Third Military Medical University 11 articles published in JoVE Immunology and Infection Cationic Nanoemulsion-Encapsulated Retinoic Acid as an Adjuvant to Promote OVA-Specific Systemic and Mucosal Responses Guo-Cheng Li1, Hui-Fang Li1, Zhe Jin1, Rang Feng1, Yan Deng1, Hao Cheng1, Hai-Bo Li1 1National Engineering Research Center of Immunological Products, Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, Third Military Medical University In this protocol, we developed a cationic nanoemulsion-encapsulated retinoic acid (RA) to be used as an adjuvant to promote antigen-specific systemic and mucosal responses. By adding the FDA-approved RA to the nanoemulsion, antigen-specific sIgA was promoted in the vagina and small intestine after intramuscular injection of the nanoemulsion. Immunology and Infection Draining Lymph Node Metastasis Model for Assessing the Dynamics of Antigen-Specific CD8+ T Cells During Tumorigenesis Yan Zhang*1, Xingxing Su*2, Lisha Wang2, Zhengliang Yue2, Qiao Liu3, Ling Ran2, Shun Lei2, Jianjun Hu2, Lifan Xu2, Lilin Ye2, Ping Ji4, Guimei Li1, Qizhao Huang5, Shuqiong Wen4 1College of Veterinary Medicine, Qingdao Agricultural University, 2Institute of Immunology, Third Military Medical University, 3Institute of Cancer, Xinqiao Hospital, Third Military Medical University, 4Stomatological Hospital of Chongqing Medical University, 5Institute of life science, Chongqing Medical University The experimental design presented here provides a useful reproductive model for the studies of antigen-specific CD8+ T cells during lymph node (LN) metastasis, which excludes the perturbation of bystander CD8+ T cells. Immunology and Infection Evaluating the Immune Response of a Nanoemulsion Adjuvant Vaccine Against Methicillin-Resistant Staphylococcus aureus (MRSA) Infection Xiaoqiang Zeng1, Hongwu Sun1, Yan Ye1, Xing Luo1, Dingyi Cai1, Yun Yang1, Ting Chen1, Cun Sun1, Shaotong Zhang1, Hao Zeng1 1National Engineering Research Centre of Immunological Products, Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, Third Military Medical University The present protocol prepares and evaluates the physical properties, immune response, and in vivo protective effect of a novel nanoemulsion adjuvant vaccine. Immunology and Infection Efficient Transfection of In vitro Transcribed mRNA in Cultured Cells Using Peptide-Poloxamine Nanoparticles Qin Xiao*1, Yuheng Liu*1, Dandan Zhang1, Chao Li1, Qihua Yang1, Dongshui Lu1, Weijun Zhang1, Joseph Rosenecker2, Quanming Zou1, Yang Li3, Shan Guan1 1National Engineering Research Center of Immunological Products, Department of Microbiology and Biochemical Pharmacy, Third Military Medical University, 2Department of Pediatrics, Ludwig-Maximilians University of Munich, 3Department of Pharmacy, Southwest Hospital, Third Military Medical University A self-assembled peptide-poloxamine nanoparticle (PP-sNp) is developed using a microfluidic mixing device to encapsulate and deliver in vitro transcribed messenger RNA. The described mRNA/PP-sNp could efficiently transfect cultured cells in vitro. Bioengineering A "Plug-And-Display" Nanoparticle Vaccine Platform Based on Outer Membrane Vesicles Displaying SARS-CoV-2 Receptor-Binding Domain Rang Feng1, Guo-Cheng Li1, Hai-Ming Jing1, Chang Liu1, Ruo-Yi Xue1, Quan-Ming Zou1, Hai-Bo Li1 1National Engineering Research Center of Immunological Products, Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, Third Military Medical University The present protocol describes the bioengineering of outer membrane vesicles to be a "Plug-and-Display" vaccine platform, including production, purification, bioconjugation, and characterization. Neuroscience Osmotic Pump-based Drug-delivery for In Vivo Remyelination Research on the Central Nervous System Xiaorui Wang1, Yixun Su1,2, Xuelian Hu1,3, Jianqin Niu1 1Department of Histology and Embryology, Chongqing Key Laboratory of Neurobiology, Brain, and Intelligence Research Key Laboratory of Chongqing Education Commission, Third Military Medical University, 2Research Centre, The Seventh