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. 2021 Feb 24;2(1):100365.
doi: 10.1016/j.xpro.2021.100365. eCollection 2021 Mar 19.

In vitro induction of trained immunity in adherent human monocytes

Jorge Domínguez-Andrés  1   2 , Rob J W Arts  1   2 , Siroon Bekkering  1   2 , Harsh Bahrar  1   2 , Bastiaan A Blok  1   2 , L Charlotte J de Bree  1   2 , Mariolina Bruno  1   2 , Özlem Bulut  1   2 , Priya A Debisarun  1   2 , Helga Dijkstra  1   2 , Jéssica Cristina Dos Santos  1   2 , Anaísa V Ferreira  1   2 , Daniela Flores-Gomez  1   2 , Laszlo A Groh  1   2 , Inge Grondman  1   2 , Leonie Helder  1   2 , Cor Jacobs  1   2 , Liesbeth Jacobs  1   2 , Trees Jansen  1   2 , Gizem Kilic  1   2 , Viola Klück  1   2 , Valerie A C M Koeken  1   2   3 , Heidi Lemmers  1   2 , Simone J C F M Moorlag  1   2 , Vera P Mourits  1   2 , Jelmer H van Puffelen  1   2 , Katrin Rabold  1   2 , Rutger J Röring  1   2 , Diletta Rosati  1   2 , Helin Tercan  1   2 , Julia van Tuijl  1   2 , Jessica Quintin  4 , Reinout van Crevel  1   2 , Niels P Riksen  1   2 , Leo A B Joosten  1   2 , Mihai G Netea  1   2   5
Affiliations

In vitro induction of trained immunity in adherent human monocytes

Jorge Domínguez-Andrés et al. STAR Protoc. .

Abstract

A growing number of studies show that innate immune cells can undergo functional reprogramming, facilitating a faster and enhanced response to heterologous secondary stimuli. This concept has been termed "trained immunity." We outline here a protocol to recapitulate this in vitro using adherent monocytes from consecutive isolation of peripheral blood mononuclear cells. The induction of trained immunity and the associated functional reprogramming of monocytes is described in detail using β-glucan (from Candida albicans) and Bacillus Calmette-Guérin as examples. For complete details on the use and execution of this protocol, please refer to Repnik et al. (2003) and Bekkering et al. (2016).

Keywords: Cell biology; Cell isolation; Cell-based assays; Immunology.

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Conflict of interest statement

The authors declare no competing interests

Figures

None
Graphical abstract
Figure 1
Figure 1
Aspect of the different layers before and after density gradient centrifugation with Ficoll-Paque
Figure 2
Figure 2
Aspect of the different layers before and after density gradient centrifugation with Percoll
Figure 3
Figure 3
Expected cytokine production by human monocytes after the induction of trained immunity in vitro and secondary restimulation with LPS Ctrl, non-trained cells; BG, cells stimulated with β-glucan at day 0; LPS, cells rechallenged with LPS at day 6.
See this image and copyright information in PMC

References

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