A protocol to detect human CD4+ T cell extracellular traps using scanning electron microscopy

Nam Phuong N Nguyen; Nicole Chizara Oparaugo; Kelsey Ouyang; George W Agak

doi:10.1016/j.xpro.2022.101932

A protocol to detect human CD4⁺ T cell extracellular traps using scanning electron microscopy

STAR Protoc. 2023 Mar 17;4(1):101932. doi: 10.1016/j.xpro.2022.101932. Epub 2022 Dec 26.

Authors

Nam Phuong N Nguyen¹, Nicole Chizara Oparaugo², Kelsey Ouyang³, George W Agak⁴

Affiliations

¹ Division of Dermatology, Department of Medicine, University of California (UCLA), Los Angeles, CA 90095, USA; Morsani College of Medicine, University of South Florida, Tampa, FL 33620, USA.
² David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
³ Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, OH 44195, USA.
⁴ Division of Dermatology, Department of Medicine, University of California (UCLA), Los Angeles, CA 90095, USA. Electronic address: gagak@mednet.ucla.edu.

Abstract

We present a protocol to detect extracellular traps (ETs) induced by Cutibacterium acnes in cultured T_H17 clones. We first describe the isolation of C. acnes-specific T_H17 clones by sterile cell sorting. We then detail the in vitro induction of ETs in T_H17 clones stimulated by C. acnes and the imaging of released ETs using scanning electron microscopy. This protocol can be applied to the study of other ETs released by other T cell subsets. For complete details on the use and execution of this protocol, please refer to Agak et al. (2021).¹.

Keywords: Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Microbiology; Microscopy.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural

MeSH terms

CD4-Positive T-Lymphocytes
Cell Separation
Extracellular Traps*
Humans
Microscopy, Electron, Scanning
T-Lymphocytes*

Abstract

Publication types

MeSH terms

Grants and funding