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Co-culture of Human Stem Cell Derived Neurons and Oligodendrocyte Progenitor Cells

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A Refined Protocol for Identifying Citrulline-specific Monoclonal Antibodies from Single Human B Cells from Rheumatoid Arthritis Patient Material

Authors: Khaled Amara, Lena Israelsson, Ragnhild Stålesen, Peter Sahlström, Johanna Steen, Vivianne Malmström and Caroline Grönwall, date: 09/05/2019, view: 152, Q&A: 0

We describe here a detailed, refined protocol for the generation of citrulline-specific monoclonal antibodies from single human B cells from rheumatoid arthritis (RA) patients. This protocol provides a detailed guide of the procedure starting from ...

Isolation of Neutrophil Nuclei for Use in NETosis Assays

Author: Dave Boucher, date: 09/05/2019, view: 265, Q&A: 0

Neutrophils are critical immune cells that protect our body against invading pathogens. They generate antibacterial DNA structures called neutrophil extracellular traps (NET). Recently we identified a new mechanism that enables NET formation. We ...

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A Refined Protocol for Identifying Citrulline-specific Monoclonal Antibodies from Single Human B Cells from Rheumatoid Arthritis Patient Material

Authors: Khaled Amara, Lena Israelsson, Ragnhild Stålesen, Peter Sahlström, Johanna Steen, Vivianne Malmström and Caroline Grönwall, date: 09/05/2019, view: 152, Q&A: 0

[Abstract] We describe here a detailed, refined protocol for the generation of citrulline-specific monoclonal antibodies from single human B cells from rheumatoid arthritis (RA) patients. This protocol provides a detailed guide of the procedure starting from single B cells of your choice and followed by amplification of the variable region of immunoglobulin ...

Isolation of Neutrophil Nuclei for Use in NETosis Assays

Author: Dave Boucher, date: 09/05/2019, view: 265, Q&A: 0

[Abstract] Neutrophils are critical immune cells that protect our body against invading pathogens. They generate antibacterial DNA structures called neutrophil extracellular traps (NET). Recently we identified a new mechanism that enables NET formation. We observed that following recognition of lipopolysaccharides, inflammatory caspases cleave Gasdermin D ...

In vitro Differentiation of Thymic T_reg Cell Progenitors to Mature Thymic T_reg Cells

Authors: David L. Owen and Michael A. Farrar, date: 08/20/2019, view: 217, Q&A: 0

[Abstract] Thymic T_reg cell differentiation occurs via a two-step process. Step one generates T_reg cell progenitors (T_regP) and is driven by strong TCR interactions with antigens presented in the thymus. Step two is initiated by activation of STAT5 via IL-2, or IL-15, leading to mature T_reg cells capable of ...

Fibroblast Gap-closure Assay-Microscopy-based in vitro Assay Measuring the Migration of Murine Fibroblasts

Authors: Agnieszka P. Looney and Mallar Bhattacharya, date: 08/20/2019, view: 348, Q&A: 0

[Abstract] Pulmonary fibrosis is characterized by pathological scaring of the lung. Similar to other fibrotic diseases, scar formation is driven by excessive extracellular matrix deposition by activated, proliferative, and migratory fibroblasts.

Currently, the two most widely used chemotaxis and cell migration assays are the scratch assay and the ...

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: 426, 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: 360, 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 ...

Isolation and Long-term Cultivation of Mouse Alveolar Macrophages

Authors: Clara Jana-Lui Busch, Jérémy Favret, Laufey Geirsdóttir, Kaaweh Molawi and Michael H. Sieweke, date: 07/20/2019, view: 976, Q&A: 0

[Abstract] Alveolar macrophages (AM) are tissue-resident macrophages that colonize the lung around birth and can self-maintain long-term in an adult organism without contribution of monocytes. AM are located in the pulmonary alveoli and can be harvested by washing the lungs using the method of bronchoalveolar lavage (BAL). Here, we compared different ...

Gentamicin Protection Assay to Determine the Number of Intracellular Bacteria during Infection of Human TC7 Intestinal Epithelial Cells by Shigella flexneri

Authors: Atin Sharma and Andrea Puhar, date: 07/05/2019, view: 627, Q&A: 0

[Abstract] Shigella flexneri is an intracellular bacterial pathogen that gains access to the gut epithelium using a specialized Type III Secretion System (T3SS). Various determinants mediating this invasive infection have been experimentally verified using the classical gentamicin protection assay presented here. In this assay epithelial cell lines ...

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: 568, 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 ...

Morphological Evaluation of Wound Healing Events in the Excisional Wound Healing Model in Rats

Authors: Lígia Reis de Moura Estevão, Puebla Cassini-Vieira, Ana Greice Borba Leite, Apolônia Agnes Vilar de Carvalho Bulhões, Lucíola da Silva Barcelos and Joaquim Evêncio-Neto, date: 07/05/2019, view: 665, Q&A: 0

[Abstract] Skin wound healing is a complex process involving different events such as blood coagulation, inflammation, new blood vessels formation, and extracellular matrix deposition. These events can be observed by using histology techniques. However, the lack of the standardization of such parameters impacts on the reproducibility of results. Here, we ...

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