Detection of Polyfunctional T Cells in Children Vaccinated with Japanese Encephalitis Vaccine via the Flow Cytometry Technique

Linlin Zhang; Meng Zhang; Mengjia Liu; Junhong Ai; Jiao Tian; Huizheng Ge; Ran Wang; Zhengde Xie

doi:10.3791/64671

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Method Article

Detection of Polyfunctional T Cells in Children Vaccinated with Japanese Encephalitis Vaccine via the Flow Cytometry Technique

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DOI:

10.3791/64671

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September 23rd, 2022

Linlin Zhang^*¹^,² , Meng Zhang^*¹^,²^,³ , Mengjia Liu¹^,² , Junhong Ai¹^,² , Jiao Tian¹^,² , Huizheng Ge⁴ , Ran Wang¹^,² , Zhengde Xie¹^,²

¹Beijing Key Laboratory of Pediatric Respiratory Infectious Diseases, Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, Laboratory of Infection and Virology, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, ²Research Unit of Critical Infection in Children, 2019RU016, Chinese Academy of Medical Sciences, ³Department of Pediatric Rehabilitation, Beijing Boai Hospital, School of Rehabilitation Medicine, Capital Medical University, China Rehabilitation Research Center, ⁴Center of Excellence (Beijing), Becton Dickinson Medical Devices (Shanghai) Co Ltd

^* These authors contributed equally

Summary

The present protocol combines ex vivo stimulation and flow cytometry to analyze polyfunctional T cell (T_PF) profiles in peripheral blood mononuclear cells (PBMCs) within Japanese encephalitis virus (JEV)-vaccinated children. The detection method and flow cytometry color scheme of JEV-specific TPFs were tested to provide a reference for similar studies.

Abstract

T cell-mediated immunity plays an important role in controlling flavivirus infection, either after vaccination or after natural infection. The "quality" of a T cell needs to be assessed by function, and higher function is associated with more powerful immune protection. T cells that can simultaneously produce two or more cytokines or chemokines at the single-cell level are called polyfunctional T cells (T_PFs), which mediate immune responses through a variety of molecular mechanisms to express degranulation markers (CD107a) and secrete interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-2, or macrophage inflammatory protein (MIP)-1α. There is increasing evidence that T_PFs are closely related to the maintenance of long-term immune memory and protection and that their increased proportion is an important marker of protective immunity and is important in the effective control of viral infection and reactivation. This evaluation applies not only to specific immune responses but also to the assessment of cross-reactive immune responses. Here, taking the Japanese encephalitis virus (JEV) as an example, the detection method and flow cytometry color scheme of JEV-specific T_PFs produced by peripheral blood mononuclear cells of children vaccinated against Japanese encephalitis were tested to provide a reference for similar studies.

Introduction

Japanese encephalitis virus (JEV) is an important mosquito-borne virus belonging to the genus Flavivirus within the Flaviviridae family¹. Many Asia-Pacific countries have long faced enormous public health challenges due to the huge disease burden caused by Japanese encephalitis (JE), but this has improved dramatically with the increasing availability of various types of vaccinations². Adaptive protective immune responses evoked by natural infection or vaccination contribute to the prevention and antiviral regulation. Humoral immunity and cell-mediated immunity are classified as adaptive immunity, and the in....