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Exposure of Legionella pneumophila to low-shear modeled microgravity: impact on stress response, membrane lipid composition, pathogenicity to macrophages and interrelated genes expression

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Abstract

Here, we studied the effect of low-shear modeled microgravity (LSMMG) on cross stress resistance (heat, acid, and oxidative), fatty acid content, and pathogenicity along with alteration in expression of stress-/virulence-associated genes in Legionella pneumophila. The stress resistance analysis result indicated that bacteria cultivated under LSMMG environments showed higher resistance with elevated D-values at 55 °C and in 1 mM of hydrogen peroxide (H2O2) conditions compared to normal gravity (NG)-grown bacteria. On the other hand, there was no significant difference in tolerance (p < 0.05) toward simulated gastric fluid (pH-2.5) acid conditions. In fatty acid analysis, our result showed that a total amount of saturated and cyclic fatty acids was increased in LSMMG-grown cells; as a consequence, they might possess low membrane fluidity. An upregulated expression level was noticed for stress-related genes (hslV, htrA, grpE, groL, htpG, clpB, clpX, dnaJ, dnaK, rpoH, rpoE, rpoS, kaiB, kaiC, lpp1114, ahpC1, ahpC2, ahpD, grlA, and gst) under LSMMG conditions. The reduced virulence (less intracellular bacteria and less % of induce apoptosis in RAW 264.7 macrophages) of L. pneumophila under LSMMG conditions may be because of downregulation related genes (dotA, dotB, dotC, dotD, dotG, dotH, dotL, dotM, dotN, icmK, icmB, icmS, icmT, icmW, ladC, rtxA, letA, rpoN, fleQ, fleR, and fliA). In the LSMMG group, the expression of inflammation-related factors, such as IL-1α, TNF-α, IL-6, and IL-8, was observed to be reduced in infected macrophages. Also, scanning electron microscopy (SEM) analysis showed less number of LSMMG-cultivated bacteria attached to the host macrophages compared to NG. Thus, our study provides understandings about the changes in lipid composition and different genes expression due to LSMMG conditions, which apparently influence the alterations of L. pneumophila’ stress/virulence response.

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Data availability

All data generated or analyzed during this study are included in this published article and its supplementary information files.

Abbreviations

NG:

Normal gravity

LSMMG:

Low-shear modeled microgravity

SEM:

Scanning electron microscopy

qRT‑PCR:

Quantitative reverse‑transcription‑PCR

H2O2 :

Hydrogen peroxide

HARVs:

High-aspect rotating vessels

RWV:

Rotating-wall vessel

CFU:

Colony forming unit

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Acknowledgements

This study was supported by the funds of Jeonbuk National University 2019, Republic of Korea and we acknowledge the Center for University-wide Research Facilities (CURF) of Jeonbuk National University for helping in SEM images.

Funding

This research was supported by Buan RIS Resource Project (Grand no. R0001102) and the funds of Jeonbuk National University.

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Authors and Affiliations

  1. Department of Wood Science and Technology, College of Agriculture and Life Sciences, Jeonbuk National University, 567, Jeonju-si, Jeollabuk-do, Republic of Korea

    Sunirmal Sheet & Yang Soo Lee

  2. School of Medicine, Anschutz Medical Campus Aurora, Aurora, CO, 80045, USA

    Yesupatham Sathishkumar

  3. Department of Biomedical Sciences and Institute for Medical Science, Jeonbuk National University Medical School, 567, Jeonju-si, Jeollabuk-do, Republic of Korea

    Satabdi Acharya

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  1. Sunirmal Sheet
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Contributions

SS and YSL designed research, SS and SA performed the experiments; SS, SA along with YS analyzed data; SS and SA wrote the draft paper; SS, SA, YS and YSL revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Yang Soo Lee.

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Communicated by Ran Wang.

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203_2023_3753_MOESM1_ESM.tif

Supplementary file1 represents the growth curve of L. pneumophila over a 24 h period in BYE broth at 37 °C (~ 90% relative humidity) under NG and LSMMG culture conditions. NG normal gravity; LSMMG Low sheared modeled micro gravity (TIF 32 KB)

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Sheet, S., Sathishkumar, Y., Acharya, S. et al. Exposure of Legionella pneumophila to low-shear modeled microgravity: impact on stress response, membrane lipid composition, pathogenicity to macrophages and interrelated genes expression. Arch Microbiol 206, 87 (2024). https://doi.org/10.1007/s00203-023-03753-z

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