and immune dysregulation. Centrifugation, as hindlimb unloading, is a good ground based-model to simulate altered gravity which occurs during space missions. The aim of this study was to investigate the consequences of a long-term exposure to different levels of hypergravity on the stress response and the humoral immunity in a mouse model. For this purpose, adult C57Bl/6J male mice were subjected for 21 days either to control conditions or to 2G or 3G acceleration gravity forces. Corticosterone level and anxiety behavior revealed a stress response which was associated with a decrease of body weight, after 21-day of centrifugation at 3G but not at 2G. Spleen lymphocyte lipopolysaccharide (LPS) responsiveness was diminished by 40% in the 2G group only, whereas a decrease was noted when cells were stimulated with concanavalin A for both 2G and 3G groups (about 25% and 20%, respectively) compared to controls. Pro-inflammatory chemokines (MCP-1 and IP-10) and Th1 cytokines (IFNγ and IL2) were slightly decreased in the 2G group and strongly decreased in the 3G mouse group. Regarding Th2 cytokines (IL4, IL5) no further significant modification was observed, whereas the immunosuppressive cytokine IL10 was slightly increased in the 3G mice. Finally, serum IgG concentration was twice higher whereas IgA concentration was slightly increased (about 30%) and IgM were unchanged in 2G mice compared to controls. No difference was observed in the 3G group with these isotypes. Consequently, functional immune dysregulations and stress responses were dependent of the gravity level.

Spaceflight-associated changes in immunoglobulin VH gene expression in the amphibian Pleurodeles waltl

Understanding why the immune system is depressed during spaceflight is of obvious importance for future human deep‐space missions, such as the foreseen missions to Mars. However, little is known about the effects of these flights on humoral immunity. We previously immunized adult Pleurodeles waltl (urodele amphibian) onboard the Mir space station and showed that heavy‐chain variable (VH) domains of specific IgM antibodies are encoded by genes belonging to the VHII and VHVI families. We have now determined how these animals use their individual VHII and VHVI genes by screening IgM heavy‐chain cDNA libraries and by quantifying IgM heavy‐chain transcripts encoded by these genes. Results were compared with those obtained using control animals immunized on Earth under the same conditions as onboard Mir. Our experiments revealed an increase in the expression of IgM heavy‐chain mRNAs encoded by the VHII and VHVI.C genes and a strong decrease in the expression of IgM heavy‐chain mRNAs encoded by the VHVI.A and VHVI.B genes in spaceflight animals. Consequently, different heavy‐chain mRNAs are expressed by spaceflight animals, demonstrating that this environment affects the humoral response. These observations may be due to a change in B‐cell selection under space flight conditions.— Bascove, M.,Huin‐Schohn, C., Gueguinou, N., Tschirhart, E., Frippiat, J.‐P. Spaceflight‐associated changes in immunoglobulin VH gene expression in the amphibian Pleurodeles waltl. FASEB J. 23, 1607–1615 (2009)

Gravity changes during animal development affect IgM heavy-chain transcription and probably lymphopoiesis

Our previous research demonstrated that spaceflight conditions affect antibody production in response to an antigenic stimulation in adult amphibians. Here, we investigated whether antibody synthesis is affected when animal development occurs onboard a space station. To answer this question, embryos of the Iberian ribbed newt, Pleurodeles waltl, were sent to the International Space Station (ISS) before the initiation of immunoglobulin heavy‐chain expression. Thus, antibody synthesis began in space. On landing, we determined the effects of spaceflight on P. waltl development and IgM heavy‐chain transcription. Results were compared with those obtained using embryos that developed on Earth. We find that IgM heavy‐chain transcription is doubled at landing and that spaceflight does not affect P. waltl development and does not induce inflammation. We also recreated the environmental modifications encountered by the embryos during their development onboard the ISS. This strategy allowed us to demonstrate that gravity change is the factor responsible for antibody heavy‐chain transcription modifications that are associated with NF‐κB mRNA level variations. Taken together, and given that the larvae were not immunized, these data suggest a modification of lymphopoiesis when gravity changes occur during ontogeny.—Huin‐Schohn, C., Guéguinou, N., Schenten, V., Bascove, M., Koch, G. G., Baatout, S., Tschirhart, E., Frippiat, J.‐P. Gravity changes during animal development affect IgM heavy‐chain transcription and probably lymphopoiesis. FASEB J. 27, 333–341 (2013). www.fasebj.org

Molecular characterization of Pleurodeles waltl activation-induced cytidine deaminase

Activation-induced cytidine deaminase (AID) is involved in immunoglobulin affinity maturation, gene conversion and class switch recombination. This protein is therefore a major actor in the creation of the antibody repertoire. We have isolated, for the first time, the AID mRNA from a urodele amphibian, Pleurodeles waltl. This mRNA encodes 198 amino acids and shares 70% and 76% of similarity with Xenopus laevis and human AID sequences, respectively. All consensus motifs necessary for AID functions are present, suggesting that AID is functional in P. waltl. P. waltl AID is encoded by five exons as in other species. However, in contrast to mammalian AID, no splice variant could be detected in that species. We also noted that AID is predominantly expressed in the spleen, the major secondary lymphoid organ of P. waltl, and that the transcriptional regulation of P. waltl AID is partially different from that found in higher vertebrates. Furthermore, we showed that AID is expressed early during P. waltl embryonic development.

iPLA2, a novel determinant in Ca2+- and phosphorylation-dependent S100A8/A9 regulated NOX2 activity.

