Capital Medical University, National Center for Children's Health

6 ARTICLES PUBLISHED IN JoVE

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.

Medicine

Dual Bioluminescence Imaging of Tumor Progression and Angiogenesis

Kaiyue Zhang¹, Chen Wang¹, Ran Wang², Shang Chen¹, Zongjin Li¹

¹Nankai University School of Medicine, ²State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University

This protocol describes the establishment of a tumor-bearing mouse model to monitor tumor progression and angiogenesis in real-time by dual bioluminescence imaging.

Biology

Rapid Antimicrobial Susceptibility Testing by Stimulated Raman Scattering Imaging of Deuterium Incorporation in a Single Bacterium

Meng Zhang^1,2, Mohamed N. Seleem³, Ji-Xin Cheng^1,2,4,5

¹Department of Electrical and Computer Engineering, Boston University, ²Boston University Photonics Center, Boston University, ³Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University, ⁴Department of Biomedical Engineering, Boston University, ⁵Department of Chemistry, Boston University

This protocol presents rapid antimicrobial susceptibility testing (AST) assay within 2.5 h by single-cell-stimulated Raman scattering imaging of D₂O metabolism. This method applies to bacteria in the urine or whole blood environment, which is transformative for rapid single-cell phenotypic AST in the clinic.

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.

Immunology and Infection

Standardization of Transfer across Labs between Flow Cytometers for Detection of Lymphocytes in Japanese Encephalitis Vaccinated Children

Huizheng Ge^*1, Linlin Zhang^*2,3, Mengjia Liu^*2,3, Xi Yang¹, Xu Wu¹, Ran Wang^2,3, Zhengde Xie^2,3

¹Center of Excellence (Beijing), Becton Dickinson Medical Devices (Shanghai) Co Ltd, ²Beijing 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, ³Research Unit of Critical Infection in Children, 2019RU016, Chinese Academy of Medical Sciences

In this study, a method was developed to facilitate the transfer of experimental settings and analysis templates between two flow cytometers in two laboratories for the detection of lymphocytes in Japanese encephalitis-vaccinated children. The standardization method for the flow cytometer experiments will allow research projects to be effectively conducted in multiple centers.