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Subcellular Fractionation of Hela Cells for Lysosome Enrichment Using a Continuous Percoll-density Gradient

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Visualizing Hypoxia in a Murine Model of Candida albicans Infection Using in vivo Biofluorencence

Authors: José Pedro Lopes and Constantin F. Urban, date: 08/05/2019, view: 446, Q&A: 0

[Abstract] Candida albicans is a leading human fungal pathogen that uses several metabolic adaptations to escape immune cells and causes systemic disease. Here, we describe a protocol for measuring one of these adaptations, the ability to thrive in hypoxic niches. Hypoxia was generated after successful subdermal infection with C. albicans ...

Precision Technique for Splenectomy Limits Mouse Stress Responses for Accurate and Realistic Measurements for Investigating Inflammation and Immunity

Authors: Shengwen Calvin Li, Anthony D. Rangel and Mustafa H. Kabeer, date: 08/05/2019, view: 393, Q&A: 0

[Abstract] Splenectomy in an animal model requires a standardized technique utilizing best practice to avoid variability which can result in adverse impact to the animal resulting in flawed physiologic responses simply due to technique rather than to the studied variables. In the case of the spleen, often investigators are analyzing the animal immune or ...

Detection of Antigen-specific T cells in Spleens of Vaccinated Mice Applying ³[H]-Thymidine Incorporation Assay and Luminex Multiple Cytokine Analysis Technology

Authors: Maria Agallou and Evdokia Karagouni, date: 06/05/2019, view: 804, Q&A: 0

[Abstract] For many infectious diseases T cells are an important part of naturally acquired protective immune responses, and inducing these by vaccination has been the aim of much research. Here, we describe a protocol for the analysis of vaccine-induced antigen-specific immune responses. For this purpose, cells of whole spleens obtained from vaccinated ...

Early Life Ovalbumin Sensitization and Aerosol Challenge for the Induction of Allergic Airway Inflammation in a BALB/c Murine Model

Authors: Kyle T Mincham, Naomi M Scott, Jean-Francois Lauzon-Joset, Jonatan Leffler, Philip A Stumbles, Patrick G Holt and Deborah H Strickland, date: 03/05/2019, view: 1121, Q&A: 0

[Abstract] The early life period represents a time of immunological plasticity whereby the functionally immature immune system is highly susceptible to environmental stimulation. Perennial aeroallergen and respiratory viral infection induced sporadic episodes of lung inflammation during this temporal window represent major risk factors for initiation of ...

Induction and Analysis of Anti-CD40-induced Colitis in Mice

Authors: Barbara Joyce-Shaikh, Daniel J. Cua and David Bauché, date: 02/05/2019, view: 2008, Q&A: 0

[Abstract] Colon inflammation or colitis affects more than 1 million people worldwide. Several pre-clinical models, including chemical-induced (i.e., DSS, TNBS) or pathogen-induced (i.e., Citrobacter rodentium) have been used to study mechanisms involved in the development and regulation of colitis. Anti-CD40 induced colitis model ...

Adoptive Transfer of Monocytes Sorted from Bone Marrow

Authors: Damya Laoui, Eva Van Overmeire, Chloé Abels, Jiri Keirsse and Jo A Van Ginderachter, date: 01/05/2019, view: 1771, Q&A: 0

[Abstract] Inflammatory Ly6C^hi monocytes can give rise to distinct mononuclear myeloid cells in the tumor microenvironment, such as monocytic myeloid-derived suppressor cells (Mo-MDSC), immature macrophages, M2-like tumor-associated macrophages (TAMs), M1-like TAMs or monocyte-derived dendritic cells (Mo-DCs). This protocol describes a method to ...

Use of Gas Chromatography to Quantify Short Chain Fatty Acids in the Serum, Colonic Luminal Content and Feces of Mice

Authors: Willian Rodrigues Ribeiro, Marco Aurélio Ramirez Vinolo, Leandro Augusto Calixto and Caroline Marcantonio Ferreira, date: 11/20/2018, view: 3630, Q&A: 0

[Abstract] Short-Chain Fatty Acids (SCFAs) are a product of the fermentation of resistant starches and dietary fibers by the gut microbiota. The most important SCFA are acetate (C2), propionate (C3) and butyrate (C4). These metabolites are formed and absorbed in the colon and then transported through the hepatic vein to the liver. SCFAs are more concentrated ...

In-vivo gp100-specific Cytotoxic CD8⁺ T Cell Killing Assay

Authors: Mahdia Benkhoucha, Nicolas Molnarfi, Elodie Belnoue, Madiha Derouazi and Patrice H Lalive, date: 11/20/2018, view: 1826, Q&A: 0

[Abstract] Cytotoxic CD8⁺ T lymphocytes (CTLs) represent a crucial component of the adaptive immune system and play a prominent role in the anti-tumor immune responses of both mice and humans.

Cytotoxic CD8⁺ T cells are responsible for the lysis of cells expressing peptides associated with MHC class I molecules and derived ...

Eicosanoid Isolation from Mouse Intestinal Tissue for ELISA

Author: Isabella Rauch, date: 11/05/2018, view: 1315, Q&A: 0

[Abstract] Activation of inflammasomes in peritoneal macrophages and intestinal epithelial cells (IEC) leads to the release of eicosanoids. To assess the amount of eicosanoids released by IEC, lipids need to be isolated from whole tissue previous to analysis by lipid mass spectrometry or ELISA. This protocol describes how to isolate lipids from intestinal ...

H1N1 Virus Production and Infection

Authors: Binbin Zhao, Jiaoyu Shan, Rui Xiong, Ke Xu and Bin Li, date: 10/20/2018, view: 1490, 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 ...

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