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. 2021 Jun 7;11:11872. doi: 10.1038/s41598-021-90458-2

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Experimental workflow. (A) Blood samples from eight healthy volunteers were collected. Peripheral blood mononuclear cells (PBMCs) were isolated and exposed to simulated microgravity (sµG) or (1G) (static control) using a random positioning machine for 18 h, then stimulated with ConA and anti-CD28 (ConA/anti-CD28) or left unstimulated. (B) The activation and intracellular signaling responses of all major immune cell subsets were quantified using single-cell mass cytometry. In parallel, the mRNA expression of a select number of genes was quantified using qRT-PCR. (C) The high-dimensional immunological dataset was visualized as an immune signaling correlation network. Cell-type-specific immune responses that differed between the sµG and 1G conditions were identified using a multivariate Elastic Net (EN) method.