MicroRNA quantitation during dendritic cell endocytosis using imaging flow cytometry

Key factors and requirements

Qiang Tong, Ying Zhu, Dandan Zhang, Qing Cai, Wenchun Qu, Christopher D.O. Cooper, Jiaqi Wang, Patrick C. McHugh

Research output: Contribution to journalArticle

Abstract

Background/Aims: MicroRNA (miRNA)-induced suppression of dendritic cells (DCs) has been implicated in many diseases. Therefore, accurate monitoring of miRNA endocytosis by DCs is important for understanding the role of miRNAs in many diseases. Recently, a method for measuring the co-localization of Argonaute 2 (AGO2)-associated miRNAs on laser-scanning confocal microscopy method was proposed to localize the miRNAs. But its definition was limited by the number of observed cells through its accuracy. Methods: In this study, a method based on imaging flow cytometry was developed to localize miR-590-5p with fluorescent probes in DCs. miR-590-5p proven to play an important role in tumor immunity. This method enabled the quantification, visualization and localization of the fluorescence intensity in 30,000 individual cells. Results: Using this method, the DCs with different endocytotic ability were distinguished. The behaviour of miR-590-5p during endocytosis under the stimulation of tumor antigen in DCs was observed, binding to its cognate target mRNA and degradation in DCs. Conclusion: This method based on imaging flow cytometry provide an additional method to study miRNA processing in DCs, which makes it a valuable addition to existing miRNA research techniques.

Original languageEnglish (US)
Pages (from-to)793-811
Number of pages19
JournalCellular Physiology and Biochemistry
Volume51
Issue number2
DOIs
StatePublished - Nov 1 2018

Fingerprint

Endocytosis
MicroRNAs
Dendritic Cells
Flow Cytometry
RNA Stability
Neoplasm Antigens
Fluorescent Dyes
Confocal Microscopy
Immunity
Research Design
Cell Count
Fluorescence
Neoplasms

Keywords

  • Argonaute 2 (AGO2)
  • Dendritic cells
  • Flow cytometry
  • FRET analysis
  • miR-590-5p
  • miRNA endocytosis

ASJC Scopus subject areas

  • Physiology

Cite this

MicroRNA quantitation during dendritic cell endocytosis using imaging flow cytometry : Key factors and requirements. / Tong, Qiang; Zhu, Ying; Zhang, Dandan; Cai, Qing; Qu, Wenchun; Cooper, Christopher D.O.; Wang, Jiaqi; McHugh, Patrick C.

In: Cellular Physiology and Biochemistry, Vol. 51, No. 2, 01.11.2018, p. 793-811.

Research output: Contribution to journalArticle

Tong, Qiang ; Zhu, Ying ; Zhang, Dandan ; Cai, Qing ; Qu, Wenchun ; Cooper, Christopher D.O. ; Wang, Jiaqi ; McHugh, Patrick C. / MicroRNA quantitation during dendritic cell endocytosis using imaging flow cytometry : Key factors and requirements. In: Cellular Physiology and Biochemistry. 2018 ; Vol. 51, No. 2. pp. 793-811.
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