Mammalia

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    Protocols in Current Issue
    In vitro Infection of Primary Human Monocytes with HIV-1
    Authors:  Patrick O’Connell, Yong-Hui Zheng, Andrea Amalfitano and Yasser A. Aldhamen, date: 07/05/2019, view: 498, Q&A: 0
    [Abstract] Monocyte infection by HIV-1 is an important component of chronic HIV pathogenesis. Following infection by HIV-1, monocytes are able to cross the blood brain barrier and set up a viral reservoir in the central nervous system. Additionally, in the setting of chronic HIV-1 infection, monocytes can become activated either directly through HIV-1 ...
    TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4+ T Cells
    Authors:  Anwesha Sanyal, Vatsala S. Rangachar and Phalguni Gupta, date: 05/20/2019, view: 995, Q&A: 1
    [Abstract] The latent HIV-1 viral reservoir in resting CD4+ (rCD4+) T cells represents a major barrier to an HIV-1 cure. There is an ongoing effort to identify therapeutic approaches that will eliminate or reduce the size of this reservoir. However, clinical investigators lack an assay to determine whether or not a decrease in the ...
    Assessment of Humoral Alloimmunity in Mixed Lymphocyte Reaction
    Authors:  Georgios Pissas and Theodoros Eleftheriadis, date: 01/20/2019, view: 1741, Q&A: 1
    [Abstract] Humoral alloimmunity remains a significant and unresolved problem that constrains allograft survival. Thus, there is a need for the development of an easy, preferably non-radioactive, and inexpensive protocol for assessing the effect of various drug treatments on humoral alloimmunity. In order to satisfy this demand, we developed such a protocol ...
    Monitoring Natural Killer Cell Function in Human Ovarian Cancer Cells of Ascitic Fluid
    Authors:  Vikash Kumar and Asok Mukhopadhyay, date: 12/20/2018, view: 1374, Q&A: 0
    [Abstract] Natural killer (NK) cells are the major effectors of the innate immune system when activated resulting in modulation of immune response of the host defense through target cell lysis and secretion of cytokines. Precise functions of NK cells are essential for the treatment outcome of different virus infections and malignant diseases. NK cells impart ...
    Quantitation of Regulatory Activity for the Complement Alternative Pathway Using an Adaptation of the AP50 in vitro Assay
    Authors:  Sheila Cabezas Falcon, David L Gordon and Jillian M Carr, date: 12/20/2018, view: 1039, Q&A: 0
    [Abstract] Complement pathways function to identify and remove pathogens and infected cells. There are three complement pathways: the classical, lectin and alternative pathway (AP). While all pathways are activated following pathogen stimuli, the AP is constitutively active and tightly controlled by activators (e.g., Factor B, Factor D) and negative ...
    Flow Cytometry Assay for Recycling of LFA-1 in T-lymphocytes
    Authors:  Katarzyna Potrzebowska, Janne Lehtonen, Malin Samuelsson and Lena Svensson, date: 12/05/2018, view: 1246, Q&A: 0
    [Abstract] To enable cells to move forward, cell surface integrins are internalized into an endosomal compartment and subsequently intracellularly transported to be re-exposed at a new site on the cell membrane. Leukocytes are the fastest migrating cell type in the human body, which express the leukocyte-specific integrin LFA-1. Here, we describe a flow ...
    Platelet Migration and Bacterial Trapping Assay under Flow
    Authors:  Shuxia Fan, Michael Lorenz, Steffen Massberg and Florian Gaertner, date: 09/20/2018, view: 1757, Q&A: 0
    [Abstract] Blood platelets are critical for hemostasis and thrombosis, but also play diverse roles during immune responses. We have recently reported that platelets migrate at sites of infection in vitro and in vivo. Importantly, platelets use their ability to migrate to collect and bundle fibrin (ogen)-bound bacteria accomplishing ...
    Immunohistochemical Identification of Human Skeletal Muscle Macrophages
    [Abstract] Macrophages have well-characterized roles in skeletal muscle repair and regeneration. Relatively little is known regarding the role of resident macrophages in skeletal muscle homeostasis, extracellular matrix remodeling, growth, metabolism and adaptation to various stimuli including exercise and training. Despite speculation into macrophage ...
    Visualization of RNA at the Single Cell Level by Fluorescent in situ Hybridization Coupled to Flow Cytometry
    Authors:  Alice Coillard and Elodie Segura, date: 06/20/2018, view: 2519, Q&A: 0
    [Abstract] The protocol described here has been developed to detect RNA at the single cell level. Fluorescent probes hybridize to target RNAs and are detected by flow cytometry after multiple amplification steps. Different types of RNA can be detected such as mRNA, long noncoding RNA, viral RNA or telomere RNA and up to 4 different target probes can be used ...
    In vitro Homeostatic Proliferation of Human CD8 T Cells
    Authors:  Hossam A. Abdelsamed, Caitlin C. Zebley and Ben Youngblood, date: 11/20/2017, view: 4573, Q&A: 0
    [Abstract] Long-lived T-cell–mediated immunity requires persistence of memory T cells in an antigen-free environment while also maintaining a heightened capacity to recall effector functions. Such antigen-independent homeostatic proliferation is mediated in part by the common gamma-chain cytokines IL-7 and IL-15. To further explore the mechanisms governing ...



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