Protocols in Current Issue
    Differentiation of Myeloid-derived Suppressor Cells from Murine Bone Marrow and Their Co-culture with Splenic Dendritic Cells
    Authors:  Giada Mondanelli and Claudia Volpi, date: 09/20/2017, view: 401, Q&A: 0
    [Abstract] Myeloid-derived suppressor cells (MDSCs) possess the ability to suppress the immune response, and to amplify the regulatory properties of other immune cells, i.e., dendritic cells. Here we describe a protocol in which MDSCs were differentiated from murine bone marrow cells, and CD11c+ dendritic cells were purified from murine ...
    Phagocytosis Assay of Necroptotic Cells by Cardiac Myofibroblasts
    Authors:  Yuma Horii, Shoichi Matsuda, Kenji Watari, Akiomi Nagasaka, Hitoshi Kurose and Michio Nakaya, date: 09/20/2017, view: 423, Q&A: 0
    [Abstract] In myocardial infarction (MI), a plenty of cardiomyocytes undergo necrosis and necroptosis due to the lack of oxygen and nutrients. The dead cardiomyocytes are promptly engulfed by phagocytes. When the dead cells are not engulfed, the noxious contents of the cells are released outside, and thus, induce inflammation, and obstruct the function of ...
    An Assay to Determine Phagocytosis of Apoptotic Cells by Cardiac Macrophages and Cardiac Myofibroblasts
    Authors:  Yuma Horii, Shoichi Matsuda, Kenji Watari, Akiomi Nagasaka, Hitoshi Kurose and Michio Nakaya, date: 09/20/2017, view: 333, Q&A: 0
    [Abstract] In myocardial infarction (MI), a number of cardiomyocytes undergo apoptosis. These apoptotic cardiomyocytes are promptly engulfed by phagocytes. If the dead cells are not engulfed, their noxious contents are released outside, resulting in induction of inflammation. Therefore, the removal of these dead cells is necessary. However, the contribution ...
    In vivo Priming of T Cells with in vitro Pulsed Dendritic Cells: Popliteal Lymph Node Assay
    Authors:  Songjie Cai, Masayuki Fujino, Lina Lu and Xiao-Kang Li, date: 09/05/2017, view: 425, Q&A: 0
    [Abstract] One-way mixed lymphocyte reaction (MLR) is a classic tool to measure how T cells react to external stimuli. However, MLR is an in vitro reaction system, which shows different response intensity compared with in vivo trails sometimes due to the lack of cytokines, tissue matrix and other immune response associated factors. The ...
    Macrophage Survival Assay Using High Content Microscopy
    Authors:  Remzi Onur Eren and Nicolas Fasel, date: 08/20/2017, view: 523, Q&A: 0
    [Abstract] Macrophages maintain tissue homoeostasis by regulating inflammation and tissue repair mechanisms. Thus, the fate of macrophages has an impact on the state of the tissue. The aim of this protocol is to quantify macrophage survival using high content microscopy and image processing software. Here, we describe a high-content image based protocol to ...
    Isolation and Analysis of Stromal Vascular Cells from Visceral Adipose Tissue
    Authors:  Jessica Vu and Wei Ying, date: 08/20/2017, view: 596, Q&A: 0
    [Abstract] The obesity epidemic is the underlying driver of the type 2 diabetes mellitus epidemic. A remarkable accumulation of various pro-inflammatory immune cells in adipose tissues is a hallmark of obesity and leads to pathogenesis of tissue inflammation and insulin resistance. Here, we describe a detailed protocol to isolate adipose tissue stromal ...
    Establishment of a Human Cell Line Persistently Infected with Sendai Virus
    Authors:  Christopher Coakley, Cara Peter, Stephanie Fabry and Saurabh Chattopadhyay, date: 08/20/2017, view: 710, Q&A: 0
    [Abstract] Interferon regulatory transcription factor 3 (IRF3) is a transcription factor that upon activation by virus infection promotes the synthesis of antiviral genes, such as the interferons (Hiscott, 2007). In addition to inducing genes, IRF3 triggers antiviral apoptosis by RIG-I-like receptor-induced IRF3 mediated pathway of apoptosis (RIPA), which is ...
    Bioelectrospray Methodology for Dissection of the Host-pathogen Interaction in Human Tuberculosis
    Authors:  Liku B Tezera, Magdalena K Bielecka and Paul T Elkington, date: 07/20/2017, view: 784, Q&A: 0
    [Abstract] Standard cell culture models have been used to investigate disease pathology and to test new therapies for over fifty years. However, these model systems have often failed to mimic the changes occurring within three-dimensional (3-D) space where pathology occurs in vivo. To truthfully represent this, an emerging paradigm in biology is the ...
    Preparation of Mosquito Salivary Gland Extract and Intradermal Inoculation of Mice
    Authors:  Michael A. Schmid, Elizabeth Kauffman, Anne Payne, Eva Harris and Laura D. Kramer, date: 07/20/2017, view: 1138, Q&A: 0
    [Abstract] Mosquito-transmitted pathogens are among the leading causes of severe disease and death in humans. Components within the saliva of mosquito vectors facilitate blood feeding, modulate host responses, and allow efficient transmission of pathogens, such as Dengue, Zika, yellow fever, West Nile, Japanese encephalitis, and chikungunya viruses, as well ...
    Analyzing the Properties of Murine Intestinal Mucins by Electrophoresis and Histology
    Authors:  Ran Wang and Sumaira Z. Hasnain, date: 07/20/2017, view: 913, 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 ...