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: 266, 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 ...
    Eimeria vermiformis Infection Model of Murine Small Intestine
    [Abstract] Eimeria vermiformis is a tissue specific, intracellular protozoan that infects the murine small intestinal epithelia, which has been widely used as a coccidian model to study mucosal immunology. This mouse infection model is valuable to investigate the mechanisms of host protection against primary and secondary infection in the small ...
    Eicosanoid Isolation from Mouse Intestinal Tissue for ELISA
    Author:  Isabella Rauch, date: 11/05/2018, view: 993, 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 ...
    Purification of Total RNA from DSS-treated Murine Tissue via Lithium Chloride Precipitation
    Authors:  Emilie Viennois, Anika Tahsin and Didier Merlin, date: 05/05/2018, view: 2357, Q&A: 0
    [Abstract] We have developed a protocol to purify RNA from DSS (Dextran Sulfate Sodium)-treated mouse tissues. This method, which prevents downstream inhibition of q-RT-PCR observed in DSS-treated tissues, relies on successive precipitations with lithium chloride.
    Adhesion of Enteroaggregative E. coli Strains to HEK293 Cells
    Authors:  Jorge Luis Ayala-Lujan and Fernando Ruiz-Perez, date: 04/20/2018, view: 2009, Q&A: 0
    [Abstract] Enteroaggregative Escherichia coli (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory ...
    Immunofluorescence Analysis of Human Endocervical Tissue Explants Infected with Neisseria gonorrhoeae
    Authors:  Liang-Chun Wang, Qian Yu, Daniel C. Stein and Wenxia Song, date: 02/05/2018, view: 2292, Q&A: 0
    [Abstract] Colonization and penetration of the epithelium is the infection-initiating route of mucosal pathogens. The epithelium counteracts infection by eliciting host cell responses while maintaining the mucosal barrier function. The obligate human sexually transmitted bacterium Neisseria gonorrhoeae, or gonococcus (GC) infects the female ...
    Oral Microbiome Characterization in Murine Models
    [Abstract] The oral microbiome has been implicated as a trigger for immune responsiveness in the oral cavity, particularly in the setting of the inflammatory disease periodontitis. The protocol presented here is aimed at characterizing the oral microbiome in murine models at steady state and during perturbations of immunity or physiology. Herein, we describe ...
    Analyzing the Properties of Murine Intestinal Mucins by Electrophoresis and Histology
    Authors:  Ran Wang and Sumaira Z. Hasnain, date: 07/20/2017, view: 8215, Q&A: 0
    [Abstract] Specialized secretory cells known as goblet cells in the intestine and respiratory epithelium are responsible for the secretion of mucins. Mucins are large heavily glycosylated proteins and typically have a molecular mass higher than 106 Da. These large proteins are densely substituted with short glycan chains, which have many important ...
    Vaginal HSV-2 Infection and Tissue Analysis
    Authors:  Marie Beck Iversen, Søren Riis Paludan and Christian Kanstrup Holm, date: 07/05/2017, view: 4229, Q&A: 0
    [Abstract] The vaginal murine HSV-2 infection model is very useful in studying mucosal immunity against HSV-2 (Overall et al., 1975; Renis et al., 1976; Parr and Parr, 2003). Histologically, the vagina of Depo-Provera-treated mice is lined by a single layer of mucus secreting columnar epithelial cells overlying two to three layers of ...
    Mouse Model of Reversible Intestinal Inflammation
    [Abstract] Current therapies to treat inflammatory bowel disease by dampening excessive inflammatory immune responses have had limited success (Reinisch et al., 2011; Rutgeerts et al., 2005; Sandborn et al., 2012). To develop new therapeutic interventions, there is a need for better understanding of the mechanisms that are ...

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