Protocols in Current Issue
    Evaluation of Mucosal and Systemic Vaccine Responses by Cyclic di-GMP (CDG)-adjuvanted Protein Subunit Vaccines
    Authors:  Samira Mansouri and Lei Jin, date: 04/20/2019, view: 289, Q&A: 0
    [Abstract] Intranasal administration of vaccine adjuvants directly deliver therapeutic agents to the lungs to induce potent lung mucosal immune responses. Cyclic di-GMP (CDG) is a promising mucosal vaccine adjuvant candidate capable of inducing protective immunity. This protocol describes an in vivo approach to induce and detect mucosal (lung) and ...
    Intracellular Invasion and Killing Assay to Investigate the Effects of Binge Alcohol Toxicity in Murine Alveolar Macrophages
    Authors:  Victor Jimenez Jr and Fernando P Monroy, date: 01/20/2019, view: 793, Q&A: 0
    [Abstract] Alcohol consumption has diverse and well-documented effects on the human immune system and its ability to defend against infective agents. While pulmonary related infections can occur in healthy humans, binge alcohol use is recognized as a major health risk factor (Nelson et al., 1991). Although binge alcohol consumption has been ...
    H1N1 Virus Production and Infection
    Authors:  Binbin Zhao, Jiaoyu Shan, Rui Xiong, Ke Xu and Bin Li, date: 10/20/2018, view: 995, Q&A: 0
    [Abstract] Influenza A virus is a member of orthomyxoviridae family causing wide-spread infections in human respiratory tract. Mouse infection model is widely used in antiviral research and pathogenesis study against influenza A virus. Here, we report a protocol in infected mice with different virus doses and strains to explore how an inhibitor of ...
    Isolation and Culture of Mouse Lung ILC2s
    Authors:  Pauline Schmitt, Anais Duval, Emilie Mirey, Jean-Philippe Girard and Corinne Cayrol, date: 10/05/2018, view: 1502, Q&A: 0
    [Abstract] Group 2 Innate Lymphoid Cells (ILC2) play an important role in immune responses at barrier surfaces, notably in the lung during airway allergic inflammation or asthma. Several studies have described methods to isolate ILC2s from wild-type naive mice, most of them using cell sorting to obtain a pure population. Here, we describe in detail, a ...
    Isolation of Microvascular Endothelial Cells
    Authors:  Kenneth C.P. Cheung and Federica M. Marelli-Berg, date: 06/20/2018, view: 3525, Q&A: 0
    [Abstract] The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. Murine endothelial cell culture is an important tool for cardiovascular disease research. This protocol demonstrates a quick, efficient method for the isolation of microvascular endothelial cells from murine ...
    In vitro Explant Cultures to Interrogate Signaling Pathways that Regulate Mouse Lung Development
    Authors:  Changfu Yao, Gianni Carraro and Barry R. Stripp, date: 05/20/2018, view: 1950, Q&A: 0
    [Abstract] Early mouse lung development, including specification of primordia, patterning of early endoderm and determination of regional progenitor cell fates, is tightly regulated. The ability to culture explanted embryonic lung tissue provides a tractable model to study cellular interactions and paracrine factors that regulate these processes. We provide ...
    Instillation of Particulate Suspensions to the Lungs
    Authors:  Etsushi Kuroda, Yasuo Morimoto and Ken J Ishii, date: 11/20/2017, view: 3319, Q&A: 0
    [Abstract] Inhaled fine particulates are thought to cause chronic pulmonary inflammation through the deposition of particulates into the lungs. To investigate the effect of fine particulates on the lungs, instillation of suspension of particulates into the lungs is required. This protocol describes direct injection of suspensions of fine particulates into ...
    Proximal Ligation Assay (PLA) on Lung Tissue and Cultured Macrophages to Demonstrate Protein-protein Interaction
    Authors:  Roberto Mendez and Santanu Banerjee, date: 11/05/2017, view: 3968, Q&A: 0
    [Abstract] In this protocol, we describe proximal ligation assay (PLA), an antibody-based detection method for protein-protein interaction. This method relies on specific binding of individual primary antibodies to the two putative interacting proteins. The primary antibodies need to have different hosts. The secondary antibodies against the two hosts have ...
    Modification and Application of a Commercial Whole-body Plethysmograph to Monitor Respiratory Abnormalities in Neonatal Mice
    Authors:  Yen-Ting Lai and Yi-Shuian Huang, date: 06/20/2017, view: 3984, Q&A: 0
    [Abstract] Proper breathing is essential for mammals to acquire oxygen after birth and requires coordinated actions among several tissues, including diaphragm, intercostal muscles, trachea, bronchi, lung and respiration-regulating neurons located in the medulla oblongata. Genetically modified mice that die early postnatally may have respiratory defects ...
    Isolation of Murine Alveolar Type II Epithelial Cells
    Authors:  Fan Sun, Gutian Xiao and Zhaoxia Qu, date: 05/20/2017, view: 5120, Q&A: 0
    [Abstract] We have optimized a protocol for isolation of alveolar type II epithelial cells from mouse lung. Lung cell suspensions are prepared by intratracheal instillation of dispase and agarose followed by mechanical disaggregation of the lungs. Alveolar type II epithelial cells are purified from these lung cell suspensions through magnetic-based negative ...

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