The neutrophil NADPH oxidase (NOX2) is a key enzyme responsible for host defense against invading pathogens, via the production of reactive oxygen species. Dysfunction of NOX2 can contribute to inflammatory processes, which could lead to the development of diseases such as atherosclerosis. In this paper, we characterize a pathway leading to NOX2 activation in which iPLA(2)-regulated p38 MAPK activity is a key regulator of S100A8/A9 translocation via S100A9 phosphorylation. Studies in cell-free or recombinant systems involved two Ca2+-binding proteins of the S100 family, namely S100A8 and S100A9, in NOX2 activation dependent on intracellular Ca2+ concentration ([Ca2+](i)) elevation. Using differentiated HL-60 cells as a model of neutrophils, we provide evidence that [Ca2+](i)-regulated S100A8/A9 translocation is mediated by an increase in [Ca2+](i) through intracellular Ca2+ store depletion. Moreover, we confirm that p38 MAPK induces S100A9 phosphorylation, a mandatory precondition for S100 translocation. Based on a pharmacological approach and an siRNA strategy, we identify iPLA(2) as a new molecular player aiding S100 translocation and NOX2 activity. Inhibition of p38 MAPK activity and S100A9 phosphorylation by bromoenol lactone, a selective inhibitor of iPLA(2), indicated that p38 MAPK-mediated S100A9 phosphorylation is dependent on iPLA(2). In conclusion, we have characterized a pathway leading to NOX2 activation in which iPLA(2)-regulated p38 MAPK activity is a key regulator of S100A8/A9 translocation via S100A9 phosphorylation.

Towards human exploration of space: The THESEUS review series on immunology research priorities

Dysregulation of the immune system occurs during spaceflight and may represent a crew health risk during exploration missions because astronauts are challenged by many stressors. Therefore, it is crucial to understand the biology of immune modulation under spaceflight conditions in order to be able to maintain immune homeostasis under such challenges. In the framework of the THESEUS project whose aim was to develop an integrated life sciences research roadmap regarding human space exploration, experts working in the field of space immunology, and related disciplines, established a questionnaire sent to scientists around the world. From the review of collected answers, they deduced a list of key issues and provided several recommendations such as a maximal exploitation of currently available resources on Earth and in space, and to increase increments duration for some ISS crew members to 12 months or longer. These recommendations should contribute to improve our knowledge about spaceflight effects on the immune system and the development of countermeasures that, beyond astronauts, could have a societal impact.

Considerations for Preventive and Therapeutic Strategies

Previous chapters have provided some insight into the regulation of immune responses under spaceflight conditions, a unique combination of psychosocial and physical stresses. This multitude of factors affecting almost all components of the immune system could lead to compromised host defense against infections and could have an immediate impact on mission performance. This chapter introduces some strategies that are potentially helpful to mitigate the risk of stress-associated immune dysfunction under conditions of (long-duration) spaceflight.

Pharmacological Countermeasures to Spaceflight-Induced Alterations of the Immune System

Opportunities for microbes to establish infections are enhanced under spaceflight conditions because space travel stimulates their growth (Chap. 15) and has a negative impact on immune functions. Indeed, it has been shown that spaceflight affects lymphoid organs (Gridley et al. 2003; Baqai et al. 2009) and induces variations in peripheral blood leukocyte subsets (Chap. 9). Neutrophil, monocyte, and NK cell functions are affected by spaceflight (Chaps. 10–12). The activation of T lymphocytes is also severely depressed under low gravity conditions (Cogoli et al. 1984) because interleukin-2 (IL-2) and IL-2receptor gene expression are modified, the delivery of the costimulatory signal to activate the B7/CD28 pathway and the protein kinase A (PKA) signaling pathway, which is a key early regulator in T cell activation, are hindered. Furthermore, a TH2 cytokine shift is associated with spaceflight. If this TH2 shift persists during long missions, it could represent a significant clinical risk for TH2-related autoimmune diseases, allergies, hypersensitivities, and disease susceptibility related to diminished cell-mediated immunity. Studies on plasma antibody levels did not reveal significant changes after short spaceflights (Rykova et al. 2008), but contradictory results were reported after long missions. Indeed, several studies (Konstantinova et al. 1993; Bascove et al. 2008, 2009; Gueguinou et al. 2009, 2010) reported increased levels of immunoglobulin while Rykova et al. (2008) reported normal amounts of antibodies after prolonged space missions. Lastly, a differential sensitivity of cellular and humoral immunity to spaceflight conditions seems to exist because it was shown that the cellular, but not the humoral, systems are affected by short periods of flight.