National Center for Children’s Health

5 ARTICLES PUBLISHED IN JoVE

Bioengineering

Designing Microfluidic Devices for Studying Cellular Responses Under Single or Coexisting Chemical/Electrical/Shear Stress Stimuli

Tzu-Yuan Chou¹, Yung-Shin Sun², Hsien-San Hou³, Shang-Ying Wu¹, Yun Zhu¹, Ji-Yen Cheng³, Kai-Yin Lo¹

¹Department of Agricultural Chemistry, National Taiwan University, ²Department of Physics, Fu-Jen Catholic University, ³Research Center for Applied Sciences, Academia Sinica

Micro-fabricated devices integrated with fluidic components provide an in vitro platform for cell studies mimicking the in vivo micro-environment. We developed polymethylmethacrylate-based microfluidic chips for studying cellular responses under single or coexisting chemical/electrical/shear stress stimuli.

Immunology and Infection

An Efficient and Simple Method to Establish NK and T Cell Lines from Patients with Chronic Active Epstein-Barr Virus Infection

Chongfeng Xu¹, Junhong Ai², Qingxun Zhang¹, Ting Li³, Xiaorong Wu¹, Zhengde Xie², Ziyuan Duan¹

¹Genetic Resources Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, ²MOE Key Laboratory of Major Diseases in Children, National Key Discipline of Pediatrics, National Clinical Research Center for Respiratory Diseases, Beijing Key Laboratory of Pediatric Respiratory Infection Diseases, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, ³State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences

A simple method for obtaining NK and T cell clones from CAEBV patients was developed with high efficiency, a small amount of peripheral blood, and a low-dose of IL-2.

Immunology and Infection

Construction of Adenoviral Vectors using DNA Assembly Technology

Xiaohui Zou^*1, Yun Zhu^*2, Chenglong Li^1,3, Yali Duan², Linlin Zhang², Xiaojuan Guo¹, Wenzhe Hou¹, Zhengde Xie^2,4, Zhuozhuang Lu^1,5

¹NHC Key Laboratory of Medical Virology and Viral Diseases, State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, ²Laboratory of Infection and Virology, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, National Center for Children’s Health, ³Henan Chemical Technician College, ⁴Research Unit of Critical Infection in Childre, Chinese Academy of Medical Sciences, ⁵Chinese Academy of Sciences Joint Research Center for Emerging Infectious Diseases and Biosafety, Chinese Center for Disease Control and Prevention-Wuhan Institute of Virology

In this study, an infectious clone of human adenovirus type 7 (HAdV-7) was constructed, and an E3-deleted HAdV-7 vector system was established by modifying the infectious clone. This strategy used here can be generalized to make gene transfer vectors from other wild-type adenoviruses.

Immunology and Infection

Separation of Immune Cell Subpopulations in Peripheral Blood Samples from Children with Infectious Mononucleosis

Linlin Zhang^1,2, Mengjia Liu^1,2, Meng Zhang^1,2, Junhong Ai^1,2, Jiao Tian^1,2, Ran Wang^1,2, Zhengde Xie^1,2

We describe a method combining immunomagnetic beads and fluorescence-activated cell sorting to isolate and analyze defined immune cell subpopulations of peripheral blood mononuclear cells (monocytes, CD4⁺ T cells, CD8⁺ T cells, B cells, and natural killer cells). Using this method, magnetic and fluorescently labeled cells can be purified and analyzed.

Immunology and Infection

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

Linlin Zhang^*1,2, Meng Zhang^*1,2,3, Mengjia Liu^1,2, Junhong Ai^1,2, Jiao Tian^1,2, Huizheng Ge⁴, Ran Wang^1,2, Zhengde Xie^1,2

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

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.