Culture, expansion, and flow-cytometry-based functional analysis of pteropid bat MR1-restricted unconventional T cells

Wan Rong Sia; Ying Ying Hey; Randy Foo; Lin-Fa Wang; Edwin Leeansyah

doi:10.1016/j.xpro.2021.100487

Culture, expansion, and flow-cytometry-based functional analysis of pteropid bat MR1-restricted unconventional T cells

STAR Protoc. 2021 May 18;2(2):100487. doi: 10.1016/j.xpro.2021.100487. eCollection 2021 Jun 18.

Authors

Wan Rong Sia^{1

2}, Ying Ying Hey¹, Randy Foo¹, Lin-Fa Wang^{1

3}, Edwin Leeansyah^{4

5

1

6}

Affiliations

¹ Programme in Emerging Infectious Diseases, Duke-National University of Singapore Medical School, Singapore 169857, Singapore.
² Department of Microbiology & Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117545, Singapore.
³ SingHealth Duke-NUS Global Health Institute, Singapore 169857, Singapore.
⁴ Institute of Biopharmaceutical and Health Engineering, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, People's Republic of China.
⁵ Precision Medicine and Healthcare Research Center, Tsinghua-Berkeley Shenzhen Institute, Tsinghua University, Shenzhen 518055, People's Republic of China.
⁶ Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, 14183 Stockholm, Sweden.

Abstract

Bats harbor viruses of global public health significance. Understanding bat immune systems may provide intervention strategies to prevent zoonotic disease transmission and identify therapeutic targets. This protocol describes how to culture and expand pteropid bat unconventional T cells, restricted by the MHC-I-related protein 1 (MR1), an MHC-I-like protein. Using multicolor flow-cytometry-based techniques, we examine pteropid MR1T cell functionality, including proliferative capacity, cytotoxicity, and cytokine production. This protocol can be adapted to aid immunological research in other bat species. For complete details on the use and execution of this protocol, please refer to Leeansyah et al. (2020b).

Keywords: Cell culture; Cell isolation; Cell-based Assays; Flow Cytometry/Mass Cytometry; Immunology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques / methods*
Cells, Cultured
Chiroptera*
Flow Cytometry / methods*
Histocompatibility Antigens Class I / metabolism*
Minor Histocompatibility Antigens / metabolism*
T-Lymphocytes / cytology*

Substances

Histocompatibility Antigens Class I
Minor Histocompatibility Antigens