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AIM2 levels and DNA-triggered inflammasome response are increased in peripheral leukocytes of patients with abdominal aortic aneurysm

  • Markus Wortmann
  • Xianghui Xiao
  • Guido Wabnitz
  • Yvonne Samstag
  • Maani Hakimi
  • Dittmar Böckler
  • Susanne DihlmannEmail author
Original Research Paper

Abstract

Objective and design

Abdominal aortic aneurysm (AAA) is heavily infiltrated with leukocytes, expressing the DNA sensor absent in melanoma 2 (AIM2) and other inflammasome components.

Methods

Using multicolour flow cytometry, we here compared the expression of the inflammasome components AIM2, NLRP3, and ASC in different peripheral immune cells derived from AAA patients with those from non-AAA patients in a case–control study. In parallel, peripheral blood mononuclear cells (PBMC) of AAA patients and controls were stimulated in vitro with poly-dA:dT or lipopolysaccharide (LPS) to analyze inflammasome activation.

Results

AIM2 expression was significantly increased in peripheral granulocytes (P = 0.026), monocytes (P = 0.007), B lymphocytes (P < 0.0001), and T lymphocytes (P = 0.004) of AAA patients. Expression of other inflammasome components did not differ between the groups. Following in vitro stimulation with foreign DNA, PBMC derived from AAA patients released significantly more IL-1β (P = 0.022) into the supernatant than PBMC from control patients. In contrast, IL-1β release upon LPS stimulation did not differ between the PBMC groups.

Conclusion

The data indicate the increased activation of an AIM2 inflammasome in peripheral immune cells of AAA patients and point to a systemic AIM2-associated immune response to AAA.

Keywords

Inflammasome Innate immunity Abdominal aortic aneurysm FACS Flow cytometry PBMC 

Notes

Acknowledgements

We thank Anja Spieler (Department of Vascular and Endovascular Surgery of the University Hospital Heidelberg, Germany) for excellent technical assistance.

Funding

We acknowledge financial support by the Department of Vascular and Endovascular surgery of the University Hospital Heidelberg, Germany. This research was supported in part by a grant of the Deutsche Forschungsgemeinschaft (DFG; No. 323488362) to MW and SD.

Compliance with ethical standards

Conflict of interest

Xianghui Xiao has received financial support from the German Society of Vascular Surgery and Vascular Medicine for attending the annual meeting of the society in 2017. Maani Hakimi is CEO of CODE Medical Frankfurt GbR, Germany. Dittmar Böckler is consultant for Medtronic, W.L Gore & Ass. Endologix. The authors have no competing interests. MW, GW, YS, and SD have no conflict of interest for this publication.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Supplementary material

11_2019_1212_MOESM1_ESM.pdf (174 kb)
Gating strategy for phenotypic characterization of peripheral leukocytes that was applied for each individual AAA patient and age-matched subject without AAA. A. Live cells gated through the FSC-A versus SSC-A plot were selected and further characterized by plotting the SSC-A versus CD14-FITC, CD3-APC, or CD19-Cy5.5, respectively. Live CD14+ monocytes, CD3+ T cells and CD19+ B cells were then selected for analysis as shown in B. Granulocytes were defined as CD14- cells with a similar size a monocytes (SSC-A &#x003E; 120k). B. Histograms of the cells gated as shown in A. The fluorescent intensities of PE (representing NLRP3, AIM2 or ASC, horizontal axis) are plotted against the number of events detected (vertical axis, normalized to mode) in different leukocyte phenotypes. Dotted lines refer to the background, as it was determined by analysis of cells incubated with a PE-labeled secondary antibody only. The solid line (grey peak) refers to the fluorescent intensities of the PE-labeled secondary antibody that was used in combination with anti-NLRP3, anti-AIM2 or anti-ASC specific antibodies. The example shown refers to a control sample. SSC-A: side scatter (reflecting granularity of detected cells), area scaling. (PDF 174 KB)
11_2019_1212_MOESM2_ESM.pdf (238 kb)
Supplementary material 2 (PDF 238 KB)

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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Markus Wortmann
    • 1
  • Xianghui Xiao
    • 1
  • Guido Wabnitz
    • 2
  • Yvonne Samstag
    • 2
  • Maani Hakimi
    • 1
    • 3
  • Dittmar Böckler
    • 1
  • Susanne Dihlmann
    • 1
    Email author
  1. 1.Department of Vascular and Endovascular SurgeryUniversity Hospital HeidelbergHeidelbergGermany
  2. 2.Institute for Immunology, Molecular ImmunologyUniversity Hospital HeidelbergHeidelbergGermany
  3. 3.Vascular Biobank Heidelberg (VBBH)HeidelbergGermany

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