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Expression and Characterization of Antimicrobial Peptide LL-37 in Dog Peripheral Blood Endothelial Progenitor Cells In Vitro

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Abstract

This study aimed to construct antimicrobial peptide LL-37 expressed lentiviral vector and explore the expression of LL-37 in endothelial progenitor cells (EPCs) from dog periphery mononuclear cells (PBMCs). The methodology on obtaining EPCs from dog PBMCs was explored and infection approach about the expression of LL-37 in EPCs was optimized. LL-37 was amplified, digested by Age I and constructed into pGC-FU vector. Meanwhile, EPCs were generated from PBMCs using endothelial growth medium. Markers including acetylated low density lipoproteins (Dilac-LDL), Ulex lectin (UEA-1), von Willebrand factor (vWF), VE-cadherin, CD34 and CD133 were used to identify EPCs. Then, LL-37 was introduced into EPCs using lentivirus infection with green fluorescent protein (GFP) as the biomarker. Finally, we used western-blot to examine the expression of LL-37 in EPCs. The pGC-FU-LL-37-GFP was successfully constructed. Viral particles existed in 10−4 μL group with titer of 2 × 108 TU/mL. Both early (round-shaped morphology) and late (pebble-shaped appearance) EPCs were observed. And these EPCs were confirmed by Dil-acLDL uptake and FITC-UEA-1 binding. An approximate 50 % of infection efficacy was confirmed by immunofluorescence. Furthermore, we successfully observed LL-37 expression in EPCs via western blot analysis. In conclusions, LL-37 expressed in the EPCs was successfully achieved. However, the anti-infection and angiogenesis of LL-37 expressed EPCs need further research in future.

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  1. Department of Urology, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuchang Area, Wuhan, 430060, Hubei, China

    Chenglong Li, Yong-wei Li, Sixing Yang, Chao Song, Wenbiao Liao & Yunhe Xiong

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  1. Chenglong Li
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  2. Yong-wei Li
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  3. Sixing Yang
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  4. Chao Song
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  5. Wenbiao Liao
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Correspondence to Sixing Yang.

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Li, C., Li, Yw., Yang, S. et al. Expression and Characterization of Antimicrobial Peptide LL-37 in Dog Peripheral Blood Endothelial Progenitor Cells In Vitro. Int J Pept Res Ther 21, 279–287 (2015). https://doi.org/10.1007/s10989-014-9453-5

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