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Lack of CD34 produces defects in platelets, microparticles, and lung inflammation

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Abstract

Lung innate immune activation results in acute lung inflammation, which is characterized by alveolar barrier disruption and accumulation of cellular lung aggregates comprising neutrophils, platelets, mononuclear cells, and microparticles. CD34 is a sialomucin, with pan-selectin affinity and recently shown to protect the endothelial barrier in a bleomycin-induced lung injury model. However, there is very little information about the fundamental role of CD34 in regulation of the lung innate immune response. We hypothesized that CD34 regulates leukocyte recruitment by promoting optimal platelet activation (aggregation and spread) during bacterial lipopolysaccharide (LPS)-induced acute lung injury. Therefore, we utilized CD34 knock-out (KO) and wild-type (WT) mice to analyze and compare the morphology and expression of leukocyte subsets from the pulmonary and systemic compartments. We utilized the chemotactic N-formylated tri-peptide, fMLP, to understand platelet aggregation in vitro, and the fundamental immune stimulant, LPS, to induce lung injury and understand platelet activation ex vivo. Our data reveal that under steady-state conditions, KO mice possess large aggregates of integrin β3 (CD61)-positive microparticles in peripheral blood. Moreover, the KO mice recruit a large number of neutrophils to lungs, which are not cleared even at 36-h post-LPS exposure. The KO mice display an increased platelet CD61 expression, which aggregates, but does not spread normally in response to in vitro fMLP treatment. The KO platelets display similar deficits in their spreading ability even after ex vivo LPS exposure. Thus, our data demonstrate that CD34 modulates platelet biology, microparticle aggregation, and neutrophil recruitment during murine lung inflammation.

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Acknowledgments

The author gratefully acknowledges the support by Dr. Baljit Singh for providing access to CD34 KO colony for further exploration of the innate immune mechanisms of CD34.

Funding

The research conducted is funded by start-up research funds from the Sylvia Fedoruk Canadian Center for Nuclear Innovation. The Sylvia Fedoruk Canadian Center for Nuclear Innovation is funded by Innovation Saskatchewan. Fluorescence imaging was performed at the WCVM Imaging Centre, which is funded by NSERC.

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  1. Small Animal Clinical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada

    Gurpreet Kaur Aulakh

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  1. Gurpreet Kaur Aulakh
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Correspondence to Gurpreet Kaur Aulakh.

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The authors declare that they have no conflict of interest.

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The study design was approved by the University of Saskatchewan’s Animal Research Ethics Board (AUP 20150018) and adhered to the Canadian Council on Animal Care guidelines for humane animal use.

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Aulakh, G.K. Lack of CD34 produces defects in platelets, microparticles, and lung inflammation. Cell Tissue Res 382, 405–419 (2020). https://doi.org/10.1007/s00441-020-03243-4

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