Affiliated Hospital of Sun Yat-sen University, 3School of Medicine, Chongqing University Demyelination takes place in multiple central nervous system diseases. A reliable in vivo drug delivery technique is necessary for remyelinating drug testing. This protocol describes an osmotic pump-based method that allows long-term drug delivery directly into the brain parenchyma and improves the drug bioavailability, with broad application in remyelination research. Immunology and Infection Analysis of HBV-Specific CD4 T-cell Responses and Identification of HLA-DR-Restricted CD4 T-Cell Epitopes Based on a Peptide Matrix Jianmei Xiao1,2, Xing Wan1,2, Haoliang Wang1,2, Guohong Deng1,2 1Department of Infectious Diseases, Southwest Hospital, Third Military Medical University (Army Medial University), 2Chongqing Key Laboratory for Research of Infectious Diseases Based on a hepatitis B virus (HBV)-derived peptide matrix, HBV-specific CD4 T-cell responses could be evaluated in parallel with identification of HBV-specific CD4 T-cell epitopes. Cancer Research Tumor Transplantation for Assessing the Dynamics of Tumor-Infiltrating CD8+ T Cells in Mice Lisha Wang1, Zhiming Wang1, Junyi Guo2, Huayu Lin1, Shuqiong Wen2, Qiao Liu1, Yiding Li3, Qing Wu1, Leiqiong Gao1, Xiangyu Chen1, Luoyingzi Xie1, Qin Tian1, Jianfang Tang1, Zhirong Li1, Li Hu1, Juan Wang4, Lifan Xu1, Qizhao Huang5, Lilin Ye1 1Institute of Immunology, Third Military Medical University, 2Guanghua School of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Stomatological Hospital, Sun Yat-Sen University, 3Shigatse Branch, Xinqiao Hospital, Third Military Medical University, 4Department of Emergency Medicine, Southwest Hospital, Third Military Medical University, 5Cancer Center, The General Hospital of Western Theater Command Here, we present a tumor transplantation protocol for the characterization of tumor-inherent and periphery-derived tumor-infiltrated lymphocytes in a mouse tumor model. Specific tracing of the influx of recipient-derived immune cells with flow cytometry reveals the dynamics of the phenotypic and functional changes of these cells during antitumor immune responses. Immunology and Infection Design of Cecal Ligation and Puncture and Intranasal Infection Dual Model of Sepsis-Induced Immunosuppression Zhihan Wang*1,2, Qinqin Pu*1,2, Ping Lin*2,3, Changlong Li1, Jianxin Jiang3, Min Wu2 1West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, 2Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota, 3State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Surgery Research, Daping Hospital, Army Medical University This protocol describes techniques to measure infectious outcomes underlying secondary hospital-acquired infections in the immunosuppressive condition, first by establishing cecal ligation/puncture mice then challenging them with intranasal infection to create a clinically relevant model of immunosuppression sepsis. Biochemistry A Western Blotting Protocol for Small Numbers of Hematopoietic Stem Cells Xiongwei Cai1,2,3, Yi Zheng1, Nancy A. Speck2 1 A standard Western blotting protocol was optimized for analyzing as few as 500 hematopoietic stem or progenitor cells. Optimization involves careful handling of the cell sample, limiting transfers between tubes, and directly lysing the cells in Laemmli sample buffer. Immunology and Infection Generation of Lymphocytic Microparticles and Detection of their Proapoptotic Effect on Airway Epithelial Cells Chun Yang*1, Wei Xiong*2, Qian Qiu2, Houda Tahiri1, Carmen Gagnon1, Guoxiang Liu2, Pierre Hardy1 1Departments of Pediatrics and Pharmacology, University of Montréal, 2Department of Pulmonology, Chongqing Southwest Hospital, Third Military Medical University Cell membrane–shed microparticles (MPs) are active biological vesicles that can be isolated and their pathophysiological effects investigated in various models. Here we describe a method for generating MPs derived from T lymphocytes (LMPs) and for demonstrating their proapoptotic effect on airway epithelial cells.
